UDLCO summary :
CUBE is a well known group of Indian school students who regularly discuss and perform science in their home labs and there's more about them here:
https://www3.hbcse.tifr.res.We participated in their discussion around culturing drosophila and discussed various research collaborative opportunities from the fly particularly focused on solutions for neurodegenerative disorders and diabetes.
[2/21, 10:21 PM] Prof CUBE : @~A @~S Please write a good summary on *how we can do research on Parkinson's Disease* in HomeLabs, *using Fruitflies*.
[2/21, 10:40 PM] student : Summary-
I have been culturing drosophila fruit flies from 9th feb and observation was done regularly in the bottle so initially there were 10-15 flies which i transferred and today approximately there are more than 50 flies in the same bottle. I have been using BRSV medium and explained the process of how i made it and how culturing was done.
[2/21, 10:40 PM] : Above i have posted the photos and video of that bottle
[2/22, 8:51 AM] Rakesh Biswas: Will be looking forward to hearing more about how you plan to utilize them as animal models for Parkinson's
Here's a cognition primer in this direction👇
[2/22, 12:05 PM] B P : @Rakesh Biswas thanks for sharing the article.Is this fly a candidate for Parkinson's mutant based on the phenotype : taking longer time to climb after being knocked down.@~Akanksha Joshi @~Rahul Sunita Kushwaha and others
[2/22, 12:05 PM] B P : All the flies are of same species and same age.
[2/22, 12:31 PM] B P : One more trial to see how far the flies can climb.As you see here too one fly is moving v slowly
[2/22, 4:14 PM] A J :
Summary
This is the video of the bottle of flies transferred to BRSV medium on 9th feb and now observation is that we can see the BRSV medium turning brown in colour upwards and the flies are more than 50 in number also there are approximately 50 pupae and 20-30 larvae in the bottle. On 9th feb i had transferred only 10-15 adult flies. Also the flies are showing upward climbing movement in the video also known as negative geotaxis.
[2/22, 4:15 PM] AJ : This is the photo of the BRSV medium which i prepared
[2/22, 4:24 PM] Prof A : Please give more details of each photo. @~Akanksha Joshi
[2/22, 7:14 PM] A S : *Potential anti-diabetic effect of certain plant extracts from the Egyptian flora on type II diabetes using Drosophila melanogaster as an animal model.*
Phytochemical screening of the tested plants revealed the presence of several constituents such as carbohydrates, glycosides, sterols and/or triterpenes, catechol tannins and flavonoids. Conclusion: The tested plant extracts, particularly A. halimus can recover and improve the symptoms of T2D in Drosophila.
[2/23, 7:48 AM] Metapsychist Number 1 Kims 2015: Interesting!
What are the symptoms of T2D in Drosophila?
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
[2/23, 10:35 AM] Ab : In this experiment, adult female insects were placed in a new vial to lay eggs on a standard diet. After two days, larvae were transferred to a high-sugar diet to induce type 2 diabetes. Then, these larvae were split into groups: one fed a regular diet, another fed a diet with plant extracts, and a third group fed a diet mixed with Glimepiride, a drug used to treat diabetes. The larvae were left to develop for 24 hours under each condition, and this process was repeated five times for accuracy. This setup aims to understand how different diets and substances affect larval development, particularly in relation to diabetes.
[2/23, 11:37 AM] Ab : Different model organisms were used to
identify the pharmacological properties of
different plant extracts such as mice (Adhikarimayum et al., 2007), Zebrafish (Giacomotto and Ségalat, 2010), Caenorhabditis
elegans (Kumarasingha et al., 2016), and
Drosophila melanogaster (Mezzoug et al., 2006). Each model organism possesses special
advantages and limitations (Panchal and Tiwari, 2017). Many features make a fruit fly the ideal
model for experimentation as it is easy to
handle, produces a large number of flies, and
the life cycle is short (Va et al., 2009; Panchal and Tiwari, 2017). This model has approximately 75% similarity with disease-
causing genes in humans (Reiter et al., 2001; Nass and Przedborski, 2011; Sengupta et al., 2016). Many studies have revealed that most biological signaling pathways are conserved between the fruit fly and human (Apidianakis and Rahme, 2011; O’Kane, 2011) Additionally, this model can simply feed on a diet to cause the disease and then transferred to another diet
mixed with the plant extract to observe the
recovery
[2/23, 4:55 PM] Rakesh Biswas: But less effective than Glimeperide.
To quote from that paper:
"The total body glucose of
HSD-fed larvae was significantly (P ˂ 0.033)
higher than that in the control (Bonferroni and
Dunn tests). However, when the diabetic larvae
fed on a diet treated with the plant extract or
Amaryl drug, the total body glucose significantly
(P < 0.033) decreased compared with those in
HSD-fed larvae. All plant extracts used were
able to recover the total body glucose near the
control level. Meanwhile, the Amaryl drug
significantly (P ˂ 0.033) decreased the body
glucose lower than the control."
[2/23, 4:59 PM] Ab : So the study is leaving a scope for us to explore that if plant extracts be made more or equally effective as compared to Amaryl Drugs? @~Akanksha Joshi @~Batul Pipewala @~SMIT LOKHANDE @~Smiti @~Dhanraj @~Theertha
[2/23, 5:09 PM] Rakesh Biswas: There apparently aren't any symptomatology yet that have been well defined as in Parkinson's well illustrated by @~Batul Pipewala 's video where she noticed inability to climb.
@~Abhijeet Singh 's shared paper simply measured the tissue or presumably lymphatic sugars, that too after gruesomely mincing their children!
[2/23, 5:12 PM] Ab : what is meaning of P<0.033?
[2/23, 5:12 PM] Rakesh Biswas: It just indicates statistically significant in the frequentist paradigm
[2/23, 5:16 PM] Ab : This is other paper
Rulifson et al. (7) performed a complete elimination of fruit fly IPCs through gene modification and found fruit fly larvae showed type 1 diabetes phenotypes such as hyperglycemia, weight loss, and developmental delay. Then Broughton et al. (48) discovered that fruit flies with partial elimination of IPCs could also develop the above phenotypes. Similarly, Zhang et al. (49) in 2009 knocked out the expression of Dilp1-5 genes, which caused fruit flies to present with metabolic defects similar to type 1 diabetes in mammals, such as stunted growth, small size, elevated glucose levels, and poor fertility.
https://www.ncbi.nlm.nih.gov/
[2/23, 5:51 PM] Prof Arunan Tata Institute: What are Amaryl drugs? @~Abhijeet Singh @Rakesh Biswas Is it like Insulin?🙃🧐
[2/23, 6:14 PM] Rakesh Biswas: Amaryl is a company name. The original name is Glimeperide and it's an agent that forces the pancreas to secrete whatever insulin it has. That's the reason it doesn't work in type 1 diabetes where the entire pancreas is destroyed. It works in type 2 where 30% of the pancreatic beta cells are preserved
[2/24, 9:45 PM] Rakesh Biswas: We'll need a drosophila gene kit?
[2/24, 10:22 PM] Rakesh Biswas: https://www.promega.in/
@Metapsychist Number 1 Kims 2015
The simple and functional design allows Drosophila researchers to drop a fly into the cartridge, turn on the instrument, and walk away.
[2/25, 9:17 AM] Prof Arunan Tata Institute: We would like to develop *HomeLab methods for isolation of DNA* from model systems, first.
[2/25, 12:59 PM] Rakesh Biswas: Let's begin to think of possible DIY solutions available for that @Avinash BMJ Elective @Metapsychist Number 1 Kims 2015
[2/25, 1:07 PM] Metapsychist Number 1 Kims 2015: Yes..
What's the model system?
Concept is simple..
1.Sample collection and disruption-Breaks down the cells and releases DNA from cells and tissues.
2.Digest the proteins-So that DNA gets unbound and can easily be extracted
3.DNA extraction-Salt to stabilise DNA.Centrifuge to seperate DNA from other cellular components.
4.dna precipitation-alcohol commonly used
5.collection and storage
Can try all these steps with household ingredients as well
[2/25, 1:11 PM] Metapsychist Number 1 Kims 2015: Soo..
1.Take the model system(any fruit/veggie)-Cut into small pieces and put in mixer or blender
2.Add some water and make it slurry..
3.add some detergent to break cell membrane.Blend the mixture till it gets smooth..
4 filter using filter paper .
5 transfer filtered liquid to another container..add little salt(less than half spoon)..so that,charge gets neutralized and DNA clumps together and precipitated.
6 we need any alcohol for this step(isopropyl alcohol in sanitizer may help)
[2/25, 1:27 PM] Rakesh Biswas: Can you find a YouTube home demo DIY video of the above mentioned steps and share the links?
[2/25, 1:28 PM] Metapsychist Number 1 Kims 2015: https://youtu.be/nq3raQX2mlA?
[2/25, 1:28 PM] Rakesh Biswas: Not this one. This was done
We need to do it in drosophila
Also we need to identify the genes and not the macro clumps alone
[2/25, 1:34 PM] Rakesh Biswas: 👆@Metapsychist Number 1 Kims 2015 For example if you have to make this gene model in drosophila how would you manipulate it's genes?
[2/25, 1:37 PM] Metapsychist Number 1 Kims 2015: We may have to use manipulation techniques
[2/25, 3:06 PM] Metapsychist Number 1 Kims 2015: We can try some frugal RNAi or gene knockout but the approach will be simplified and may not achieve same level of specificity and efficiency as lab methods
[2/25, 3:07 PM] Metapsychist Number 1 Kims 2015: For DIY methods also,we need RNAi kits
[2/25, 3:42 PM] Rakesh Biswas: Read this article now!
One of the feeling driven realization I had was traditional Chinese medicine could be same as Ayurveda!! @Metapsychist Number 1 Kims 2015
Amazing potential of the fruit fly though
[2/25, 3:53 PM] Metapsychist Number 1 Kims 2015: 'The known anti-diabetic active ingredients of traditional Chinese medicines include polysaccharides, flavonoids, saponins, and alkaloids (77–79). The studies based on the Drosophila diabetes model have notably enhanced this standpoint. However, the components of traditional Chinese medicines are complex, and only a little number of bioactive compounds from them have been well-elucidated. Thus, traditional medicines may also have potential adverse effects besides potential benefits.'
👆🏻Yes..reminded the other day's ayurveda formulations article..
Fruitfly is amazing!
As mentioned in this article,
'The fruit fly has obvious advantages as a diabetes model: (1) it is small and can be easily raised in large quantities in the laboratory (8); (2) it has a short life-cycle (10–12 days) and thus it is the data results of certain studies based on it that can be obtained quickly; (3) it has complex neuropils and neural circuits and rather complicated behaviors; (4) it only has four chromosomes in relatively smaller genome size, but those can match 75% pathogenic genes of human (9); (5) its genome is well-annotated in complete sequence; (6) dozens of powerful genetic tools are developed on fruit fly, which can provide convenient approaches to achieve various genetic manipulations, including UAS-GAL4, LexA-lexAop, QF-QUAS, and mosaic analysis; (7) The reduction of the cost of experimental research.'
The kits mentioned here are for gene knockout or RNAi
[2/25, 8:08 PM] Prof N : Let me study
Drosophila BTW has been studied by Lakhotia et all for screening g of Ayurvedic drugs
[2/25, 8:12 PM] Rakesh Biswas: Made me feel that the ancient Ayurvedic wisdom that was lost in India due to invaders may still be surviving in China!
Yu tu got a Nobel for Qiinghaousu now block buster Artemesin, which I'm sure is based on the principles of Ayurveda
[2/25, 8:26 PM] Rakesh Biswas: The lost Ayurvedic wisdom is not just in the type of herb but the way it's prepared and consumed?
To quote :
"As Tu also presented at the project seminar, its preparation was described in a 1,600-year-old text, in a recipe titled, "Emergency Prescriptions Kept Up One's Sleeve". At first, it was ineffective because they extracted it with traditional boiling water. Tu discovered that a low-temperature extraction process could be used to isolate an effective antimalarial substance from the plant;[12] Tu says she was influenced by a traditional Chinese herbal medicine source, The Handbook of Prescriptions for Emergency Treatments, written in 340 by Ge Hong, which states that this herb should be steeped in cold water.[13] This book instructed the reader to immerse a handful of qinghao in the equivalent of 0.4 litres of water, wring out the juice, and drink it all.[3] After rereading the recipe, Tu realised the hot water had already damaged the active ingredient in the plant; therefore she proposed a method using low-temperature ether to extract the effective compound instead."
Above under CC left from : https://commons.m.wikimedia.org/wiki/File:Fruit_flies_mating.jpg#mw-jump-to-license
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