Energy Metabolism - Science topic

Dear Getu Habte

I found this article, that I also have added above, as I think it is highly relevant.

There is a chromatography-spectrophotometric method. At the beginning, a protein-free esetract with perchloric acid is prepared. Then it is neutralized with potassium carbonate. Then ATP, ADP and AMP are separated by current layer chromatography. Macroergs are eluted with hydrochloric acid and subjected to UV spectrophotometry.

Thank you very much for your answers Yu. I'll look more into the mass spectrometry.

Have an excellent day !

Thank you sir for your kind response.

Dear Elizaveta Klimanova,

Thanks all for your valuable informations. :)

Super nutrition - You mean Food Suppliment like Vitamines / Minerals / anti-oxidants .

Hi,

Please, have a read:

Not sure that it's promising to seek it this way. I would start from Linkedin (enriched profile with all information would help) and Indeed - both can collect positions automatically. Monster is not really useful, since it acts mainly through recruiters. Official academic job aggregating sites (e.g., zeit academic jobs for Germany) are extremely useful since universities have to advertise all their open positions there. Aiming cover letter and well structured CV would improve your chances. I've found my posdoc position twice using this approach. Good luck!

Dear Michael, I think you are looking for the paper of Costa et al. 2015:

Costa S, Ogilvie D, Dalton A, Westgate K, Brage S, Panter J. Quantifying the physical activity energy expenditure of commuters using a combination of global positioning system and combined heart rate and movement sensors. Prev Med 2015;81:339-44. https://ueaeprints.uea.ac.uk/55193/4/Manuscript.pdf

Best wishes from Germany, Martin

@Sarmistha Sarkar : Please which enzyme in particular do they target in the bacterial energy metabolism?

Thank you for the article. I have familiarized myself with Abstract of this article which was studied role environmental matrix of cancer cells and metastatic niche via hydroxyliated collagen, i.e. modificated collagen but not production collagen. This interesting work does not reject above presented cancer metabolism mechanism as "aerobic glycolysis" of Warburg effect.

Dear Samsher,

Unfortunately (or luckily), ROS are complicated and a collective name of many different reactive molecules. I don't work on mammalian systems but plants. Similarly, ROS bursts can occur upon entering hypoxia, but may decline (depending on the type of ROS) upon reaching anoxia depending on the type of ROS and organelle/tissue studied.

I would always suggest looking at their methods and check how and when they did their measurements and draw careful conclusions from there.

Best of luck

I'm just reading about the scandal too. Many thanks and best wishes.

Hi Rasmus,

Thanks for your answer. We have tried some of the inital pipeline for 16s sequencing using the Omnigut kit from DNAgenotek and the Power fecal kit from Qigen/MoBio. We also tried (only once though) to run a total bacterial PCR for the V3-V4 region and quality checked it on the bioanalyzer and it looked acceptable.

Do you think that working out the qPCR will give us an idea of how well the samples will work for sequencing and a general idea of what to expect from the run or should I spend my time making sure all the samples get past the first PCR and clean-up stage?

Hello Uwem, I suppose you are done with your metagenomic sequences and the need purifications and trimmings have been satisfied. Please see the attached documents. I am sure you will find what you need.

Type IIx is intermediate between type I and IIb fibers, they have intermediate numbers of mitochondria and oxidative potential resulting in moderate strength and improved resistance to fatigue , and usually considered as less oxidative than type I fibers in term of metabolic profile, have either higher or similar PGC-1α content compared to type I fibers.

Duan Y, Li F, Wang W, Guo Q, Wen C, Yin Y. Alteration of muscle fiber characteristics and the AMPK-SIRT1-PGC-1α axis in skeletal muscle of growing pigs fed low-protein diets with varying branched-chain amino acid ratios. Oncotarget. 2017;8(63):107011-107021. doi:10.18632/oncotarget.22205.

Gouspillou G, Sgarioto N, Norris B, et al. The Relationship between Muscle Fiber Type-Specific PGC-1α Content and Mitochondrial Content Varies between Rodent Models and Humans. Schuelke M, ed. PLoS ONE. 2014;9(8):e103044.doi:10.1371/journal.pone.0103044.

Dear Faizan,

I refer you to this paper by Oppelt et al. 2016:

Best,

Robin

it is mean Conversion into melanin, or an increase in the concentration of melanin within (a tissue or organism)

please check: http://www.eolss.net/sample-chapters/c17/e6-58-09-08.pdf

Yeast fermentation of glucose will generate heat energy without burning off the glucose by direct heat.

I will be interested in knowing all the interpretations of 'sex' as in reproduction or for         destressing and the related but forgotten notion of 'gender'. Good luck with your work. i think it will be very important.

If there is any carbon dioxide retention taking place, subsequent to perhaps some acid accumulation, there can be low RQs. In goldish, low RQs  were observed during recovery phase after a hypoxic exposure.

I'm glad I could help.

Hills, A. P., Mokhtar, N., & Byrne, N. M. (2014). Assessment of Physical Activity and Energy Expenditure: An Overview of Objective Measures. Frontiers in Nutrition, 1, 5. http://doi.org/10.3389/fnut.2014.00005

You may find this article helpful. The MET is used to categorize PA intensity. It is useful for describing and prescribing exercise of different intensities, but is not an accurate or precise measure as body weight and body fat varies between individuals.

Hi Susan- this is a question that I get a lot and have though about extensively.  The bottom line is what does the clinician want to know- the basal homeostasis, or how the patient is doing under their current, and perhaps life-long conditions of supplementation?  For the former, I believe that such a disruption of metabolism, if real, would take at least two weeks (perhaps more) to get back to a true steady state- ideally one would test for baseline prior to initiating any supplement protocol.  The time to "re-homeostasis" depends on the pharmacokinetics of each supplement (fat soluble having the longest half life of course), and importantly the half life/turnover time of the primary enzymes involved. Takes a while to get to a new steady state, and I'm not sure that the labs have truly evaluated that.  Then there is the potential for supplements, and specific food s/additives to directly impact the assay and/or analyte levels. For example, I have a real problem with the claims about urine propionate (PA) levels as being indicative of Clostridial overgrowth- BS, a couple of folks have done pilot studies comparing the OATS-derived PA levels to Clostridia abundance by REAL molecular techniques- no correlation.  As you probably know, PA is on the GRAS list and is a common food additive!!!  So the latter approach- assessment of metabolism under current stable supplement conditions.  That I prefer because it reflects reality- true some of the supplements may not be necessary, but more importantly one gets a sense of how the clinicians intervention is working...or not.

Just one man's opinion, I know that you will give my bloviating careful thought and add it to your thoughtful processing.  Best,  David Quig, PhD

We use this one in culture:

You can also use the same insulin that humans use (i.e. Humulin R-100) which is usually around 700 uM stock. We actually have compared both at multiple concentrations in a number of assays, and they give exactly the same results.   

I hope the attached table will be of help to you.

You can inhibit creatine kinase with fdnb (fluro-dinitro-benzene) but you will find the muscle goes into rigor pretty quickly!

I've just read the whole question and realise you are talking about field testing, so probably fdnb wouldn't be the best thing to try!  I don't know of any way you could assess this in a whole animal/person.  NMR studies always show ATP to remain very constant during exercise which suggests that ATP hardly acts as a store at all but is mainly a means of passing high energy P from PCr to myosin, or wherever it is needed.

For accurate monitoring of total energy expenditure (EEtot), DLW or calorimeter is highly recommended. The method of DLW needs sophisticated equipment and facility (radiation control) and is expensive to set up. Even budget is available, it will take considerable effort to get it done right. Calorimeter (closed-circuit respirometer) is less expensive, but it only houses one animal at a time. Many calorimeters are needed if all animals are monitored at the same time.

If more than 30 animals in a group by randomization procedure and they live under the same situation (diet, ambient condition, etc.), I would argue that the effect of other activities is considered under control. Even there may be possible difference in the mean EEtot between groups; it would be minimal comparing to the much larger EE change associated with exercise training. In addition, if energy balance is a concern, the amount of food consumption can be measured by weighing method.

@Mr. Asturiano: thank you very much..

@Mr. Kelany: it is interesting

Hi there,

Glycerate kinase activity can be easily coupled to pyruvate kinase and lactate deshydrogenase. GK activity will result in the stoechiometric consumption of NADH measured at 340nm. For more info this paper should help :

Acetyl-CoA carboxylase (ACC) is a bonafide AMPK target in multiple tissues.  Many commercial antibodies that recognize the S79 phosphorylation on ACC by AMPK are available and provides a good readout of AMPK activity.  For greater throughput CisBio offers a HTRF based assay to screen for phospho-ACC.  Detecting AMPK activity in serum would be much more complicated.

Fexaramine reduced diet-induced weight gain, body-wide inflammation and hepatic glucose production, while enhanced thermogenesis and promoted browning of white adipose tissue (WAT) in diet induced obesity (DIO) mice. Its effects in humans are not known yet.

although most of research on obesity is concenrating on white adipose tissue and the development,differentiation ie hypertrophy and hyperplasia of adipocytes and pfeadipocyytes in WAT and role of inflammation and macrophage infiltration directly or conversion of preadi[ocytes,metabolic endotoxemia,gut microbiota lreading to increased LPS absorption from increased GIT permeability with altered gut microbiota is some of the aetiopathogenesis with infiltration of Treg cells,cdt4. CHT8 cellsiNKT CELLS th17 all have ben found but importance is being given simultaneously to the microrna which affect the differentiATION OF BRITE AND BROWN ADIPOCYTES AND IMPORTANCE OF MIrna'S 26,155,27a,b,130 a ,miR155 etc in adipocyte differentiation and how further roles of FGF21 may affect thermogenesis and natriureteic peptides

you can to use Luciferase Assay since PGC1 alpha is Transcription factor co-activator

, pgc1-a participate in a lot of  gene transcription... so by Luciferase Assay you can see if its still have this ability...    

Umar:

Perhaps, the paper I reference you below (which uses "ultrasonic telemetry" in fishes, can be of interest for your research problem.

"The ultrasonic telemetry of cardiac rhythms of wild brown trout (Salmo trutta L.) as an indicator of bio-energetics and behaviour"; by:  I. G. Priede† andA. H. Young.

Journal of Fish Biology,  Volume 10, Issue 4, pages 299–318, April 1977

Antonio

Hello, T4 is much better than T3 because thyroid gland release 90-95 % T4 and 5-10% T3. Hence, T4 is the best indicator of thyroid function. Too late, T4 is converted to T3 in the inner of the cells by deiodinase type II. Good Lucky.

This paper might be useful:http://www.ncbi.nlm.nih.gov/pubmed/?term=rothman+1991+liver+glycogen

Seems to stabilise at 40-50 h of fasting in humans.

IMPACT OF BLOOD TYPES ON ENERGY METABOLISM IN MUSCLE OF PIGS

In pigs the locus Hal for malignant hyperthermia susceptibility, Phi for phosphohexose isomerase, H for blood group system H, and Pgd for 6-phosphoglucogenate dehydrogenase, are closely linked to each other. Pigs posessing the Haln gene are stress susceptible and react with post mortem muscle hypermetabolism causing inferior meat quality characterized as pale, soft and exudative (PSE). The linkage disequilibrium Hal-Phi-H thus results in an effect of the H and Phi systems on meat quality, and the two systems may explain up to 38 per cent of the additive genetic variance in meat quality. In Danish studies (Fogd Jørgensen 1981) pigs possessing type PhiBPhiB have a low meat quality index (KK-index); pigs with PhiAPhiA a high and pigs with type PhiAPhiB are interntermediate. Within Phi types the meat quality depends on the H type. In pigs of type PhiAPhiB and PhiBPhiB the lowest index is found in H type HaHa, the highest in type H-H- with type HaH- being intermediate.

In studies of Norwegian landrace pigs (Frøystein & al 1981) it was shown that the halothane sensitive (Haln/Haln) genotype almost exclusively found among pigs with H blood type Ha (HaHa & HaH-: 42 of 46; H-H-: 4 of 46). Furthermore halothane sensitive were almost exclusively PhiBPhiB (38 of 40; PhiAPhiB: 2 of 40). Both genotypes carrying the Ha allele (HaHa and HaH-) had markedly increased incidences of PSE meat (as determined by a rapid pH decline early pos mortem and large degree of sarcoplasmic protein denaturation) compared to the H-H- genotype.

References:

P. Fogd Jørgensen (1981) Blood types and other biochemical markers for stress-suscepibility and meat quality in pigs. In proc. Porcine Stress and Meat quality - causes and possible solutions to the problems. (Eds. T. Frøystein, E. Slinde & N. Standal). Agricultural Food Research Society, Ås, Norway. pp.146 - 159.

T. Frøystein, S.O. Nøstvold, M. Brænd, A. Storseth, K.A. Schie (1981). Halothane sensitivity and blood types in the Norwegian pig breeding stock and the associations with production and meat quality traits. In proc. Porcine Stress and Meat quality - causes and possible solutions to the problems. (Eds. T. Frøystein, E. Slinde & N. Standal). Agricultural Food Research Society, Ås, Norway. pp. 161 - 176.

Hi Junric,

I have a paper under review that details a rapid and easy method to test glycolytic and mitochondrial contribution to cellular energy production. Essentially, I culture my cells in media containing either glucose, pyruvate, or glucose and pyruvate as the major carbon energy sources. I then treat my cells with nanomolar concentrations of glycolytic (2DG, IAA) or mitochondrial (rotenone, antimycin, oligomycin) inhibitors for 4 hours. I then add a mixture of Cell Titer-Glo (promega) and CyQUANT (Invitrogen) directly to the cells. After a 10 minute, room temp incubation, luciferase and fluorescent measurements give me ATP and cell number measurements, respectively, for a combined "relative ATP/cell" index. Comparing control values in each media condition with the metabolic inhibitor treatments tells me the relative contributions of each metabolic program (glycolysis vs mitochondrial oxidative phosphorylation) to energy production.

These reagents will likely also work on protozoans, and if not, all you need to do is find the appropriate detergent to combine with the above protocol. The protocol I have generated is able to accurately define bioenergetic programs down to a minimum of 50 cells.

If you are interested in a more detailed protocol, send me a message and I will give it to you.

Best,

Nate

It is difficult to account with precision the ratio of carbohydrates and fat utilization. Probably, as mentioned by Cory Champagne, isotopic tracers + respirometry will help. With isolated mitochondria I have found that the usual usage of substrates: sucinate+rotenone; glutamate+malate; pyruvate+malate is not informative, particularly when comparing mitochondria from different organs. Usage of succinate + rotenine is particularly stupid. Unfortunately, to this day I did not published in full my data on physiological mixtures of substrates, except for brain and spinal cord mitochondria (see my ResearchGate account). Though, in my book "Practical Mitochcomdriology. Pitfalls and Problems in Studies of Mitochondria" I presented polarographic evidence of sharp increase in the efficiency of oxidative phosphorylation with heart mitochondria simultaneosly oxidizing palmitoyl-carnitine and a substrate originated from carbohydrates (pyruvate, succinate) or aminoacid (glutamate). Simultaneosly, there is a several-fold increase in production of ROS. This means, that under situations of increased oxidation of fatty acids (metabolic syndrome) the primary danger is not in development of atherosclerosis, but in increased Oxidatiove Stress. Thus mitochondria in each organ have particular mixture of substrates. Bouever, this problem is very poorly studied.

You can try a French Press - it works well for bacterial cell walls - it is a system that first pressurizes the culture to high pressure and then releases it thru a small orifice relieving the pressure rapidly - the result is like popping balloons. See this link for more information:

During night that is a resting phase for muscle cells, the energy stores have to be restored from the nutrients supply during evening. So, the AMPK and catabolic pathways should be down regulated. The low cardiac activity maintained during sleeping period do not correspond for an acute energy demand. So, I agree with you : AMPK is probably not activated during night that is conversely characterized by anabolic process. (sorry for my english that is niot very fluent...)

Valérie

HI Bridget. Thanks for that. Yes, I have read that article and while its terms are similar the context is different. "passive mechanisms" in nest thermoregulation refer to things such as nest site choice to capture environmental heat, nest modifications to reduce heat loss and supplement brood warmth. My question was more to do with the heat produced by the colony's brood and massed workers, without them actually "actively" increasing the heat production by such behaviour as shivering or wing muscle vibration. I think i have managed to sort out the problem and am hoping to resubmit the MS soon. Just waiting on some statistical support...