A great adventurous constructionist paradigm in education and research indeed!

SMITI SAY’S

:sparkles: Okay so as I’ve completed my 2nd year in BSc and in final year we have to submit a thesis, we discussed what topic i can work on and my interest is in Diabetes
:sparkles: So Diabetes is basically a metabolic disease which is cause due to lack of sufficient insulin or when configuration of receptor changes (acc. to me these are 2 possibilities) as reception is a protein and like enzyme and substrate bind to perform a process in metabolism to form product here insulin needs to bind with its receptor so that cell will get a signal to open glucose channel so that cells can take glucose to perform further process like glycolysis and kreb cycle to get energy
:sparkles: Here we also discussed about how insulin is produced from INS gene in pancreatic cell and then transferred to blood
:sparkles:Now in both the scenarios, either when insulin is less or it can’t bind with it’s receptor, glucose will not get transferred into the cell and it will stay in blood causing problems as it increases blood sugar level
:sparkles: Here the question is are there other possibilities through which diabetes is caused? And what causes diabetes type 1 and type 2?


:sparkles: Then @kiranayadav4894@gmail com SKYPE ma’am asked if we can study diabetes in model organism i was working it that is drosophila and it reminded me of an incident that happened that time
:sparkles: On 8th Oct 2021 i trapped and made my 1st culture successfully but media had some issue we when i joined cube we discussed on how to make media and developed BRSV (Banana Sugar Rava Vinegar) media which I used in my home lab
Here we also compared homelab media with standard corn meal agar media used in labs
:sparkles: Like how we make media using ingredients available at home by discussing what we use in standard media and why is it important and then replace them with what we have and would work in similar way like instead of agar we use rava here to solidify media and instead of propionic acid and orthophosphoric acid we use vinegar which is an acetic acid to prevent contamination i.e bacterial and fungal growth here
:sparkles: I used this media to culture my flies for about 8 months and when i trapped flies i was looking for drosophila melanogaster - The 6 time noble prize winning organism!

:sparkles: For identification of flies and to see if i have drosophila melanogaster in my culture, i made single line cultures but lost most of them due to mite infestation… Now here single line means when we isolated 1 gravid (pregnant) female fruit fly and culture her in fresh new media bottle and all her progeny are from same species
:sparkles: So when mites started destroying my culture by killing flies, i called Saida di one of my senior to ask what i can do to save my precious single lines and she told me that when similar thing happened with Aswathy di she used to transfer flies early and later on used larve from old bottles to save her culture
:sparkles: So i can use larva for saving my culture by adding 20% of sucrose solutions in the bottle and when larve come up i can isolate them
:sparkles: But that time i didn’t understand it much and i thought if we wash it sucrose solution then mites will go away so i had a bottle that didn’t had flies and larve were there so i took them out with bursh and washed them with sucrose solution and didn’t washed it again with water cause i didn’t knew that i should and after taking them out from solution I kept them on cotton and placed that cotton on new media
:sparkles: After 2 days i noticed that larve were becoming fatter or obes and i reported that in C2C and while discussing Saida di told this could be due to diabetes cause i dipped them in 20% sucrose solution i.e 20 gm table sugar in 100ml water
:sparkles: These larve took too long to become pupe around 6-7 days and were in form of pupe for like 2 weeks after 2 weeks the fly that i got looked very different then my fly and i thought this was some kind of mutation or diabetic flies might look like this but then when i posted a video @Batul CUBE ma’am told it’s a different fly from some phoridae family
:sparkles: So if my flies took around a week to complete it’s whole life cycle these fly took a month to complete it and when i came to know this i thought we can’t study diabetes in fruit fly and we were wrong but yesterday we came to know that IT IS POSSIBLE TO STUDY DIABETES IN DROSOPHILA!! as they have 7 insulin like peptides and different insulin pathways and some are similar to us I guess and i got to know this from references shared in chatshaala… Given below :point_down:

REFERENCES

The Drosophila insulin-like peptides (ILPs) also have nonredundant functions [18–20]. The ILP2 peptide has the highest homology to the vertebrate insulin gene and is synthesized together with ILP1, ILP3, and ILP5 in the IPCs of the brain, and their synthesis depends on ILP3. ILP3 expression also activates the insulin pathway in the fat body [21].

Drosophila as a Model for Diabetes and Diseases of Insulin Resistance
P. Graham and L. Pick1

Drosophila melanogaster as a Model for Diabetes Type 2 Progression https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5941822/ Drosophila melanogaster has been used as a very versatile and potent model in the past few years for studies in metabolism and metabolic disorders, including diabetes types 1 and 2. Drosophila insulin signaling, despite having seven insulin-like peptides with partially redundant functions, is very similar to the human insulin pathway and has served to study many different aspects of diabetes and the diabetic state
The pancreas produces an exocrine secretion (pancreatic juice from the acinar cells) which then enters the duodenum through the main and accessory pancreatic ducts and endocrine secretions (glucagon and insulin from the pancreatic islets of Langerhans) that enter the blood Anatomy, Abdomen and Pelvis, Pancreas - StatPearls - NCBI Bookshelf Anatomy, Abdomen and Pelvis, Pancrea

2 Likes

BELOW IS THE ARCHIVED CUBE CONTEXT 2 CURRICULUM WHAT’S APP GROUP CHAT OVER THIS INTERSTING FINDING OF SMITI 'S AND HER COLLABORATORS

Drosophila melanogaster has been used as a very versatile and potent model in the past few years for studies in metabolism and metabolic disorders, including diabetes types 1 and 2. Drosophila insulin signaling, despite having seven insulin-like peptides with partially redundant functions, is very similar to the human insulin pathway and has served to study many different aspects of diabetes and the diabetic state. Yet, very few studies have addressed the chronic nature of diabetes, key for understanding the full-blown disease, which most studies normally explore. One of the advantages of having Drosophila mutant viable combinations at different levels of the insulin pathway, with significantly reduced insulin pathway signaling, is that the abnormal metabolic state can be studied from the onset of the life cycle and followed throughout. In this review, we look at the chronic nature of impaired insulin signaling. We also compare these results to the results gleaned from vertebrate model studies. Drosophila melanogaster as a Model for Diabetes Type 2 Progression - PMC
Fruit fly as a model for Diabetes. Wonderful frontier topic being discussed in Chatshaala.

How do we get insulin mutants in Drosophila? What are the symptoms which can be studied?

Interesting and Surprising at the same time @Jai kishan CUBE @kiranayadav4894@gmail com SKYPE and @SMITI CUBE !!!:grinning::grinning:

So @SMITI CUBE @kiranayadav4894@gmail com SKYPE what is this Diabetes and Drosophila story exactly?
Can we have a brief summary about it?
"The flies larvae harvesting from the culture medium bottles of fruit flies was discussed. @Smiti talked about isolating larvae from fly bottles using Sucrose solution. She explained how flies need to be transferred from one bottle to another so that we can get staged larvae as flies will get only sometime to lay eggs and the larvae born will be of a similar age. These larvae can be used for several studies. @magpie talked about fruit flies and their larvae can also be used to study Diabetes.

How to get a Drosophila insulin mutant from nature is the question which came out of discussion. It was very interesting to find that Drosophila also has insulin signalling pathway like humans."

In diabetes the body is unable to process sugar because the sugar processing chemical insulin isn’t produced as per the requirement amount.

This is the understanding of Diabetes that i have.
How does Insulin ‘process’ Sugar? Let’s discuss this to understand Diabetes. Are all types of Diabetes Insulin dependent?
I’m not exactly sure how the insulin acts on sugar i mean ofcourse sugar is a carbohydrate and it would be something related to glycolysis but where does insulin comes in the picture?:thinking:

About the types of diabetes there are Type 1 and type 2.

In type 1 i think the insulin is produced insufficiently and in type 2 it’s the combined effect of insulin insufficiency and sugar not being able to enter the cells for their metabolism or in simple term processing to start.

I shall find out more on this to be concrete.
Insulin, a hormone made by the pancreas, helps the glucose in your blood get into your cells to be used for energy. Another hormone, glucagon, works with insulin to control blood glucose levels.

In most people with type 1 diabetes, the body’s immune system, which normally fights infection, attacks and destroys the cells in the pancreas that make insulin. As a result, your pancreas stops making insulin. Without insulin, glucose can’t get into your cells and your blood glucose rises above normal.

I think this reference solved the issue.

The insulin helps sugar to enter the cells for their metabolism to start.

And when there is not enough insulin then sugar can’t enter the cells.
@Rahul CUBE
Yes, insulin is a signalling molecule. It binds to Insulin receptors like it’s said in the reference to signal the cells about the free glucose in the blood. Without this signalling, glucose can’t be uptaken by the cells to put in the glycolysis pathway. Hence, leading to continuous increase in blood glucose levels.
What are the causes of diabetes? What causes the immune system to destroy insulin producing cells?
Is diabetes genetic disease?
Or is it related to consuming more sugar?
These are the frontier questions that i think we need to address whilst @SMITI CUBE develops the connection between her work she did earlier and with diabetes in drosophila. @SMITI CUBE @SMITI CUBE please reply on this. What do you think about the question posed by @Jai kishan CUBE
:sparkles: Okay so as I’ve completed my 2nd year in BSc and in final year we have to submit a thesis, we discussed what topic i can work on and my interest is in Diabetes
:sparkles: So Diabetes is basically a metabolic disease which is cause due to lack of sufficient insulin or when configuration of receptor changes (acc. to me these are 2 possibilities) as reception is a protein and like enzyme and substrate bind to perform a process in metabolism to form product here insulin needs to bind with its receptor so that cell will get a signal to open glucose channel so that cells can take glucose to perform further process like glycolysis and kreb cycle to get energy
:sparkles: Here we also discussed about how insulin is produced from INS gene in pancreatic cell and then transferred to blood
:sparkles:Now in both the scenarios, either when insulin is less or it can’t bind with it’s receptor, glucose will not get transferred into the cell and it will stay in blood causing problems as it increases blood sugar level
:sparkles: Here the question is are there other possibilities through which diabetes is caused? And what causes diabetes type 1 and type 2?
It’s an Astonishing Hypothesis for average biologists to connect Insulin with Drosophila and to use an insect like Drosophila as model system to address research in Diabetics.
*Congratulations":+1::+1::+1: @SMITI CUBE @Khushi CUBE and @kiranayadav4894@gmail com SKYPE for innovative thinking in yesterday’s CUBE Chatshaala.:+1::+1::+1:

@SMITI CUBE please post the path breaking references on how Drosophila is used to address research in Diabetics. @SMITI CUBE @kiranayadav4894@gmail com SKYPE
:sparkles: Then @kiranayadav4894@gmail com SKYPE ma’am asked if we can study diabetes in model organism i was working it that is drosophila and it reminded me of an incident that happened that time
:sparkles: On 8th Oct 2021 i trapped and made my 1st culture successfully but media had some issue we when i joined cube we discussed on how to make media and developed BRSV (Banana Sugar Rava Vinegar) media which I used in my home lab
Here we also compared homelab media with standard corn meal agar media used in labs
:sparkles: Like how we make media using ingredients available at home by discussing what we use in standard media and why is it important and then replace them with what we have and would work in similar way like instead of agar we use rava here to solidify media and instead of propionic acid and orthophosphoric acid we use vinegar which is an acetic acid to prevent contamination i.e bacterial and fungal growth here
:sparkles: I used this media to culture my flies for about 8 months and when i trapped flies i was looking for drosophila melanogaster - The 6 time noble prize winning organism!


:sparkles: For identification of flies and to see if i have drosophila melanogaster in my culture, i made single line cultures but lost most of them due to mite infestation… Now here single line means when we isolated 1 gravid (pregnant) female fruit fly and culture her in fresh new media bottle and all her progeny are from same species
:sparkles: So when mites started destroying my culture by killing flies, i called Saida di one of my senior to ask what i can do to save my precious single lines and she told me that when similar thing happened with Aswathy di she used to transfer flies early and later on used larve from old bottles to save her culture
:sparkles: So i can use larva for saving my culture by adding 20% of sucrose solutions in the bottle and when larve come up i can isolate them
:sparkles: But that time i didn’t understand it much and i thought if we wash it sucrose solution then mites will go away so i had a bottle that didn’t had flies and larve were there so i took them out with bursh and washed them with sucrose solution and didn’t washed it again with water cause i didn’t knew that i should and after taking them out from solution I kept them on cotton and placed that cotton on new media
:sparkles: After 2 days i noticed that larve were becoming fatter or obes and i reported that in C2C and while discussing Saida di told this could be due to diabetes cause i dipped them in 20% sucrose solution i.e 20 gm table sugar in 100ml water
:sparkles: These larve took too long to become pupe around 6-7 days and were in form of pupe for like 2 weeks after 2 weeks the fly that i got looked very different then my fly and i thought this was some kind of mutation or diabetic flies might look like this but then when i posted a video @Batul CUBE ma’am told it’s a different fly from some phoridae family
:sparkles: So if my flies took around a week to complete it’s whole life cycle these fly took a month to complete it and when i came to know this i thought we can’t study diabetes in fruit fly and we were wrong but yesterday we came to know that IT IS POSSIBLE TO STUDY DIABETES IN DROSOPHILA!! as they have 7 insulin like peptides and different insulin pathways and some are similar to us I guess and i got to know this from references shared in chatshaala… Given below :point_down:

The Drosophila insulin-like peptides (ILPs) also have nonredundant functions [18–20]. The ILP2 peptide has the highest homology to the vertebrate insulin gene and is synthesized together with ILP1, ILP3, and ILP5 in the IPCs of the brain, and their synthesis depends on ILP3. ILP3 expression also activates the insulin pathway in the fat body [21].

Drosophila as a Model for Diabetes and Diseases of Insulin Resistance
P. Graham and L. Pick1

Drosophila melanogaster as a Model for Diabetes Type 2 Progression Drosophila melanogaster as a Model for Diabetes Type 2 Progression - PMC Drosophila melanogaster has been used as a very versatile and potent model in the past few years for studies in metabolism and metabolic disorders, including diabetes types 1 and 2. Drosophila insulin signaling, despite having seven insulin-like peptides with partially redundant functions, is very similar to the human insulin pathway and has served to study many different aspects of diabetes and the diabetic state
The pancreas produces an exocrine secretion (pancreatic juice from the acinar cells) which then enters the duodenum through the main and accessory pancreatic ducts and endocrine secretions (glucagon and insulin from the pancreatic islets of Langerhans) that enter the blood Anatomy, Abdomen and Pelvis, Pancreas - StatPearls - NCBI Bookshelf Anatomy, Abdomen and Pelvis, Pancreas
So what reasons made you accept that your fly was a Phoridae fly?Did it have black eyes?Did the fly run instead of walk along the bottle?@SMITI CUBE
Why would your fly be a diabetic mutant?Can giving 20%sugar solution to the larvae mutate them?
Do you still have that video on which @Batul CUBE commented that it’s another fly?
Yes they were bigger in size and looked a little weird like with spindle shaped abdomen… The ran in a crisscross manner and had big black eyes

And about mutation i didn’t knew much about it as I was in first year that time and things were new so i thought if we give them different and extreme bad conditions like here dipping in high sucrose levels will cause mutation
It was there in my phone and i don’t have my phone right now with me… They say the data is gone and it won’t work again so i need to get a new one i hope I get all my data back
How would a diabetic mutant fruutfly look?@SMITI CUBE would it have black eyes and big size?if yes why?
I really don’t know cause i haven’t seen one. But I think it won’t have much to do with appearance at very 1st stage like as Diabetes is a metabolic disease so first there would be changes inside and we might observe bottle turning brown early… i think for this we should have a control to compare with and they also might eat more media so that too needs to be compare

I’m saying this cause for diabetes there are symptoms like feeling hungry and having frequent urination issues so they will eat more and excrete more than usual that can be noticed
Why would the bottle turn brown?
Brown in a sense when we culture fruit fly they are packed in a bottle containing media and they have media as it’s their food and then they fly around in bottle and walls performing different activities and they also excrete on the wall so after like 10 days we can see wall of bottle turning brown and that brown substance is excreta of fruitfly which is more on the neck region of bottle
Nothing can stop a cubist in doing research…
Even @SMITI CUBE’s phone is not working, she uses her sister’s phone to get us updated
Your approach is much appreciated!
If we perform an experiment with one species of fruit fly (single line) we can divide them into 2 batches or groups so we can take one group of fruit fly and dip them in 20% sucrose solution and see the results like their metabolic activity and compare it with other batch of flies we have that are from same species and same mother in general
True!.. Thank you sir :blush:

I meant we can dip the larve of those flies in sucrose solution and not the fly :grimacing:
What is the color and form of fruitfly liquid waste?
Do larvae going obese explain diabetes?
Do all diabetic patient go obese?
@SMITI CUBE
It’s more like liquid or semil liquid and has brown to black colour, brownish orange sometimes
Or do they loose weight?@SMITI CUBE
No not all diabetic patients are obese but one of my uncle and a neighbour who have diabetes type 2 have obesity too so that made me think that was a thing also in chatshaalaAgain not much sure about weight is loose or not… People i have seen mostly gain weight and by doing exercises and diet they try to lose it

It can be like in thyroid people say that some thyroid patients can’t gain weight and are too slim or they are too fat and can’t lose weight at all… So Diabetes can be of 2 types too but still i don’t think that weight would be a major thing too look here as I’ve recently came across discussions they say diabetes targets eyes, kidney, heart and foot told that obesity in larvae is due to diabetes and that
made sense so ya…
What effect would you see in their(fly) eyes?
In humans it’s that the vision get blurry and we need to have glasses on all the time but in fruit fly I’m not sure about how do we check if their vision too is getting bad and blur as time passes by
Drosophila melanogaster as a Model Organism for Obesity and Type-2 Diabetes Mellitus by Applying High-Sugar and High-Fat Diets
Nieves Baenas1,† and Anika E. Wagner2,*
Vladimir N. Uversky, Academic Editor

I was looking for if there is weight gain or lost in diabetes patients… So if i summarise what I’ve read till now is… When insulin is not produced in sufficient quantity then cell does not recieve glucose to get energy then our body starts burning fats and muscles to get energy and people lose weight

https://www.diabetes.co.uk/symptoms/unexplained-weight-loss.html#:~:text=Diabetes%20and%20sudden%20weight%20loss,reduction%20in%20overall%20body%20weight.
But for people who take insulin injections or have bad lifestyle choices such as sitting at one place and working with not much body movements or having a lot of sweet and high sugar containing stuff or caffeine and alcohol may gain weight

Insulin and weight gain: Keep the pounds off - Mayo Clinic.
P.eople with diabetes are more likely to gain weight faster. In addition, unhealthy lifestyle choices such as eating a lot of sugar, caffeine, and alcohol and not getting enough physical activity can increase the risk for weight gain.

Diabetes: Why Am I Gaining Weight?.
Interesting !!!
This i never thought about (TINTA Moment ) Fruitfly is a model system for understanding Diabetics !

So basically there are two types of Diabetics we are familiar with, One is because of the over production of insulin by Beta cell of pancrease and other by defective insulin receptor !? Is it !? Correct me if i wrong @SMITI CUBE @Abhijeet CUBE @Batul CUBE @Rahul CUBE @+91 94837 16348 & others

If that is the case, do Diabetics is a hereditary disease !?
Which type of Diabetics are hereditary !?

Coming to Smiti’s interest of study in famous 6 time Nobel prize winner Fruitfly ,
Do some aged Fruitfly that there in our surroundings are already diabetics patient ?!? @SMITI CUBE @kiranayadav4894@gmail com SKYPE @Abhijeet CUBE & all others
A very interesting experience of @SMITI CUBE of her home lab experiment. She described how we can make a nutritional media at home … she made a BRSV medium​:partying_face: :v:t2:

:point_right:t2:So BRSV medium stands for banana , rava , sugar and vinegar

:point_right:t2:take one banana + 25 g of rava + 2.5 g of sugar
:point_right:t2:add this to 100 ml of boiling water

Then add some vinegar in it

In this media sugar is source of glucose
Rava is used as solidifying agent
Vinegar to make media acidic so that media is free from microbial contamination
This media was really effective in flourishing the growth of fly
Yesterday was totally amazing…
I was astonished to know that fruitfly has insulin like peptides…I now realise why it’s been popularly used for research topics …and no wonder drosophila has been 6 times Nobel prize winner :trophy:
“Female and male flies were subjected to HSD and HFD for 10, 20 and 30 days. The obtained results showed clear differences in the effects of both diets on survival, glucose and triglyceride accumulation and dIlps expression, being gender and age determinant. The present study also suggested that weight gain does not seem to be an appropriate parameter to define fly obesity, since other characteristics appear to be more meaningful in the development of obesity phenotypes. Taken together, the results demonstrate a key role for both diets, HSD and HFD, to induce an obese fly phenotype with associated diseases. However, further studies are needed to elucidate the underlying molecular mechanisms how both diets differently affect fly metabolism.”
Drosophila melanogaster as a Model Organism for Obesity and Type-2 Diabetes Mellitus by Applying High-Sugar and High-Fat Diets - PMC.
“Thus, the aim of the present work was to investigate and compare the effects of HSD (30% sucrose) and HFD (15% coconut oil) on symptoms of metabolic dysfunction related to obesity and type-2 diabetes mellitus, including weight gain, survival, climbing ability, glucose and triglycerides accumulation and expression levels of Drosophila insulin-like peptides (dIlps). Female and male flies were subjected to HSD and HFD for 10, 20 and 30 days. The obtained results showed clear differences in the effects of both diets on survival, glucose and triglyceride accumulation and dIlps expression, being gender and age determinant. The present study also suggested that weight gain does not seem to be an appropriate parameter to define fly obesity, since other characteristics appear to be more meaningful in the development of obesity phenotypes.”
Drosophila melanogaster as a Model Organism for Obesity and Type-2 Diabetes Mellitus by Applying High-Sugar and High-Fat Diets - PMC.
There was no gain or loss of weight for me Siddika. Annual check up post retirement revealed Type 2 diabetes.
Yesterday’s detailed description of the experiment by Smiti was very interesting. Never thought that little fruitfly which I usually drive away from kitchen is such an important source of medical research!
interesting question :+1::+1::+1:by @Theertha CUBE

Are there many elderly Drosophila suffering from Diabetes, as in the case of human beings?

Age and diabetes in Drosophila, an interesting area to pursue.:innocent::face_with_monocle:

Is there a behavioural assay for determining Diabetic Drosophila?
What is drosophila diabetes ‘mutant’? @Batul CUBE ?

The drosophila which is diabetic is a mutant? What’s your understanding about it? Coz I’m not able to get it.
From what im able to grasp is that it’s a general understanding among the masses that mostly obese people are at the risk of getting diabetes because it includes unhealthy food choices, lifestyle, and etc.
Here is a research article, perhaps would help us answering the query raised by @Abhijeet CUBE

In this perspective, model organism research is crucial. This review traces an overview of the genetics of diabetes and mainly focuses on Drosophila as a model system, describing how flies can contribute to diabetes knowledge advancement. The Genetics of Diabetes: What We Can Learn from Drosophila - PMC
From the references smiti quoted:-

WHEN NO EXTERNAL INSULIN IS TAKEN.
Insufficient insulin (either type 1 or 2)-> Sugar not entering cell-> body not able to produce energy → body starts metabolising it’s food reserve and muscles → person loses weight.

WHEN EXTERNAL INSULIN IS TAKEN. (With unhealthy lifestyle and food habits)
Insufficient insulin → Sugar not entering the cell-> body not able to produce energy → External insulin is taken-> more intake of calories to maintain weight-> more sugar received by cell-> obesity.

3 Likes

How do we make mutants? What are the factors causing mutation?