CharonY

Please clarify this to me. In order to measure recovery do you have quantified (beware, did it in a rush and may get numbers wrong): A) Original concentration (without separation). Assuming you started with the 0.32 ml , your concentration would be Xmg/ml and multiplying it with 0.32 ml the original concentration is 0.32X mg total (X being 0.5 in your example) B) After diluting it in a final volume of 5 ml you therefore start with 0.32/5=0.064X mg/ml (as you have 1 ml injection). Effectively you concentrate the sample to double the concentration, if you captured everything (final volume 0.5 ml) resulting in 0.128 Xmg/ml as concentration, assuming you used the same volume of your eluate to run the assay as in A). However, since you only have 0.5 ml total, you still only got 0.064 total load. C) Now you load that amount on the second column. Here you should also take into account what final volume your eluate is now (which I cannot find in your posts) prior to running the assay. E.g. if your eluate is again 0.5ml you should yield the same amount (again, assuming no loss). Any other volume would increase or decrease the concentration accordingly. D) Now if you run your original sample undiluted and loaded the whole 0.32ml you start off with five times the amount (with an 1 ml injection), assuming you got the same elution volume. If you do not get that, there is a chance that you may overloaded your column, especially as other components in your sample can interfere with the analysis. Diluting the original sample and run a SEC alone is a possibility, though a dilution series could help in assessing whether you are actually still in the dynamic range of the separation method.

None of which would matter if you start your treatments with different amount and types of bacteria. The best explanation at this point is that you transferred different amounts of bacteria to the plates (though it is hard to tell as you have not specified how you measured growth). In other words, you first need to know the characteristic of your untreated condition and get that reproducibly. Only then can you be sure that differences because of whatever kind of treatments are really due to the treatment and because of how you set up the experiment. The interpretation comes after that. Finger swabs or similar techniques are quite unsuited for assessing bacterial growth, rather you can only assess the contamination you got at the moment of transfer (limited to what is able to grow on the plates, of course). A simple exeperiment would actually be growing e.g. a pure culture of E. coli and measure growth (or just titre at a given selected point) and do the treatment there.

Well, if you used your own fingers, how can you be sure that each print has the same inoculate and titre?

I find it hard to visualize what OP envisions, but the closest I can think of are localized increased in viscosity by polymerization of parts of the cytoskeleton as part of certain mobile cells. In almost all complex organisms calcification and other rigid structures that I can think of are used as a frame only serve a function if they are, well rigid and localized.

Indeed, it basically demonstrates that perception is a highly active and integrative process and can be readily modified by learning and/or habituation. With regard to the video, I assume that it is easy enough with google translate, but if needed I, I could translate some passages.

If I recall correctly, after about a week (longer is some cases) the subjects perceived vision as normal while wearing the goggles. This effect is accelerated when cues are given (e.g by touching objects) to which side is the right way up. Also interestingly, strongly focusing on cues while seeing things the "correct way" but focusing on the fact that they are inverted may make them inverted again. In other words, the brain uses cues to figure out which way is up and help you interpret it correctly and it is actually not a exclusively hard-wired effect. Once cues are missing, however, one could expect that the brain uses the information relatively raw (conceptionally speaking). As such it is not inconceivable that an infants perception may be incorrect, but there is really no way to tell (other that they are able to distinguish direction at a certain age). Once they can relate the relative position to their vision it would correct itself, assuming the mechanisms are similar to adults. Edit: while searching for the original research (gotta love the classics) I found this. Unfortunately it is in German but it actually describes some of the effects I mentioned.

Considering that it is split now, I have posted a few papers a while back. Some of the older ones that I remember date back to the 80s where only large concentations of caffeine were found to have a diuretic effect. I suggest that you check some reviews instead of taking the first (obscure) paper that seems to support your assumptions. For example, specifically looking at athletes (which are under higher risk of dehydration) no difference in fluid retention was found between caffeinated beverages to water (Armstrong 2002 Int J Sport Nutr Exerc Metab). There is another good review that does a meta-analysis, but I cannot recall the author and would have to search for it, in case you are interested.

The arthropod eye is nothing like mammalian eyes with very different developmental and evolutionary histories. Conflating those serves no purpose. What you refer as "simple eye" most likely refers to the ommatidium which in itself is vastly different from the far more complex mammalian eye. That being said, in some ways arthropod eyes are in some way the consequence of certain developmental decisions (so to speak) including having a convex bulge as an optical unit. Of course, in evolution there is little sense to try to predict complex traits and even with seemingly simple ones it is typically not quite possible (except, perhaps, under highly controlled conditions with very finely-tuned selective pressures). But it one can probably state clearly that at least in animals that have developed eyes there is little reason to assume that they will change to a completely different anatomical unit. The developmental changes required would be too massive (presumably).

Concentrations found in standard caffeinated drinks (such as coffee or tea) are not diuretic (compared to equivalent water intake). Caffeine is only diuretic in large doses in non-adapted individuals (i.e. if deprived of caffeine for a couple of days). Also, IIRC as Strange mentioned in hypertensive individuals caffeine intake can result bursts of blood pressure that at least have been associated with cardiovascular events (obviously these were not controlled tests). Though it is generally assumed that in well-controlled hypertension patients caffeine consumption is of little risk.

Yeah, according to a recent pew poll ACA got an approvable rating of 54% - the highest it ever got as well as the lowest disapproval rating (43%). Presumably now that having had it for a while and being at risk of losing it make people change their mind. Perhaps unsurprisingly the support among republican voters remained low, but among independent there was a large jump (from 36% in 2010 to 53%). They should have named it the Patriot Care Act. Or, given the current climate "Tremendous bigly win care".

Actually, experiments with inversion goggles showed that people actually do get used to inverted vision and function normally after a while (classic experiment by Kohler and Erismann in the 50s). That being said, I do not see lit to suggest that an inversion of vision has been demonstrated in infants. In fact the big issue is that initial vision in infants is poor and only little testing can be done. There is for example evidence that infants as early as 4 months may struggle to recognize a face if it is held upside down (though with inconsistent results), but that in itself does not tell us which way around they perceive a face. If everything was perceived perfectly the other way round, one would also not necessarily expect a mismatch with motor responses (such as eyes tracking the wrong direction) as no experience of the "correct" way would lead to a misinterpretation of direction. As such an experiment would actually be tricky, I imagine. In fact, prior to 4 months infants tend not to focus on certain hallmarks of the face (such as eyes) but scan more wildly around, which would also make the identification of a direction difficult.

I assume you are just not taking all dilutions into account. From what I read you are doing a colorimetric assay after capturing the respective fractions. What dilution did you use there (note that you have to take the elution volume into account) ? If you take those into account as well as the initial dilution (and assuming no loss) the calculations should be quite simple. It is easier if you actually calculate with final concentrations than just with ratios. From your description it is also not quite clear to me at which step you actually do the quantitation and which values you compare to each other. Originally I assumed that you used the traces of the chromatographic runs, and missed that you measured the eluates.

In contrast to eukaryotic cells in most bacterial cells we do not distinguish between cell cycles, mostly because there are, with few exception, no easily traceable events that would make it useful. Perhaps I should specify that in older works (~70s-80s) it was more common to time bacterial cell cycle based on how far DNA replication progressed. But that is hardly being used anymore. Rather, most characterization is done on the population level. That being said, the expression mostly depends on the promotor (obviously) which results in a constitutive expression. I.e. it is being expressed throughout the life-cycle of the cell. There is also evidence for a feedback inhibition which maintains it a low level.

I don't think that is correct. I assume you refer to Canadian common-law unions? From what I read it appears that they fall under provincial rights with quite some differences. One example I found was Quebec where certain protections do not exist for partners (e.g. spousal support). From a legislative standpoint the issue is also that while they may have the same rights de facto, they can changed independent of laws that affect marriages.

I am still not sure what you really mean. Do you mean why there is so little variety in soft-bodied organisms? There already are several answers to that, including that a) a lot of soft-bodied animals exist (such as annelids, certain molluscs, cnidaria, animals with few or single cells etc.) , but without some for of external or internal body shapes there are, at minimum, mechanical limitations especially outside of water. This issue scales with size. I.e. a very large worm can only grow in length but not too much in girth, otherwise it would be too massive to move. But even at small sizes the ability to move differs significantly. Just compare the movement types found in a variety of annelids and contrast that to e.g. equally sized caterpillars.

Just to clarify, you used affinity purification and got two fractions (target and unbound). And on the SEC you did three independent injections (total sample, target fraction and unbound fraction)? And if so, what is the injection volume in each case (did you use an autosampler or manual injection)?

If you read on the various topics, where do you think would you like to invest time in? Then get a basic textbook on that topic and read that and work through the exercises. Still fun? Consider studying it.

No, what I am saying is that your idea of what a career entails is likely too vague and you need to get more insights. I.e. take a look what an academic job really is like and what jobs outside of academia could be had e.g. with marine biology as background. Meanwhile you could just study what you enjoy.

In Germany it is very piecemeal, last time I looked. There is a civil union for same-sex partners, but there were many aspects that were different from regular marriage. Some rights have been amended over the years (such as income splitting sometime around 2010 ish) some remain lacking. Things are still different (afaik) include adoption rights, some weird legal aspects and compensation in certain jobs though much is likely to get amended at some point. So instead of stating it is the same, it is considered different but often after legal battles rights are getting added. The big issue is that same-sex civil union is not protected by the constitution as regular marriage, meaning there is no constitutional right of having the same legal rights per se. I think in the US, before marriage was allowed, the respective same-sex unions only had very little rights which were specifically enumerated and different on a state-by-state basis. To answer the first part of the question: In Germany as well as US marriages confers a set of rights and responsibilities. Check here for the US. I do not have an English source for the German rights but it is part of the family right and is listed as part of the BGB. But some of the more obvious things include benefits, salary for those working in public service as well as tax rights. I don't think there are specific tax breaks just for marriage but via income splitting you can benefit from getting into different tax brackets and/or one can claim a partner as dependent if their salary is very low.

Actually it is worse. If you really want an academic career your time is split between teaching administration, proposal/paper writing, people management and a tiny fraction of it is actually doing analyses. If one is interested in performing experiments, a technical position is more suitable, as after postdoc the time dedicated to that part sharply declines. There are few exceptions such as being staff scientists e.g. at national labs or research prof at unis. However these positions are relatively rare (and thus highly competitive) and the research prof positions are also often badly financed. Even if one is alright with the little time dedicated to research, faculty positions are highly competitive and for that, badly compensated. So one should really inform oneself about the details of scientific careers. That being said, being in college is also about figuring out your interest and not only building a career. As such I would recommend following interests to some degree and use that to build general (e.g. problem-solving) skills that can be applied to a broad range of careers.

Short answer is: yes it seems to be the case see this review, for example:

The actual author Tony Schwartz has given interviews about Trump and the book. One may want to read those.

To maintain shape a certain level of rigidity is required and mechanically more is required to support larger body sizes. However, when considering octopus as soft, why are snails considered to be different? Edit: or do you mean amorphous?

Your question was freezing, and as we established that does not happen. Tissue damage due to frostbite usually happen at ~ -4C. There is non-freezing damage above that temperature, but that is typically caused by inflammatory processes and/or undersupply of extremities with blood. Usually, that takes quite a while and obviously, organs have ischemic issues either way. How long they survive is very different. Hearts are maintained at 4C and it survives for about 4h at that. Longer than that being separated from the bloodstream negatively affects function. Kidneys on the other hand are usually preserved in a hyperosmotic solution that allows storage for up to 48h. There are efforts to increase those times in order to be actually able to store organs. However, I do not know how successful the latest developments are.

As an exercise, it forces you to read up on concepts that you are completely unaware of. While it is a struggle, especially at the beginning, it does train you to gather such information effectively. After all you are unlikely to be familiar or even interested in all questions and tasks you may face later in life.