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A type 2 diabetic rat model was developed with High fat diet + low dose Streptozotocin. High fat diet included a 2:3 mixture of coconut oil & partially hydrogenated vegetable oil (10ml/ kg body weight per oral) IN ADDITION to normal pellet diet(56% carbohydrates; 18.02% protein; 3.95% fat; 7.85% ash, 8.75% moisture, 5.45% fibre, 1.6% Ca and 0·85% P, w/w)
How much % of total energy of diet is given by the fat in the HFD?
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Caroline Fernandes-Santos provided an excellent method that you can use to calculate % energies from dietary components.. you can plug in your consumption numbers and calculate the energy % values. Memorize Caroline's formula and use for all aspects or dietary energy calculation.
best regards to Caroline and Syed.
Dr. Walls
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Do they include extracted ingredients like oils, fat, isolate amongst others and products from processed whole insects like flour, paste, meal etc.?
OR
Are they extracted ingredients only?
Thank you
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Also, kindly check:
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Is there perhaps a way to convert mg FAME/g fish performed on GC-FID to g lipid/100g fish? We had enough sample to analyse fatty acids but not enough to measure total fat. Unfortunately I do not have the retention factors of the equipment. Sample weight in 30mL CM was 50 mg. 10 000ul of this solution was transmethylated. We used 100ug C17 internal standard.
Any recommendations are welcome
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Apologies, the question should read:
Converting (g) fatty acid methyl esters to g/100g fish?
Not %
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Hi,
I want to include Crude Fibre, Crude Protein, and Crude Fat analysis of my plant sample (pulverized). If any lab (Indian) can perform these without charging, I shall provide co-authorship in the relevant manuscript, which I intend to send to an SCI/SCOPUS indexed journal.
Thank you, and looking forward to your responses.
Regards,
Soumi Paul
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Thank you, Sir Gaurav H Tandon
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In partial hepatectomy models, fat accumulation occurs during hepatocyte regeneration. Whether it is needed is controversial. Though it was initially thought to be the "fuel", later experiments have shown that proliferation can proceed even if fat accumulation is disrupted. My question is, what triggers this accumulation and what is its function.
Thank you
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Hepatic lipid accumulation after partial hepatectomy is a consequence of enhanced fatty acid mobilization and metastasis to the liver, as well as increased hepatic lipogenesis and decreased secretion of very-low-density lipoprotein.
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I am currently working on analyzing the association of dietary intake and microbiome with type 2 diabetes (T2D). I plan to validate my findings using publicly available databases and was wondering if there are such studies/cohorts that enrolled T2D cases and controls and have macronutrient (protein, fiber, carbohydrate, and fat) and/or gut microbiome data. It would also be great if the study focuses on an Asian population.
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Has anyone developed or found specific anthropometric models for this population in the scientific literature?
I'm looking for this to solve a problem in my clinical practice..
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Dear Marcio,
there are many indicators that can impact body composition. using the mathematical formula for these studies is not suitable for journal publications. in addition, wheelchair users are suffering from sarcopenia which significantly makes bias in the composision. Do you have any specific aims?
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Hi,
I want to extract amino acids and nucleosides from samples with high salt/buffer, protein, fat and cellular debris. Amino acids and nucleosides will then be analyzed by LC-MS.
Do you know how I can isolate amino acids from my sample? And how I can isolate the nucleosides? (two different analyses).
We are modelling the digestive process so there is digested food stuff (blended before mixing with digestive enzymes) in the substrate that varies depending on the experiment and the substrate used.
The digested substrate is in 40 mL of liquid.
I would like to avoid MWCO filtration if possible.
Thanks
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Hi, u can check this link its will help
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We're trying to perform a finite element analysis (FEA) simulation on the application of force to the skin of a human limb. In order to set up the simulation we are in need of damping coefficients for two of the layers - skin and fat. We've searched for but cannot find separate values for those two tissue types. Can anyone point us to references for these?
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I do not think you will find a specific value for the damping as it depends on the type of skin, thickness, and age of the human. Furthermore, the skin itself will have a varied damping value for the same person from one spot to another (due to the boundary conditions).
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In lipid extraction with Folch method, from two phases (upper & lower) which phase will be used for further fat analysis and why?
Regards,
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Lower Phase
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Fat corrected milk (FCM) Kg contains 0.4 (kg milk)+15 (kg fat)
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Like I would like to get help with the steps in writing a practical report
Regarding the above question.......
Thank you
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for practical report, it includes the following standard steps
F o r d e t e r m i n a t i o n o f m o i s t u r e c o n t e n t , t h e s a m p l e i s d r i e d t o a c o n s t a n t w e i g h t i n a n h o t a i r o v e n w h i c h d o n o t d e c o m p o s e a t 7 0 - 1 0 0 o C ( a b o u t 70 o C f r u i t s a n d 1 0 0 o C v e g e t a b l e s ) .
R e q u i r e m e n t s :
F o o d S a m p l e , H o t a i r O v e n , W e i g h i n g b a l a n c e , M o i s t u r e d i s h e s ,
D e s i c c a t o r , e t c .
P r o c e d u r e :
1 . T a k e k n o w n a m o u n t ( 30 - 50 gram ) o f t h e f r u i t / v e g e t a b l e / f o o d
s a m p l e
2 . D r y t h e m o i s t u r e d i s h a n d l i d i n h o t a i r o v e n a t 7 0 - 1 0 0 o C a n d c o o l i n a d e s i c c a t o r s .
3 . T a k e t h e d r y d i s h a n d n o t e t h e t a r e w e i g h t .
4 . K e e p t h e s a m p l e i n d i s h .
5 . T h e n k e e p t h e s a m p l e s i n t h e o v e n a t 7 0 - 100 o C f o r 1 2 h o u r s .
6 . T a k e o u t t h e s a m p l e f r o m o v e n , p l a c e i n a d e s i c c a t o r s a n d c o o l .
7 . A g a i n w e i g h t h e d i s h e s a l o n g w i t h d r i e d s a m p l e .
8 . A g a i n d r y t h e s a m p l e t i l l c o n s t a n t w e i g h t i s a c h i e v e d .
9 . N o t e d o w n t h e f i n a l w e i g h t o f t h e s a m p l e ( a s h )
10 . M a k e c a l c u l a t i o n s .
C a l c u l a t i o n s :
% M o i s t u r e = F r e s h w e i g h t ( W 1 ) – D r y w e i g h t ( W 2 ) x 1 0 0
F r e s h w e i g h t ( W 1 )
% D r y m a t t e r / T o t a l s o l i d = D r y w e i g h t x 1 0 0
F r e s h w e i g h t
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I want to dissect inguinal white fat in mouse, but I don't know where exactly it is. And I am confused whether posterior subcutaneous adipose tissue is the same with inguinal adipose tissue.
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perturbations and that body fat distribution is an important determinant of obesity-related complications. Individuals with increased upper-body adiposity are disproportionately burdened by obesity-related diseases, compared to lower-body obese individuals. Thus, it is paramount that studies continue to elucidate the pathways linking various adipose pads and depots in relation to health and disease, as well as the mechanistic underpinnings dictating how body fat is distributed in order to answer fundamental questions. Rodents are commonly used to model features of human metabolism and obesity, yet it is unclear to what extent rodent fat pads are a suitable model of human fat depots. Here, we have highlighted examples of both shared and divergent traits among rodent fat pads and human fat depots. Given some of the stark differences in adipose tissue location and function among species, we urge careful consideration in experimental design and interpretation when attempting to draw definitive parallels between rodent fat pads and human fat depots.
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Phosphorus applications in black cumin increase the seed oil rate. A similar situation is observed in other medicinal and aromatic plants.
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Dear Eric Nunes Gomes;
Thank you very much for this wonderful contribution.
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I have been wondering why when using a model of high fat diet the control diet has to be the 10% fat instead of normal chow. Does anybody understand the rationale behind this?
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Hello.
The main reason is that you don’t really know the composition of the normal chow diet. As it is a maintenance diet, providers use different batches of cereals and the compositions varies a lit bit between lots. Using a marching diet you ensure all micronutrients are balanced between the diets, and all macronutrients are always the same amounts since the providers use the same sources according to your specification and use specific calculations to get the % you want.
Hope it helps.
Best
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We are starting soon an in vivo trial with broilers fed diets supplemented with microalgae and insect full fat larave. We will collect blood samples and I'd like to have your suggestions about the best parameters to be analysed on blood to evaluate the oxidative and metabolic status
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Kindly check also the following useful RG link:
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When I set transesterification reaction with eggshell Cao and Beef fat/ palm oil, biodiesel was produced. But, When I set the reaction with the mixture of beef fat and palm oil (waste cooking oil or virgin palm oil), the reaction was never completed, I got only a homogeneous product of fat and oil only. even I have tried by varying the parameters like temperature (60, 70, 75), speed (500, 1100 rpm), reaction time (3, 4, 5, 5.30 hr), oil to methanol ratio (1:9, 1:12, 1:15, 1:20), catalyst loading (2.5%, 5%, 7.5%, 10%).
Please suggest to me what is the reason behind this and what would be the solution?
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Pas Cao mais cest CaCO3 voir mes publications
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In my study on Awassi sheep, it was concluded that adding turmeric to the ration led to diverting the path of fat accumulation from the parts of the carcass to the broad tail, with significant weight increases in the treated lambs.
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Nice and useful study
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Hello everybody
I am facing an issue in fish gut digestion using 30% H2O2. After the digestion process, I am getting a fat oil layer that might trap plastic particles. Kindly help me out with this to digest the oil layer or separate particles from it.
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Thank you sir for your response C.A. (Kees) Kan
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Post mortem sample.
Lungs, 100x, red-oil-O staining.
Regardless of the patient's history, would you consider the finding as a fat embolism?
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Agree with Sarah's comments.
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I want to centrifuge yogurt in order to pellet bacteria and yeast cells. But of course all the fat pellets as well. I tried a mixture of EDTA and TE buffer as used in a journal article I found, but the fat solids do not collect on the top as indicated in the paper.
Could tri-sodium citrate be used to keep the solids from pelleting, and if so, what ratio would I use? Or is there another method that would allow separation of the solids without discarding the cells?
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The material pelleting cannot be fat, since fat is lighter than aqueous solutions and will float on top; it is probably casein, the major milk protein.Like EDTA, citrate is a chelator that will remove free calcium from solutions. Since calcium is necessary for the formation of casein aggregates, this is probaly why your protocol includes EDTA to solubilize casein. However, milk contains about 25 mM Ca2+, thus, you will need at least that much EDTA or citrate to chelate Ca2+ completely. Changing chelators will probably not affect the result provided that you add enough of them.
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Do you think there is a correlation between diet and the spread of the SARS CoV-2?
Maybe a diet based on fish, rich in fat, omega-3 and vitamin D is better ?
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Upon reading following paper: https://pubmed.ncbi.nlm.nih.gov/31727409/, I saw that the power analysis is based on a two-sample t-test, while the primary endpoint is based on ANCOVA analysis (with two groups: placebo vs treatment and baseline liver fat as a covariate).
Is it OK to perform a power analysis for a two-sample t-test while not using the same statisitical approach afterwards for the primary endpoint?
Secondly: is the p-value of 0.05 valid for the calculated power when there was an amendment of the protocol (two dosing groups became one dosing group)?
==> the authors write
"Around 117 patients, randomly assigned to 80 mg (two-thirds) or placebo (a third) treatments, and given an estimated treatment difference of 30% change in hepatic fat fraction from baseline to week 12 between any dose of resmetirom and the placebo group and a common SD for the percent change in hepatic fat fraction of 35%, would provide 90% power with a two-sample t test to achieve a significance of 0·025 for a two-dose multiplicity (after protocol amendment, the significance level was reset at 0·05 because a singledose group was used)."
Thanks in advance!
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(you may need to copy by answer and paste it with fixed-width font to read it, sorry!)
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I want to extract lipid and protein from mice whole serum followed by quantification of Sialic acid in total fat. I found many methods but I am wondering, is there any kit available?
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Because it is a nonionic detergent i do not think it will effect the protein.
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Hi, i was looking for some research papers on junk food advertising restrictions/bans in outdoor spaces to reduce the exposure to advertise of food and beverages that are high in fat, sugar and/or salt. AND Any pilot/project evaluations of implementation of advertising bans at regional and local levels?
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Thanks everyone 👍🏼
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Hello,
I need a list of patents about the production of fat from cells.
Could anyone please help me?
Many thanks for considering my request.
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Hi Asghar
like this one:
you know, google is a powerful search engine that can produce all expected results with selected words, but it's a pleasure to share here
all the best
fred
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Dear all,
I'm looking for informations about vitamins stability (both water&fat soluble).
I've done several HPLC analysis in different kind of dietary supplements (tablets, powder, liquid, ..) and I've found that, for example, Folic Acid is less stable in liquids than in powder (at same storage conditions). I'd like to know if, in your opinion, this is a common behaviour for this vitamin.
I'm also looking for the paper "A study of folic acid stability in solutions of the B complex vitamins" by Alfred R. Biamonte,George H. Schneller or similar.
Thank you!
Antonella
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Vitamins are of great help in growth of fishes. I have worked in formulating fish feed and have added vitamins after the protocol of heating, boiling etc, so that vitamins are not denatured in the process and are available to fish through formulated feed
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Dear colleagues,
I would like to cryosect aortic plus perivascular adipose tissue from obese mice. Unfortunately, at -23°C, with a new knife, embedded in Tissue Tek and very careful handling, the tissue is too unstable and it tears when cutting.
Thicker cuts have shown no improvement.
Do you have any tips on how to cut the tissue better?
Also maybe someone who has only cut fat tissue?
I am very glad for help:)
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Thanks for the tip. Until now, I have not fixed the tissue in succrose before embedding.
I will try that and also cut in colder temperatures:)
Kind regards
Jasper
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I have seen everywhere that mitochondria in brown fat produce heat as UCP1 diverts protons from ATP synthase. What I can’t find is whether these mitochondria synthesize any ATP at all. In other words, which part of the energy (and protons) goes through UCP1 and which through ATP synthase?
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BAT mitochondria are permanently uncoupled. They contain UCP i. e. UnCouplerProtein. Their function is to generate heat to maintain body temperature. Hence rather than synthesizing ATP the energy is used in the form of heat.
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#research #biotechnology #spectroscopy
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Molecular fluorescence spectroscopy is one of the most sensitive and highly selective spectroscopic methods, which can detect extremely low amounts of chemical substances. In the milk industry, it is used for the determination of vitamins, fatty acids, residual amounts of antibiotics, and the identification of different milk species in dairy products.
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Hi, may I know how to determine the crude fat in liquid sample? Does the method is same like milk product ?
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how to crude fat analysis in liquid sample by using hexane & separatory funnel
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Hi,
I will be grateful if any one could give a detailed description on their experience/precautions to be taken with RNA extraction from breast tissue (stored in -80). I am planning to use Qiagen RNeasy mini kit and homogenize the tissue with help of crushing in mortor and pestle with liquid nitrogen followed by homogenizing with lysis buffer. I was wondering the normal breast tissue which is usually with fat, will homogenize in the same way? Any information related to this will be greatly appreciated.
Thanks!
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Qiagen has a RNA extraction kit specifically for tissue with a high lipid content (RNeasy Lipid Tissue Mini Kit) that might help maximize your RNA yield.
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Hi everyone! I have had good luck asking questions before so I thought I should ask this. Has anyone else ever noticed that fat globules surrounding secretory tissue seem to prevent cells from emerging from the tissue? These are plated secretory tissue from bovine mammary glands. Fibroblasts and mammary epithelial cells emerge from tissue but I've noticed that there are none where there is a sort of "halo" of milk fat globules. Not sure if this is just a coincidence or maybe I just don't notice them as much when there are cells there.
I have the milk fat "halo" pictured and then a picture of emerging cells on another piece of tissue. Thank you.
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Unfortunately, I do not have information in this field and I think that you can obtain information in your field of specialization by searching in scientific sites interested in this field. I wish you success and success.
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I am working on the differentiation of MSCs to adipocytes . I would like to know if the fat droplet-containing cells are terminally differentiated adipocytes or are they preadipocytes? are these cells post-mitotic?
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MSCs committed to adipogenic lineage are post-mitotic and terminally differentiated.
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Hello, I'm trying to analyze the proteome of some tissue by mass spectrometry. After lysing the samples and centrifuging them, sometimes there is quite a bit of fat floating on top of the supernatant.
Even assuming that I have enough material to take only the "clean" part, I wonder: should I? Or should I rather leave it all there and hope that it gets discarded when I precipitate the proteins (e.g. Acetone)?
I'm thinking that I might be losing lipophylic proteins (e.g. membrane transporters) if I remove the part of the supernatant which floats on top. But I also don't want to risk having fat messing around with ...anything. How do you go about it?
Thanks in advance.
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Exactly
You can extract membrane proteins using a lysis buffer that contains 2% TX-114 then precipitate them using 1:30 Aceton(Wash 3X), Afterwards you can run a SDS-PAGE to separate them. After commassie staining you can cut out each lane in 10 parts and prepare them using standard protocol for LC-MS analysis. SEC fractionation would be another alternative.
You can load the gel with 20 µg total protein.
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Hello.
I am debating the best way to normalize indirect calorimetry data when comparing normal mice (about 30g) vs obese mice (about 60g) Using body weight is not correct since fat has a very low energy expenditure (EE) so obese mice would present a lower EE/g than the real value. On the other hand, I consider that normalizing the data only with the lean mass is also incorrect. Normal mice present about 10-15% of fat and obese mice around 50%, I think it would be a mistake to consider that half of the body weight would not participate in the energy expenditure. There is the option of showing EE data without controlling for the animal weight, but bigger mice would have higher EE. I was looking for a formula combining lean and fat mass for data normalization but I could not find it. Have you heard about any consideration about normalizing indirect calorimetry data when comparing normal/lean mice vs obese mice?
Thank you very much.
Best regards.
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Actually using the same scaling exponent on total weight would be problematic because, as you said, fat is metabolically "inactive".
I would rather apply a correction to use the structural mass. I mean removing the fat mass for all individuals and then apply the exponent (0.66 or 0.75) to correct energy expenditure by body mass.
Not a perfect solution, but without specific data exploring the scaling exponent of mice with different fat% i believe that it would be the best way to make comparison
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I am Performing the pomegranate juice evaluation, So Please help me with finding lipid or fat percentage or provide the link of a related paper or website.
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Dear all experts,
I has calculated the fat oxidation during exercise every 5 minute till 60 minutes, and found the negative fat oxidation when the RER is reached up more than 1.00 (using the Perronet's equation). How should I do with the negative result? Should I cut that result before calculating the total fat oxidation or calculate the total fat oxidation by using all results which are positive and negative?
Best regards,
Parimon.K
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Hi Parinom.
I am not familiar with Perronet's equation and I would recommend Frayn's stoichiometric formulae for calculating the oxidation of these energy substrates, as they are the most widely used in the literature.
With regard to the negative values in lipolysis when RER>1 is reached, this is due to the total dependence on carbohydrates from this point, so that fat oxidation is non-existent or 0. Therefore, although the formula shows a negative value, this will be 0 whenever the RER is greater than 1, as Frayn himself explains in his article ( ).
Best regards.
Jordi Monferrer-Marín
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Hello everyone,
I'm required to do a histology experiment to determine fat accumulation after an HFD and to compare it to a normal diet. I planned to utilize red O oil staining, however, the equipment for this experiment was not available at my university. Besides, I am unable to go to other campuses owing to the lockdown, which seems to be taking an extended period of time. Thus, I wonder if H and E may substitute oil red O stains
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Dear Khaled Benchoula
The lipid will be dissolved with H&E staining therefore I suggest to use sudan or Oil Red dyes.
Good luck
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I am stuck in the result analysis of the physiology lab report. I am looking for the effects of ketone ingestion on fat oxidation rate. The same participant was tested before and after the ketone drink ingestion and measurements were taken at 4-time points. which test could be used to see the effects of ketone ingestion on fats oxidation rate.
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Use Least Squares analysis
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Does anyone have experience in extracting proteins from rat white adipose tissue?
I would like to analyse them in Western Blottingand I couldn't remove all fat cake from samples.
Thanks!
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Dear Sylvie, the following paper may help you: Diaz Marin R, Crespo-Garcia S, Wilson AM, Sapieha P. RELi protocol: Optimization for protein extraction from white, brown and beige adipose tissues. MethodsX. 2019;6:918-28. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6500908/pdf/main.pdf
Best wishes from Germany, Martin
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What percentage of trypsin should be used for mouse fat mesenchymal cells? These cells are difficult to separate from the bottom of the plate. What can I do to increase the growth of my cells in the flask?
Are fat mesenchymal cells divided into two groups?
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There is range of concentration but maximum is 0.25% Trypsin that works with most of cells. May be you can try when cells are only partially monolayers (say 60%). You can get idea from the enclosed sheet also, not necessarily you should use this company.
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Hello!
The results of my study are leaving me a little baffled. I would appreciate any insight you could give me.
What I expect: Larger objects (living or non-living) should heat up slower than smaller objects
My results: Heating rate is increasing with size, larger beetles heat up faster than smaller beetles
I tested for: The amount of water absorbed during the rehydration process does not play a significant role.
I am wondering: Could it be related to differences in fat/protein contents?
Thank you!
Emilie
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Thanks for the interesting question. Indeed, the larger the insect, the more it heats up (but I studied the content of proteins / lipids of different insects), but there is no clear pattern. In what it really can be traced it in the development of the trachea. At the same time, insects with a low temperature threshold (those that are active at 0 degrees), for example, boreus, have a high lipid index. In general, I am also interested to know the opinion of colleagues on this issue.
Regards, Sregey
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β oxidation requires sufficient oxygen when glycerol is converted into dihydroxyacetone acid, which enters the sugar metabolism process; fatty acids are converted into fatty acyl-CoA, which enters the tricarboxylic acid cycle for complete oxidation. But now we try to envision that E. coli can use fat in the intestines.
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This article may be your help. Process can be anaerobic too, but the proteins/enzymes involved are different.
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As part of my master's degree, I am working on a research project to cultivate fat lobules from explant tissue. We would like to study cell viability and functionality of fat cells over a period of 14 days in vitro and are looking for easily feasible light microscopy methods that are not as labor intensive as the preparation of sections.
I am looking forward to your answers!
Michael Ruegenberg
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Osmium Tetroxide Paraffin Procedure for Fat
I. The purpose of the stain: Demonstration of fat by a method that allows paraffin embedded of the tissue.
II. The principle of the stain: Osmium tetroxide chemically binds with the fat blackening it in the process. This is the only method for demonstration of fat on paraffin sections.
III. Fixatives: 10% NBF.
IV. Sectioning: Stain the wet tissue with the Osmium tetroxide, then process the tissue. The section must be no thicker than 2 mm or the osmium will not penetrate.
V. Controls: Most tissue contains fats, normally no control is needed.
VI. Reagents:
1. 1% Osmium tetroxide solution: osmium tetroxide & distilled water.
2. 0.5% Periodic acid solution: periodic acid & distilled water.
VII. Procedure:
1. Trim formalin fixed tissue to 2 mm thickness, then wash thoroughly for at least 30 minutes.
2. Rinse the tissue well in distilled water.
3. Place the section in a small quantity of Osmium Tetroxide solution and leave for at least 1 to 2 hours.
4. Rinse slides in two changes of distilled water for 15 minutes each.
5. Differentiate slides by placing them in the 0.5% Periodic acid solution for 30 minutes.
6. Wash slides in tap water for 30 minutes.
7. Process routinely, beginning with 70% alcohol, embed as usual.
8. Cut paraffin sections 4-5 microns, pick on slides and dry as normal.
9. Deparaffinize and hydrate as usual.
10. Counterstain with hematoxylin, Masson Trichrome.
11. Dehydrate, clear and mount slides as normal.
IX. Results:
Fat .............................................................. Black
Other tissue elements ................................. According to method used
View
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I have gone throughout literature but there are several methods
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Thank you for your nice question. There are several ways for the extraction of fat and water soluble vitamins. You may visit the following link:
file:///C:/Users/User/Downloads/molecules-23-01484.pdf
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Hello.
I am working with a diet induced obese model and run my mice on indirect calorimetry cages. I am search for the best method to adjust the energy expenditure (EE) results but there are some considerations to have in account.
It is assumed that fat does not importantly impact EE and people usually adjust EE to the lean mass (NMR based). However, my high fat diet (HFD) mice have 3 time more fat that the low fat diet (LFD) mice, and with it probably more brown fat also, that is very active metabolically. In this study is not completely correct to adjust neither by total body weight nor by lean mass, and absolute EE might be harder to analyze since BW differ a lot between LFD and HFD.
Is there any combined formula that is accepted to use? If so, can you reference the work?
Thank you very much.
Best regards.
Andre Cabral, PhD
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Thank you
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In Plants Sample
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Combined analysis of Fat Soluble and Water Soluble vitamin analysis (at low concentrations like in plant samples) by HPLC is rather difficult. Please check this review. https://www.chempap.org/file_access.php?file=593a202.pdf
Or do mean a combined work up, with separate analysis for FS and WS ?
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Fish specimens fixed in formalin are stored in 70% Isopropyl alcohol and 1% Glycerin solution. After some time, a fat layer was observed on the surface of the container. But when they were stored in formalin this never occurred. What might cause this effect?
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I used to extract RNA from epididymal fat(Using Takara RNA extract kit),approximately 900 mg epididymal fat tissue can get ideal RNA concentration. While the weight of brown adipose tissue of a C57/BL6J mouse is about 60 mg. So how can I extract RNA from brown adipose tissue (BAT) for qPCR?
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Hi Chao,
We routinely assess transcriptional changes of brown adipose tissue in the interscapular region.
You can refer more of our method in this recent publication:
For tissues with high fat content, you can centrifuge your sample at 12,000 x g for 10 minutes at 4C to isolate and remove the fatty layer. However, this might not be necessary for Interscapular brown fat as they contain less lipid content as compared to the epididymal fat. A typical brown fat mass (i.e., 60 mg as you have) should yield sufficient RNA for q-RT-PCR.
Please feel free let me know if you have any more questions or ideas to discuss.
I hope this is helpful.
Hunter
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Can the consumption of extra fat (2%) in the diet of prepubertal gilts cause an increase only in the weight of the ovaries? The rest of the uterine structures are not affected (no CL, CA..)
Why does this occur?
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But do you know how it can affect? I cannot find literature that relates the increase in dietary fat with the increase in the weight of the ovary
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I am working on mouse adipose mesanchymal stem cells. After removing the mouse peritoneal fat and extracting, I do the primary culture, I used DMEM F12 medium with 15% FBS, but with this, the growth rate of these cells is not good and they seem to grow slowly.what am I suppose to do?
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Hello, Mahtab!
Please You look at this article:
Materials and Reagents
Procedure
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How much metabolizable energy is need to producing a one-kilogram broiler and poult live weight with the lowest fat carcass?
How to calculate the energy requirement for a special purpose of weight and carcass increasing body weight in different weights?
Are there any differences between breast and thigh energy requirements for increasing weight in broilers and poult?
Is exist a preference for particular nutrients in the breast and thigh in the broiler and poult?
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a lot of references to be see like wise.
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Hi,
I am studying Treg using a mouse model. In order to get a high number of splenic and thymic Treg, I combine 10 spleens and thymus and digest both tissues using collagenase type 4 for 30 min. After digestion, tissues are filtered using 70 um and 40 um strainer and centrifuged to get a single-cell suspension. And, I found a huge amount of fat or mucous-like materials in the pellet. I tried to remove this by filtering again, but it didn't work well. Downstream procedures are affected negatively, leading to failure of enrichment of mouse CD4+CD25+ cells. Actually, I can carefully get rid of the fat or mucus using the pipette and blue tip.
Have you guys ever happened to this issue and solve this?
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What are your suggestions for reducing the crude fat content in fish meal? Excluding alcohol.
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Doing the fermentation.
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Hello, everyone.
I’m doing a meta-analysis on several continuous variables (fruits, vegetables, whole grains, fat, and so on) of the same outcome (dietary intake).
Can I put all the different variables in the same forest plot (as if they were subgroups)?
Thanks so much in advance
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Yes of course. The key issue is the use of an appropriate methodology of the conducted research, which will enable the implementation of an appropriately complex, multi-faceted meta-analysis. A higher level of meta analysis means a correspondingly greater amount of data obtained from the forest ecosystem and processed, for example, on Big Data Analytics platforms.
Best wishes,
Dariusz Prokopowicz
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For evidence, a few snapshots from any Scientific/Medical Conference are enough.
Physicians are obviously not good enough to heal themselves, and, do not practice what they preach.
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This is mildly Ironic. However, let's look at where most of these experts live. The United States and Canada are highly respected and cited. 75% of North Americans are overweight. Few areas like Colorado have obesity rates below 20%
and the national average is 40%. see:
Therefore it is possible that this is just a coincidence.
At any rate, the fact that they are fat is linked to where they live n'est pas?
(France and the rest of the modern world is fast catching up) So why can't our most eminent & established medical and scientific experts heal themselves but the American Quarterback Tom Brady seems to do that quite well?
The General Principle
The French natural philosopher Brillat-Savarin—once said “Tell me what you eat and I shall tell who you are”. So let's accept t hat the problem is FOOD, industrial food consumption. I am not going to concentrate on how much sugar has been added to American junk food, because I hope everyone knows this, and Marion Nestle once told me hidden sugar is our fat problem. This sounds really good and is certainly covers a great portion of the problem, but would not explain our medical experts' enigma. They would know sugar and act accordingly you would hope.
Hence back to food:
Our Daily Bread
Bread is certainly not an industrial product... that's food, right? Well, that has not been true since about 1939 when the American government made it mandatory to add iron powder to wheat products and then in the 1950s to rice and corn items. Yes, this health supplement experiment was without the consent of the test subjects. Even if you were eating naturally or "bio", would you avoid corn, whole wheat, and rice products? Thus, even if you know your science or medicine you are probably unaware that the largest unproven medical experiment is ongoing in your countries.(USA, Great Britain, Brazil, Australia, Mexico, Canada, etc.
Incidentally, iron and its deficiency are linked to obesity in many ways here is just one example
not to mention iron apparently adversely affects aging
Plus, many Southern States add niacin or nicotinic acid to bread which seems to amplify iron utilization.see
BTW Tom Brady eats gluten-free and effectively avoids most magnetic cereals, muffins, and bread.
Leaky Gut and Obesity
Your microbiome affects your body weight
And iron can perturb such an ecosystem
Meat
Let's eat meat every day, three times a day! This was not how Americans ate at the beginning of the twentieth century when obesity was not a problem.
Worse, many fast food outlets and supermarket items add sugar to their meat. Sugar added to iron-enriched or iron-rich food is throwing gasoline onto a fire.I do agree with M.Nestle.see
Nutrition advocacy in action: the politics of sugar vs. fat
I will let you guess... Tom Brady doesn't eat meat much and sugar is not in his diet.
Vegetables
So let's examine eating vegetables out of season say tomatoes. Well, that would be hydroponically grown. Great yields, but depending on what metals, say iron, are in the hydroponic fluid then because of osmosis this will be reflected in the fruit or vegetable. Coincidentally TB12 does not eat tomatoes;-)
How about some nice ripe black olives? Would you believe me if I told you those olives are softened with lye and dyed black with ferrous glutamate?
I can't cover all the various ways minerals have entered our food chain here but be aware that it is happening all the time.
Milk and Dairy Products
Again milk production has increased so much since the 1950s that many small dairy farmers have ceased to exist. Because cows are so overmilked, that cows udders need treatment with iodine, plus odd ways of increasing yield dairy products are now the major source of iodine for most people instead of seafood.
We have added meat to bovine diets with disastrous results, How do we get such increases in milk production?
Interestingly enough Tom Brady doesn't seem to eat milk cheese.
Pregnancy
Weirdly pregnancy is a risk factor for obesity in women with women becoming more obese with each birth at least in Peru. see
It is a common medical practice worldwide to give expectant mothers folic acid and iron.
could iron explain these results?
To sum this up.
What are the health effects of too much metabolites like iron, folic acid, iodine, and so on? Is this road of good intentions a path to obesity and poor health? We just don't know.
but I think
It is mostly about the iron stupid.
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We routinely use the iDISCO+ and Adipoclear techniques followed by imaging on a light sheet system and I find very bright punctate labeling around the outside of tissues such as brain or fat as well as along the inside surfaces of ventricles. We filter our secondary antibodies through a 0.22 um filter as described in the protocol but that does not seem to be enough. Does anyone else have this problem or any suggestions that we can try and reduce it?
Thanks!
David
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Thanks, Ra. Unfortunately, that method only uses small molecule dyes for labeling, not antibodies for immunostaining. I also notice little to no background if tissue is stained with dyes such as ToPro, PI, etc.
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Hi,
Was wondering if any of you conducted an Oil Red on AML-12 cells, if so, after how much time did you executed this?
Some studies call for 24hr and some for 18hr.
For fat content analysis.
Thank you.
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When a person is put on a diet consists of low carbohydrate and high-fat, the body starts using ketones from fat for energy instead of glucose. This situation forces the body to use fat stores as fuel, which may lead to weight loss. Acceptable macronutrient distribution ranges (AMDR) are 45-65% calories from carbohydrate, 20-35% from fat, 10-35% from protein. What could be the long term effect on overall health if a person is put on low carb and high fat diet?
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Couple of months before, I helped some medical students in data analysis of similar research and have mentioned some of the research outcome in the answer. I will check with them if they are willing to share their research outcome with you. Though the sources are not mentioned, still you can also check the following link for detail: https://www.webmd.com/diet/guide/high-protein-low-carbohydrate-diets
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We are doing research with C57BL/6 mice to induce obesity using High-fat diet (D12492-60% kcal% fat). 
After 2-week observation, the high-fat diet intake mice have a wavy hair problem and also sample treatment group showed the same. We are curious without any sample treatment mice also having a wavy hair problem.
Please help me out! Is there any problem with the high-fat diet or not?
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Reason 1: The dirty cage, mixed up with diet , bedding material makes mice uncomfortable and induces stress.
Reason 2: The diet you are using have high fat makes mouse skin to secrete more oil from body and makes it greasy.
Reason 3: the piloerection, High fat diet, specially the diet you used increases body fat(http://www.epsekishin.co.jp/cn/lsg/service/researchdiets/pdf/NAASO%20poster%202005.pdf)..
increased brown adipocytes acts through UCP1 to regulate thermogenesis. It may nonshivered thermogenesis or thermoneutrality.
the imbalance in protein/carbs/fat makes mice unregulated thermogenesis .
you may check mice body temperature to find it.
good luck for your research.
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I have a dietary intervention experiments on diet-induced obesity animal model and the results demonstrated that the intervention reduced body weight and fat mass and increased lean mass with statistical significance.
Although weight loss ≥ 5% is considered meaningful, what percentage changes in fat mass and lean mass is known as clinically meaningful?
If you have answer for this question, please also provide the reference so that I can cite it in my paper.
Thank you!
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Agree with Mousa Numan Ahmad .
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I want to apply regression analysis using SPSS. Kindly help as to how to apply the regression model in the study where i have recorded Weight, BMI, Total body fat mass, insulin resistance and nerve conduction velocity in type 2 diabetics.
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I want to do scoring for macrovesicles and microvesiclular fat diposits in Histopathological Liver sections. Can anyone suggest me a software and discuss a suitable protocol to achieve this. I want to check for the development of Hepatic steatosis or fatty liver condition.
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Previously I used a semiquantitative method for fat accumulation assessment in hepatocytes. the following sentences of our paper (2017) describe the method:" For each section, four to six unbiased counting frames were sampled in a systematic random fashion. Fat accumulation in the liver was graded as previously described (Brunt et al. 1999). Briefly, Grade 0, no fat was found; Grade 1, <33% (light); Grade 2, 33–66% (mild); and Grade 3, more than 66% of hepatocytes affected by fat droplets "
- Brunt E M, Janney C G, di Bisceglie A M, Neuschwander-Tetri B A and Bacon B R (1999) Nonalcoholic steatohepatitis: a proposal for grading and staging the histological lesions. American Journal of Gastroenterology 94 2467–2474.
Hope this will meet your concerns,
Mehran
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WHO recommends a fat intake of no less than 15% of the daily caloric requirements (https://www.who.int/dietphysicalactivity/publications/trs916/en/).
However, imagine a catastrophic situation in which a population somehow lost access to fat-rich foods for several years, while still having access to a good supply of carbohydrates, protein and micronutrients. Is there a point at which nutritional problems would arise if the consumption of fat was near zero? Would people survive if this was the case? what if the consumption was 5% or 10% of the total caloric intake?
Could the body survive via its own capacity to synthesize fats from carbohydrates?
-Please assume that essential fatty acid consumption (linoleic and alpha linoleic) is covered in this scenario-
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Current Dietary Fat Intake Recommendations for Adults by WHO (Fats and fatty acids in human nutrition Report of an expert consultation. FAO Food Nutr Pap. 2010;91) -
Total: 20-35%, Saturated: <10%, Trans: <1%, n-6 PUFA: 2.5-9%, n-3 PUFA: 0.5-2%.
Lichtenstein & Van Horn (1998) defined "a very low fat diet is defined as one in which ≤15% of total calories are derived from fat (33 g for a 2000-calorie diet, 50 g for a 3000-calorie diet) with fat calories distributed approximately equally among saturated, monounsaturated, and polyunsaturated fatty acids. Approximately 15% of total daily calories consumed should be derived from protein and ≥70% from carbohydrates."
In contrast, at this time, no health benefits and possible harmful effects can be predicted from adherence to very low fat diets in certain subgroups. In addition to young children, the elderly, pregnant women, and persons with eating disorders should not attempt a very low fat diet. Persons with insulin-dependent diabetes mellitus, elevated TG levels, and carbohydrate malabsorption illnesses should also avoid such a diet.
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We are trying to extract RNA from liver samples that came from animals that were either fed a high fat diet or a low fat diet (Using Qiagen mini kits). Should we use a lipid mini kit for both liver samples, a lipid mini kit for the high fat diet livers and a regular mini kit for the low fat diet livers or does it not really matter?
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I don't think that using two different kits it would be a good idea. You will introduce variability to the sample preparations and you could find unreliable results. I also use the qiagen lipi tissue kit for RNA extraction from both tissue samples and differentiated cells in vitro and I always find good also with non-fatty samples. I think that the spin columns are optimized to not interfere with lipids but it is not a problem if you start from a non-fatty sample.
All the best.
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Am looking for the best method to extract and analyse water insoluble organic cpd from animal fat tissue
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you may extract it by organic solvent such as EA, DCM and evporate the solvent, at last analysize it by UPLC
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I'm trying to compare several different surfactant efficiencies for Swage Fato Oil and Grease emulsification into water. I don't think that methods for fat determination in food for example Ethanol extraction after the emulsification won't really work in this case due all sords of different impurities into the FOG material.
Maybe you have faced similar problems.
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There are several ways to compare the stability of emulsions. First of all, you can get a visual impression of the stability by just storing both emulsions for a certain period of time and observing the degree of different destabilization phenomena, like creaming to sedimentation and compare. The second way could be any microscopic technique (optical/TEM/SEM)depending on the size of the droplets, you might get an idea, how fast Ostwald ripening or any other things happening and compare. Static or dynamic light scattering can also give good information. Ultracentrifugation technique or even rheology analysis can give you an idea depending on the droplet size or property of your emulsion.
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hi, i want to ask something about weight.
i provide a high fat diet(around 45% of fat) to my mice and did a gavage sth( one of herbal thing)
but one group of mice eat lesser than other but gain more weight.
Concentrate what i did a gavage was less density,
the other group is more higher density, but wasn't like that.
even the group only provide high fat diet ate more but gain weight less.
is it possible eat little amount of high fat diet and gain a weight more than other mice group i describe?
Have any way to gain a weight in healthy way while they take high fat diet?
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You can also use the:
The basic diet contained: 54% corn, 14% wheat bran, 13% al-falfa meal, 10% cotton meal, 6% fish meal, 1.5% vitamin and mineral, 1% limestone, 0.3% sodium chloride and 0.2% dicalcium phosphate (Rodent diet datasheet, 2016).
The High-fat diet contained: 69.5% basic diet, 15% pork fat, 15% sucrose and 0.5% pig bile (Tolouei Azar J, et al. 2020).
In recently, we have had good success with this diet.
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I am dealing with an internal combustion engine, <500kW rated power, 1500rpm rated speed, for stationary application in CHP configuration. The engine, designed to operate with neat vegetable oil, operates a Diesel thermodynamic cycle and it is directly fuelled with animal fat. An excessive amount of Calcium (Ca) was detected through a chemical analysis after several failures occurred. In particular, fuel filters clogged several times, and I am wandering if the excess of Ca could have caused those failures. In particular, I am curious about the possible precipitation of Ca-based solid particles, also combined with a wrong thermal management of such a viscous and chemically unstable fuel.
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I have no experience with Ca as factor in fuel, but see similarities between animal fat and other bioDiesels. Filter plugging is a real issue because of the lower thermal stability of those fuels, causing oxidation and sludge formation. And so filter plugging. See for instance this EMA warning: https://www.biodiesel.org/docs/default-source/fact-sheets/ema-statement-regarding-use-of-raw-vegetable-oil-or-animal-fats-in-diesel-engines.pdf?sfvrsn=e72a4047_14
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Hi... Currently I am developing RTD milk coffee. As I read on article, the low molecular weight surfactant can replace protein from the surface of fat droplet. Indeed it can reduce the fat droplet size better than protein in the beginning, but the stability of the emulsion will be less during storage. Then this several question arose in my mind:
1. if we have enough protein in the system, does it mean that the additional LMW emulsifier is not necessary?
2. In case there is no enough protein, does the addition of protein (such is in the form of sodium caseinate) is a better choice than LMW emulsifier?
Thank you in advance for your answer! :)
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I once made a stable flavor emulsion based on the formulation of Bailey's Irish cream, so caseinate as a stabilizer with sodium citrate to complex calcium and ethanol to prevent microorganisms. It worked really well. Small droplets too. It was successfully produced in 100's of kg quantities. So low molecular surfactants are not always necessary, but as Ulf said, it depends on your exact constraints, as always.
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We are testing various physical parameters of both fish and tomato samples, such as acidity, color, texture, fat content, etc. What is an appropriate sample size for these types of tests? References are always appreciated.
Best
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I believe you mean by sample size, the quantity needed of the sample material? This is usually mentioned in the standard procedure tests. So, usually small to medium amounts are needed .... say less than 2 g. BUT what's more important that this small amounts be a correct representative of the bulk sample materials. So, all the materials (for example tomatoes) from a particular treatment and replication, after processing, should be mixed thoroughly, and sub-samples be taken by methods such as coning and quartering technique .
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I am Ruminant Nutritionist, we bought commercial calcium salts of fatty acid (By-Pass Fat), we tried to analyze the fat contents by soxhlet method, it gives less than 3%, Actually, it contains 84% fat
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Read this article and surf the web
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As I know Vitamin D is diluted in fat. But I want to try it for cell culture medium. Any help will be appreciated.Thank you.
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Long time back I did all my vitamin D stocks in ethanol. Should work even these days...
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hello
I am trying to test my scheduling algorithm in cloudsim. However I am trying to create fat tree data center that consists of computing servers, storage servers and switches. Any help regarding how to start creating this topology is appreciated.
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Prof. Ayad Ghany Ismaeel Thanks for the response. Actually I am working on cloudsim as a simulator for my algorith. I have worken on it before using brite files, but currently I have a brobkem creating a specific topology “ Fat tree” for my datacenter, is that applicable using Brite generator?
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I am using Ethanol 95% for fat extraction from Soxhlet & sometimes extraction comes up and sometimes not with same setup. The water boils properly but the solvent is not actually boiling but the vapor seems come down. What could be the issues? Can anyone help me with the solution?
Thanks in advance.
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First of all, your Soxhlet device may need servicing...
then you need to choose the best solvent that extracts your material.
You need to know the boiling point of your solvent (e.g. ethanol 78-79C°) and set the temperature of the Soxhlet according to the protocol.
Ps: I used 70% ethanol for the extraction of medicinal plants and I found these problems too
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How to increase serum HDL level at the same time avoiding fat containing food?
What are the dietary sources of HDL?
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