Science topic

Equidae - Science topic

Equidae is a family of hoofed MAMMALS consisting of HORSES, donkeys, and zebras. Members of this family are strict herbivores and can be classified as either browsers or grazers depending on how they feed.
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I have been researching on bullet proof military technology and wanted to asses the vulnerability of human body on the battlefield. I would love an layout and explanation of this question.
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This is not a simple question. You do not sacrifice mobility for protection from blast and/or projectile wounds. There are trade-offs. The development of body armor is a science. I suggest that you read about the development of recent body armor systems of the United States Armed Forces (e.g., Improved Outer Tactical Vest, Interceptor Body Armor). Wikipedia does a decent job describing these two.
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In my imaging of mouse brains, there are fluorescent dots that appear in the image from confocal microscopy which are not any of the target cells in my experiment. I was wondering if using a secondary antibody from a donkey and a normal donkey serum in the blocking could potentially cause this?
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You should avoid using blocking serum from the same host species as your primary antibody that is detected by a secondary antibody against the same species (immunoglobulins in the blocking serum will compete for the secondary binding).
I agree with Yulia Panina. Washing is an important step. Washing after each application of antibody or other fluorescent probe eliminates antibodies with lower binding affinity present in the sample and thus reduces non-specific signal, or cross-reactivity. Washing for a few minutes in PBS with at least two buffer exchanges will help to eliminate unbound and loosely bound antibody from the sample.
If you are using donkey secondary, then you should use donkey serum for blocking which you have already been doing. Instead of donkey serum you may also try using using BSA (3-5%) for blocking.
Best.
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I am working towards to design a study to asses the efficacy of different LRRK2 inhibitor compounds in neuron. I have been meaning to ask if there is any gold standard to check LRRK2 enzymatic activity.
Thank you
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You may consider the detection of phosphorylation of LRRK2 substrates via immunoblotting:
1. Autophosphorylation at S1292
2. T73 Rab10 phosphorylation
3. to a lesser extent, T72 Rab8 phosphorylation
You may also consider Phostag assay for Rab10 phosphorylation, as published by the lab of Alessi doi: 10.1042/BCJ20160557
Good luck
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I know that PSQI is used as a self reporting questionnaire to evaluate and score the sleep quality of a person. I am currently trying to test the effects of different conditions on sleep quality.
My question is: Can I use this questionnaire each day where participants will be filling it after waking up from a specific condition night? Conditions I am testing is separated by nights and will continue for 3 to 4 weeks with random order.
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This is an older questionnaire, but it has been evaluated for validity and reliability more recently. You might want to review those studies to see if the stats are acceptable for your population. You need evidence to show that one night is enough for subjects to use a specific condition and get a significant change.
Also, what is the impact of subjects taking the same tool every day for up to 28 days? Does anyone suggest rotating the order of the questions in the PSQI for that long a use?
Great idea! Good luck with this project.
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I am looking for a measurement tool for to asses antisocial behavior in university students and i did not find any good measurement tool plz if someone know plz let me know as soon as possible
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Dears,
Do we need exploit further the genetic robustness aroused from distant hybridization? Mule is an excellent example in this regard.
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Good luck.
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I did ApoTox-Glo Triplex Assay to asses viability/cytotoxicity and apoptosis on three prostate cancer cell lines (LNCaP, 22Rv1 an PC-3) stimulated with high concentration of docetaxel and campthotecin in my pilot study.
I observed no differnece between control and research sample, moreover cytoxicity was higher in control (LNCaP)
Could I ask for any suggestion what to improve? Did anyone work on this lines? Could you reccomend an alternative for ApoTox?
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MTT assay and FACS
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I'm working on my research proposal about Epistemological belief and I need help in finding an instrument that could asses students' epistemological belief specifically in physics. Your help will be greatly appreciated
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Dears,
I a bit confused. Does a mule has a 2N+1 chromosome or a 2N-1 chromosome?
Best regards,
Takele
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Mule has 32 chromosomes from Horse and 31 from Donkey
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A group of my students would like to asses the effects of video games, play station, mobile games etc on sleep quality in medical students. Hence, they are looking for a validated questionnaire that would facilitate their project.
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can we design a questionnaire for video gaming and sleep?
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What is your experience in using runoff models in small gaged and ungaged catchments (less than 50 km2)? How did you asses the accuracy of the model you used? What was the input data used in the process?
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I suggest ATHYS and HEC-HMS, Both models used DEM and Land cover and other inputs data as Rainfall, CN...
You could use Nash and Sutcliffe, Percent Bias coefficient (PBIAS), and RMSE-observations standard deviation ratio (RSR) to evaluate the output of these models.
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Hi there,
I am comparing the activity patterns of zebra and wildebeest in order to see which animal is more active, and which forages for longer over a 24-hour cycle using camera trap images. Since the population sizes differ vastly, comparing counts won't help, and I'm concerned about compounding errors associated with standardising the sample sizes. Another option I considered was comparing the proportion of images collected in which foraging behaviour was observed, with the proportion of images in which all activities other than foraging were observed. I am worried however, that given foraging is typically a "slow" behaviour relative to running and walking for instance, and the speed of these activities differ interspecifically, that it would be underrepresented and wouldn't allow for interspecific comparisons to be made.
Thank you so much,
Ryan
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Dear Ryan,
Yes. it is possible. You can use the package Overlap to determine patterns in diel activity in any species. However, the increased effort of trap-nights and number of detection (of your targeted species) will produce more reliable results.
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my project is to asses the birth preparedness and complication readiness among pregnant attending antenatal clinic in West Malaysia.
i have emailed most of the author of the related study but non of them answering my question and non of them willing to share those questionnaires
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try to rewrite questions, answers
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Planning to use an intervention. I am trying to understand the way we can take feedbacks from the field experts about the efficiency, appropriateness, and relevance of a particular intervention.
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Interesting question.
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I'm currently experiencing more diverse signs for Rabies virus in animals-particularly in domestic dogs and donkeys. But when we send samples to the laboratories, the results are more often positive.
Can anyone help me with the rationale behind this, please?
🙏
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i am planning to do my research on attachment and instant digital gratification among adolescents. if any of you have some materials related to it or scales kindly pass on to me and help me in this process. i appreciate in advance your generous heart.
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Hi Arul
You might find this reference useful as a starting point if you have not already looked at it:
  • Chou, C., & Hsiao, M. C. (2000). Internet addiction, usage, gratification, and pleasure experience: the Taiwan college students’ case. Computers & Education, 35(1), 65-80.
Keith
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I am doing Love wave tomography for the region of India and Tibet. Curious to know is there any method or way to solve the smearing in the tomographic checkerboard test.
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Thabk you very much for your reply @HrvojeTkalcic
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Hi everyone!
I am trying to use STATA to create a funnel plot to asses publication bias using proportions from studies for a large meta-analysis (having used the metaprop command).
I"m having some trouble formatting the funnel plot to give me a meaningful result.
Any help would be much appreciated! Thank you!
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Thanks for these suggestions, I was wondering if the Doi plot could be applied also to prevalence studies, when you have e.g., just numerator of incident cases and denominator population. Thanks
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Hello,
As the tittle says... Since tamoxifen induces DNA recombination, is it possible to detect this recombination using specific primers? I am also assessing recombination through immunohistochemistry, but I was looking for other methods.
Thanks
Cristina
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Yes, you can. You need to remember that WT1 is not expressed in all cells. If you do not isolate YFP positive cells, you will have some contaminations from other cells. You can FACS your cell before genotyping to avoid the problem.
Best way to do a qPCR. You can check the level of expression (at cDNA level) for your gene of interest, and compare the expression level with your control groups (Cre without TAM induction, Flox controls and etc).
Good luck
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My subject animal is the zebra finch, with which I can typically only obtain small volumes of blood (resulting in only about 20 to 40 uL of plasma). The kit I am using (Enzo Life Sciences) has a small volume protocol that requires 10 uL of plasma. I used this and found that baseline corticosterone levels were undetectable. There is nothing wrong with the kit because all of the standards and controls are fine, and the kit detects stressed-state concentrations.
How can I increase sensitivity without extending the standard curve or using more sample volume? Does anyone have experience keeping the plasma samples chilled while the rest of the kit is room temperature so the steroid displacement reagent can work more effectively to increase free CORT? Would longer incubation with the plate-bound antibodies work?
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Laura West this information may help to increase the sensitivity: Little other way of handling the sample. Before using the samples, they must be fully thawed and keep samples on ice [because we know corticosterone binding to corticosteroid binding globulin (CBG) decreases as temperature rises]. (Further reading: https://academic.oup.com/jcem/article/95/10/4689/2835223)
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Hi, I am a student from the Bocconi University, I have decided to write an Emphirical Thesis on smartwatch. I am interested in actual smartwatch users, but I have a very unsolvable doubt: can the Technology Acceptance Model be applied to actual users of any technology or the sample population must include also potential users (i.e also people that currently do not own or use a smarwatch)?
In the actual definition of the TAM said that "technology acceptance model (TAM) is an information systems theory that models how users come to accept and use a technology" but I have not yet found a research applying the TAM on an actual users, even if the definition stated that can be used for asses the use of the technology.
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Thank you all for your kind support!
So, as I can understand, the sample population must include also potential users.
I think that I will apply a revised TAM, both on actual and potential smartwatch users: it will help me to better understand the differences between these two groups of consumers, as I can evaluate not only users' perceptions but also their actual use of the technology.
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I am interested in using the Postpartum Partner Support Scale by Dr Cindy Lee Dennis in my final academic research project. If anyone can please guide me or help me about whether we require permission to use the scale, or anyone can suggest me any scale to asses Partner Support related to Postpartum Depression Thank You!
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Hi! In such attempts, try finding her contact details in public websites first and e-mail her directly. She might even be active here in ResearchGate (although I haven't checked). Is this her: https://bloomberg.nursing.utoronto.ca/faculty/cindy-lee-dennis/?
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I'm working on a retrospective study to asses the QOL in adhered rheumatoid arthritis patients, I didn't have any issues with measuring the adherence using PDC Retrospectively but im planning on assessing the Quality of life prospectively by giving the patients questioners in their next follow up , any ideas on how to do that and is it a correct way or there are other preferred methods.
also is there a way on how can I measure the QOL retrospectively ?
thank you
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Abdulsatar Mathkhor Thank you very much for your help , but if our disease is rheumatoid arthritis and we want to asses the QOL depending on ADHERENCE , what do you suggest as a method ? we have the tools such as PDC,AND WHOQOL-BREF questionnaire But we are having a problem on how can we utilize these tools, when do we give them , how we will follow up , and in your answer do you mean depression as an example? but how it will correlate with adherence ? since it is our focus and our objective is that we aim that people with better adherence have a better QOL.
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I want to demonstrate that there is grow of primary follicle to a secondary stage in vitro not only with histological analysis but also with qPCR, any ideas on which genes could I analyse that have different expression in primary and secondary follicles?
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growth and development
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Using formalin fixed brain sections. Sections are 50μm thick and fixed on the slide.
Was using 1 hour antigen retrieval in Citrate Buffer (ready made, sigma one) at 90C temperature.
Then washing 3 times for 10 minutes with 1% PBS-Triton-X100 and blocking (for 1 hour) with in the same PBS Triton with serum of Goat and Donkey, each 0.5%. I am not washing the sections after blocking. Just throwing out the blocker and then diluting the primary antibody in the same blocker.
Then Primary antibody (1:1000) overnight at 4C and then washing with PBS-Tween-20 (1%) and making of secondary antibody (Alexa Fluor) (1:1000) in PBS-Tween20 (1%). after the secondary antibody, washing with PBS. Using VectaShield as mounting medium.
Please tell me:
1. Should I increase the antigen retrieval time or temperature? or both?
2. Should I change the blocker? I am using 0.5% Donkey and 0.5% Goat serum in 1% PBS-Triton X100
2. Should I increase the PBS-Triton-X100 concentration from 1% ?
3. Should I increase the PBS-Tween-20 concentration from 1% ?
4. Should I change the mounting medium?
5. Should I increase the washing time. Now I am washing thrice for 10 minutes after each step.
OR
should I change the whole protocol ?
Thanks!
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I agree with Vahid M. Harandi that 2.5 or even 5% BSA, Goat, or Donkey serum is preferable. Also since sections are fairly thick I would suggest blocking overnight at 4C. You also missing the permeabilization step before blocking for your section thickness it would be about 30 minutes in 0.5% Triton X it might not be required if you are looking for the cell surface proteins. I also think your time in citrate buffer is too long 15-20 minutes is usually sufficient. Keep in mind that brains are fairly autofluorescent at 488 nm so your secondary should avoid this wavelength the best is 647nm/CY5. I believe by increasing blocking solution concentration and using 647/Cy5 secondary you will get publishable quality data.
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I was offered to become a part of this journal editorial board - and encouraged to publish in it
I am suspicions.... you have to pay a substantial fee for publishing , the journal does not appear in SCIFINDER
If you can help me to asses this journal - I will be grateful
thanks
best
Yosef Scolnik
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In addition to the good points made above, any journal that charges a "substantial fee for publishing" is a predatory journal. Nice trick, " We'll put your name on the editorial board and take your money for each article you publish."
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Hello all,
I am trying to asses the viability of stem cells in GelMA (gelatin methacryloyl). Most articles make their own GelMA. However, our team bought a commercially available one. The thing is : the product is fluorescent-green under the microscope, so it is difficult to evaluate the cell viability with the available staining kits, which show live cells also in green.
Thank you,
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Dear João Luiz Monteiro,
I believed your current finding on GelMa is auto-fluorescence with green color thru specific wavelength. To my knowledge you could diminish or differentiate the auto-fluorescence thru the software and identify true wavelength for your vianlbility kit. It is totally challenging but could do that. Otherwise, you could using live cell tracker with different color as I believed your fluorescence color for dead cells is red in color? You could justify your procedure for some limitation, that's research.. Best of luck!
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Experimental and comparative studies suggest that the striped coats of zebras can prevent biting fly attacks. Is there a practice in meat industry that can be fusion with the zebra striping taking advantage of both techniques?
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I think there is use and studies on genetics, but in a limited fashion
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I would like to do an immunostaining on mouse brain. I have a primary raised in goat and an one raised in rabbit. For secondaries I have a donkey anti goat and a goat anti rabbit (I do not a have a donkey anti rabbit). Is it possibile to do a serial incubation for the secondary (first the donkey anti- goat; wash, block, then the goat anti rabbit)? Will this prevent the cross- reactivity of the donkey anti goat to bind to the secondary goat anti rabbit?
Thanks a lot for the help!
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While cross-adsorbed secondary ABs are good, you can also use TSA and use citrate between staining for each epitope.
If you develop TSA-based protocols you can go into the multiplex environment, adding more stains/colors even using primary ABs from the same species.
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I would like to asses the psychological distress among August 2018 flood-affected individuals.
Can I use GHQ-12 in 2020?. Because there is a time gap of 1 and half years after the floods, does it give an accurate result?
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My first thought is there might be a better measure for psychological distress that's less general than the GHQ-12. But also, regarding you question about the past hurricane and measuring psychological distress now, the only way you might justify that study would be to compare your hurricane survivors with a benchmark of control subjects who are of similar age/descriptions, etc. Because otherwise, there is no way to conjecture whether any current distress results from the hurricane or not. Another option to consider might be to construct (or find) a survey or questionnaire specifically relating to effects of disaster/hurricane survival.
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Currently working on ASS affected Ramsar Wetlands.
I'm particularly interested in the effects of sediment on migratory wading shorebirds, and on the invertebrates that lay eggs into the sediment.
What are invertebrate eggs called, how long can they persist in sediment, and does any particular geochemistry effect the integrity of the egg's shell? Or is it only the acid produced by ASS that's an issue? And if so, is there research that quantifies the effects of the acid on the invertebrates, or their eggs?
Do wading birds get sores from acidic pore water on mudflats? Does the geochemistry of sediment affect them in any way, at any stage, or is it simply the threat of ASS producing acidic water?
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Regarding birds, have you read Blancher's papers?
I didn't find much on invertebrate life-cycle... And I suspect that you already read what I have.
You have picked a very singular research line. Naturally, there will be lack of literature.. You'll have to search throughout other papers and pick up bits of information from here and there.
We don't treat invertebrate eggs by any other name. Is well known that nematode eggs are very resistant, and they are abundant in wetlands... Would be a very interesting subject to investigate, I think.
Cheers!
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suggest alternative cell lines for HaCaT cell lines for the assessment of antipsoriatic activity
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Check this recent article:
Lysosome Alterations in the Human Epithelial Cell Line HaCaT and Skin Specimens: Relevance to Psoriasis
Int J Mol Sci. 2020
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Dear Corrosion Experts,
Is there any research paper comparing various stainless steels ASS, DSS, MSS at different pH, Cl- ion concentration and Temperature? please share if someone has it.
Sincerely,
Abdulkadir Godil
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I recommend a publication by my colleagues from BAM Berlin, who have carried out a project to compare alternative stainless steels (for civil engineering) and their corrosion behaviour.
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Hi everyone,
I am writing this to gather some suggestions for my thesis topic.
I am a student of MSc Quantitative Finance. I am in need of some suggestions from the experienced members for a research topic in Portfolio management.
My expertise are in statistics and empirical analysis. I believe that I will be able to present some good work in field portfolio analysis. Currently, I am researching for some good topics where I can apply machine learning or machine intelligence e.g. for forecasting portfolio performance or may be use it to asses portfolio optimization strategies.
I will be very grateful for you suggestions and guidance. If it suits you, you can also email me on narendarkumar306@gmail.com
Regards.
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Topics
  • Behavioral Finance ...
  • Derivatives. Options ...
  • Factors, risk premia. Analysis of individual factors/risk premia ...
  • Fixed income and structured finance. ...
  • International Investing. ...
  • Legal/regulatory/public policy. ...
  • Long-term/retirement investing. ...
  • Mutual funds/passive investing/indexing.
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Assesing interaction by use of Parflow alone can lead to appropriate result?
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I think you can use HYDRUS 3D and modflow
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My cells are cultured primary neuron from fetal rat, transfected with GFP-HA-double tag surface protein at DIV 7.
IF was preformed at DIV10-12, fixation with 4% PFA 15min and block with 10% Donkey serum. 594 fluorenes marked HA tag.
below is my picture captured with 60X microscope. The first picture is kind of OK but most of the rest is quite ugly. I am new to this and want to know what happend, would you please help me figure out?
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You have to try with more dilution in secondary antibody
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The sections are about to mount. As far as I know leaving them too long in PBS (can correspond to too extensive washing) might weaken the signal.
(I have Dylight 488 goat anti-rabbit and Cy3 donkey anti-mouse sec. antibodies.)
Thank you.
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Thank you for your responses.
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Hello everyone
I am trying to asses the inflammation in mice feet but I am a bit confused about the best way to measure the inflammation if I can use any device .
please help me with your valuable suggestions
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I am looking for a way to asses animal welfare in a zoological institution on a longer term. I was wondering if anyone has used AWAG before, and what their experiences were?
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Take a look for animal welfare in zoos:
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Please, English article
thank you Doctor
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To all scholars.
your inputs are welcome
1. How to identify a worthwhile opportunity and asses the identified opportunity using the PESTEL AND Porter’s five force model?
2. How to position the opportunity in the Ansoff’s opportunity matrix. decide growth paths from the initial quadrant to other quadrants when the company capture more opportunities.
3. How a company benefits from the BCG Matrix. What would you advise to a company which occupies a weak position in the “Dog” quadrant? What would be the generic strategy for a company in “Star” quadrant??
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You may a chose a company which has various brands/products and operates in several markets (e.g. Unilever, ABF). You can pick a market and analyze its ecosystem and 5 forces (e.g. Australia, Turkey, USA for Unilever).
For Ansoff, you need a multi-brand and multi-market company so you can analyze existing product/existing market/new product/new market strategies.
For BCG, Dog position need to be closed if there is cash drain and retain Star position for greater market share until it becomes a cash cow.
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Will using Bovine serum albumin(BSA) as a blocking and dilution agent with donkey anti-sheep secondary antibody yield significant background?
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I routinely use 1% BSA routinely for immunohistochemistry. I have not had significant background problems. You should be fine.
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i want to asses oxidative stress in hepg2
is it right to measure lipid peroxidation and oxidized and reduced glutathione in cell line and what is the procedure and protocol
please with reference
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Heba Essam you are right that lipid peroxidation and oxidized/reduced glutathione can be used to measure oxidative stress. However, as a note, you may also wish to consider other oxidative stress biomarkers such as 8OHdG, 3-Nitrotyrosine and protein carbonyls as well as other tests including DNA oxidation. I highly recommend the book "Free Radicals in Biology and Medicine" by Barry halliwell and John M. C. Gutteridge. This book is highly regarded in the free radical biology field.
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Hi Flow Folks,
I've got a puzzle I need your help with. I've developed a flow assay to stain for intracellular neuronal markers. The protocol uses either 2%PFA fix with 0.1% Triton X-100 perm or the Thermo eBioscience FOXP3 kit. With either kit I also add 10% donkey serum to the perm/wash staining buffer. I also maintain the perm condition through the primary and secondary stains since I'm using indirect staining methods. I've used this method to titrate a total of 6 unique intracellular antibodies. I have good staining index for these markers.
Here's where it get interesting. To test the antibody specificity I want to use negative control cells that do not express the markers of interest. Using Human Protein Atlas I selected BJ, PC-3 and Reh cell lines. However, when I run this protocol I get positive staining on my negative cells! This has me quite puzzled because we have validated these antibodies for use in ICC and know that they localize correctly. Take a look at the attached image for one marker (MAFB) to see what I'm talking about. Secondary only and also isotype plus secondary give negative staining so I don't believe this is just simply blocking. What do you think?
-Mike
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Hi Mike,
I see the pattern you describe with MAFB. Is it the same with the other antibodies you're testing? With the 6 unique antibodies you've worked out already- were those tested against negative control cells as well, but did not do this?
Rich
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To asses the success of of prepared adsorbent what isotherm it should follow? I mean there are lot of isotherm studies describing different methods of adsorption out of them which is considered as the best so a to manufacture an adsorbent commercially?
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By comparing isotherm at lab scale you can not decide on the performance of the absorbent.
Besides the aforementioned criteria (production and operational cost, regenerability, selectivity toward the desired compound) physical and chemical stability should be considered non of which are shown in isotherm. In my part, the isotherm might only be useful in giving and insight into adsorption capacity which differs in lab and industrial level.
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Hi
i have a set of users that are located in different Ass (Autonomus System) and I want to find for a target user list of thier neighboor (closed location ) , i find works that say As -level topology has been widely used to measure distance between internet users , i want to have more information like how to measure distance between two As ?
thanks
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C K Gomathy thank you so much
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Mice were perfused with PBS and 4% PFA
we use a rabbit anti mouse ASB4 AB in PBT buffer with 5% normal DS on at 4°C followed by a 2h RT incubation with donkey anti rabbit Alexa fluor 568
wash steps are with plain PBS 3 x 10'
even in our negative control (only stained with secondary AB) we get a slight background film which makes it hard to see the true positive cells.
Any ideas what could be the reason for this?
I've added a similar protocol in attachment
thnx
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I agree with Renee, the secondary antibody incubation is too long. I always do half an hour at room temperature. It may also be that you are using a highly concentrated secondary antibody solution in combination with long incubation. Have you tested a concentration range of this antibody? I highly recommend to do so. Also the primary antibody seems quite concentrated in the protocol. Does your staining solution also contain blocking buffer? This is important.
You say you get a background 'film' so I believe your deparaffinization is not very successful. Leave the slides 5-10 min in the xylene and then 5 min in each decreasing alcohol solution. Also, try using 1% BSA in your blocking solution.
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Hello,
I am looking to do some simulation on liquid crystal. What I am trying to simulate is a system like this. Let say I have an electrode which on top of it has a polymer layer that works ass an aligment layer for the liquid crytals on top. How would the direction of the liquid crystals would change as I change the voltage applied to the electrode.
Thank you
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It depends on the complexity of the problem that you wish to simulate. In short, you can use LCD Master but I would recommend COMSOL.
If it is only what you described, then you can use LCD Master — this is specialised software for simulating liquid crystals and it is widely used in industry (so is COMSOL, for custom models). However, from my personal experience, I would NOT recommend this software. It caused me lots of problems and after switching to COMSOL, I never looked back. If you want to use the order tensor representation or defects, you will need to use something other than LCD Master.
If you are okay with writing your own equations, I would recommend using COMSOL. It doesn’t have a liquid crystals module, but it does have an easy to use interface and a very good solver built into it. You can create a custom geometry, write your own equations (by using Euler Lagrange minimisation on the free energy integral) and have lots of control over your model. You can also use any other FEA software that allows you to create custom models, such as MATLAB (you will need the FEA package for this) or Python (SfePy package is free I think).
If you have access to COMSOL you should try it with a simple 1D or 2D model with a single elastic constant (I would say that 2D is easier to construct and visualise when you are starting out). Otherwise try using MATLAB or python (a bit harder to use but not too bad once you understand how to use it). I think that the following book has the Euler Lagrange equations for the director in 3D in it “The Static and Dynamic Continuum Theory of Liquid Crystals: A Mathematical Introduction”.
Edit: I would recommend getting a trial version of COMSOL and trying it out before purchasing it. I don’t k ow if LCD Master offers trials, but it’s worth asking them.
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Hi everyone. I am interested in asses biological activity of purified reelin in cultures, so I have decided to analyze phosphorylation of Dab1 by immunoprecipitation and WB against p-Tyr residues, as other authors have done. Could anyone recommend me any good antibody for immunoprecipitation of Dab1? Thank you so much
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Thank you Russell. Unfortunately, there are no good reviews for the Dab1 antibodies you suggested. All the best
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Hello
For the past few months, I've been trying to knockdown a gene in HEK293 cells using siRNA. I've tried different transfection reagents and have recently started using RNAiMAX as I had heard that it's very effective.
I have noticed that it kills my cells and that when I'm doing RNA extraction, the RNA concentration of samples that contained Lipofectamine (even the mock samples without siRNA) have very low RNA concentrations. I then do qPCR to asses knock-down efficiency but I don't know how trustworthy the qPCR results are given the quality of the cells.
I'm using a 24 well-plate and cells are about 50-60% confluent at the time of transfection. I create a mastermix of Lipofectamine and OPTI-MEM as instructed. In the end, each well contains about 1 μl of Lipofectamine. Is this too much? I used to use more than that but since it's been killing my cells I've been gradually decreasing the concentration but still no success.
Does anyone have a successful protocol for mammalian cell line siRNA transfection using Lipofectamine? Any help will be greatly appreciated. Thank you.
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In my hands, the RNAiMax reagent works great if you swap out the media at 24 hours. You don't need the transfection reagent in there for that long, it has already done its work if its going to work, and we see enhanced cell viability if you change the media.
Additionally, make sure your media that you are transfecting in does NOT have antibiotics in it, this also enhances cell viability.
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Describe the problems,consequences and asses the performance of the projects
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I have an interesting presentation about: The Potential Role for Hydro-Economic Projects in Sustainable Development.
Please check the link:
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in a cross of male horse with a female donkey fertilization takes place inspite of different genomes, but the offspring's produced are sterile.
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In order to asses muscle performance in MDX mice, I have performed the four limb hanging wire test. According to the literature this test should be able to discriminate between MDX-mice and C57/Bl10ScSnJ mice. However the majority of C57/Bl10ScSnJ mice deliberately jump off the grid. None of these mice were able to hang for 600 seconds, however they are able to hang for 100 seconds. Maximum hanging times of C57/Bl10ScSnJ are very similar to those of MDX mice. I have followed the protocol that is described in SOP's.
I have tested 4, 6, 8, 10, and 12 week old mice. During my first set-up mice hung at a height of +/- 27 cm. In order to improve the hanging time, I have increased the height untill +/- 40 cm, however this does not seem to have a lot of effect on the max. hanging time... Any recommendations? Should I increase the height even further, however I do not want them to harm themselves.
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I am performing the four limb hanging wire as described in http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/DMD_M.2.1.005.pdf.
Yes, I know this test is also dependent on the motivation of mice. However, as described in literature and in this SOP, this test should be able to discriminate between healthy and MDX mice. In addition to the four limb hanging wire, I will also perform electrical stimulation of muscles in vitro. However, I want also an in vivo test to assess muscle strength...
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I found that to asses effect of intervention we usually use one group pretest posttest design. but that design mainly uses primary data while I'm in a position where I can only use secondary data
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Since the analysis would be identical, regardless of whether the data was primary or secondary, I don't see any problem here.
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any one use biological methods to asses the vaible form of heavy metals in the soil intade of chemical one
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I will isolate different fungi and see if they produce any metabolites with antibacterial properties against E.coli and Staphylococcus aureus. I want to use 3 different media and grow the culture in both solid and liquid substrate.
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If you want to check antimicrobial metabolites of fungi better to use liquid media. But also take into count that several antimicrobial activities can be inducible which means that activity will observed in your liquid media only when bacteria of your interest exist in same media and you will not see activity in liquid media of fungi when it grow alone.
Hope this info will help you.
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I have found mixed nematode infections in donkeys sampled from a local community in northeastern Nigeria. I am looking for the best way to explain my findings in terms of epidemiological significance.
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The epidemiology significance is that there are mixed infections which could either boost and negatively impact the immune system of the donkey. Depending on the immunomodulatory effect of these worms in donkeys. Howbeit, the public health significance is also very important, since donkeys are often used by humans in those areas. You may want to know of some of these worms are zoonotic. It is also important to know the infective stages of these worms and how they impact the life of the animal in question. It's an interesting finding that could be broadly explained. You may also want to conduct some molecular studies on the worms.
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Dear Researchers/Scholars,
Suppose we have time series variable X1, X2 and Y1. where Y1 is dependent on these two. They are more or less linearly related. Data for all these variables are given from 1970 to 2018. We have to forecast values of Y1 for 2040 or 2060 based on these two variables.
What method would you like to suggest (other than a linear regression)?
We have a fact that these series es have a different pattern since 1990. I want to make this 1990-2018 data as prior information and then to find a posterior for Y1. Now, please let me know how to asses this prior distribution?
or any suggestions?
Best Regards,
Abhay
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Let me play the devil's advocate:
You have data for the past 50 years. However, you say that there is a mayor break or change in the pattern around 1990, so that you want to use only the more recent 30 years ... to predict what will be in 30 or 50 years in the future?
I doubt that this makes any sense. Toss some dice. It will be as reliable as your model predictions.
If "phase changes" like around 1990 can happen, they can happen in the future, too. Additionally, many other things can happen that we are not even aware of today. The uncertainty about such things must be considerd. Further, as you don't have any model that might be justifed by subject matter arguments, there is a universe of possibilities, again adding to the uncertainty of the prediction. If you consider all this, you will almost surely find that the predcition interval 30 or 50 years ahead will be so wide that it can't have any practical benefit.
You can surely grab one possible model, select some subset of your data, and neglect anything else, then you can make a possibly sufficiently precise forecast, which applies to this model fitted on this data, assuming that nothing else happens or can impact the dependent variable. Nice. But typically practically useless. It's a bit different when you has a model, based on a theory. Then you could at least say that this theory would predict this and that. But if you select a model just because the data looks like it's fitting, you actually have nothing.
It's important to think about all this before you invest a lot of work and time in such projects! It may turn out, in the end, that your approach is still good and helpful. But many such "data-driven forecast models" I have seen in my life have benn completely worthless, pure waste. Good enough to give a useful forecast for the next 2-3 years, but not for decades.
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In order to assess the risk of confined space working area, we need to identify all the hazard first. After we identify the hazard, we can determine the risk assessment by risk matrix. In order to reduce the risk matrix, we should determine the risk control by hierarchy control (qualitative). So, the question is, how we could obtain the value of each control quantitatively ? so we can reduce the previous risk assessment.
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A rule of thumb is that hazards evaluated between 20-25 (Likelihood * Impacts, scale 1-5) are ranked first fro treatment, even stop the work till to mitigate or eliminate them. After that, hazards evaluated between 15-20 must also be treated and so on. Nevertheless, bear in mind that hazards which might directly threat employess' lives are of first priority to treat them, even though its evaluation are not lying between the aforementioned limits.
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I am studying 6 behaviours in 2 separate groups of zebras in a zoo, and comparing them. I want to compare how long the 2 groups (as a unit) spend showing each behaviour. I will be observing them for 5 weeks and by the end should have a table like this -
Avg Duration (mins)
Behaviour Gp 1 Gp 2
forage
rest
social
vigilance
move
other
I'm not interested in within-group comparisons (e.g. Gp 1 forage compared to social), but I am interested in between-group comparisons. However not only would i want to compare foraging between the 2 gps, resting between the 2 gps and so on, but i would also want to compare e.g. gp1 forage with say gp 2 social - between categories of behaviours between the 2 groups.
Now I was thinking ANOVA is partly about whether 1/more variables of various levels have a effect on another variable, but the only variables i have will be time and behaviour. I don't especially think one will have a dircect effect on the other - it's purely just observational. So would I have to do multiple t-tests, since I'd only be looking for differences between 2 means at a time?
Cheers!!!
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I would use a chi squared test (Null: frequency of behaviours is the ~same across groups). I would start with all behaviours and, if there are significant differences, analze one behaviour at a time. You can do this with or without time as a factor (former is more complicated). e.g., Analyzing Frequencies chapter in Quinn & Keough (older versions of the text are available for free online). Let us know how it turns out!
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We would like to know some information about this animal and whether it is genderless or productive.
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Mules are a cross between a female horse and a male donkey. Since mules are cross between species, therefore they are not fertile. Mule is an ideal example of "hybrid vigour".
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Dear,
I have a question about how i could analyse my data.
My question for my thesis is:
what is the effect of a game on the social skills of people with ass, and is this effect differ in gender, age, work and education level.
In my analysis i did a paired t-test, which allows me to check if the average on the pre-test differs from the average on the post-test.
But how can i do a paired test which analysis:
wheter the effect on the pre-post test differs in gender, work and education level?
Could someone help me, please?
My analysis are done with SPSS
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Where is your supervisor? You are doing a postgraduate degree – you should have people to teach and advise you. If you are here, on ResearchGate, it means that they are not doing their job.
Please insist on getting proper statistical help from your institution. You need to sit down with a statistician who you can explain your research to, and who will help you.
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The Delphy study or technique is a survey method designed to structure group opinion and discussion, generate group consensus, and quantify the judgments of experts, asses priorities, or make long-range forecasts (Waltz, Strickland, & Lenz, 2005). If we use this technique to quantify the judgments of experts, how many minimum sample size for this study?
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I think that this article can be of help to you.
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Hi, I am working with Hela cells and my treatment condition is always UV light. One of the assays that I work all the time is ELISA on UV photoproducts, more specifically cyclobutane pyrimidine dimers. The issue is that i cant get to make the assay robust. It seems that it was always unstable in the lab and the way it has been coped with was to basically bombard the bench with a million of experiments and hope that some will work out fine with all the controls behaving as expected and then basically consider your other samples. I am afraid this is not a possibility anymore, I am running out of time and I just need to make sense of this situation as very often than not I just spend weeks on getting my samples and then all this work goes to trash. Anyways, I am pretty confident that the cells are fine, that the irradiation step is correct, and the ELISA per se is fine. I always run 3 technical replicates per condition and they are always satisfactorily similar. I additionally trust my pipetting and the fact that the technical replicates are fine prompts to think that whatever the problem is, it is in the tubes/samples at some point in the process. The hypothesis I am now considering is that maybe UV irradiated DNA is not stable in the sense that, freezing and thawing is able to break the DNA with a articular affinity to the sites of UV photoproducts. this would explain the situation really beautifully, as I can easily see how I am loosing my epitopes in the tube when they break in a somewhat "stochastic" way. It additionally aligns perfectly with the notion that the higher the DNA yield I obtain, these samples behave better, as there is this rule of thumb rule that the higher the concentration of a chemical the more stable it is. I am of course testing this possibility empirically but I would like to know if anyone noticed this or if this is an actual notion in the field. I have tried to find the answers online but cant get any such studies and I am just so baffled that if this is the case how it is possible that no one ever told me before. Additionally, since I am thinking this way could it be that the vortexing is also doing the same thing? this would be a massive pain in the ass, as I handle many samples/experiment and i need to be super precise with the amount of DNA i load, and avoiding vortexing would make the whole thing far more time consuming and additionally could introduce variability due to the loss of precission in my nanodrop quantifications and when I prepare my dilutions from the samples.
Thanks a lot
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Thanks for your answer! but I am not sure if I get what you mean, I extract DNA from the cells using an standard QUIAGEN kit and those are the samples i work with.
Additionally UV induced DNA damage is far more than well established, despite the damage it can also cause to other types of molecules, UV does damage DNA
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I am doing free floating immunofluorescent staining on my mouse brain sample.
I've been having difficulty using rodent host (ie. mouse, rat, guinea pig) primary antibodies, and I don't know why. There is no detection under confocal microscope.
Even for the popular antibodies (ie. NeuN from Millipore etc) don't work on my sample.
My samples were previously exposed to 4% PFA (perfusion + overnight post fixation) then washed and stored in 1XPBS with 1% sodium azide.
Tissues were embedded into LMP 3% agar and sliced (50 um), using vibrating microtome.
So far I've tried...(***blocking solution contains 0.1% tritonX-PBS)
  • 10% FBS blocking with donkey anti-mouse secondary
  • 4% BSA blocking with donkey anti-mouse secondary
  • 5% horse serum blocking with donkey anti-mouse secondary
I'd really appreciate if anyone could suggest me protocols/references.
Kind regards,
Yuka
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Yuka,
I have a couple of suggestions. First, we typically use a 4 hour post-fix for CNS samples. However, most antibodies will work with an overnight post-fix. After the post-fix, transfer your samples to a 30% buffered sucrose solution. The samples should remain in this solution until they sink to ensure proper freezing (this may be why your samples wrinkled after transfer). Once the samples sink, they are ready for freezing and cutting. Once tissues are cut, follow this protocol:
Day 1
  1. Wash 3 x '3' in 1x PBS
  2. If using Biotin/Avidin, use a strepatavidin/biotin blocking kit such as Vector SP-2001. If not, proceed to step 3
  3. Protein Block (3% normal sera matching host of your secondary + 0.3% Triton X-100 in 1x PBS) for 30-60' at RT
  4. Incubate primary antibodies in antibody buffer (1.5% normal sera + 0.3% Triton X-100 in 1x PBS) overnight (16-18 hours) at 4 degrees Celsius (you can do RT for 1 hour before putting in 4 degree as well).
Day 2
  1. Wash 3 x 3' in 1x PBS
  2. Biotin secondary incubation (if using biotin, 1:500 in 1x PBS) for 30" at RT. If not using Biotin, skip and go to step 6
  3. Wash 3 x 3' in 1x PBS
  4. Avidin/Strepatavidin Incubation (1:500 in 1xPBS) for 10' at RT.
  5. Wash 3 x 3' in 1x PBS
  6. Secondary Incubation (1:500-1:1000 in 1x PBS) for 1 hour at RT.
  7. Wash 3 x 3' in 1x PBS
  8. Coverslip
CNS usually does not require antigen retrieval. However, the type and length of fixation can affect this and mask your antigen. I think overnight with 4% PFA may be causing a problem masking your antigen. I can suggest some antibodies that are tried and true for us as well if you would like. Alternatively, you could try using a method of antigen retrieval to try unmasking your antigen from the paraformaldehyde. I would also try to run a positive control and a negative control with your IHC each time as it can help you answer some questions about what is going on. Hope this helps.
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I am currently tryig to do double immunfluorescent staining in myotubes but I could not be able to see a clear negative control.
As you may see in the presentation, I used two different kinds of negative control. 1) no primary antibody 2) Another cell line that should not be expressing desmin (fibroblast). In both of my negative controls I still have strong staining, especially localized in nuclei that I could not get rid of.
%4 PFA is my fixative agent. I am using donkey anti-rabbit (AF488) and donkey anti-goat (AF568) secondary antibodies in 1/5000 dilution.There is not any negative staining in Texas red but in FITC. I do all the washes with %0,5 PBS(T) and 15 minutes. I used 1% FBS and 5% BSA blocking but there was no change. Moreover, I changed the cell type and do the experiment in C2C12 myoblasts but still the same result. Finally, I did desmin staining on its own but negative control was not clear again with donkey anti-rabbit (AF488) but when we do double staining in skeletal muscle tissue, the result was satisfying; no staining observed at negative control.
What could have gone wrong with the cells or antibodies? Any ideas?
Thank you for your kind answers.
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Dear Seyda,
One problem may be autofluorescence from Paraformaldehyde, which is typically seen in the FITC (488) channel. If you take some cells run them through your protocol, but do not apply any antibodies at all (either primary or secondary), do you still see a light green signal?
If so, there are methods to quench autofluorescence. I usually use 0.1 M glycine in PBS, pH 7.4, as a 30 minute rinse, before adding my blocking agent, on 11um-20um tissue sections. If you don't have any glycine, lysine works too.
Otherwise, try using a different wavelength fluorochrome. Jackson ImmunoResearch has a large variety of Donkey anti-rabbit secondaries that are useful for double-labeling (note: no financial interest).
Good Luck!
Jill
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Hi I have two dose response curves fitted on aseveral data points made with different drug concentrations. I would like to somehow asses whether the two fitted curves are significantly different.
Could someone propose a method, and if possible a statistics package where it is availalbe?
Thank You in advance
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Hi everyone
I am looking for some studies or books in (metal or coal) pit lakes that have summarized (if possible) the combined effect of oxygen physical entertainment into bottom thermal zones (eg monimolimnion) and the impact of biogeochemical processes (eg methane, ammonia and hydrogen sulfide oxidation).
Most studies seem to assess these two processes separately or by hydrodynamic modelling. While the latter helps, modelling requires continuous validation. I would be interested in (field) studies integrating both mechanisms to asses the oxygen dynamics (i.e oxygen supply vs oxygen demand).
A great example of what I am looking is:
Thanks – hope to get an answer soon
Daniel
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What do you recommend your patients after rotator cuff repair concerning return to work?
- Do you give general recommendations?
- Do you differ between physical vs. non-physical workload?
-Do you asses the capa to return to work by time and/or functional testing?
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Dear Tim
The ability to return to work after a rotator cuff repair depends a lot on what type of work you do.
This article may be useful for you: Farhad O. Moola MD, Inc. Orthopedic Surgery University of British Columbia Trauma • Hand Surgery • Shoulder & Elbow Reconstruction
For most sedentary jobs, I recommend taking one to two weeks off work.
When you return to work your arm will be in a sling (for 4-6 weeks after surgery), but you should be able to manage as long as you do no lifting, pushing, pulling or carrying.
Light duty work involving no lifting, pushing, pulling or carrying may begin within four weeks after surgery.
Work at waist level lifting 5-10 pounds can begin 3-4 months after surgery.
Work at shoulder level can begin 3-6 months after surgery.
Heavy lifting or overhead use may require 6-12 months before full recovery and work return
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Hi!
I am trying to detect incorporated BrdU in PPFF liver tissue sections of zebra finch. I am using abcam kit for this purpose
I am looking for someone to comment my first stainings, if this staining is only artefact or it is acceptable for collection of data.
My biggest concern is about high signal from end parts of tissues but not in places where tissue were cut off. Is it possible that BrdU is there because of diffusion (BrdU was administrated intraperitoneally 1h before dissection).
I am looking forward any suggestion and comments,
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Question about diffusion was my “wild speculation” but now I know it is an artefact I have to work on. For these samples I have very limited possibility to try different fixation protocols because samples are already collected but I saved this suggestion for future.
So my next step for now I have to try to modify concentration of antibodies, and try to understand what is happening with positive/ negative controls.
Thank you very much for such informative discussion.
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I would like to make an elisa sandwich with this microplate 1) mouse hybridoma supernatants as capture antibodies, 2) a rabbit monoclonal antibody in detection, followed by a donkey secondary antibody
Does anyone have experience with this? Is there a risk of nonspecific binding?
thank you in advance
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This might work, if the antibodies are selected carefully. A general answer about non-specific binding is not possible. However, you should prefer high-affinity antibodies and use low reagent concentrations. Since you plan to use 4 different antibodies, from which two are polyclonal, potential species cross-reactivities need to be examined in detail.
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Any validated exercise protocol for PCOD?
How can asses the outcome ?
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These articles may help; Exercise and reproductive function in polycystic ovary syndrome: protocol of a systematic review
  • Isis Kelly dos Santos, Romilson de Lima Nunes, Gustavo Mafaldo Soares, Tecia Maria de Oliveira Maranhão and Paulo Moreira Silva DantasEmail author
Systematic Reviews2017
6:264Quality of Life Research
October 2017, Volume 26, Issue 10, pp 2593–2605 Quality of Life
Does exercise training augment improvements in quality of life induced by energy restriction for obese populations? A systematic review Daniel J. van den Hoek, Clint T. Millerm, Steve F. Fraser, Steve E. Selig,John B. Dixon
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Dear Colleagues
I have a complete genome sequence of a s.aureus isolated, and is looking for a quick way to asses whether it is identical to one of the 324 s.aureus complete genomes already registred in refseq - if any one have a good way of doing this I would be pleased to know of it.
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Hi Marius,
You would probably need to download all the RefSeq sequences and apply simple phylogenetics to determine the genetic relationships (there is an automated way to do this). There are many ways to do this, you can either use reference based approach such as Snippy, or core gene approach such as Roary pipeline and get the sequence alignment to build a tree. Some other alignment free approaches e.g. CVtree, but more or less the tree will be pretty much similar in topology.
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in case of equidae family
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When you remake the tree with only the sequences used in the original source you are referencing, do you get the same result? Are you using the same model and parameters as used to generate the original reference? Assuming replicating the original gives the same answer, then the changes come from adding more data, which is unlikely to actually be a problem. As long as the extra data is not bogus, the addition of data often changes our results and interpretations somewhat.