 Ladies and gentlemen, good morning and As-Salaamu Alaikum. Welcome to the first session of the fourth day of the STEM education training program funded by British Council. Let's start the session with the name of Allah Almighty. Ladies and gentlemen, today we have with us Dr. Noreen Latif. She is assistant professor at Center of Excellence in Molecular Biology University of the Punjab. She did her PhD from the same institute and joined as post-doc fellow and later as assistant professor. Her area of research is regenerative medicine for the repair of damaged organs. She has produced 19 ampoules and seven PhD scholars. Today she will deliver her talk on STEM Cells Research Bands to Clinics. Welcome, ma'am, on the behalf of Virtual University of Pakistan, please proceed for your presentation. My name is Dr. Noreen Latif. I am from CEMB Center of Excellence in Molecular Biology University of the Punjab. It is a state of art institute in the field of molecular biology. And we are producing ampoules and PhD scholars and we are proud that our graduate students are serving around the country as well as around the world in the prestigious institutes. And one of my students, Dr. Azam, is here and he invited me for to give a talk on urgent basis and yesterday he asked me to give a lecture. So in urgency, I prepared something for you. And my today's lecture is about STEM Cells Research from Bands to Clinics. The topic I selected is based on my area of research. My area of research is STEM Cells Biology. We in the center of excellence in molecular biology are the pioneers of the STEM Cells Research in Pakistan. And we started in way back in almost 2000 or 2001. And now different other departments and other universities as well are doing research on the STEM Cells as well as different clinics are importing STEM Cells and they are telling their patients that they are importing STEM Cells from different international companies and they are transplanting the STEM Cells in the patients for the treatment of the different diseases like osteoarthritis, spinal cord injury, infertility, baldness and skin regeneration, as well as for burn patient, et cetera, et cetera. So I will not comment on that whether they are truly transplanting the STEM Cells in the patients or not and they are taking a lot of money from the patient as well. So we are working on the grounds and we have published many, many international publications in international journals in with good impact factors, as well as we are in collaboration with different hospitals in Pakistan and around the world. And we are moving from bench to the clinics and currently we are in collaboration with Lahore General Hospital, Lahore Jinnah Hospital, LRBG Hospital, Gurki Hospital, et cetera. This was the brief introduction of mine. So I come to the point that what are STEM Cells and what are their importance and how a little bit about the research that we are doing there as well as what are the clinical aspects of the STEM Cells and what people around the world are transplanting or implanting the STEM Cells for the treatment of different chronic as well as acute diseases. But first of all, maybe you all know or might not don't know that what are STEM Cells. STEM Cells are the cells. I actually I was laughing because many familiar faces are here. I know Azam Humaira, Samina and one of the person sitting behind I don't know his name. I know many people now. So STEM Cells are the specialized kind of cells. They can. STEM Cells are special kind of cells. Okay, that has unique capacity to divide and produce cells like themselves and specialized type of cells also. STEM Cells are special kind of cells that can renew themselves as well as they can produce other kind of cell means they can produce cells which can be differentiated into other kind of cells or they are specialized cells. As they are uncommitted cells, they remain uncommitted in our body until an analyst, they get some kind of signal and they get differentiated into the specialized kind of cells like they can be differentiated into heart cells into the liver cells into the cartilage cells into the heart cells. So they are uncommitted, but they can get specialized or differentiated when they get some kind of differentiation signal and they can be differentiated to specialize tissue and then later on into the organ. So STEM Cells are basically a revolutionary idea that it is now prevailing into the medicine that in medicine we try to regenerate the organs or replace the organs. The idea behind the STEM Cells regeneration potency or regeneration science is that we try to to repair the organ by itself or regenerate the organ by itself so we do not get the need to transplant the organ. For example, if the liver is damaged inside the body, there is a natural phenomena of regeneration but if that phenomena is not properly working and we are taking the complementary medicines as well, but we need something as that we do not want to replace that liver. Then we can transplant the STEM Cells, we can isolate the special kind of STEM Cells from the body and then we can either differentiate those STEM Cells into the petrocytes which are the cells of the liver and or either we can even transplant simple STEM Cells into that liver and then the STEM Cells will divide there. They produce some special kind of paragraph effect means they produce some special signals which help the transplanted cells as well as the cells of the liver to get mature, to get divided and so they can produce some factors and the regeneration process starts and the organ is not needed for that to be replaced. Cells has the properties that they are self replicating, they are unspecialized cells, they have a capable of generating specialized doctor cells or progeny and they are plentiful in embryonic development but in the later stages they are less in numbers and in the children they are more active but with the passage of the time as we go older then they are less effective and less effective you can say that. So in that case we need the isolation of the STEM cells or the same body or from any other organ or organism or the human being and then we can grow in the lab and then we can transplant them back into that human. They have the capacity for self renewal and but that self renewal capacity increases with the time or aging, they can be categorized as embryonic or adult STEM cells and I will discuss it in the later slide. They have specific markers by which we can recognize them we can characterize them so whether they are true STEM cells or they are some other kind of mature cells or not. They exhibit varying plasticity depending on the environmental and commitment level. When the STEM cells they are two kind of the VN one is called as the symmetrical VN and other one is called as the asymmetrical VN. In the symmetrical VN when the ones are divided it produces the same kind of cells they draw on and the daughter cells we divide and people produce the same cells later on also. But in the asymmetrical VN, because the STEM cells has the asymmetrical kind of cell reveal, when the STEM cells divide it produces a cells like the parent cell which is called as the STEM cells and the other kind of cell which is called as the progenitor cells. The progenitor cells has the ability so that it can be differentiated when it gets some kind of signal or some kind of markers or some kind of information that you need to be differentiated. Then if that progenitor cell will, the progenitor cell will be differentiated you can see that the blue color is for the STEM cells and the red color is for the progenitor cell. So the STEM cells has the ability of asymmetric is divided by the asymmetrical VN and it produces the STEM cells like itself as well as the other cell which is called as the progenitor cell. What are the progenitor cells? You can see that STEM cells divide and produces the STEM cells and one other cell which is called a specialized cell but the progenitor cell which is some kind of specialized cell it will only divide to produce the specialized cells so that it cannot produce any STEM cells. So where the STEM cells are present or where the STEM cells reside? You all know that the life starts with the fume of the sperm and egg. When the egg divides after the fertilization when the zygote divides then first there is a one cell stage then two cell, four cell stage and so on. So when there is a zygote then which is the fume of the sperm and egg then it has the cells which when it is at the stage of five to seven days of embryo you can say that the five to seven days of zygote then it has all type of original cells that can produce the all human beings or all any other organism later on. So the cells present at the five to seven days of a zygote or an embryo that is called as the embryonic STEM cells. These cells are totally potent and they can produce the whole organism. But after that, you can say that five to six days of seven days of fertilization there are cells which can divide and produce different organs. They are called as the embryonic STEM cells and they are very potent and they cannot produce the whole organism but they have the potency to differentiate into many organs and it can help in the regeneration of the many organs later on also. And the pluripotent cells they are present inside the embryonic germ layers and they have the three germ layers which are the mesoderm, endoderm and actoderm structures layers and you know that these three layers are important part of all the organisms. So what are the difference between the totally potent, unipotent and multi potent STEM cells? Totally potent are the cells they are capable of any fate means they can generate all cell type necessary for the development of any organism, which I told that they are present inside them. We can isolate these embryonic STEM cells which are totally potent at the five to seven day of the fertilized embryo or zygote. But the pluripotent STEM cells they are cells or STEM cells which have the possibility to produce any other tissues and organs but they have some restrictions and limitations that they can produce many kind of cells or organs or tissues but they cannot produce the whole organism. Multi potent they have limited to low number of specialized cell types in differentiated tissues like the we can isolate the STEM cells from ourselves also we have STEM cells in our eye we have STEM cells in our teeth we have STEM cells in our hair follicles even in our fat in the umbilical cord which is get wasted after the child delivery in the umbilical cord blood as well as from the liver tissue from the any other tissues which are present from the heart as well. But all these organs have STEM cells but they are limited in number because they are developed organs they have limited number of STEM cells but they when there is an injury or any disease than these STEM cells get some signals from our body because there is a process of homeostasis that body try to heal itself by first and then when there is not healing and by the body then we should take we need to take some medicines to heal our body. So the stem cells are present in the mature organs also but they are limited in numbers and we have to isolate them by passing the special kind of procedures in the lab and then we have to characterize the cell whether they are stem cells or they are the part of the mature cells or mature organ. So they are very important for example if I want to isolate the stem cells from our fat which I will discuss in later we have abundance of fat like me including me so we are very God is very helpful and he has provided stem cells even the fat also. So we can isolate the stem cells from the fat and then we can characterize it by different markers which are present on the stem cells and then either we can transplant those stem cells back into my body in the disease organ or we can differentiate those culture stem cells in the lab and then we can specific kind of tissues or specific kind of cells differentiate itself and then we can transplant them back into the disease organ and then when we transplant the cells in the disease organs then there is a paracrine effect. That it proves the different kinds of growth hormones and different kinds of protein different kinds of products towards that transplanted or disease organs where we have transplanted the stem cells, then those signals, those proteins, those growth factors will help in the regeneration of that organ and the stem cells they are present there they can also get differentiated there into the specific organ. And then they can start the process of regeneration but in the adult cells they have limited potential to be different to get differentiated so they have we can call them they are multi potent stem cells. This is the image. The equipment that shows that what are the sources of the stem cells and what are their attributes or the potency. You can see that these are the stages of the human development, this is the single set embryo, then three day embryo or five to seven day embryo. When this stage is achieved which is five to seven days then we can isolate the cells from this stage also and these cells will be called as the embryonic stem cells. They are two deported. Why because they are the region of the development of the whole human being. So they have the potential to get differentiated into any kind of cells. So there is a debate on the use of these stem cells. Being a Muslim, we also have some ethical issues as well as the European or international society they also debate on the use of these embryonic stem cells. So there is no permission around the world in most of the countries by to use these embryonic stem cells. So, mostly people are working on these fetus stem cells and adults stem cells. We in the camera also working on the fetus stem cells or the adult stem cells, mostly, and they are called as the adult stem cells, and we are not working on the embryonic stem cells, because in our religion we believe that if we destroy this embryo that it will be the killing being which is not permissible in our religion, so we do not go for this. As you know how and how the people will can isolate the cells from this embryos. As you all know, there is a procedure of IVF transplantation, which is called as an in vitro fertilization when there is a fertility issue, then there is a facility available to that we can go for the IVF also. In the IVF procedure, there are many numbers of the embryos of the eggs and the sperm, and some of them are used later on and the other get wasted. And the doctor will see the healthy embryos or fertilized egg and transplant into the ovary of the patient and the other embryos or other fertilized egg they are getting wasted. So, the people think that they are getting wasted, so we can isolate the stem cells from them and then we can use it for the research purposes and for the benefit of the human being to cure different diseases, etc. But we are not working on it and the fetal issue and the other tissue which is our cells which are called as the embryonic germ cells and they are pre-ported in nature, we can isolate it from the six weeks embryo and we are not working on those tissues or cells and the fetal issue which are pre-ported or multi-ported, they are called fetal because they are isolated after the fetal delivery life. There is a baby delivery and there is a placenta and the embolical cord by which the baby is attached to the mother's womb, then they get wasted but that embolical cord or the blood which is present in the embolical cord which is called as a cord blood or even the placenta around the baby, they are very good source of stem cells because they are close to the embryonic thing. But they are not embryonic because as the baby is born, it has a big capacity to develop it into a human being and big regeneration potential also. So the tissues or the samples which we take from the hospital, as I told you that we had collaboration with Jinnah Hospital, LGH Hospital, Dungaram Hospital, then what is the purpose? First purpose is to take the embolical cord from there. We take the embolical cord which is get wasted after the baby delivery, there is no use of it. We take that embolical cord and then we use it for the isolation of the stem cells. And the other thing we are in collaboration with them from private clinic and there is a procedure which is called as the liposalarm section to get away from the fat from your body. So it is also a waste product. We take that lipospirate and then we isolate the stem cells from that lipospirate. Lipospirate is some kind of mature or added tissue. The fetal tissue which is the embolical cord is more close to the embryo but it has a potency is multi-ported or some kind of pluripotent and it produces the cells in a lot of number and it can be transplanted into pure different diseases. And in the edel you can see that we can isolate the stem cells from the cord-based stem cells or placental stem cells and in the human being we can isolate the stem cells from any other portion like bone marrow was the first source for which the stem cells are isolated but it is a difficult procedure, it is a more intensive procedure. So we do not go for that bone marrow isolation nowadays. We have switched towards the lipospirate and the embolical cord now. This site shows what is the difference between the embryonic stem cells, embryonic stem cells and adult stem cells. The main difference is the source from where we are isolating it, that is inner cell mass, the parmodial germ cells, or conical region of the 5 to 10 big fetal and it is the adult organs. What is the adult organ? Bone marrow, blood, cornea, retina of the eye, brain, skeletal muscle, dandelion, liver, skin, lining of the gastrointestinal tract and pectia. As you can see that there are a lot of organs in our body where the stem cells are present and they can produce stem cells later on if we have some disease and we can go in lab and transplant it back into the human being. It has the potential unlimited production of the cell that it has unlimited number of symmetrical deviants without differentiation. It can be divided into for the few years. It will produce the cell like itself so it will not get differentiated. The growing cells will not get differentiated because they are more to the embryo like. They are pluripotent and the other thing is the moded number of symmetrical deviants without differentiation is 7 to 80 dubbing population. They have 22 years of dubbing time and they have 72 to 80 dubbing time. If you do it more than this, then it starts going on the stage of the maturation. Whereas in this you have a limited number of deviants, you have cycles like we grow the cells in our lab. When we grow it on a special number, we detach it from it again and it goes to the next stage. It is called massaging after transplantation. We usually use P2P3 cells because they are the mixers of cells that are unwanted cells that are not pure. And it has a good proliferation potential. We can go to the fourth stage but we try to stay there. The more we go, the more maturity they have. They have less potency and their dubbing time increases and their number of cells is less. So this is because of their less potency and their limited time and less plasticity. But there are many types of cells because there is unlimited growth. One major drawback of this is that carcinoma and cancer is also a limited role of any tissue or cell present in our body. So there is a chance that when you transplant it, it becomes unlimited growth of the cells and it becomes cancer development. So this is the main reason why it is not used. It is not here and it is not here either. And the oct4 and other factors, these are all present in it. There are also specific markers in it. There are more markers in it. There are more markers of mature and fetal. The markers are that you have to recognize everything. I have to recognize that I have a special condition. I have a special condition that it is not like this. But when you recognize cells, you don't have cells in fat. You have adipocyte in fat because of your fat tissue. All the cells you have to isolate, they are not stem cells. So the specific markers of stem cells are on the surface, which are called as the meson, maybe called as the meson stem cells. The marker is CD190, 105, 173 and something like that. When we check the positive, we check the negative too. These markers are not in it. Most of the metabolic markers, the blood markers, they check that they are not markers, they are pure cells. And then we can take it. This is the card pool just to summarize the water stem cells. That is where stem cells, grouping centers ask the question that can you divide it yourself? He says yes, but he says that I am still here. But after that, you can see that I am un-specialized, but my brother, which is like me, it can be specialized and it can be differentiated when some get some signal or some proteins or some information to get differentiated. So my brother or the daughter cells, like the wisdom stem cells, it can become a skin cell, liver cell, but any other kind of cell. And he specialized to form tight junctions to his neighbors. You know that cells have some junctions or tight junctions and they can attack themselves with each other. So just a nice card to show you that what are the stem cells. They are un-specialized, they produce some kind of cells also, cells like themselves as well as other cells, which are called progenitor cells, and that they can be differentiated, that progenitor cells can be differentiated in the specific order. What is human and chronic stem cells? You can see this is the summary of them, on the previous slide that with the ideal treatment, the native diet, which is not no longer required, we can isolate that stem cells or embryonic stem cells at the 5 to 7 days ambulance stage and then we can culture that cell in our laboratory and then we can use that cells from different purposes like research, normal development and understanding the disease, procedure, molecular biology of the disease and developing therapies to treat different diseases as well as we can use that for the discovery of different type of drugs also. And other things that we can do with these embryonic stem cells that, for example, if there is any kind of gene which is mutated and which is the main reason for the cause of different diseases in that order of human being and we do not want that disease to get transferred into the next progeny that we will want to replace that gene. In that case, we can do the gene therapy or procedure which is called as the somatic cell nuclear transfer. In that procedure, we take the daughter cell as well as the any other eye and then we take them and they will move that disease nucleus in which the DNA is present, mutated DNA is present, mutated gene is present. Then we replace that nucleus with the nucleus from any healthy donor and we just call it the somatic cell. Somatic cell means we have taken the nucleus from a somatic tissue like skin tissue or any other tissue and we have replaced that nucleus from the somatic tissue into the eye and then it will divide and then the morula or blastula stages will go on and there will be development of a human being or foetus which will not carry that specific disease which is present inside the head of the mother. The other tissue kind of cells which I told you that are called as the tissue specific cells they are also called as the adult cells but mostly we call them adult cells but they maybe they are more likely to be tissue specific cells because we isolate from the adipose tissue we isolate from the liver tissue we isolate from the cartilage tissue from the heart tissue from the eye etc. so they are called as the tissue specific cells they are involved in tissue hemistasis and repair they are generally mighty potent and they because I told you that they are less in number in the organs so they are more difficult to there is more difficulty in isolation of the stem cells from that tissue or adult tissue but we are very expert in that in the use of the protein stem cells as you know that adult tissue are unipotent or biotent or some you can say that multi potent but they are not pluripotent they cannot get differentiated into multiple organs or many organs there is another kind of stem cells which are manipulated to be like pluripotent these tissues or these cells are called as the induced pluripotent stem cells as the name depict that they are induced to get pluripotent means we can take the cells from the patient like mostly the first experiment was on the human skin fibroglasses and there was a Nobel prize for this procedure which is IPSC Yamanaka from Japan was nominated and get the Nobel prize for this research for the production of the induced pluripotent stem cells for example we take a skin tissue from the patient and we take the fibroglast from the skin tissue and we grow that skin biopsy in the culture dishes in our lab and then we provide that fibroglast tissue with some transcription factors these four transcription factors are very much important CMEX of IV, KL4 and SOX2 the combination of these four transcription factors causes the reversal of these mature fibroglast cells toward the pluripotent stage and then these pluripotent stem cells which are patient specific we have taken it from the patient and then we can do multiple of things with that induced pluripotent stem cells we can either use gene targeting to repair the disease gene in that patient or either we can differentiate that pluripotent stem cells with the help of some signals, some compounds and some other materials so that we can use some specific drugs and then we can transplant that back into the patient so you can see that we have moved the gene targeting and repaired the IPSE cells and we have then differentiated into the skin cells and then have transplanted back into the patient so means we can take the skin cell if there is a disease of the skin and it is not possible to cure the disease with the help of some kind of drugs etc etc because the cause is the gene mutation and gene is disease so what we do we can take the disease from the patient and then we can revert it back into the pluripotent stage and then we can repair that gene and then we can differentiate it back and then transplant it back into the patient and the other thing that we can culture in and then we can use the different compounds to differentiate it back into the skin cells and then we can use different kinds of drugs and therapeutic compounds and then we can treat it back tissue back into the disease human beings so what was the features we can create a cancer directly from the patient it can be maintained indefinitely in the laboratory and then pluripotent with similar properties to embryonic stem cells so if we cannot use embryonic stem cells but we have one solution to that problem that we can revert it back into the pluripotent stage and then we can put it back into the patient so what are the different attributes of the embryonic stem cells, IPSEs and tissue cell stem cells the source is as you all know now and the rate of proliferation is very high in these cells and it is very low in the adult stem cells the availability is high but it is low here and spontaneous differentiation is high and it is very low in tissue specific or adult stem cells and they can produce diverse cell types and high in the IPSEs and ESCs and low in the adult stem cells so you can see that all these properties are very similar to each other so we do not need embryonic stem cells so that we can transplant it back into the patient and we can rejuvenate the tissue of the organ of the specific patient so why these stem cells are important because they are present in the muscle, neuronal and rheumatopartic cells and we can use any kind of stem cells from different purposes one is good, one is somehow not good that much but there is no specification or no limitation that we use that only kind of stem cells and we can use only pluripotent stem cells and if there are no pluripotent stem cells then we cannot transplant into the patient and there is no regeneration process in the later last slides I will show you that people around the world are using normal adult stem cells and other kind of stem cells and vital cord stem cells for the regeneration of different chronic diseases in the patient around the world and they are getting benefit from their transplantation also so Neutral can be discovered with the help of these stem cells we can use it in the genetics, molecular biology, biological control, tissue growth between ecosystems and cell therapy stem cell transplantation or cell therapy gene therapy they can also be done with the help of the stem cells so there are three to four slides I want to give you some introduction what the research at them can accurately we are doing on this is the building of our tissue tissue care as we told you that we do not use embryonic stem cells we use only adult stem cells in start we started from the bone marrow and bone marrow of the rats we have a very good state of the art animal house we have rats there we have farms there and I told you that we started the research there around 2000 and 2001 and at that time there was no know how of the what are the stem cells so that CMP is the pioneer of the stem cells research in start we started with the bone marrow of the rat or mice and we actually did the bone marrow stem cells from that because around the world the main source of the stem cell circulation at that time was the bone marrow but it was a very busy procedure and we also took one sample from the Dr. Amirasee is very non orthopedic and spine surgeon but the procedure is very invasive and so we do not go for that and later we switch to the other tissues also so the other issue is that nowadays we are working on the basically amdical cord as well as leprosyperin so you can see that this is the amdical cord the cord by which the baby is tied to the mother's womb and it is a mean route of the infusion of the different nutrients and the excretion of the mother and with the help of this amdical cord baby gets all the nutrients and all the base materials are treated out with the help of this amdical cord we take that amdical cord from the patients basically C-section deliveries because normally there is a chance of contamination so we reward that and then we take that amdical cord in a very sterile previous solution and then it is taken back to the amdical cord then after different washing and other things we make it, we chop it Dr. Sana is sitting here and she has worked a lot on this amdical cord and you can see that we chop it with the help of the scalper, with the help of the knife and there is a vein inside that amdical cord and we remove that vein and after chopping and mincing we add some also enzyme in it and get the collagen inside that amdical cord gets digested and after that we strain it and then we spin it and there is a pallet and then we grow that cells in our special medium and with some kind of growth factors and FPS and then after some days, within almost 1-10 days you can say confluency that we can use it for the even transplantation procedure in the human beings also and then you can see that this is the culture of SARS and after that after almost 15-20 days there is a colony formation of the issue of the cells and then we do different factorization procedure to make it confirmed that the isolated stem cells are the amdical cord cells the cells are stated from this amdical cord which is the mesenchymal stem cells and we use different mesenchymal stem cells marker to make it confirmed that these are the mesenchymal stem cells the other source is leperspirate there is a funny picture that as I told you that we have abundance of fat in our cell because of our lifestyle because of the very less active lifestyle also we get stituted all the tears and then we have lactobacillus infertopus and we have burgers and etc. there is a lot of source of stem cells in our cells also so this picture only shows the fat issue but now we are not using the fat issue but we are using that the picture on the top of the right side is the leperpirate we wash it we wash it to take it from the hospital and then we extensive washing and other things there is a long procedure and then we take the pallet and then we cultured the pallet in the cultured class and then we grow the amdical area leperpirate which is called the adipose drive mesenchymal stem cells in our lab you can see that these are the very nice picture of the mesenchymal stem cells in our cultured class or spindle shaping structure and we get very happy to see these cells coming out in our cultured class then we use different stem cells markers we use immunostatic procedure we use antibodies and then we strain with the fluorescent dyes and then we check it under the fluorescent microscope and check that there is a process of the died or immunostated cells or there is a procedure which is also called as the FACTS analysis by which we sort the cells that whether they are died to the specific antibodies which are mesenchymal specific like CD4090 positive 4090, 73 etc. but there are also negative markers like CD34 etc. and there is also another procedure that is I told you that three mesenchymal stem cells has the nature to get differentiated into the three germ layers epidermis, endodermis and mesodermis so we can also differentiate that these original MSCs into three lineages so that we are sure that the cells that we are working on are the mesenchymal stem cells and there is no impurity in that cells besides just a brief outline of the research that is being done by me and by the micro so we are basically working on three organs one is the cartilage and whether it's a spinal cord and third one is the cornea so the main theme is we take the samples from the human tissue and glycal cord and leprosy and then we use different modalities or different procedures we use different compounds and so that we can either improve the potency of the stem cells also so what is the logic behind them for example we are planning to transplant the cells into any human beings or even in the rat model of any disease first of all the first procedure is to take the, tryponize the cells, palliate it down and then count the cells and then inject the cells into the specific disease organ but you know that when we do the injection there is many chances that many of the transplanted cells get pasted with all the circulatory system with the injection and etc etc they may not reside there for a longer period of time so that they can do their function properly so for that purpose initially we were transplanting the stem cells into that into the any disease organ still we are transplanting it but there is also a need that there should be some kind of SCEP4 there should be some kind of bedding that we can grow the stem cells on it and then we can transplant it back into that disease organ this is the reason of using different kinds of SCEP4 and for the purpose of transplantation so me and my other colleague we are working on the stem cells and stem cells as well as on the SCEP4 also and we are working on spinal cord, osteoarthritis cartilage disease for the skin burn patient as well as for the liver regeneration procedure liver cirrhosis as well as for the diabetic foot ulcers also diabetic there is there is a disease in which diabetes there is ulcer formation mostly on the patient's foot does not get cured and there is a very problem with the patient and in some time patient has to cut his knee or foot so to get rid of that disease part and it is a very painful procedure so we try to help the patient to for the treatment of that diabetic foot as well as for the burn patient as well as for the osteoarthritis as well as for the for the damaged spinal cord injury etc. so we are working on SCEP4 we are working on different natural compounds as well as the biopharmaceutical compounds to increase the potential of the stem cells as well as we are working on the phyto extract and I would like to share that recently I got an overview of a project from the HSE on the phyto extract in combination with the stem cells so that as you know that there is a science which is HICMAT science or they are plant-based science that are being used from the centuries and they are currently being also used locally also so the plants have flavonoids and other phenolic compounds they are used for the inflammation for the coronary diseases and for other diseases so we are using these phytochemicals or extracted from them different plants mostly from the northern area of the Pakistan and then we are using in combination with the stem cells and then we are using it for the cure of the basically osteoarthritis and we have seen that one of my PhD students getting a degree on the Monday and his work is also on the phyto extract stem cells and the work is very promising and we have seen that if we the basic purpose to use this is that plants had anti-inflammatory as well as anti-oxidant compounds the stem cells also has anti-inflammatory and anti-oxidant properties but if we merge these two properties together then we can increase it many force and then we can use it transplant it because in any disease there is a procedure of inflammation the liver disease there is inflammation in the garter disease as you know that there is inflammation, swelling and pain and because there is a damage there is a production of rectifoxins species there so when we transplant the cells into that disease or inflammatory conditions then there is a chances that the stem cells get wasted they get damaged and they cannot survive there properly so we provide the stem cells with these anti-oxidants compounds and then we transplant it into the disease so there is a more chances and survival of the stem cells there so after that we are also working on nanoparticles also we make animal model and then we transplant the cells and then we do different molecular biological asses to check whether the disease is reword back there is reversal of the disease there is a regeneration of that specific tissue or organ or not this is one of the publications in which we use adipose stem cells which were differentiated into chondrocyte which are the cells of the cartilage and then these we have seen that the differentiated chondrocyte transplanted and ADSY transplanted these were more close to the normal as compared to the OA which is osteoarthritis and we did the histology also and the other wakitani and other management stores and we saw that there was a regeneration in the transplanted adiabas we have I will also see discuss in the later slide as I told you that there is a placenta around the baby and there are different layers of the placenta which are covered by membrane which is called as the amniotic membrane is very much important structure around the placenta it has very high tensile strength and it has very important growth factors also so it can help it can be used as the scaffold for the culturing of the stem cells also so we are we have used that amniotic membrane also for the culturing of the stem cells and even the differentiation of the stem cells and we have published it internationally around the 4.3 impact factor and then we have seen that it helps in the culturing or growth of the stem cells very well so this is the image from that publication that shows that the plastic surface also supported the growth of the cell as well as the amniotic membrane it sported more than the plastic surface as in previous slide I tell you that amniotic membrane is used for the culturing procedure but from many years around 20s, 1940s this amniotic membrane is being used for the ophthalmology procedures doctors or ophthalmologists use that amniotic membrane on the eye of the patient and it helps in the reduction of the inflammation in the eye of the patient as well as for the cure of some kind of diseases there so we are in collaboration with LRBT hospital mostly but many other hospitals like Aftaseh medical hospital and there was other hospital new hospital, Adil hospital Evesina hospital etc they take that amniotic membrane from us and we take it from the patient we do all the screening virus screening of the patient that there is no virus in the mother then we make the amniotic membrane and then we freeze it at minus 80 degrees E and when there is a demand the patient comes and then he takes the amniotic membrane from us and then the doctor transplants on the patient eye and there is help to the patient we do it with the no cause to the patient and we use our facility and our all resources for the benefit of the masses other thing which is being done with the help of this with the help of this amniotic membrane is the limbless transplants transplantation what is the limbless transplants there is a layer of a limbless tissue around the this cornea of the eye and it helps in the movement of the cells from this part on the cornea but when there is a damage on this limbless structure then the cells from this adjacent layer get get over the cornea and then that cornea is covered and there is opacity on the lens of the patient what the doctor do if the one eye of the patient is having this limbless sensor's efficiency and it has opaqueness on it then what the doctor do they take the limbless tissue very small limbless tissue from the healthy eye of the patient and then they provide us and then do them it has the limbless transplants in it so we can isolate the limbless transplants from it and then we grow it in a large number in certain stages we characterize it and then we provide it to the doctor in the form of injection and doctor put it back into the damage eye of the patient and that stem cells will help in the healing of that eye and we have seen that there was cure and there was regeneration and there was a very good real improvement, equity improvement in that damage eye of the patient so whether it was a burn patient an ugly burn patient an acid burn patient we have transplanted on a 10 years child a way back many years ago also that he has a damaged eye due to some car battery acid and his eye was improved very well and he was improved also so we also provide it but the doctor take the nautic membrane from us and we have a bank of that nautic membrane this picture just shows you the structure the place from where we take the nautic membrane and we then after procedure reserve it, preserve it at minus 80 degrees centigrade in the frozen form and then we provide it on the demand of the doctor this is the image showing the open force of the patient having the chemical injury of the bow thigh and you can see that the counting finger 1 and counting finger 1 was improved to the counting finger 5 meter it means the patient was able to see or counting can count the finger from the 1 meter before the transplantation and after that from the 5 meter distance he was able to count the fingers this is also the same for the another procedure and this was the basic research and later on the clinical side in which we were providing the nautic membrane to the a different hospital now coming to the clinical side of the stem cells so science is moving from the drugs to the personalized medicine means my me is basically the source of my cure also the cells inside me which are the stem cells they can be used any time of my life for the cure of many diseases in my body there are countries very advanced countries even in India also and in Pakistan some international companies taking a lot of a lot of money what they are doing they are in India also and other countries they are teaching people they are giving awareness to the people that when there is a child delivery then we are here you donate your mlycal cord or mlycal cord dirt to us we will take a lot of money from you and then we will freeze or we will preserve that mlycal cord or your cord dirt in for the lifetime and when there is a need or any disease in your children then he can use that stem cells, preserve stem cells and take from us and then we can cure it they are saying that this is the most precious gift that you can give to your children because remember it is a very costly procedure and they are using liquid nitrogen and other all the things they take it initially money from it and on the monthly basis they take money from the patient but people are doing this very well financially very good patients or persons they are doing this but if we are not able to do that in our country but we can help the patients also in another way what is that procedure we can make some good facility a large facility where we can isolate the stem cells and then we can provide it in the bulk for the treatment of the patient there are two sources of the stem cells one is called as allogenic and other one is called as autologous, allogenic is from the other patient other person autologous is from the myself only so we can use allogenic source like we can use mlycal cord mlycal cord is more like to feed us so it does not have any immune rejection if we transcribe into the patient so we can use that mlycal cord but if we are not willing we want to use our cell zones only then we can take the liposperate and then we can isolate the cells and then we can provide to the doctors and clinicians, many surgeons and many private clinics they are saying that they are importing the stem cells from other countries and many private companies international countries are working evenly now in Pakistan and they are charging 5 lakhs 10 lakhs, 2 lakhs, 3 lakhs all based on the patient pocket so they are giving to the patient I am not saying that all are corrupt they are all wrong and they are all illegal in the sense it does not have anything maybe even if it happens even if it happens and then improvement comes but I am not sure it does not have anything so now we because it is a basic research but we want to help the patient and we want to move from the bench to the clinic we want to help the clinic and help the patient later on so for that purpose there are different authorities and different agencies from that we have to take permission etc etc for the research purpose there is no need of that permission but for the other purposes or the clinical or the commercial purposes we need some kind of permissions so we are passing through that which is called as the current good manufacturing practices lab it is a lab like the OT operation theatre of the hospital that it does not have curves particle count is that much and there is no contamination etc etc so there is no chances of the any contaminations and we can provide the cells without any contamination to the different doctors so we have in collaboration with the NABAN centre also and we have a very small set up there which is CGMP approved there and we are caching samples here and we are providing for the research purposes only I will mention that and my colleague Dr. Azra she is working on facial reconstruction and as a diabetic foot ulcers and she is providing to doctors for the treatment of that disease and one other project which we have done is also the treatment of the patient with the COVID-19 patient with the help of this and black and cold cells in the last slide I will also show you another publication around the world and also thank you I am saying so much I was not able to finish this I am just showing that you have a CGMP facility where you can either culture the fresh cells and then provide it to the hospital as well as you can grow the cells and then you can freeze the cells and then you can provide it to the hospital for the transplantation purposes these are the different diseases around the world they are transplanting the cells like Alzheimer's disease Parkinson's disease, spinal cord, cartilage injury heart disease, fear burns and diabetes also they are being currently in clinics for the treatment of the disease and one is for the baldness like they take the stem cells fat tissue and then in the clinics and then they in the tube they digest it, they spin it and then adipocytes are also mixed many aesthetic people are doing this and they transplant it for the facial reconstruction if there is some kind of someone's neck is lying here along with our old man we also go up here so you can make it fat like you know everything else so you can make it fluffy and if you don't have hair then you can put it on your hair and if you don't have hair then you can put it on your hair that is for the beauty but if you have a disease then you can help it it is useless so for the bald patients you can put it on your skin patch because the grafts that come from outside are so expensive so much grafts are it is 100,000 rupees if someone has a lot of baldness then in a country like us you can't give it 50 plus 50-60 percent 1 patient is also survived because of facilities the amount of good money is not done here so if we can help them then it would be very good for the patient and also for us there are different diseases which are metapartic stem cells or bone marrow stem cells which have been transplanted and its data is around the world Lisa Roy she was diagnosed with some blood cancer and she was treated with the help of the stem cells transplantation and now Mashala she is recovered and now she is the brand ambassador of the stem cells therapy in India these are some of the papers around the world you can see that neural stem cells transplantation from the spinal cord injury I will mention that there is a very good non-hospital doctor he is the chairman and we are planning for the research purposes only and we are planning to transplant the viral cord stem cell on the spinal cord injury as well as on the osteoarthritis patient adipose type stem cell there is a knee injury or any sports injury with the passage of time and I think that they get the problem of these knees and there is a lot of pain in them there is a lot of weight there is a lifestyle there is also a transverse there is a problem in that you transplant it and a lot of clinics are doing this they are taking money they are saying that we are transplanting and I know one that he came to our clinic he was very happy and he is saying that his life has changed and the second HIV you know that HIV is a brown-eyed dream that we have been hearing since childhood but now it is taking some treatment and one of the main treatment was done in a patient he was a positive patient and he was seen to be negative because he was able to get some other blood cancer a donor transplanted in which the CCR5 and 32 were mutated in that patient when transplanted he was seen to be remission because he is a receptor that is your AIDS virus it attacks me and it grows in the cells and this patient is one in which his remission is seen Stem cells, I talked to you about COVID-19 we have done some patients we have taken their data they said there were moderate results you know there are severe cases if there is any cardiac arrest chances do not survive but we have seen the different intelligence there have been changes and improvements compared to the control patient this was recently I read in BBC news recently I read that there is a patient who is not published but it is from the news he was ill and it was very painful because in blood, the oxygen gets cells moved so it was a lot of procedure but then he found out that there are stem cells and there will be some changes and then he got the transplant and now he is very happy he wants to read an article I have given the link what was the procedure that he had bone marrow the cells in the bone marrow the disease that could make red blood cells sicker he isolated the stem cells and he turned the IgD-C11A gene and because of this the link stem cells were extracted stem cells were genetically modified and stem cells were given back to the Jimmy and these stem cells make the healthy fetus the basic idea behind it was that this cell 11 gene because in fetus there are some other genes that turn on and turn off and when the fetal gene is more he activated the gene that it should be active and it should be normal and it should be mature he did this procedure and he transplanted it and this is a very good case and he is very happy thank you very much I want to ask a question I am super excited that you are getting more and more after transplantation yes where does it take more time to transplant it takes more time 6 to 2 years but sometimes you are getting more and more and you are getting more and more and you get more but what effect does it have it is good to see that you need to follow it the studies of stem cells it takes 2 to 3 years to follow and that patient doesn't follow these studies so it is different but nothing happens there is no procedure there is no treatment but there are some advances there is research there are improvements there are more patients so everybody's body has a different response that is why there is a good improvement there is a moderate improvement that is why it is coming their shots are different after 3 months and 6 months there is another shot but again the internal body is going towards aging it is losing its activity so ultimately you have transplanted it it loses its activity so regular shots are required but obviously recommended regular shots are 4 to 5 years so no one is taking care of it so the aging factor is it will be a revert back but you can stop it from transplantation as you are taking drugs you are taking mydewyters so you should not take it like you are taking drugs you are taking it again and again to make your wrinkles so you have advised them that there is no such thing that it will not even end it will be your improvement because it is a paracrine effect so it has to recruit the effectors so do not listen to too much it has to be done by victory or it will be differentiated or it is a paracrine effect because of this paracrine it was in treatment but if an organ was going towards transplantation and you were very painful and you did not do it then atleast after 2-4 years you have to do a correction so you have to take care of it so it will be better you are taking somatic stem cell sorry taking somatic cells then revert back so what is the need to take stem cell when we can take stem cell see its limitations we are working with adult stem cell so it will not be more legal that we revert back first then we differentiate it you have to differentiate it after that you have to take it so there are same properties in stem cell but we are depending on the factors we are giving those it is very effective there is an added property when we convert stem cells into IPCs they can be used specifically for the personalized medicines and normally when we go towards allogeneic in our country and anywhere else allogeneic is completely bad if we want autologous transplantation it allows it does not allow there is no permission so when we want to get away from allogeneic and we want to get rid of multi-potency so that we can go towards a complete reconstruction so IPCs are for that you can take it from an individual because the HLA's and all surface markers related to person they are intact stem cell properties you can understand that thank you very much for a beautiful talk and very informative I have a question that when you apply the phytochemicals so it is a great chance of contamination into the stem cells so what criteria you adopt for the introduction of these phytochemicals yes but then if we have slated the phytochemicals then we dissolve in special kinds of medium like ethanol extracts water mint is all available it has a solution for that to remove the PCB from the material we filter it according to the string filter and in this way we do the stem cell culture and it supports our UV and then our sterilization procedure so our phytochemical extract also comes in combination with the combination we have more chance because we take samples from hospitals we take an anti-calcic cord we take some exposure or we have a lot of smoke and then we take it or we take the human dignity classes in July and June in the fungal rule in our individual we have some American domination so that the phytochemicals are used but not only the phytochemicals extract we use a multiple of other compounds phytochemical compounds vitamin E alpha4 Ease and the perfume and other different compounds also and then we mix it in an environment needs to be able to solution and then we string filter it so that it's okay there is no chance of elimination with that Thank you so much ma'am Thank you so much for joining us at very short notice Thank you ma'am Thank you for joining us at very short notice and it was very Thank you so much So nice of you Ladies and gentlemen, let's take a tea break and we'll resume the second session at 11.45 Welcome back Ladies and gentlemen, our next presenter is Dr. Sumera Anjum She is assistant professor at department of biotechnology Knade College for Women, Lahore She is a young and enthusiastic early career researcher in the field of nanobiotechnology She always remained a brilliant student during her education career and also proved herself as an excellent researcher in a very short period of time She has published many research articles in internationally recognized journals with an accumulative impact factor of 112 and more than 550 citations She has synthesized a large variety of metallic and metal oxide nanoparticles and evaluate their biomedical Today, he will deliver her talk on an effective simulator of plant mediated biosynthesis of nanoparticles and their biological applications Welcome Dr. Sumera, please proceed Assalamu alaikum First of all, I would like to say thanks to virtual university for inviting me this talk and I am feeling honored because my all senior faculty from Knade College is sitting here. Thank you so much To start, I have distributed a question This is just to check your basic background about the nanotechnology Please don't google it Just I am circled the correct option and after the lecture, we will again solve this questionary to check the either you have gained anything from my lecture that is usually the basic questions of nanobio technology You have five minutes for the questionary, then we will start the talk Thank you I think all the audience have done with it Today, I am going to talk about a research-based topic that is light and effective stimulator for plant mediates, senses of nanoparticles and their biological activities This is basically a research-based topic but obviously before going into the details of this topic, I will briefly give a short introduction of nanotechnology What is nanobio technology What is the classifications of nanomaterials and why nanomaterials are important What are the sources of the nanomaterials What are the properties of the nanomaterials What are the general applications of the nanomaterials and then what are the biomedical applications of the nanomaterials and at the end, we will going to discuss the specific topic of research, light as an effective stimulator for plant mediated senses of nanoparticles and their biological activities I would like to start the talk with the famous quotation of Sir Eric Russell It's always the small pieces that make the big pictures These quotations so beautifully clarify the concept of nanotechnology It's really the small things which are going to the next big things The nanotechnology is basically derived from the word Raphism, like nano means small, so basically the all beauty of the nanotechnology lies in its small size that is the main feature of the nanotechnology and in the current era the nanotechnology has revolutionized the every field of science engineering and technology and it has impacted the field of science to such an extent that it has changed the thinking of the researchers The researchers have started thinking in new ways that how these medicines can be targeted to a specific target There are so many biomedical applications of nanotechnology in life sciences So Next we start Sorry What is nanotechnology? First, this is a pure specific definitions of nanotechnology I am not going into the reading of this definition Just giving you an idea basically the nanotechnology field in this field is an interdisciplinary field in which we coin two basic sciences one is the science and the other is the engineering to fabricate different kind of devices and also to synthesize different kinds of materials and the materials which are synthesized by using the nanotechnology are then characterized and then they applied for their applications So basically nanotechnology is the synthesis and fabrications of different kinds of devices and materials which are then characterized and used for various applications in different fields of sciences But the materials either we are synthesizing how we can say that it fall in the category of the nanotechnology or either it relates it's our nanomaterial then basically the main key point that any material falls in the field of nanotechnology it is that it should be at least in one dimension in nanometer scale and what is about one dimension one dimension mean to say if you have synthesize and any nanomaterial either it should be in its length in nanometer scale either it should be in width of nanometer scale or either it should be its height in nanometer scale in any one parameter it should be in the nanometer scale it means if you have a rod that is one meter long but it has a diameter in nanometer scale then it falls in the category of the nanotechnology it's basically a nanomaterial but then the main point is that it should fall in nanoscale in at least one dimension now what is nanoscale you all know that one nanometer is equal to 10 days per 9 meter that is a very very small so you can have an idea about the nano things from this figure like this particles of the sands are not the nanomaterials they fall in the scale of millimeter polyuns from the plants in micrometer red blood cells are even very small but not they are the in the scale of the nanomaterial similarly the bacterias microorganisms are not in the nanometer scale yes the viruses started from the nanomaterials these are in the dimensions of the nanoscale similarly some proteins are there which are in nanometer scale DNA is approximately 2 nanometer in diameter similarly the carbon atoms the colorines are also in the nanometer scale and the atoms are also in the nanometer scale like carbon, hydrogen, oxygen then their protons, electrons all fall in the nanometer scale now as I already mentioned nanotechnology is an interdisciplinary field like some people have this concept of working in this field like we are from the life sciences we think that the nanotechnology is all about nanobiotechnology and if you belong to the physics they said nanotechnology belongs to the physics it is the domain of the physics and the chemist said nanotechnology is the domain of the chemist but it is not true basically nanotechnology is an interdisciplinary field and if we talk about the basic concept of history basically the region of nanotechnology was just started in 1969 in 1969 Sir Richard Feynman which is called as the father of the nanotechnology he gives an idea of small things Sir Richard Feynman basically delivered a lecture in University of California with the title there is a plenty of room in the bottom in which he gave the idea of small things but he didn't use the word of nanotechnology at that time the idea given by Sir Richard Feynman didn't gain such popularity or didn't get much intention until in 1986 Sir Eric Dexler published a book with the name of creation of engines in which he discussed the nature and he correlated the small things with the nature like Allah has created us our cells in nanometer scale and our organellies, mitochondria, golgi bodies they are in the nanometer scales and he said Allah has beautifully created these organellies and they are performing so many complex functions then humans can also engineer such types of materials which can perform different kinds of big functions so basically he was the first Sir Eric Dexler he used the word nanotechnology he has introduced the word nanotechnology but he was also a philosopher he was not the scientist so he just gave the idea of nanotechnology and after 1986 till 2022 almost there are 36 years only 36 years and nanotechnology has boomed every field of science it's a big market and it's the largest market currently I was seeing a paper in which that was published on the market value of the nanotechnology like in European countries it's going to be 3 trillion US dollar industry in 2024 and in Europe it's going to be 2 billion dollar industry in 2024 and in Asian countries it's going to be 5 billion US dollar industry in 2024 it means that the current industry is the highest booming industry that is nanotechnology and it has just boomed in 35 years periods of time so basically it's belong to all fields of sciences, chemistry, physics, biomedicine computer sciences, electronics and it's basically an interdisciplinary field and it's an individual subject it's not a part or property of any other subject now we said that in nanotechnology we manufacture different kinds of devices or either we synthesize different kinds of materials now what kind of materials we can synthesize by using the nanotechnology basically nanomaterials have been classified into three main categories one is called as the organic nature, organic type second is classified as the inorganic type and third one is called as a hybrid if we further divided them then the organic nature belong to polymeric nanoparticles liposome or lipid based nanoparticles these are belong to the organic nature these kinds of nanomaterials basically are used in nano medicines for drug loading like nanomers you can see have as a branch structure due to its branching it can load a variety of drugs and a high quantity of drug can be loaded next inorganic nanoparticles are usually metal based nanoparticles metal based can also be classified further into two classes like metallic nanoparticles are like silver nanoparticles gold nanoparticles, titanium nanoparticles copper nanoparticles and also the metal oxide nanoparticles metal oxide like zinc oxide nanoparticles, titanium oxide copper oxide these are all of the inorganic nature and metal based the other class with the inorganic material is carbon based nanomaterials the carbon based nanomaterials are further classified into carbon nanotubes which can be either single wall carbon nanotubes which are further used in various applications of electronics and also for biomedical applications and the quantum dots the quantum dots are really really small and the most amazing type of nanomaterials which are although currently manufacturing is very expensive but they have been commercialized due to their quantum confinement principle you all have might be seen a car that automatically change the colors of the surfaces in which the quantum dot technology has been used just to give a push of light or a source of light the color of the car is automatically changed either red, green, white the materials used are the quantum dot materials due to their optical excellent optical properties they are currently using in different types of devices surfaces coating and diagnostic imaging so this is brief what are the classification of nanomaterials now why nanomaterials this is the main thing why nanomaterials are so much important why we study nanotechnology so basically the nanomaterials as we have discussed they are important the first property that is their size they have very small size and anything we all know that if it has a small size then automatically its surface to area volume increases as you cut down cut down and then become to a smaller level its surface area become increasing the main attractive property in nanomaterials that is first of all their small size which correlates to the high surface area to the volume ratio and when it increases the high surface area to volume ratio then you can load a lot of drugs and other different kinds of materials can be loaded functionalized and they can be more reactive as compared to their bulk material this is the major one other than that why nanomaterials are important they have unique thermal properties like thermal properties thermal conductivity they have very low momentum and low size of the nanomaterials therefore they diffuse heat so rapidly and therefore they are having extra unique thermal properties and being used in different diagnostic purposes they have the unique optical properties rather than their bulk material like they have unique reflection, diffraction, absorption transmission properties which are basically using in diagnostic imaging bioimaging may use for this the optical property next is the mechanical property they also shows the unique mechanical properties these are like their hardness they differ in hardness they differ in elasticity they are sort and style they are lighter in weight and they can be so thin in structure that you can craft thinner weight or lighter weight aeroplanes in space energy they are using this property similarly they are using this mechanical property they are coating different substances with the lighter weight like in automobile industry they are coating with the nanomaterials to lighter their weight and to reduce the fuel consumption and to increase the speeds of the automobile next is the unique same electrical properties some of the nanomaterials are conductors some are semiconductor and some are insulators you can modify or you can manipulate the electrical properties of the nanomaterials by different techniques and you can use in different applications and the last one is the unique property is the magnetic some nanomaterials are ferromagnetic means they are completely magnetic and some are diamagnetic either they are repelled by the magnetic field and some are paramagnetic they are attracted by the magnetic field so by manipulating these magnetical properties you can use them in different electronic devices like nano chips nano barcodes nano biosensors so these are the unique properties of the nanomaterials that why we prefer nanomaterials than their bulk materials next what are the sources of nanomaterials the sources of the nanomaterials basically three types of sources one is the natural sources that is created by nature and the second one is the incidental or you can say that accidental sources and the third one are called as the engineered or intentionally generated nanomaterials in natural resources if we talk then our blood is the best example of nanomaterials milk is the best example of nanomaterials is the suspensions of different kinds of nanomaterials similarly used by the silk mite shock by formamine are the examples of the naturally occurring nanomaterials and if we now talk about the accidental nanomaterials that are not created by our intention like mining like different industrial processes different waste from the industrial processes containing the heavy metals which are then accumulating in our environment contaminating our underground waters and you can air contamination or different industrial processes like grinding corrections of the metals are generated the fumes or dust of the nanomaterials which are not created by intentionally and the last one is the intention this is the main class in which we researchers are working these are also called as the engineered nanomaterials because they are made by intentionally by the human beings and different kinds of nanomaterials are currently being synthesized and being synthesized by using different methods of synthesis that we are going to discuss in the next slide now coming towards the what are the roots of synthesis of nanomaterials that how we can synthesize the nanomaterials so basically the synthesis of the nanomaterials can be carried out by using two approaches there are two main approach one is called as the top down and other is called as the bottom up approach as you can see in the figure in this one in the top down approach you use the bulk as a starting material and then you cut down cut down and cut down and comes into the nanoparticle level while in the bottom up approach you use the starting material lesser than the nanometer scale like atoms, atoms are smaller than nanometer scale and then you get assembly of them into the clusters and then created into nanoparticles so basically these two main approaches are used for the synthesis of the nanomaterials these two main approaches are either bottom up or top down then by using these two approaches different methods have been formulated like there are three main methods that are used for the synthesis of the nanomaterials one is called as a chemical synthesis or chemical methods other one is the physical methods and the third one are called as the biological methods if we specifically talk about the chemical synthesis then in chemical synthesis we used the metal precursor salts in which you want to synthesize the let's suppose nanoparticles if you want to synthesize the silver nanoparticle let's suppose then you have to need a metal precursor salt that contain the silver like you can use silver nitrate you can use silver coloride which containing the silver ions and then you have to take another chemical which should reduce the silver ions from that solution and that region that is using for reduction is called as a reducing agent and different kinds of reducing agents are being used for reduction of different kinds of matters or different kinds of materials are currently being induced and after reduction the nanoparticles have been synthesized then you need an extra stabilizing or capping agent or capping agent to care for stabilize the nanoparticle because the nanomaterials or nanoparticles produced by the chemical methods are not capped until you have not capped or stabilized them by adding an other extra chemical so basically chemical methods are not the preferred one methods because they involves a lot of chemicals that further contaminate our environment and also we cannot use them for biomedical applications because we are introducing these nanomaterials into our living systems either animal models or either human beings then they are not recommended one nanoparticles that are synthesized from the chemical method but there are uses of chemical methods like chemical synthesis very refined nanoparticles that can be used in electronics that can be used in construction in construction industry that can be used in food packaging but we cannot introduce them into the living system or biological applications and the different types of chemical methods are currently in use like co-presentation method one of the simplest and the basic method one part reaction for synthesis of the nanoparticles the other famous method is the hydrothermal method and the most common method is the sol-gel method that is the most efficient in terms of its yield and in terms of the assembly of the nanomaterials the next class is the physical methods in physical methods as the name shows that we didn't use any kind of chemical but we use the physical equipment like we use the laser light we use a source of energy to evaporate the metal like different kinds of physical methods are currently in use for the production of different kinds of nanomaterials like laser ablation method is one of the most famous method that is in currently used for physical generation of the nanomaterials in which we use a pulse of laser light to evaporate our target metal and once it's evaporated it's evaporate in the form of atoms and these evaporated vapors are then collected into a cold finger and then scrap out from this cold finger or you can say from the substrate the main advantages of the physical method is that you can produce a nanomaterials in bulk amount but assembly is not controlled they are not well assembled they are not uniform in their size but yes you can synthesize in the bulk therefore if you want to control senses of the nanomaterials you have to control different factors we are going to discuss in the next slide the third methods are called as the biological methods biological methods are also called as green senses of nanomaterials the green senses of nanomaterials is not basically the senses only from plants this is not the senses only from plants as we generally assumed at green greener plants and only the green senses therefore green senses is not the only senses from plants this is the senses from all biological sources and there are mainly two types of biological sources are used for green senses of the nanomaterials one is the main class microorganisms like bicaria, viruses, fungi which contain a lot of oxido reductase enzymes and different proteinases which can cause the reduction of the metal ion from the precursor salts and the other main class is the plant you can use phytochemical and enriched plant extracts can be used for the senses of the nanomaterials the phytochemical rich plant extracts can be any medicinal plants which contain rich amount of phenolics or phlemonides or other kinds of scandium metabolites which basically can act as a reducing agents now currently the biological methods of senses and specifically the plant mediated senses of nanoparticles is currently being more practicing technique for senses of the nanoparticles especially for biological applications because they didn't contain any hazardous chemical on their surface and the one major property and one major point which is the plant mediated senses that is you didn't need any external capping or reducing agent you just have to take the plant extract and there is also no boundaries for the plant extract you can take the leaf extract, you can take the stem extract, you can take the root extract you can take the seed extract just you have to verify either it contain enough amount of the phytochemicals that can act as a reducing and stabilizing agent if yes this plant can't contain the enough phytochemicals that can cause the reduction and capping of the nanopaterials then yes of course go for it and this is the simplest method of senses and these are nanoparticles produced by the biological methods or knives you can use as such into the biomedical applications now I have discussed in the previous slide what is the green senses of nanomaterial that basically involves the two main roots from microorganisms and either from plants and basically the green senses we have seen in the term so anything not in the field of the life sciences anything generated in chemistry in physics, in electronics if it follows the 12 set of principles of green chemistry then it falls in the category of the green chemistry these 12 rules of the principles were given by American chemical society there are set of 12 rules that the methodology adopted in their senses are eco friendly they are safe, bioproducts are safe the methodology used is cost effective less source of energy used if they follow all 12 set of principles either in the creation of electronic devices, chemical devices or either in the creation of the nanomaterials they fall in the green chemistry but in case of nanotechnology we specifically talk about green chemistry from two sources organisms, microorganisms and other one is the plant now these both as I already told you the plants contains various phytochemicals basically the phytochemicals like alkylides phenolics, flavonides every plant contains certain amount of phytochemicals and different variety of phytochemicals these phytochemicals are actually acting as a reducing agent plus also acting as a stabilizing agent it means you have no need to add any chemical agent for reduction of the metal ions and you also need not any capping or surface agent to cap or stabilize the nanoparticles by using the green methods or green senses of nanomaterials you can synthesize the controlled or engineered nanoparticles of your own choice on which you can engineer you can formulate your types of nanomaterials by working or by controlling some factors which factors effect the senses of the nanomaterials like temperature temperature effect the senses of the nanomaterials it effects the morphological crexates of the nanomaterials if you are using the same plant source same molarity of the stock solution of the metal precursor and one senses on 30 degree centigrade temperature and other is the senses on the 60 degree centigrade temperature and other is on let's suppose 90 degree centigrade temperature temperature will surely effect the morphological properties they will differ in their size they will differ in their shape which means the temperature is one of the most factor you can optimize the temperature for your own choice of plant whatever you are working on you can optimize the temperature to attain the desire or acquireable sizes which effect the senses of the nanomaterials is the pH you can change the pH you can optimize the pH most of the time the lower pH have created have been seen that they impacted the smaller size created the smaller size nanomaterials if you increase the pH then the sizes of the nanoparticles also increases similarly you can modify the incubation period incubation period mean to say if you are going to synthesize let's suppose silver nanoparticles and you have prepared the one millimolar stock solution of let's suppose silver nitrate and you have prepared your plant extracts of the leaf of any plant which is let's say 10 gram per 100 ml and you are mixing them in a definite ratio like one into one, one into two one into four this is all your own optimization you have to go through a series of optimization for senses of the required size type or morphology of the nanomaterials so then you can also change the incubation time if you give 10 minutes the size of the nanoparticles may be different for those 30 minutes incubation time then 150 minutes incubation time incubation time basically increase the contact time which increases the rate of reduction and sometimes the incubation time also negatively impact the senses of the nanomaterials they reverse back the oxidation reduction reactions and the nanoparticles generating are not stable then again you have to optimize the contact time according to your plant according to your type of the nanomaterial and the last one is the centrifugation centrifugation timing also matters either you are speed and timing giving for a larger time or shorter time larger nanoparticle will be sedimented or the smaller nanoparticles will be sedimented now my area of specialization is plant mediated senses of nanoparticles I am being a biotechnologist and my field of expertise are also related to the plant biotechnology and I have worked in the plant mediated senses of the nanoparticles and basically I am working on the formulations of different kinds of nanoparticles then why plants are the best contenders if we talk about green senses then there are microbes in green senses and there are plants in green senses then why we prefer the plants in plants one it is a simple technique second that is a disadvantage of the microbial like their maintenance you have to require a lot of effort and money to isolate the pure cultures and to maintain the pure culture of the microbes while in case of plant there is no need to maintain the culture plants are available everywhere being a agriculture country in Pakistan almost every plant is present and easily bio available environmentally eco friendly cost effective just you need the metal precursor salt and you will get free of cost next single pot reaction just you have to mix the metal precursor salt with the plant extract in single pot you can synthesize the nanoparticles next moderate temperatures you can synthesize them even at room temperature you just go for modifications of the temperature or increase or decrease of the temperature if you are not gaining results or if you are aiming to develop different kinds of shapes or sizes of the nanoparticles otherwise most of the time nanoparticles are synthesized at the normal room temperature and pressure in case of plant dated of nanoparticles and the most important no additional surfactant and capping agents are required while in case of the chemical methods you need the extra surfactant or capping agent and same in the case of the microorganisms mediated microorganisms can reduce by using their oxido reductase enzyme the reduction of the nanoparticles but they cannot cap them in microorganisms mediated you again needed the external capping or surface agent but in case of the plants the same phyto chemicals which have reduced they can also cap the nanoparticles after their formulation and making them stable this is the main point that in plant mediated instances you need no any extra capping or surfactant or capping agent next no toxic solvent is required like all plant extracts are made in water and water is everywhere and water is the most best solvent or that is environmentally friendly easily available so all plant extracts can be made in the water low toxicity or biocompatible so these are the main points or the reasons that why we prefer plants for senses of nanoparticles now what is the chemistry behind plant mediated senses of nanoparticles basically the senses of the nanoparticles in plant mediated case take place in three stages or three cycles one we can be called as the reduction process second is called as the growth phase or growth process and the third one is called as the stabilization or capping process in the first step that is called as the reduction step up now what is the basic chemistry behind that how we think that we took a salt precursor and mixed it with plant extracts or just mixing say nanomaterials are made actually what is the chemistry you are using first initially a metal precursor I will again show you the example of the silver nanoparticles there are thousand papers have been published on the senses of the nanoparticles by plants for example you have taken the silver nitrate as a metal precursor salt of the silver you have to prepare of any definite molarity stock solution of the silver nitrate like you can prepare one millimolar you can prepare two millimolar you have to optimize according to your reaction let's suppose you have taken 10 grams leaves from the wild plant and then wash it and then grind it and then simply boiled in distal water and after boiling you have filtrate it that is a 10 gram per 100 ml water the stock solution of the plant extract then you have to mix them in a definite molarity in a definite ratio like you can take one ml of the silver nitrate stock solution and you can take one ml of the silver nitrate stock solution and you can take one ml of the plant extract solution when you mix them with a specific ratio then what actually happened actually the silver nitrate when salt is when dissolved in the water it dissociate into iron silver ag plus ions plus nitrate NO3 minus one nines of these silver ions that are the ketines ag plus one nines these are unstable and we have to reduce these ions into the zero valent form we have to convert ag plus into ag zero and the ag zero that is the zero valent form is the nanoparticle so basically the simple chemistry involved is the reduction you have to reduce the cationic ions into its zero valent form and the reduction is carried out by the phytochemicals present in the plant extract these phytochemicals actually no ma'am they are just inorganic inorganic natures like zinc oxide, silver, titanium they can be but they are not stable like the zinc that makes a dimer with oxygen it is in the form of oxygen we cannot do pure zinc so not all the matter but some most familiar matter whose nanoparticles have been synthesized then after zero valent ion they are unstable and they start to agglomerate with each other then again the phytochemicals present in that plant extract that might be different it is not necessary that the phytochemicals have reduced the phytochemicals they will do their capping plant extract are diverse resources of phytochemicals in single one ml of the plant extract there might be thousand phytochemicals different nature of the phytochemicals you can also work on isolated phytochemicals we are working as a whole plant extract but you can also work on the isolation of a single metabolite and can work on that synthesis so these metabolites can also cap the nanomaterials which are basically stabilizing them and preventing them from agglomeration the stability of the nanoparticle is the main factor for their applications either you are using in biological applications or either using for electronics chemistry or any other applications you have to stabilize it if they are not stable they will agglomerate and they will loss their unique properties therefore stabilization process is very important basically there are three stages one is the reduction second is the growth phase and third one is the termination or stabilization or capping phase and in this figure also the we have given the structure of a phytochemical in case of silver nanoparticle synthesis which are acting as a reducing agent or bioreductant and reducing the silver into zero valent form which then grow from the silver nanoparticle and these nanoparticles are then again stabilized by these scandal metabolites now after synthesis if you have synthesized the nanomaterials then we cannot use them into any kind of applications superior to their characterization it's me any that you have made nanoparticles you have just made you didn't know about its phytochemical properties you have to know about all its phytochemical properties mainly these properties may have been interested in biological fields like size what is the size of the nanoparticles what is the shape of the nanoparticles what is the crystalline nature of the nanoparticles what is the surface morphology or surface topology of the nanoparticles what is the surface charge on the nanoparticles these are some basic phytochemical properties that we should know before applications of the nanomaterial and the applications of the nanomaterials are also dependent on these properties some properties some properties are size-based applications are size-based like size matter if you are looking for smaller size if you are looking for smaller size then those applications are specific if you don't know what is the size of the nanoparticles then you cannot use it if you didn't know about its applications what applications you can use after since this is the next step in green chemistry of the nanoparticles is their characterization and for characterization there are different techniques which are used for characterization of the nanomaterials firstly one is the UV visible spectroscopy in UV visible spectroscopy you basically find out the optical properties of the nanomaterials optical properties like surface plasma resonance and in case of the inorganic metals like silver nanoparticles UV visible spectroscopy is the first confirmation technique which confirms yes the nanoparticle has been synthesized because every kind of the metal give a specific surface plasma resonance in a specific range of wavelength like in case of silver it give absorbance peak in the range of 400 to 480 nanometer in case of gold it give peak in the range of 500 to 600 nanometer in case of zinc oxide nanoparticle let's suppose it give the peak of surface plasma resonance in the range of 300 to 380 nanometer to the first initial confirmation technique which confirms yes the nanoparticles has been synthesized is the UV visible spectroscopy next is the transmission or scanning electron microscopy that is basically microscopic techniques used to find out the basically shape of the nanomaterials basically the nanomaterials basically the nanomaterials there are many other applications that are used in physics and chemistry but basically as a nanotechnology nanotechnology is if we are getting nanomaterials transmission time or SM then we are interesting to find out the shape of the nanoparticles like either they are spherical in shape either they are rod type or shape of the nanomaterials that is the basic information that we get from the transmission electron microscopy the second information that we can get from the transmission electron microscopy is the size of the nanoparticles you can also calculate the size of the nanoparticles by analyzing them either SEM or TEM but for precise calculation of the size SEM and TEM are not the ideal techniques but you can have an idea about the sizes of the nanomaterials the main information in which we are interested is the shape by which SEM or TEM if we talk about either SEM is preferred or either TEM is preferred and if you have a choice between both of these techniques then TEM is preferred it has a high resolution it can generate 3D images it can generate bright and contrast images and gives you a clear picture about the shape of the nanomaterials as compared to the SEM this is the XR diffraction analysis XR diffraction analysis is basically used to find out the crystalline nature of the nanomaterials the basic information you are getting by analyzing this technique is the crystalline nature of the nanomaterials either they are face centered cubic either they are monocleaning either they are orthorhombic either they are hexagonal system which class they belong to in their crystalline properties by analyzing them the XRD technique the next is the energy dispersive X-ray analysis energy dispersive X-ray analysis is in billet property of the SEM and TEM that is always used in conjunction with SEM and also with TEM in case of nanomaterials we are interested in the EDX analysis to find out the elemental composition of the nanoparticles like we have synthesized silver then we have to confirm either silver is present in that so this is the elemental similarly let's suppose we have synthesized zinc oxide nanoparticles then the EDX analysis which tells us zinc and oxygen both are present in it and other than zinc or oxygen either any other elements are present and if they are present then in which ratio by mass and by atomic number that all information are given by the energy dispersive X-ray analysis next is the dynamic light scattering analysis dynamic light scattering analysis also give about the uniformity of the sizes what is the distribution of the nanoparticle sizes like might be there is a thousand number of nanoparticles that have been prepared in your sample and from thousand might be 100 particles of 20 nanometer size the next 300 nanoparticles can be of 18 nanometer sizes to find out the uniformity or particle size distribution we do the dynamic light scattering analysis the next is the Fourier transform infrared spectroscopy abbreviated as FTIER analysis the FTIER analysis is basically carried out in case of nanomaterial there are many other functions that make chemistry either they have synthesized a novel compound so they analyze it from FTIER spectroscopy but as a nanotechnology if we are going to done the FTIER analysis then we are particularly interesting to find out which biomolecules are responsible for reduction and capping of nanomaterials and if you want to know which biochemicals are capping the nanomaterials then you have to do the FTIER analysis which gives you informations about all kinds of bonds and all kinds of atoms that have been capping by your nanomaterials like carbon-carbon oxygen carbon-carbon bond, carbon-carbon double bond carbon-oxygen bond that vibrate with a specific set of wave energy that given to a histogram or a graph where we will discuss last is the zeta potential analyzer that also gives about surface charge and also the size of the nanoparticles now after characterization what are the applications of the nanotechnology or nanomaterials you have synthesized either from any of the source from biological sources from chemical sources or either from physical methods so the main applications these are in the all areas of life like nanomaterials are currently being used in agriculture industry in agriculture industry we basically specifically talk about the plant so there are different kinds of nanomaterials that are currently used for crop movement like many nanoparticles used as a nano fertilizers as a nano pesticides as a nano herbicides as a nano fungicides as a nano biosensors as a nano encapsulations for delivery of the micro and micronutrients to the plant that have overall improved the crop improvement similarly if you talk about the food industry in food industry nanotechnology is also being used and there are so many applications in the food industry if you specifically talk about food industry then there are the two main applications or there are two main spheres in which nanotechnology is being used one is in food processing and other is in food packaging in food processing different kinds of nanomaterials are used as a antimicrobial agents during their processing they protect the food from contamination from destroying and some of them are using as a anti-caking agents that prevent the fallation or emulsions in the frozen foods and other kinds of reserve food similarly if you talk about the food packaging then there are nanotechnology has introduced three different kinds of packaging in the food industry one is called as active packaging, smart packaging and intelligent packaging in active packaging the packaging packaging of the food is carried out by such material which contain the antimicrobial agents and in the smart packaging the nanomaterials are introduced which are actually acting as an indicator either food is safe to use or food has expired simply these nanomaterials change the colors and you can check the expiry date of the nanomaterials by their packaging either this food is expired or either this food is fresh to use so different kinds of packaging have been introduced by the nanotechnology the next is in electronics or computing if we specifically talk about electronics or computing we all know that the electronic industry has a lot of revolutionized smart watches, digital watches smart mobile phones desktop screens which have different kinds of semiconductor materials like quantum dots, carbon tubes flexible display sheets nano chips for different diagnostic purposes nano transistors that all have been synthesizing by using the nanotechnology next is the catalysis catalysis basically belong to the chemistry nanotechnology has also revolutionized the catalysis field like I will like to give the examples of the two major industrial processes of the chemistry like one is the synthesis of the ammonia I know that you all know that we basically study in matriculation synthesis of the ammonia by hybrid process and why we synthesize ammonia why it is a big industry because ammonia is the main source of fertilizer in the synthesis of fertilizers ammonia is used and conventionally the ammonia synthesized by using the hybrid process is used different kinds of convention like like iron based catalyst which has a low efficacy low speed and also prolonged durations different kinds of nano based catalyst have replaced this industry which have speeden up the production of the nano ammonia synthesis and also they are biocompatible or eco friendly they have speed up the synthesis of the ammonia by the hybrid process more than two times similarly another industrial process that is the hydrogenation of the ghee or oil we make ghee is saturated or oil unsaturated yes the formation of ghee we are hydrogenating the ghee that is also an industrial process in which we also use the nickel as a conventional catalyst which sometimes recent studies have shown that they are accumulated in our organs if you are consuming the ghee nickel accumulates in the ghee therefore the nano technology has given a replacement of the nickel catalyst with the nano catalyzed which have speed up the hydrogenation reactions more than the five times and with no toxicity or with no toxic effects then the automobile industry I have already discussed different kinds like bucky ball sheet papers carbon sheet papers have been designed which are so lighter in weight that if we talk about the flight space industry they are reducing the flight times like a recent study was published in which carbon bucky balls and carbon sheets and graphene sheets were painted in the aircraft industry the outing was covered and then travel the jets of spacecraft were checked that from earth to moon those two times the flight time was reduced by using these bucky balls they have basically reduced the friction of the air and also they are very lighter in weight as compared to the steel or other conventional kinds of conventional kinds of materials next you can use nano technology in defense and in military in defense and military different kinds of textiles they have generating the textiles of different colors that can be camouflaged like you can electronically gated textiles or dresses which can change their color you can get it green if you want to camouflage it you can change into the skin or dust color these are the main which can be used in defense other than that IT field intelligent devices have been devices by using the nano technology for defense purposes or for military purposes similarly different kinds of biosensors which can detect the biological weapons which can also increase the defense of any country then textile industry is the same as I am discussing different kinds of camouflage clothing and in a recent study the scientists of the Harvard University have made a nano surprise these nano surprise are simply the emergence of the nano materials with the cotton conventional cotton which we use in cloth industry or textile industries they have mixed nano particles and they spray just on the body after drying they will be converted into the cloth that will be just like same flexibility and you can dissolve it again they are re-invable after that you dissolve them in the solvent then again the spray is ready you can do this and you can carry it with you in your bags for any kinds of clothing you can design similarly in energy generation nano technologies giving a huge bump like in nano fuels, nano batteries nano generators, nano transistors there are different applications in which nano technology is playing its positive role next is the construction industry like lighter weight materials have been produced other than nano paints nano fillers giving that strength required strength to the construction and also different kinds of the nano paints which are dust repellent, water repellent which are being used in the construction industry the last one is the main domain that is the domain of life sciences that is the health care industry the next we are going to discuss the applications of the nano materials in the health care sector the nano materials different kinds of nano materials have been used and currently in use like dentimers, liposome, quantum dot meslis, nanospheres, nanoparticles polymeric nanoparticles and carbon nanotubes which has been used for different kinds of biological applications like for gene therapy for treating different kinds of genetic disorders they can like if we talk about the gene therapy I am not the molecular biologist I don't know so much details but these applications know basically what we use for delivery of the genes sometimes they are degraded by the nucleases enzymes to protect them from these nucleases degradation we use the nano material as a nano carriers of the genes they one protect from the degradation and secondly acting as for a target delivery of the genes by surface functionalization of the nano materials similarly different kinds of nano materials are used in drug delivery if we specifically talk about the health care sector the main or booming field of nanotechnology is drug delivery in drug delivery they have revolutionized the word with the word of targeted drug delivery nanoparticles can deliver the drug to your target more precisely as compared to the conventional drug systems basically nano materials can act as a nano drug and they can also act as a nano carriers I will briefly differentiate the nano materials sometimes we are a concept player that nano materials act as a nano drug yes they are working and sometimes they are itself not working as a drug but acting as a carrier of the drugs so nano materials can act as a nano drugs and also they can act as a nano carriers for example in case we can say if nano particles are acting as a nano drug then what is the example cancer one of the most leading infectious disease causing death worldwide cancer treatment in nanotechnology is a massive role in every aspect either from diagnostic to the treatment of the cancer during the cancer treatment nanoparticles can be used for the treatment of the cancer in which the two main conventional therapies have been coupled within nanotechnology one is called as a photodynamic therapy and other is called as the photothermal therapy these are the two main therapies which are used are conventionally used for the cancer treatment but now it has been coupled with the nanoparticles in which different kinds of nanoparticles like they are called as the photosanitizers they are sensitive to the light they have been administrators systematically through our systematic system to the person and when those nanoparticles surface functionalize where our cancer cells are basically we attach them to any kind of surface membrane receptors or antibodies or specific epitomers that our specific cancer cells target them when they target and accumulate the main source is when we expose them these nanoparticles have the property to absorb light when they absorb the light they start to vibrate and once they are vibrated they transfer their vibrational energy into the heat energy and the cancer cells are very sensitive to the heat and the area of the cancer cell is heated up which causes the apoptosis or you can say the death of the cancer cell that is coupled with convention photo thermal therapy in which nanoparticles are acting as a nano drugs this was the example in which they are acting as a nano drug now talk about the nano carriers nanoparticles also can act as a nano drug can also act for the targeted drug delivery if we talk about the targeted drug delivery then you all know that the targeted drug delivery is also of two types one is called as the passive targeting and other type is called as the active targeting in both kinds of targeting either it's active targeting or either it's passive targeting nanoparticles have been used like the passive targeting works on the retention time in which we increase the retention time of the drug in our cancer cells and in the active targeting we are working on the specific targeting to specific cancer cells I will give an example of this in active targeting and passive targeting nanoparticles and nanotechnology how it helps like the nanomaterials and the nanoparticles they pass through your circulation system they pass through your circulation system you can functionalize their surfaces like you want to target to the cancer cells then you can make the antigens of the surface of the cancer cells and every cancer cell has a specific surface receptor or antibodies you can functionalize or core the nanomaterial surfaces with that kind of antibodies and you can load the drug in the core of the nanoparticles either through incorporation method or either through the adsorption method and once you are taking either systematically or orally that medicine so these nanocarriers that are carrying the drug will goes towards your target they will not open up anyone to the cell they are specialized surface specialized with the antigens of the cancer cells so once they will find the cancer cells through your circulating blood circulation then they will open up or their matrix is biodegradable it will be dissolved and it will release the drug to the target this is the active targeting now what is the passive targeting in passive targeting we didn't functionalize the nanomaterial surfaces with the surface receptor of the cancer cells or any other disease in which just we work on the retention time specifically in case of cancer like we all know that the cancer cells they are fast growing cells and when they are fast growing then their vascular system they are poorly developed they are not fully functional because they are dividing after every minute they are so fast their application or multiplication their vascular system is very poorly developed poorly developed so what we do in that too nanoparticles that are using nanocarriers that load our drug either you need score or we absorbed the drug on the surface we further cap it with hydrophilic molecules like one of the best example of hydrophilic molecules is the polyethylene glycol that is a highly hydrophilic molecule it can absorb 99% water that is the same when we coat our water we take it to our circulation system and our body also has water it absorbs the water so the nanocarrier swells when it swells the circulation system is not easily passed out by veins and RT the normal tissues are functionally developed but when the circulation system comes through cancer cells they are poorly developed so they get stuck when they get stuck because their diameter is not enough to pass out the swell substances so nanomaterials reside there for longer time as compared to the normal cells and if they are residing for a longer time in the cancer cells then they release the drug there and at least reside for a specific time in which the drug can show their effect this is the kind of the passive targeting in case of the cancer so nanoparticles are currently using not only the treating the cancer for also the delivery of the different central nervous system diseases like Alzheimer's diseases Parkinson's diseases and many other diseases in which it's delivering the drug to their target cybernetic so basically the scientific names but the mechanisms behind them are the main chemical theories and that of the PPT and PDDP now the market launching names that can be different are not in my knowledge next they are also using for to treat ocular diseases like conventionally simple eye rashes or any microbial infection and their conventional disadvantage is that you put an eye drop and it leaves there is no retention time and if there is no retention time then the drug doesn't work so basically nanotechnology has worked on the retention time of the drugs that if you are taking then they can increase the retention time and if the retention time is increasing it means that the frequency of the dose is decreasing and they are the most effective and they like to increase the angiogencies and to remove the plague from the arteries and veins different kinds of nanomaterials have been formulated similarly the stunts that are used to treat cardiovascular diseases the stunts have been coated with different kinds of nanomaterials which are the strongest antimicrobial agents and prevent from infection after post surgery these are the some biomedical applications of the nanomaterials this was all about what is nanotechnology what is the main roots of the senses of the nanoparticles and what are their general applications and what are their biomedical applications now coming towards our specific topic research based topic that is light and effective stimulator for our plant mediated senses of nanoparticles and their biological applications those five minutes okay that was the main topic actually I could not proceed without introduction because I think so that audience is not from the nanobiotechnology therefore all should have some little bit information about nanotechnology basically this is my research topic I am working on light light is an external effector like I just discussed that there are different factors that affect the synthesis of nanoparticles that are important factor of light I have seen that there was not much work on light then I thought we should explore the potential of light different intensities and different wavelengths of light how they can affect the synthesis of the nanoparticles and in terms of their control synthesis that how they control the shape of the nanoparticles how they control the size of the nanoparticles then I have done a series of experiment on lights short of time one was to check the effect of UVC light this paper is published in international journal of molecular sciences impact factor 5.9 in 2021 in which we have studied the effect of UVC light what is the impact of UVC light on the synthesis of monometallic one was the silver nanoparticles and other was the zinc oxide nanoparticles and also on the synthesis of the biometallic nanoparticles in which the nanoparticles silver, bi-zinc oxide nanoparticles were synthesized in two different ratios and UVC light a strong impact on the shape and sizes of the nanomaterials like just to give I had to discuss all these papers in detail but I have just give to you an idea about these are the characterization techniques XRD to find out the crystalline nature edX results to find out the elemental composition of the particles these are the same images of the synthesized nanoparticles like in figure 9 A, B, C and D A is representing the control silver nanoparticles that have been synthesized in dark without any exposure of light and B represents the synthesis of the nanoparticle under UVC elimination for 2 hours and you can see a difference in the morphology of the nanoparticles nanoparticles silver nanoparticles which have been synthesized under the influence or radiation of the UVC are more dispersed no aglomeration with smaller size and clear spherical shape as compared to the irregular shape of the control silver nanoparticles similarly the C and D represents the zinc oxide monomaterallic nanoparticles in C the control zinc oxide nanoparticles which have been synthesized in the dark and in D it represents the UVC radiated zinc oxide nanoparticles UVC has changed the shape of the nanoparticles and very well defined orthorhombics zinc oxide nanoparticles have been synthesized after the exposure of the UVC light similarly in the case of the bimetallic nanoparticles A represents 0.1 by 0.1 ratio of silver and zinc oxide nanoparticle in control conditions and D represents the under UVC condition these ratios didn't show so much promising results as compared when we change the ratio of 0.1 to 0.5 as in case you can see the C and in the D in the D figure you can say there are two different types of material one is needle type that is silver and one is other again orthorhombic type that is the zinc so that have clearly differentiated the both types of matter present in the nano materials in which when we further characterize them after XRD analysis then they shows the zinc oxide was the most dominated in case of bimetallic nanoparticle and it has masked the effects of the silver this is the size of the nano materials calculated by the XRD data in which you can see that there was a pronounced effect of UVC on the synthesis of UVC mediated silver nanoparticle was smaller in size than the control that was 91 millimeter while in the case of the UVC mediated nanoparticles it was only 22 and these impact was seen in all of the UVC mediated nanoparticles were smaller than their counterpart control nano materials after synthesis of the nano materials and after their characterization these monometallic and biometallic nanoparticles were evaluated for different kinds of biological activities like anti-diabetic activity was evaluated by checking the inhibition of the two enzymes one was alpha glycosidase inhibition and alpha myelase inhibition which are involved in the breakdown of the carbon hydrogenation of the glucose if you inhibit these enzymes then it means that you can control the production of the glucose in body the nanoparticles shows very promising results in inhibiting the these both kinds of enzymes like in figure 11a alpha myelase and alpha glucosylate similarly the anti-glycation activities was evaluated in which the advanced glycated end products inhibition was monitored like vesperine like and pentosidene both types of advanced glycated end products inhibition was monitored by exposing to a specific amount of the nanoparticles and the UVC mediated nanoparticles shows the highest rate of inhibition either in case of anti-diabetic enzymes and either in the case of anti-glycation enzymes and at the same time their biocompatibility or toxicity was also evaluated by performing two asses one was the percentage hemolysis of the human red blood cells and other was the LC-50 value of the brand-streamed LRY asses and in both cases they show very least toxicity to the red blood cell let's out of 100 only 3% hemolysis of the human red blood cells was observed it's mean they were 97% cell bar red blood cells were surviving and nanoparticles are biocompatible you can use in your in vivo systems for delivery of different kinds of drugs at the end anti-cancerous activity of the silver nanoparticles of both types monometallic and biometallic was evaluated by using their by evaluating their anti-cancerous activity against the hepato-carcinoma cell lines, hep G2 cell lines in which we have worked on the 5 factors firstly we have evaluated the viability of the cancer cells then we work the production of the reactive oxygen species and reactive nitrogen species then we have also checked the mitochondrial membrane potential of the cancer cells and then finally what is the mechanism how a proxies of cancer cells is done by the nanoparticles by checking the gene expression of caspace 3 by 7 gene and also evolving its activity and in all these results nanoparticles that were synthesized by UVC light due to their smaller size and unique morphology proven to be the best as compared to the controls next after that experiment I worked on the effect of UVA and C both first I evaluated the effect of UVC we have also done the experiments with UVB but there was no result of the UVB therefore we have skipped and we published this paper in 2021 in pharmaceuticals with impact factor that is the effect of UV radiation A and C in which biocompatible zinc oxide nanoparticles were synthesized and their different biomedical applications were evaluated due to short of time I will not discuss the results just on the same pictures in which the A and B represent the control zinc oxide nanoparticles C and D represent under UVA elimination and E and F represent under UVC elimination and in this research study again the UVC was the most dominating light that can impact the morphology of the nanoparticle in terms of their shapes and also in terms of their sizes this is the characterization and this is the anticancerous and biocompatibility evaluations of these nanoparticles and now currently I have completed our startup research program of HEC in which I have evaluated the effect of LEDs light in which 5 different LEDs green, blue, red, yellow have been checked on the synthesis of the silver nanoparticles like this is the different ratios first we go through the optimization of the synthesis of the silver nanoparticles then characterization by UV visible spectrophotometry then LED mediated we took control of it without any exposure of light and then give the 30 minute exposure to different LEDs like green white, yellow, blue and red and the UV visible results shows that initially we have set the time for 120 minutes for 120 minutes UV light exposure, sorry LED light exposure to check the stability when they tolerate and stay stable they maintain their morphology to 1D and D, red was the blue and red the peaks become distorted they become too unstable therefore we have decided the 30 minutes was the approximate time in which all of the LEDs light were stabilizing their nanoparticles now there is the FTIR analysis SEM analysis, silver nanoparticles of all kinds of LEDs almost shows the spherical nature one type almost spherical there were some refined like in the case of B picture that was the blue LED mediated silver nanoparticle was amongst the smallest size of nanoparticles synthesized among all LEDs as compared to the others this is the XRD analysis this is the size calculation next EDX analysis then their biological activities anti-cancerous activity biocompatibility, anti-glycation and next is the summary, so basically I am working on the light, how different sources of light can affect the morphology of the nanoparticles and then further these modified or controlled nanoparticles how can work differentially with the biological applications of the nanomaterial, so as a whole we can say that nanotechnology is a booming field, it can boost the economy of any country, new therapeutic solutions, it has given new therapeutic solutions to co-op diseases and open new frontiers for research in every domain of science nanoparticles can be engineered by controlling different parameters in which light prove to be a very good physical factor for trailing of nanoparticles these are the references thank you any question thank you I am Jahan Zayb, serving as a lecturer here in department of biotechnology thanks for such wonderful talk and wonderful articles in high perfected journals thank you no doubt nanowire technology is a paradigm shift in the area of molecular medicine but let's talk about some facts thousands of papers have been published on the toxicity of the nanomaterials nanoparticles but you see due to high EPR effect of nanoparticles and due to their size they get accumulated in tissue and organs and creating lot of toxicity and damaging the organs so what can be the strategy to resolve the toxicity problem and to make them capable for enhanced therapeutic effect generally we talk about nanomaterials if there are more advantages and disadvantages no doubt thousands of papers have been published on the toxicity of the nanomaterials causing in vitro like they can accumulate in our body so one solution on which the scientists are working is the formulation of biodegradable nanomaterials like lipid nanostructure materials lipid conjugate nanomaterials which are biocompatible to human body they can be degradable and if they are degradable they will decompose in the body and will not cause any potential toxicity to the human body but still they are causing toxicity to the environment and researches will find the solutions to cope the toxicity of the nanomaterials thank you thank you yes sir lipid nanoparticles yes, nanostructure lipid carriers as a carrier we use or as a drug we use as we read that reactive oxygen species can accumulate reactive nitrogen species yes sir, absolutely oxidative stress causes and lipid species can accumulate so are these lipids which are coming through nanoparticles also cause the stress on the cells yes sir, they can cause I have not so much knowledge about this but I think so they can cause because these nanoparticles when we are introducing in the metallic nature then obviously if they dissolve in the body they will regenerate the oxidative stress either in the form of reactive oxygen species or either in the form of reactive nitrogen species but if they are covered in the biodegradable materials might be this toxicity will be reduced and my second question was that you use the UBC what was the reason to use the UBC it is a shorter wavelength and it is a dangerous one sir, it is a dangerous one basically the main purpose was to evaluate the range of UVA or wavelength as compared to both B and C or UVC so basically my aim was to explore either what light or what intensity or what frequencies of the light affects the shapes or sizes of the nanopaterials like I have skipped the mechanisms involved in the light in which we have discussed the shorter wavelengths being the more energetically absorbed or captured more effectively to the nanoparticles and the valence electrons of the nanoparticles are excited to the conduction bands and they affect their surface or optical properties or surface plasmon properties that might affect the shapes and sizes but what is the accurate, this was the proposed mechanisms that we have proposed but the accurate mechanism I have didn't find how light accurately or precisely affecting the morphology of the nanomaterials Thank you sir Thank you so much Dr. Sumeda for such a comprehensive lecture Ladies and gentlemen this brings us to end our second talk of the day let's take a lunch break and resume the session at 2pm Welcome back ladies and gentlemen we will now resume our second training session of the day for this session we have invited Dr. Mariam Javed she is young and prominent researcher in the field of animal genomics she is serving as associate professor in institute of biochemistry and biotechnology university of veterinary and animal sciences ultimate benefit and sole purpose of her research is to raise the living standards of focal farmers she has identified various novel genomics regions which provide useful insight into the unique genomic architecture of our indigenous animals she is contributing substantially to the field of animal genomics in Pakistan today she will deliver the talk on climb animal narrative a verge of balance between devil and deep blue sea Welcome Dr. Mariam on behalf of virtual university of Pakistan please proceed welcome first of all thank you very much for giving me this opportunity to stand in front of you and share a few of my work with you actually it's not going to be a typical lecture or presentation that I have prepared today it's the kind of interaction with all of you guys I'll share a bit of what I have done already and I'd like to listen from you what you actually are doing so maybe together we can develop a collaboration or we can find the solution of the problems that we'll identify in next few slides few years back I visited I visited Netherlands and the theme of the year was the climate change and global warming and all that before that visit I never took that topic seriously in my life and I was like it's okay fine we all are doing fine so what is that hype about the climate change but once I attended that conference I came to know that this problem is real and we need to find a solution to that and since then I have become an advocate of the problem and today my topic is quite relevant to that the topic of my presentation would be climb animal narrative and we'll try to identify the verge of balance between devil and the deep blue sea so this figure would show the population rise and challenges we are facing so far for the provision of safe food in developing countries specially Pakistan in first figure you can say that how you can see that how the population has been increasing day by day and since like 5 years or 6 years in past 5 years or 6 years you can see that there is a very sharp trend of increasing the population in our region and in second part you can see the trend of the population that is increasing in different provinces of the Pakistan but if you compare the figures that are present on the right side to the figure that is present on the left side do we have developed the resources to cover the needs of the increasing population the answer would be no definitely with increasing the population there is increase in the pressure in terms of food in terms of supply of the safe food towards the population to make them healthy but this is what we are fail to do and this is the area which we need to focus in our future research according to an estimate the demand of the livestock products by 2050 is expected to increasing drastically if you compare it with the years before you can see that we don't have much dependency on the animal based proteins but with the increasing years with the passing years that the population is increasing we are more dependent on the animal based proteins and unfortunately or fortunately we are part of the society that we love to eat meat and we are one of the nations in the world which love to eat meat in their you know daily their daily food and it's not only meat we are also dependent on the animals in terms of other products as well like we eat yogurt, milk, egg and there are many many products which are which we are dependent on to get energy and that is why in recent years our demand for the animal based products and animal based proteins has been drastically increased and it is going to increase even more in next 25 or 30 years so it's a big room, it's a big area that we need to develop our strategies in such a way so we can develop the new animal species which can work the way through and we can develop or we can cope up the need of the population by increasing our production capabilities this is the world map in terms of total cattle population because this is the major species which we are focusing for the production of meat and you can see that US is contributing a lot in terms of 15% of the total cattle meat produced in the region and then is the Brazil then is the other European Union and China and Africa and India and somewhere we are actually existing in the rest of the world portion which is 13% although we are having the best animals in the world although we are having the best species in the world, best breeds in the world but still our contribution in the global market is very limited I think this is because of the lack of strategy, lack of research, lack of genomic exploration of our animal species that's why we are lacking behind and this is the area that we need to focus in our future research endeavors the contribution and importance of livestock sector in Pakistan cannot be denied, we all know that the share in total GDP is much more than many of other countries like it's 53.2% so livestock is actually contributing in GDP about 53.2% and the share of livestock in the export of different goods it's about 8.5% and the families in Pakistan which are dependent on livestock in one way or the other they are 8.5 million families because Pakistan is an agrarian country and many of the rural families sell their animals and they actually deal these animals as a expensive commodity if they are in the need they will sell their animals and they will get the cash or they will sell their products and then they will run their families in terms of finances so they provide raw material for the industry as well and then social security for rural poor families and then security against crop failure in Barani areas but what challenge do we face once we talk about the rearing of livestock this is to increase the food production but never compromising the climate that we are affecting actually in the face of climate change so we need to develop the strategies that we can increase the production of the animals but not on the face of climate change which is already affected severely by our various activities while reducing the carbon cost of the forming or carbon footprints that animals actually are delivering in our environment with lower intensity and taking more land because there is not enough if you travel in the premises of Lahore you will see that many of the agricultural lands have a board outside and which is written that it means that we are changing our trend as an agrarian country to the residential we are changing the land which has been restricted and which should be used for the agrarian activities and we are now shifting those lands towards the residential plots and maybe it's real estate is a big business in Pakistan now and what would happen if we keep going like this like in 10 years or in 50 years in coming years what would happen that we lost all the lands that we have already designated for the agriculture facilities and we will be dependent on the other nations to get even the basics that we are actually growing and we can grow in our country so challenge faced by livestock this is what is on the other side the one thing is on the one side the benefits that livestock can contribute but there are different challenges as well which livestock is also facing contrary first of all is expending demand and supply and trading okay do we have developed our species and breeds so well that they can cope up the need and feed food energy competition do we have prepared them accordingly the livestock diseases do we have develop the strategies or vaccines or treatment policies to prepare on our animals for the future infections or safe food supply safe food supply is another very big concern because adulteration issue in Pakistan and in the developing countries is very increasing day by day and last but not the least is climate animals are interacting with the climate and climate is affecting to the animals so both are trying to find their ways to mingle in the middle and to find something that works for both impact of livestock on the environment but it could be how livestock can impact the surroundings that you live in what do you think do you have any idea that would they affect or impact on the environment yes they do actually they are consuming the food they are consuming the fodder that land can be used to grow more of the cash crops but still you are forced to use that for the livestock and in terms of methane emission they are doing a lot they are doing secretions and their burp their fart and all the gases that they are separating the environment they are increasing the temperature day by day and it will cause the global warming one day so carbon dioxide methane and nitrous oxide these are three of the main players which are actually contributing negatively towards the environment and this issue needs to be addressed by research and thorough study it has been estimated that livestock alone is responsible for 18% of all the global anthropogenic GHG emission and you can see in the what happened inside of the animal that from the behind from the mouth everywhere the methane is actually increasing in the surroundings especially in those areas where animals are being reared or animals are being formed so what is the situation of forms which are having animals and how their environment is actually differing from the environment in surroundings the water we are getting in that environment where animals have been reared it is more polluted its indexes are very high the animal waste from the factory forms have been have been polluting actually the underwater resources the livestock produce a large manure each year and it is a very big challenge that how we are going to use that and what this can do with your environment and livestock pollution these skills fish and contaminates the drinking water and this not only affecting this is not only affecting the animals but as well as the human health some big numbers which are quite big concern is that 20 billion domestic animals they do exist globally and 45% of earth surface actually occupied by livestock system it means that we are only having 55% of the earth surface where we live in the 33% of the global crop land used for feed production and that feed is basically consumed by animal so this is a very big share in total cash crops if you counted and 8 to 18% of the global greenhouse gas emission is because of the livestock gases and 72% is the reason of the deforestation and 30% of the global freshwater consumption has been suppressed or has been compromised because of the pollution that is caused by the mineral the other side of the balance as we discussed earlier that if they are contributing negatively but still we cannot deny their importance in our environment that Pakistan is one of the countries who are actually quite reliant upon the protein based source which is coming from the animals if you see in the graph the maroon line depicts the USA and the very light line depicts India even still you can see that many of the developed countries they are dependent on the animal based products and demand of the meat it's very high in those countries and that figure shows that contribution of total meat production is about 37% and buffalos are contributing about 21% and sheep and goat they are contributing about 26% and 21% and rest of the meat is coming from other sources so what are socio-economic benefits of the livestock livestock is significant global because of value-added products and meat 1.4 trillion dollar industry and livestock industry is organized in long market chain that provide income and employ at least 1.3 billion people so it's a very big number and livestock GDP which we have discussed previously as well it's about 30 to 40% and in case of Pakistan it's even more which is 53.2% livestock important as a risk management tool for the poor and livestock is also key for nutritional security about 17% of the global kilo calories and 33% of the protein it is coming from this source so these are also deliverables in Ethiopia, New Zealand, India, Brazil, Netherlands, China and United States where how much protein actually it is an input-output table that how much protein we are giving to the livestock and how much we are getting from it and finally we have calculated our ratio of both giving an input and output and you can see that in many of the countries the output is very high and input is very low right it means that they have successfully established a forming system where they are getting more investing little so how we can steer a balance of both sides that we can get maximum benefits out of the livestock but without compromising the environment and climate which they are affecting negatively so this is a big question that what we can do in this it arises over the year is can we steer a balance between livestock good and bad and the answer is yes and how we can do that there are several ways I'm not going to read this out we'll keep it safe for next few minutes but few of the methods that I would like to share it with you it's artificial meat artificial milk or international or global market is actually shifting their focus towards these two entities and then it's n fixing serials which are being under trials and then germinants producing less methane this we can do by using the advanced genomic methods so I'll like to give a break here to you all and we have actually identified the solution in our first half so I would like you to give me a solution for that we have identified a problem that livestock is actually how they are contributing and what are different problems that we are facing and how we can develop the suitable research based strategies this is what I want to listen from you so I would like you all to get a piece of paper and try to give me a research idea which can be used to solve these issues can you do that for me can we do this because till now we have all the blame I'll start in Urdu till now all the blame we put on the animals as humans haven't done anything we are running cars so we will consider the standard or aesthetic and we are taking the factor of livestock and we will try to develop a project or we will try to develop a research based strategy with which we can bring some impact and make the climate better so I would like to listen from all of you you have 5 minutes on it you are talking about a specific project but if somebody is doing a research of the project and you are working on it how can you develop a strategy based on research and based on the of the challenges that you are facing we have been searching for some research which I think is quite audible we are searching for some research which I think is quite audible what are your suggestions about this project hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello hello The same was I was going to share that, no, I'm from G.C. Women's University, Fesla Bar. So I think we can cultivate the bacteria utilizing the methane. Methane can also be used as a biofume. So I think some interconnected technology must be used so that these waste can act as a substrate for some future technology that, yes, so recycling, you know, recycling we can put these methane and carbon dioxide to recycling so that the other things can get energy as a source of carbon and then we can increase some biomass and we can make some useful products. So the microbes there are also we can use them as a protein source. There's the feed additives. So by utilizing the methane, we can grow the methanotrophs, the bacteria, which can use this methane and then that biomass can serve as a energy and carbon source as a feed for the poultry, for the fish, and I think the puff nose as well. Thank you. Assalam-o-Alaikum, I'm Dr. Salaam. Okay, there could be many genetic solution to improve the livestock production, but as your topic was related to environment. So I think the main feed that the livestock take is from comes from the agriculture crop and pesticides and herbicides are one major factor that may cause toxicity or may cause a reduction in the cattle or livestock. So one matter, cure, not cure, one remedy could be that instead of using chemical-based pesticide or herbicide, we can use bio fertilizers or bio-based material so that they can be biodegradable, less toxic to the animals and improve the livestock. Also, we can develop, we can use cell culture to develop artificial meats, like we can culture chicken cells in lab and we can use that as artificial meat. And I think this has been done in some country in which there started serving cell culture based chicken in their restaurant. I don't remember that restaurant exactly, but this is doing, this is done there. Yes, this is happening, but not very proved kind of the methodology. Ma'am, we need to go to the waste management that the animals, because of the waste that is in our environment, wherever there are farms or any place where animals are being treated, waste management can be done there too, like those people who are less dangerous or you can say less toxic, they can be done by using some microbes like nitrogen-fixing or something like that, so that they can consume the manoeuvres that are present in them and deplete it. This is a very big concern actually, because a lot of the bulls are investing in where to find their trash. And few years back, there was a petition in the court, because a resolution passed in which we preferred chasing the trash from Australia and we are giving some of our deserts because they don't want to use their land for that. So it's a very petition we are talking about which is probably, but still there are a lot of plants countries to hand over, maybe it's for now, because they are doing it, they want their animal-based trash or general trash whatever they are having. We are transferring that into the developing countries or countries which are to thrive and they are providing resources to the quality of protection. Ma'am, it's not the time yet, right? We are at the Elisabeth stage, Inshallah, we will come on time. Assalamu alaikum ma'am. Ma'am, I read a little bit about the facilitated cows. The facilitated cows are there, you will know better. It is said that artificially fermentation takes place in them. The result is that methane production also reduces, the second is that milk production also increases. The second commonly used is that the dung is collected by a big biogas, then it is taken to the biogas, then the spheri that comes out later is used as a fertilizer, then the dung is finished, because during biogas production, fermentation is done, then everything is cleared. The third can also be done is that we feed the cows, the people who are related to the village, they have seen a culture of farming, so the cows and the animals, the livestock, it is broken down. There is a lot of carbon source in it. These ultimately become cows for methane production. So we can make a modification in four, we can make the component that ultimately becomes cows for methane, or those things that are environmentally friendly, it can be replaced. There are four modifications in it. These two or three are very simple and basic. Solutions are to work this problem. Which can work for both hands. You can increase the production of the phenomal. You can reduce the carbon footprint that they are causing. Excellent. Thank you. Anyone else wants to do it? Or you want to share that you are already doing it? Assalam-o-Alaikum. My name is Abraha Yusuf from Banaji Department. I read an article two years ago. Actually, they used the gut of the fish to produce alkaline protein. I mean, they used it as a substrate. The alkaline protein is an industrially important enzyme. Yes. Similar studies, I think in Juwas and maybe in Punjab, they used the chicken gut to produce alkaline protein or any other protein. They synthesized it. If you are talking about the cattle, I don't know if it's an oxo or not. But it can be used to use the waste. Because it has protein in it. I am not talking about methane. If we throw the gut where it is, which is causing pollution, we can use it as a substrate which is an industrially important enzyme. Your point is that instead of wasting all the things we are getting from cow, we need to use them in a way that we can get something good from it. Excellent strategy. Thank you. Anybody else? Sure. Is there anybody practically engaged with a project which is dealing with the environment or climate or all that? Okay, madam. Definitely, you have heard about silage. It is a fermented type of fodder. Given to the cattle, it is consumed in lesser quantity. Obviously, it will produce less waste. In Pakistan, it is ongoing. My husband himself is ongoing cattle production. You are not talking about marketing. Yes. I am not going to market the product. We have purchased silage. He told me that it is a fermented fodder. It is consumed in lesser quantity. You don't need to give the food in tons or in many kilos. It is given one or two times per day. It gives less waste. These kinds of technologies may help to produce less waste. If less waste, it will be environmentally friendly. Thank you so much. Thank you, everybody, for sharing your research experience and your thoughts with me. Okay. We have one more. If you go to the biotechnology or molecular side, you can do a lot of things. You can go to the genetic engineering side and modify the bacteria. But I think we should first see why we need livestock. We need it as a source of food. If we want to reduce the burden of consumption or livestock production, we need to reduce the role of it. If we want to use meat and milk, and go to the vegetarian side or vegetables side, we can control it on a basic level or initially. We can do the same on crisper, genetic engineering and fermentation. But first tell people how much production you need to do, whether you want to do it or not. I think it will be better. You can leave the food. This is excellent. If we go to the vegetables, there are a lot of diseases due to meat. This transition is actually very hard to induce. In many of the advanced countries, they are working on these trends for being vegetarian and removing the meat totally from your diet. There are many specific diet plans which are actually encouraging what you are saying. It's really hard to convince people to do that. But this is again one of the things that we should do. Let's hear from the back first. You were going to add something in the back. The sources are present. You don't take protein, right? Yes, yes. It's a protein source. If you start taking protein from plants or those algae species with high protein content, you can easily replace it. Yes, we already have alternatives which we can go for. If you are in Europe or America, I will give you a small example. There is an algae, a blue-green algae spiral. Check its protein content. It's more than 55 percent. It's more than your broiler. It will be more than that. It's very easy to get rid of diseases. It's very easy to get rid of diseases. Anyway, this is a debatable issue. The benefits and pros and cons of it will be a long debate. Alternate means that it's present. What you are talking about is that this is a debatable issue. It can be thought over that people have to convince people to come this way. But this is not an issue that can't be done. It can be done because it hasn't started yet. People are using it in a tablet form. People are making cakes and eating it. They are using it in a vegetable. If we start, it will be easier. My comment wasn't that it can't be done. My comment was that it's really hard to convince people to follow the trend and create. It's important to cook the chicken when we have guests. It's important to cook the chicken on the wedding. So, meat is a very essential component of our social setup. Meat is an essential component of our diet. But diet is left behind. So, what we are serving is what we are. So, yes, we need a little frame shift. Otherwise, alternative resources are there. Madam, this is also here. So, like our normal flora, we introduce it in the lifestyle of livestock. You can use meat as their food source. Excellent. There are different strategies if we compile it. Some people cut it down at the initial level. Which is actually implementable. We can get benefits out of it. Some people say if there is a mission then how can we take it towards biofuel. It's an excellent strategy. We don't need to work on it in the coming 50 or 25 years. We will be deprived of it. Many of your natural biofuels or your natural sources of fuel are reducing. The world is on the verge of scarcity. Some people say we need to introduce certain elements which can reduce the emission of the meat. If we conclude the session. Excellent talk. Thank you for participating. What I have done actually it's an indirect approach. Indirect means that in US what we are trying to do is enhancing the genetic potential of our animals so that they would be producing more milk. They would be producing more meat. They would be very friendly towards the environment. If we look into the sub-tropical zone of the world in which Pakistan is actually also relying. There is a high humidity around the year. In terms of extreme temperatures in these regions, the temperature is very high. There is frequent precipitation whenever there is high temperature precipitation rate would automatically increase. There is high humidity again due to the high precipitation. Four seasons. These are blessings as well. But still the span of the summer is much larger than the summer season. Then diverse flora and fauna and finally populace. These areas are quite populace. The population of animal. The population of human. Both is very high in this area. It means that demand and supply is also very high and we face a lot of challenge in feeding the mouths that we have here. Extreme weather cause food scarcity. High humidity provides excellent environment for infection as the hot and humid environment is very good and high population load requires nutritious dietary sources to be explored so that we can feed the high population that we are suffering. But whenever there is a problem the nature has always blessed you with a solution you just need to find out. Nature has blessed this region with the kind of animals which are well adapted, which are efficient which are active and which are productive. And we can call these animals a survivor. Another list is buffalo. I have put this animal on the top because I am working on this animal but actually buffaloes are quite unique among all because they have the ability to convert poor quality food roughages into the excellent products and in terms of meat, in terms of milk the production potential of buffalo stands out among all the animals that I have put down. The next is cattle. In Pakistan we have 16, 17 cattle breeds which are quite unique in the Pakistani cattle here which is considered to be the ancestor of all the cattle in Asian region and in the region of Central Asia specifically. Then we have sheep breeds goat breeds and then is the camel and in Pakistan actually we have flavour of all. If you see USA, if you see Australia there are few species that are existing there. I mean camel is in Egypt Australia and New Zealand are more of the goat and sheep kind of countries in the US you will see in Texas especially you will see the cow but in Pakistan we are having all of these species and we are getting benefits from this. The camels they are the animals of the future because they are well adapted to the warmer environment and finally is the yoke yoke is well adapted in or hilly areas. River buffalo which is an animal of choice for my research and animal of choice for this region as well why because it is highly adaptable to the warmer climate condition and it is carrying the high production potential and it can consume the poor quality roughages you don't need to feed the animal with high concentrates or high quality feed even if it can give you high production in terms of milk yield itself or in terms of the fat percentage that is present in the milk and it can give you valuable products like milk and meat. If you see the table which is depicting the data you can see that buffalo milk stands out in terms of calorie count and in terms of fat percentage and in terms of calcium and mineral count and if we will see the meat composition table you will see that fat percentage is again very high but these are good fat because cholesterol is lowest in the bufflos and secondly we are having iron content which is very high in buffalo meat so due to the productive potential this animal is called black gold of buffalo and we are lucky to have about 42 million heads in terms of population of river buffalo or Neeli Ravi buffalo and Kundi and there are different breeds and it's contribution in total milk produced in the region is about 68% it means that major milk which is coming into the market is from buffalo and in terms of meat it's contribution is about 24% so now question here is if the production is good potential is good then where does the problem and why these animals are still suffering or why we cannot get the maximum benefit the reason is that bufflos are poor in their reproduction they are shy breeders and their reproductive performance of fertility is quite pure poor in terms of Easter cyclicity in terms of age of the maturity bufflos are called late mature because they mature late if you compare it with the cattle and their Easter cycle is not very regular if you compare it with the cattle and the animals are repeat breeder in terms of their reproduction and fertility and their Easter signs they are not very visible not very prominent so these are some of the challenges that farmers are actually facing to get the maximum to get the next generation from these animals so if we cannot maintain the chain of the cyclicity in terms of Easter cycle then it's very hard to maintain a viable buffalo form second main problem is the recurrent infections in the animal and the main out of them is bovine tuberculosis this is one of the most widespread infectious disorder in Pakistan and in recent year FAO, WHO and OIE they have classified Pakistan as one of the nations where tuberculosis is neglected and it has been estimated then in Pakistani bufflos the prevalence of tuberculosis it's about 33.7 and it means that every 3 out of 10 bufflos they are affected by this and what possible reason can it be that it's really hard to draw a line we like to interact with our animals if we visit the villages you will see these scenes very frequently and we like to interact with the animals physically and that's why if animal is infected then they are crossing the barrier and the issue of zoonosis is there and infection is actually traveling from bufflos to the human side so this is the red zone there are some of the scenes from the slaughter house and what would happen that animal with tuberculosis would come into the slaughter house and people they will remove the lung they will cut that part which is affected and then they will sell the rest of it and we don't know that what is happening and what type of meat is actually there which we are eating right so this is a very big problem that animals are infected and we are consuming the meat of infected animals so two main problems first of all is the poor fertility and this is the overall poor status in terms of health but there are good things as well that our river buffalo is quite unique in its genomic architecture first of all is the high heterozygosity many of the low side that we have studied in university of veterinary and animal sciences so far their gene structure actually supports the high heterozygosity many of the regions are highly polymorphic many of the genes are highly polymorphic they give you actually a barrier for the selection of the superior regions and then there are certain mutations which are transversions then there are certain mutations which are transitions both have been reported in buffalos definitely transversions are less, transitions are more and then we have also identified certain residual truncations in proteins which are actually affecting the function of the proteins and then diversity in certain genes is medium to high and some of the low side they are monomorphic it means that they are less informative and most of the mutations they are in line with Hardy-Wainberg equilibrium which gives you an excellent opportunity to put the animal through the selection process to get superior animals through marker assisted selection so genomic investigation in US actually comprises of dairy, meat, fertility and disease resistance traits and these are different genes that we have explored so far and in dairy you can COLR1, CYP 11B1, SCD all of these genes majority of them are related with the fat percentage in the milk majority of them some of them are actually responsible for the overall yield of the milk in meat this is for the increasing the number of cells for fertility mainly we are actually targeting the silent estrus behavior and in disease resistance these genes have been explored in animals with bovine tuberculosis so success of buffalo production system is primarily dependent upon the efficient reproduction system and for this we need to manipulate the environment as well as the genomics in environment it comes nutrition season age, week and disease and in genomics what we have different genes I have mentioned few of the candidate genes this is the data of the AVP which is arginine visopressin gene which is precursor gene important for the production of estradiol and interestingly we have identified a simple you know deletion in one base and due to that deletion actually the functional moiety of the gene which comprises of 9 amino acid it has been changed and you can see that last two residues have been shifted and due to that shift actually the attachment of the protein to its receptor has been reduced and due to that reduction there is low production of estradiol and if there is no estradiol it means that estrus cyclicity would be highly compromised this is the population genetic analysis this is of the same gene AVP its paper is still submitted and total 5 polymorphic sites were being identified but kaif square value have been depicted that mutation number 2nd, 3rd and 4th they are in line with Hardy-Wainberg equilibrium it means that these mutations are quite fixed in the population they are showing the trends which are not changing generations after generations it means that they are good marker for the association analysis and this is their association analysis the score of significance of these polymorphic sites it shows that polymorphic site which is showing the deletion which I mentioned earlier it is actually very high it means that it is impacting the trade in a very big way so we have identified 3 genotypes the wild type genotype then we have mutant form and then we have heterozygous form and you can see that score of heat it is quite different in all 3 genotypes so it gives us a selection marker where we can choose the animals with the better trade this is the three-dimensional protein structure analysis actually and it is indicating the structure of the protein and you can see that secondary structure elements are quite different like alpha helix and beta strands and there is an absence of the TM helix in the mutant form so that helix is actually responsible for the binding of the protein and due to the change in those two amino acids which we saw previously the attachment of the protein with the receptor is hindered and due to that there is no production of estradiol that is why animal is suffering from silent estrus behavior this is the analysis of comparative genomics in which the river buffalo has been compared with swamp and mediterranean and you can see that there are certain regions which are very unique at two places you can see the deletion in total in swamp and mediterranean but in mura which is quite near to the swamp and mura is from India but they are having very unique region there this is the analysis of phylogenetic relationships exon distribution in gene and motive patterns and you can see that in cattle there are pattern which are different and the same pattern has been followed by swamp and mediterranean but in terms of mura buffalo you can see that pattern is totally different and conserved domain region and especially see motive pattern at sea position the motive pattern the protein is highly diverse and protein is changed if we talk about mura so this is the truncation of the protein right but it includes the 9 amino acid moiety and there is a variation in that as well and this is BB genotype in buffaloes the rest of the animal species they are following the same pattern as in the standard the second work we have done in the tall like receptor one these gene actually was being explored for bovine tuberculosis and its immunity natural immunity in the animal and we identified total 5 SMPs even in this gene and there is also a stop codon isoleucine was converting into a stop codon and due to that stop codon there was again truncation of the protein which has been observed in the cattle it is the genes that we have selected so far it is quite common that you find some of the regions that deleted and then or maybe they are converted into the stop codons so unique genomic variations have also been identified in interferon gamma gene in this gene we have identified total these SMPs and you can see that proteins are also being affected functionally due to the presence of these proteins this is the association analysis and this analysis indicates majority of the regions they are obeying the Hardy-Wainberg equilibrium means that association analysis shows that they are actually affecting the basic immunity of the animal so if we select these markers to select the animals with superior immunity trait it means that animal would survive the infection in upcoming life CYP11B1 gene this gene actually is associated with the milk fat content the fat content actually in milk is measured in two ways you can calculate the total fat percentage in the milk or you can actually calculate the total fat in kilograms as well the CYP11B1 gene was actually studied in Neeli Ravi buffalo for fat overall and there are different SNPs which are being identified and due to these SNPs you can see that there are different genotypes wild mutant and heterozygous genotypes and this provide us the opportunity that we can select the best out of it and if you see the scores the polymorphism this one this one this one this is highly significant and the fat percentage is up to 7.9% it means that if you select the mutant form of this particular dosai the animal can give you very high fat percentage OLR1 gene is also responsible for the fat content in the milk these are different variations some are transitions some are transitions which are being identified in the gene and here also you can see the first one and again the first one and the second last one these are the two SNPs which are actually showing great variation towards the trait then is the SCD this gene is also responsible for the milk fat content these are all actually production trait genes in this also novel polymorphic sites were being identified in the river buffalo and you can see that association analysis provides us with the high values in which we can identify those dosai which are showing more of the fat content and then we can select those animals for future breeding program the same polymorphic sites were actually put through the three dimensional protein configuration in which the protein structure was in silico constructed in silico and you can identify a certain transmembrane helix then you can identify secondary structural elements in the protein and you can see the sites or you can actually identify the sites of your mutation that how functionally they are interacting in the biological environment in OLR1 gene we have identified two domains one is steroid binding domain and second one is the okay these are two different motives and two different domains and you can see that your mutation is also actually existing in those domains it means that it is affecting the binding sites of the protein this is the SCD gene result and you can see that your mutation actually is existing in the transmembrane helix region so if we conclude these results being most diverse among the livestock species specially bufflos river bufflos need extraordinary attention in terms of research and policy to get the maximum benefit out of this astounding and phenomenal animal and you can see the diversification of this animal and its existence globally its existence globally green color depicts the presence of river buffalo in different regions the yellow color depicts the swamp buffalo which is more in china and where it is orange color it means that both of what type of bufflos are existing in those regions so this is some of the basic research that we have done so far in which we have identified certain candidate genes which are responsible for controlling different traits of economical importance in livestock and if we select the animals on the basis of these traits it means that we can make these animals better and these animals would produce better and these animals would survive in our environment in better way so what comes next is to use this knowledge to design more of the advanced projects that we actually are doing in US for the selection of those animals or for making those genes viable which are otherwise non-functional in bufflos so we have planned certain projects on the stem cells then micro RNA then we are also working on the genome editing and this is a lipofectamine that we are going to make a complex and then we are going to use it to treat the animal one of the research project that is in the progress it is with the collaboration of royal veterinary college and the Francis Crick Institute UK and student is also here it is sponsored by commonwealth and our title is study of the role of Kata 3 gene in cell lineage and differentiation and we would be able to silent those genes which are responsible in the poor fertility of the animal by using CRISPR Cas9 system. The second research project is about the variant transcriptome analysis in Neeliravi buffalo breed this is actually funded by ALP which is agricultural linkage program by Pakistan Agriculture Research Council Islamabad and they are also supporting this project and many of the students have completed their thesis in it one of the PhD student is still here who is working on this. And the last one is study of circulatory micro RNA as a novel remedial approach for the issue of silent stress behavior in Neeliravi buffalo this is also a funded project in which we what we are going to do is that we are going to isolate the circulatory micro RNA and then we are going to make a complex of these RNA with lipofectamine and then we are going to use that complex for the treatment of the animal and that micro RNA would actually affect the expression of the gene and we'll get the expression of our own will and finally the production and the fertility status of the animal would be enhanced so today's activity is actually related to this project what we are going to do in our next session is that we are going to use that database that how you are going to select your micro RNA if you are entrusted in a project like that that there are different databases just like genes where you can select the sequence of the gene just like that you can use different databases for the selection of relevant micro RNA as well so my student is going to demonstrate you in next 15 years so I hope you will copy Ajay please, Tikra if you have any questions so far you can ask or we can have the question and answer session in the end Thank you very much Thank you very much towards that slide in which you are showing us the effort was 7% so when we we were student and we were reading I am a student of economics so we know that cow has 3 to 5% fat percentage and buffalo is plus minus 5 to 6 so I was really happy to see the result of the 7% fat in the nearly Ravi buffalo and most probably at your Ravi campus you might have that buffalo available with you and you are in the process of fertility and you are as I was just listening to your cloning and other things so I wish this 7% fat percentage buffalo's genes and through embryo transfer technology or whatsoever the technology you know better because you are the expert on it so that our farmers can get the buffalo's having good quantity of fat in that milk because in our villages still they we love to have a thick layer of cream on the milk so we wish and you are the only university leading university of Pakistan on livestock side so we wish you people should give us a gift to the Pakistan with having the 7% fat buffalo thank you very much sir we are doing what we can actually and fat percentage is quite desirable trait you are right in Pakistan because in some of the nations people are quite concerned over the fat content and they are health conscious and they are moving towards the skim milk but not in Pakistan so yes it's not just one animal actually once we collected the samples we collected more of like 150 or even more samples and then we got some ratio so that ratio was very low there were very few animals which were touching the 7% or 7.5% majority of them they are like 5, 5.2, 5.1 you are right but thank you very much sir for your comments thank you so much so now we have Iqra she is going to demonstrate you about the use of the micro RNA software that how you can collect the data from that thank you ma'am please stop it you are the source of the internet okay Assalam-o-Aliakum my name is Iqra Khursar and I am a PhD scholar in the field of molecular biology and biotechnology under the supervision of Dr. Mariam Javed at university of veterinary and animal sciences Lahore the purpose of my presence here is to tell you about the software that I use for the micro RNA analysis actually micro there is a long list of micro RNA software for the micro RNA analysis available online they are freely available if I name some of them such as Mir-Base, Mir-Path PICTAR, Target Scan Mirvok and many more today I will tell you about some of them so let's start with the Mir-Base software it is freely available online no issue regarding its access I will write the name in the search bar okay this is home page for the Mir-Base database it is a biological database and it contains information about more than 1000 non published micro RNA sequences in various species if we talk about some of its main features is that micro Mir-Base database it provides a way to retrieve the sequence of mature micro RNA as well as the stem loop precursor which is its premature form because we know that the micro RNA has a mature form and its mature form is in the stem loop or the hairpin precursor so this software provides an opportunity that you can retrieve its premature form and you can also retrieve the mature sequence which is quite simple you have to mention the name of micro RNA in the search bar you can also use keyword or any other reference if we talk about some other features of this Mir-Base database it provides a consistent naming system for the micro RNA for the micro RNA and its naming system is just sequential it means that if we talk about mouse species if it has a large published micro RNA its name is 2 to 4 so the next published micro RNA will be named as 2 to 5 so the naming of this sequence and if you want more information you can visit the help pages for more information about the naming service other than these features this Mir-Base database it also provides experimental evidence means that it provides an open access to the papers in which this micro RNA is reported so it's a way that you can verify it you can verify your micro RNA you can also open access to the papers in which this micro RNA is reported apart from this if we talk about another feature it provides a link to the other databases this micro RNA will be reported so this way through this software you can go to the databases and you can verify the results of your sequence this database is managed by University of Manchester now let's see how we can retrieve the sequence by using this database simply I have selected micro RNA 132 so I will write its name here M-I-R 132 the next page will be open if you see it here it says your micro RNA 132 returned with 211 results so we have reported the micro RNA in various species if I scroll down you can see how long the list is and you can see further whatever your interested species is you want to see this micro RNA I have selected the first three letters D-R-E the first three letters represent the species name D-R-E D-N-O-R-E and M-M-U I will select this as I already mentioned you want to retrieve your STAMLOOP sequence if you are interested or you can retrieve your mature sequence actually this page it contains three information STAMLOOP sequence or mature sequence if we talk about references it provides experimental evidence system the references are telling you if we go back to this and see what kind of information we can obtain from here so basic information its accession number its symbol, description gene family literature search 178 open access papers in this micro RNA mouse species if we click from here it will access your papers these are papers it provides a way to verify you can verify deep sequencing information is also available here confidence its confidence level is high its confidence level is high means you can rely on the authenticity of this micro RNA if you go to the genome context if you go to the genome context if you go here the gene related information it is located in promise on number 11 its starting position ending position on the positive strand you can get this kind of information from here then we have clustered the clustered means that set of two or more micro RNA data transcribed by the micro RNA genes that are physically adjacent to each other means those genes which will be physically adjacent which are physically adjacent so the micro RNA they are in a cluster form like our targeted micro RNA which is 1,3,2 in the mouse species this cluster is 2 and 2 this means that these two micro RNA genes are physically adjacent to each other and this is why they are making a cluster then its further information on the positive strand and the starting position and the ending position okay as I already mentioned it provides database links through which you can see this sequence if you see here we have an interest gene if I click on it it will take us to the NCBI page and the NCBI page where we get a format information okay official symbol full name what is the primary source what is the organism what is the lineage if it has synonyms it is mentioned we can retrieve the sequence from here and we can also retrieve the sequence in the faster format from this software okay if we say in this software all the micro RNA which is reported here published we will also get NCBI so its not like that only the micro RNA which will be reported on NCBI will be referenced here my saying is that if I take any other micro RNA then its not necessary that I will also be given this same software in the database links the same software will be provided here means you can retrieve all the micro RNA from NCBI if it does not have a sequence then you cannot retrieve it from NCBI similarly you can retrieve the sequence from these databases the stem loop structure which is its pre-mature form after that we have a sequence of mature form mature sequence if we notice the micro RNA which will be written as a small r then this small r will represent its pre-mature form okay when micro RNA MIR will be written with small r so it indicates its pre-mature form but when we see with capital r then it represents its mature form and the mature form here we have 2 arms the result is 5p and 3p 5p means the sequence is retrieved from the 5 prime arm of the stem loop structure and 3p means the sequence is retrieved from the 3 prime arm of the stem loop sequence and means for every micro RNA these 2 sequence are not shown here here the 5p arm and 3p arm which is shown here is its purpose because the predominant state of these 2 arms is not determined if any micro RNA is pre-dominated if the form is determined then only that form is shown here but for this micro RNA 5p and 3p which state is pre-dominated or which is the abundance if we don't know then these 2 sequence are shown here and this information as we have seen in the stem loop structure this is the accession number which is replaced with a new ID if there is any micro RNA then it will be shown here for sequence we will click on the get sequence option and we can easily retrieve the sequence from here okay, deep sequencing information is also available here evidence as I said earlier you can verify your result by providing experimental evidence basically the information of the evidence comes from these 2 sources first one is experimental evidence which is experimental evidence like published papers and second is predicted homologues for the micro RNA and homologues are predicted on the basis of the sequence similarity folding properties of the stem loop structure on this basis the evidence is provided here and for example if there is any predicted homolog and it is not verifying the experiment then that information is not given here so the same information comes from the predicted homologues which is experimentally verified and the database links along with the other databases are provided predictive target the database through this we cannot retrieve the sequence we can predict the target because we know that micro RNA has to bind its target messenger RNA in order to downregulate it so how will we know that it is micro RNA and in which gene it has its target which gene it will bind and it will downregulate it so that information we can obtain from this software because it predicts the target actually this software is provided with the other databases so here for predicted target I click on this option MirDB is another online tool which is used for prediction target and here MirDB is a separate database but this software has a link we can access it from there we can access it separately here we have 30 predicted targets for this micro RNA in this database and here we have a table before we see what kind of information we can retrieve from this database we can see what information this table is providing target ranking target score mirna name the gene symbol which gene has its target the gene description the target score because the target score if you see all the targets the target score is from 50 to 98 its range is normally 100 so in this the higher the score it means you have more statistical confidence in the prediction result the higher the score you can rely on the result I will select this one further we have a page open in which we provide information what kind of information is micro RNA name micro RNA sequence we can retrieve it from this software as well previous name target score ncbi gene id if we click here we will get all the information of this gene gene symbol gene description but the important thing here is seed location what is seed location or what is seed sequence we say that micro RNA has to bind with its target messenger RNA in order to downregulate it but the binding actually between micro RNA and messenger RNA it takes place between in the seed region so you need to identify the seed region and this is the gene target and this is the location where the micro RNA will bind and the messenger RNA will either destroy it and its translation will inhibit and the gene expression will downregulate we can take that information from here and this software also tells us the location of this gene there are two seed locations and starting from 303 position and the other one starting from okay so this is our information you can retrieve the sequence you can get seed location information from this software okay then we have the mature sequence for the 3p arm there is a session number previous id sequence you can retrieve the sequence deep sequencing information the experimental evidence is the database links in the same way predictive targets so here since I have used a separate database for 5p arm so here I open target scan this is a separate biological database and here the table we have shown that these are target genes their transcript information can be obtained from here what is the gene name the number of 3 prime sequence which was used for the construction of the 3 prime profiling link to sites in utr if we click on it then we get this type of information in front of us the white color sequence highlighted region is white you can see these are conserved sequence this is evolutionary conserved sequence among various species like mouse, rat, squirrel, rabbit human chain so in all these species these are the sequence this is evolutionary conserved and these are the sequence with which our targeted micro RNA can bind further if we scroll down there are two tables one table is conserved and the other is poorly conserved so here since 2-1-2-3-p is shown because it is made with this information and this is their conserved this is their seed sequence which has binding which is shown and what is the type of site if we want to get this information then we will click on it and we get that information that what is the site its definition is an exact match to position 2-8 of the mature Mirna followed by an A this is seed sequence actually it is located at the 5 prime end of the micro RNA and when we have complementary binding then we call it as 8 mer site and further information which is a 7 mer site we can get this information and this is references information to verify and this is the database we have opened it we can also open it separately okay sir thank you so this was a small demonstration about the micro RNA software which we can use when we have to plan the micro RNA study how we can retrieve the sequence apart from this we can see the seed location information and the gene targets can be predicted so these are the softwares we can work through them so thank you so much thank you so much Dr. Mariam for the nice presentation okay so there could be two possibilities if you wanted to detect the MI RNA from any source either we can detect the mature MI RNA or we can go back to the immature form which is the men group form which form is better for a diagnostic purpose or for to characterize it actually both these forms exist the first form which is called the mature form it is that one in which there is no stem loop it is that sequence which is in fast A format and if you want to synthesize it artificially like we design primers and then we send the commercial facility or sequence and then ask them to make those primers and send us back this is how we use the fast A format of the MI RNA but if you look at the functional consequences inside of the cell it is not existing in the linear form it is in the form of a loop so that software actually is depicting both type of formats for you if you want to predict the functional consequences where it is going and how it is going to attach with your DNA you can have that form as well and you can have the fast A format as well if you want to synthesize it artificially what about the circulating MI RNA if you want to detect the circulating in our project actually we are more of exploring the role of circulatory MI RNA and they are premature form and once they reach to their destination then they got their mature confirmation and then they got bind with their target thank you so much thank you so much everybody thank you so much Dr. Maryam for the wonderful presentation and thank you Ms. Sikram for hand on practice on the software and thank you so much all the participants to our fourth day of the training program and see you tomorrow the last day of the program thank you