text:
What causes the yellow and brown spots on our sour cherry tree?
So at the end of spring (maybe at the beginning of summer) our sour cherry tree started producing these yellowish/brownish spots on its leaves. Can you help me identify the couse of this? What can we do against it? I cannot see any bugs on it, my best guess would be some kind of a fungus, but I am way far from being an expert. Thank you in forward for the answers!
Edit: I am located in Hungary. The brown spots are not bigger than my nails, but the yellow area can cover even the whole leaves. Also, the capillaries visible on the leaves can turn red, as you can see on the pictures. Unfortunately, the exact species of the cherry is unknown to me.

how rich is the soil? is do other plants get big or do they stay small and pale white, like for a limestone soil and sandy mineral soil? brown spots in plants is an indicator of phosphate deficiency. phosophorous deficiency also produces light veins. check a deficiency leaf chart. https://www.pinterest.nz/maryinnz/phosphorus-deficiency/ it could be something else. https://www.google.com/search?q=deficiency+leaf&tbm=isch

it may also be dryness. where I live, cherries go thirsty and have messed up leaves after june, but they come back strong again the year after. they are early summer plants in hot countries and keep sugar in the tree for next spring. it can be root damage also or top soil dryness or defficiency or other causes.

also to note is that there is an inviasive species on european cherries from aisa which is a bug which lays eggs in every single cherry and causes early cherry rot. perhaps hungary freezes enough in the winter to keep their numbers down. Asiatic cherry fly (Drosophila suzukii)

The tree is most likely sick. Plant diseases take years to develop versus diseases in animals caused by either bacteria, viruses, or parasites. This is because the tree has thick cell walls that can be a huge barrier hindering viral infection. Another thing is that trees lack a circulatory system and rely on partial water pressure from the air to draw water from the roots up to the leaves.
Your tree might have a virus that is slowly killing it. Which insects are endemic to your region? We can deduce the virus if we know the main types of bugs living in your area.


text:
What is this plant with distinctive leaves?
Location: Vienna, Austria
Leaf size: two big thumbnails
Plant height: large thumb

I assumed it might have come from the seeds of some fruit I ate.  I consulted these websites:
https://www.baumkunde.de/baumbestimmung/
http://www.blattbestimmung.de/blattform-herzfoermig
but found nothing. Perhaps it is neither shrub nor tree.
Edit:
This is how it looks now, ten days later:

Seedlings are often tricky to determine, but I think this may be the common weed Chenopodium album (from the family Amaranthaceae). This is common across Europe, and the leaf shape, growth pattern and the silverly underside seen on the top leaf in your picture fits well.
Here are pictures of a seeding and a larger plant:

(from flickr.com)

(from https://plants.ces.ncsu.edu)

@blabbath Where you write "settle with the family", are you referring to the actual family (Amaranthaceae) or the genus (Chenopodium). In general, I agree though, which is also why I phased it as "may be". It could well be another species in the genus, even though C. album is most common in many parts of Europe.

source: Stack Exchange

license: Creative Commons - Attribution Share-Alike - https://creativecommons.org/licenses/by-sa/4.0/

sort: votes


text:
How to form biological metallic tissue?
There are numerous popular articles claiming that the wasp Apocrypta Westwoodi Grandi features an ovipositor tipped with zinc. You can find such an article here. You can watch an impressive video here. Beyond that, I have found information on the genus to be sparse.
Yet my question is, how metallic tissue can be formed. The popular article claims actual zinc was detected. It seems unlikely to me, these creatures consistently find a source of zinc to feed from, so as to deposit zinc in their tissue. Even so, I don't understand how this zinc can be made to deposit at a specific point. The article clarifies:

With an x-ray detector and the electron microscope, they discovered the presence of zinc. It was only on the tips of the parasitic fig wasp’s ovipositors. Gundiah said, “We see it very consistently only at the tip and not anywhere else.”

I am no biologist, but, to my feeble understanding the only alternative is that the so called zinc is actually synthesised by the body as a protein normally is. Surely no atomic reactions could take place in the wasp! :-p  Even teeth are mostly Ca5(PO4)3 OH, Calcium is found in plenty of our foods. Phosphorus is present in meats. But zinc? I totally don't understand it!

I'm not aware of body synthesising ions de-novo.

I don't know about non-trace amounts of zinc but iron is common, and Iron reinforced tissues exists, things like beaver teeth and scaly foot snail shells which are made of iron reinforced enamel and iron sulfide respectively.

@Ludi You may not be aware but zinc is an essential mineral for all plants and animals (not certain about all bacteria), as are several other metal ions.

@BryanKrause exactly. Of this I wasn't aware. Now, due to Gillian's answer, I did some research. Before I thought it was extremely rare. Embarrassing, since I was a physicist, but what can I say...

First and foremost, it looks like the original article is slightly less dramatic:

Parasitoid ovipositor tips have a higher content of the transition metal zinc compared with the cuticle elsewhere on the ovipositor

So a 'higher concentration' isn't quite the same as 'tipped with'. It looks like they are describing a higher numbers of metal ions, rather than pure metal:

The presence of transition metals in insect cuticle and mandible is hypothesized to increase material hardness to permit cutting through hard substrates with minimal wear

There are many examples of organisms using inorganic chemistry to make materials. The overall topic is called biomineralization and you even mention teeth - which is hydroxyapatite (not really organic chemistry!).
However, a pure metal like zinc is unusual, I think. There are examples such as the snail that has an iron pyrite 'armor' or even the iron nanocrystals in ferritin.
More spectacular examples include the bacteria that produce gold particles

As a result, Au
  detoxification is mediated by a combination of efflux, reduction, and
  possibly methylation of Au-complexes, leading to the formation of
  Au(I)-C-compounds and nanoparticulate Au0.

Where Au0 is pure gold. I don't know of any other examples of pure zinc in organisms, but there is a nice review here of zinc biochemistry.
When you say 'synthesise' I assume that you mean 'deposit'.

I am not a biologist or chemist so my (faulty?) understanding is that the wasp has to eat zinc to deposit it. Otherwise it has to somehow produce it, which I cannot imagine. My other assumption was that substances based carbon are called organic, with some historically motivated exceptions.

@Ludi - Sure, they certainly have to eat it, as organisms are not known to synthesise elements! You are right that most compounds with carbon in them are 'organic', but "Ca5(PO4)3 OH" is a calcium compound, with no carbon.

@gillean oh, gosh, what a stupid mistake! Anyway, it is clear we can get Calcium and Phosphorus from some foods. But where the heck would a wasp get Zinc?

@Ludi Most foods will have small amounts of elements that we tend to think of as rare - zinc, molybdenum, iodine, etc. It's actually quite common to find zinc in biochemistry, as they are used in a "zinc finger" that binds to DNA - of course, that's just a single atom

Maybe you put that in the answer, unless I am the only simpleton who doesn't know it ;)

@Ludi: Just to make things clear: the wasp only needs to consume some substance that contains zinc atoms, not actual bits of metal, right? So the molecule containing a zinc atom doesn't need to behave like pure or even concentrated metal at all. From the Wiki article on Zinc Compounds: A very large number of metallo-enzymes contain zinc(II). Also many proteins contain zinc for structural reasons.

license: Creative Commons - Attribution Share-Alike - https://creativecommons.org/licenses/by-sa/3.0/


text:
What kind of crow is this?
Years ago, when I lived in Greece, I saw crows from a close distance, but never with this peculiar colouration. Today I saw two individuals, both with these gray parts.
Location: Austria, Vienna. Pretty much in the city centre. Specifically Altes AKH Hof 2 near the rear yard of the department for East Asian studies.
Time: I am not sure. Probably before my lecture (around 10am) or after it (12am). I'll see, whether I find the timestamp of the picture anywhere.
Size: Now, at this I am really bad. I would say roughly the length of my boot, which is a 47-48 European continental size.
Behaviour: nothing interesting. Two of them came, stalked around the benches, did not make sounds and as far as I remember were gone quickly. They were not afraid of people.

My guess is Corvus cornix , but I have always been bad at identifying birds.
EDIT:
Unfortunately, I didn't see the linked answer. But had I seen it, I would probably have assumed my bird wasn't Corvus cornix. The plumage  in the linked answer looks significantly more colourful. Whether you want to mark it as duplicate or not, I am fine with your decision, since I have little practice in identification. However, I find the answer here more detailed when it comes to differentiating C. cornix from other corvids. The linked answer only states the results. The answer here teaches us what to look for!

Its a hooded crow or grey crow. Their common in the uk and they are a plague in Ireland.

Can you add some more details to the question - habitat and location where you saw it (more specifically than 'Austria'), what time of day you saw it, and its rough size?

@AlwaysConfused interesting! To me it looks like C. Cornix but I can't pinpoint the difference!

I'd say you guessed correctly. This does appear to be the hooded crow (Corvus cornix).

In German, it's called Nebelkrähe (meaning "mist crow").

 A Hooded Crow Corvus cornix in the garden of Belvedere (Vienna). [Source].
Description: ashy grey bird with black head, throat, wings, tail, and thigh feathers, as well as a black bill, eyes, and feet.
C cornix is a Eurasian species that would be present in Austria:

Similar but incorrect species:
The only other similarly grey crow species I know are the grey crow (Corvus tristis), which is only present in New Guinea, and the western jackdaw Corvus monedula. The jackdaw is present in europe, but it can be ruled out because of its grey eyes and because of the grey feathers that creep superiorly from the birds nape onto its head (unlike your specimen).

Note: I exhaustively checked all commonly recognized species of the Corvus genus

all_licenses: Creative Commons - Attribution Share-Alike - https://creativecommons.org/licenses/by-sa/3.0/


text:
How can fennec foxes eat scorpions?
I just watched a video of a small desert fox (fennec) that toys and eats a scorpion. Although I am unsure of the exact type of scorpion, it is apparently quite able to sting. Another online instance of the same video states that the scorpion is Androctonus Australis. Not only is this breed severely venomous and claiming, as its name suggests, human lives every year, but it is also well armoured. You can hear it when the fennec chews on it...
Fennec Fox eats Scorpion
So, my question, which might be stupid: how can this fox take the scorpion in its snout again and again? I notice that it lets the scorpion go many times. Is it able to avoid being stung? Is it extremely resistant to venoms? I read in the Wikipedia article, that it eats insects, but was unable to find out more!
I was able to find some information about hedgehogs and snake venom, but nothing similar about canines. In the case of hedgehogs, a partial immunity is apparently due to a protein called erinacin in its muscles.
After a user kindly corrected my spelling, I found some comments beneath YouTube videos, claiming it was nearly immune to scorpion venom. Sadly without additional information.

Very carefully.  First remove the stinger, the rest is relatively easy.  In your video, you see the fox quickly bite at the stinger, then back off.  Over and over until the scorpion's abdomen was loose as a rag, unable to launch the stinger at 3:08 minutes.  3:32 - you can see the stinger is almost off.  3:40 - stinger gone, she can now eat it more calmly.

I have to admit it's hard for me to see if she gets stung. But even with these techniques, they must get stung every once in a while, right?


text:
RNA-Seq: Is it conventional to use a threshold RPKM value to determine if a gene of interest is expressed in a sample?
I am interested in finding out at what stages of development a gene of interest is expressed in the human brain. I am using the Developmental Transcriptome tool from the Allen Brain Atlas to find this information. The heatmap showing the gene expression levels across the different brain samples (taken from donors of different ages) was generated from RNA-Seq data.

From the official documentation on how the RNA-Seq data for the Developmental Transcriptome was normalised, RPKM (Reads Per Kilobase Million) were the gene expression units that were used in the RNA-Seq data analysis. The gene expression values were then transformed into a log scale (log2 RPKM) for easier visualisation on the heatmap.
From visually looking at the heatmap, you can see at which stages of development the gene of interest is expressed, with brighter colours (light blue, green) indicating a higher level of expression compared to dark blue regions of the heatmap. However, I do not think that visually looking at the heatmap colours is an unbiased way to determine at which stages of development a certain gene is expressed.
I am completely new to analysing RNA-Seq data. This might be a basic question, but is it a convention/practice in research to use a "threshold/cutoff" RPKM value to determine whether a gene of interest is expressed in a sample? E.g. If a sample has a log2RPKM above the threshold value then you consider that gene to be expressed at a sufficient level.
Any advice is appreciated.

Please specify what"sufficient level" means. You are trying to convert something that is continuous into a binary — IMO this makes unsupported assumptions. 2) This also assumes the level is a constant! 3) Is your gene of interest protein coding? If so, to really know whether the expression meant something you would need to do something like show it was also translated and then confirm that the protein meant something — e.g. by observing an effect after preventing the protein from acting at that stage. ——— Bioinformatics is great, but it is not a substitute for wet experiments ...

The proportion of reads containing the mRNA of your gene is all what RNA-seq can tell. What else do you expect from it? The relevant cutoff probably depends on the function of the protein, and the exact methodology of the RNA-seq experiment.

possibly a better fit for bioinformtics.stackexchange.com. I think that the real question here is about what the "limit of detection" (LOD) is for bulk RNA-seq of some depth. One example of an assay doing something similar is here: https://pubmed.ncbi.nlm.nih.gov/29929942/. There are some statistical procedures used to estimate this LOD, which would ideally include a spike-in control of standards of some kind. RPKM is hard because it is an inherently relative/compositional measure; LOD RPKM will vary from expt to expt based on e.g. depth.

I checked and your resource's documentation does contain information about data processing and spike-in controls, I'd suggest consulting that: https://help.brain-map.org/download/attachments/3506181/Transcriptome_Profiling.pdf?version=1&modificationDate=1382036562736&api=v2

all_licenses: Creative Commons - Attribution Share-Alike - https://creativecommons.org/licenses/by-sa/4.0/


text:
When analysing phosphoproteins via Western blot, why is total protein level of the target protein recommended as an internal loading control?
I am analysing the expression of a protein kinase via Western blots, and it is a phosphoprotein. I have labelled my membrane with antibodies against the phosphorylated form of the protein (using phospho-specific antibodies) and also for total protein (using pan-specific antibodies).
I have read that when studying phosphoproteins, you should normalise the phosphorylated protein signal to the signal for total protein. In this journal paper, it states that the Journal of Biological Chemistry recommends that “signals obtained using antibodies specific for phosphorylated epitopes should be normalized to the total protein level of the target protein”.
I have seen in other journal papers analysing the expression of a phosphoprotein via Western blots that they also normalise the phosphoprotein signal to total protein signal (of the target protein).
However, I am not sure why the total protein level of the target protein can be used as an internal loading control?
I have read in this handbook that internal loading controls are endogenous reference protein(s) that are present in all samples at a stable level and unaffected by the experimental conditions.
I want to compare the expression of my phosphoprotein between samples of different experimental conditions. I know that normally total protein stains such as Ponceau S or housekeeping proteins are used as an internal loading control and for normalisation.
But why is it recommended that you should normalise to total target protein (when looking at phosphoproteins), because one does not know whether the expression of total target protein will be stable between different experimental conditions? Any insights are appreciated.

It is because you don't know how your treatments affect the expression of the total specific protein; perhaps it down- or up-regulates the amount of protein being made, but the relative proportion of phosphorylation doesn't change. For example:
Imagine you have 4 samples from different treatments of which you have loaded an equal amount of total protein (lets say 50 ug total, note that this is too much to load on a WB for accurate quantitation; usually you want less than 10 ug). You run a western blot and find the following numbers (completely made up BTW) for the specific protein. In this scenario, 1 is untreated control.

10 ug
20 ug
10 ug
20 ug

You then try the phospho-specific antibody and get the following

1 ug
2 ug
3 ug
1 ug

What does that tell us about the 4 treatments?
Well, it tells us that 2 is up-regulating the total protein, but the relative proportion of phosphorylation is the same. In 3 phosphorylated protein is up-regulated (3x) but not total protein and in 4 total protein is up-regulated but phosphorylated is down-regulated (0.5x).


text:
Why are these firebugs going inside those fruit?
These insects (Pyrrhocoris apterus) seem to love these dry hollow fruit. I think that those fruit are of Koelreuteria paniculata and mildly toxic to humans. I think that the Kolreutetia intertwines with a yew tree, giving the peculiar impression of those “lanterns” on a yew.
The insects seemed pretty immobile and it was around dusk. Are they going there for shelter?


text:
Can any glider incapable of powered flight strike its prey by descending on it?
TLDR
I was not able to find the precise definitions of “true flight“, “sustained flight“ and “gliding“, that are often used on Wikipedia.  My layman’s understanding is that true flight is the same as “powered flight” and means the animal can utilise muscles to generate life force that exceeds its weight under a wide range of air flow conditions. That is, it does not need upward air currents such as thermals
By contrast, gliding is defined here as:

falling at an angle less than 45° from the horizontal with lift from adapted aerofoil membranes

You are welcome to suggest more correct definitions.
Question
Can any animal that glides but is incapable of true flight (of course hawks may glide) descend upon its prey?
I am aware of this question not being very well defined, since animals such as sugar gliders can jump on their prey and  perhaps utilise a tiny amount of lift to prolong their jump. It is difficult to rigorously separate this from gliding. Let’s require that the animal glides a distance more than 10 times its length from a height more than 10 times its height.
Inconclusive information
Bats
Don’t count because they are capable of powered flight. But they can do it.
Sugar gliders
Can “leap out to catch flying insects“ (source), but that’s all I found.
Flying squirrels
“harvest fruits, nuts, fungi, and birds' eggs”(source), so they may not even prey.
Colugos
Appear to be strictly vegetarian (source).

Sure, but it would be very situational. True flight = powered flight = sustained flight (although an albatross soaring the waves indefinitely could be technically be sustained but does not follow the spirit of the term).


text:
What is this fungus growing on tree stumps?
Location: Austria, Vienna, Ottakring Forest
Time: 2019, Feb 2, 16:02 local time
Size:

From below; lamellar structure:

Some more angles:

I found many tree stumps nearby covered by moss, but only this one displaying the fungi. I am not knowledgeable enough to determine the species of the tree.
I first suspected a Ganoderma, but none of the species listed on Wikipedia looked very similar.
A better bet might be Lenzites Betulina although the pics on Wikipedia always seem more richly coloured than mine. You can check many pics of L. Betulina candidates on mushroom observer. The L. Betulina in this Video also seem to have more marked rings.
Edit:
After some searching I am also considering Trametes Pubescens, as Wikipedia remarks:

Unlike most other turkey tail-like species of Trametes, the cap surface lacks strongly contrasting zones of color.


text:
What is a function-triggering antibody?
I am reading a journal paper on the effect of a neural cell adhesion molecule on neuronal development and in the abstract of this paper I have come across the following:

The autophosphorylation and activity of Pak1 were enhanced when
isolated growth cones were incubated with NCAM function triggering
antibodies, which mimic the interaction between NCAM and its
extracellular ligands.

In this paper, the NCAM protein becomes activated when it is exposed to NCAM function triggering antibodies. I understand that antibodies are molecules produced by the immune system and bind to antigens. However, I am not sure how incubating growth cones with NCAM triggering antibodies would actually trigger/activate the NCAM protein itself.
Isn't the purpose of an antibody to bind to an antigen  so that it can be removed from the body? I am not sure how exposing an antigen its respective antibody would trigger its function. I have read several other papers about neural cell adhesion molecules where antibodies were used to activate their function. I have searched function-triggering antibodies on Google but I have found no useful results regarding this. Any insights are appreciated.

The term "function-triggering antibodies" is not standard, these are more widely known as cross-linking antibodies. As you may know, an antibody has multiple "arms", with the antigen binding region or Fab (Fragment antigen binding) at the end. IgG antibodies have 2 arms, IgM can have 10. The Fab portion is generally identical on all the arms, although it's possible to engineer them differently. This allows the antibody to bind to multiple antigens at the same time, drawing them close together.
NCAM is a homophilic binding protein, with an extracellular domain that binds to other NCAM molecules, both on neighboring cells and on the same cell. That binding induces structural changes in the intracellular domain which promotes the binding and activation of kinases, proteins that phosphorylate (and thus activate or inhibit) other proteins. This starts a signaling cascade that induces changes in the cell, typically by controlling the activity of transcription factors in the nucleus.
Receptors are generally not spread out evenly over the surface of the cell, they often exist grouped together in membrane structures known as lipid rafts. When drawn close enough together, NCAM molecules can begin their signaling cascades on their own, as adapter proteins bound or recruited to the intracellular domains get activated – the kinases and their substrates are in close proximity as well. To induce this type of signaling in the lab, cells can be treated with cross-linking antibodies that draw the receptors together. This in effect triggers the function of the receptors, beginning their downstream signaling.

Source

Thanks for your reply. You stated that receptors are grouped together in lipid rafts located in the membrane. However, I just want to clarify whether NCAM binds to receptors in lipid rafts, and that it is the close proximity of NCAM molecules that can initiate signaling cascades?

Yes. NCAM is a homophilic receptor, meaning that one of its ligands is itself. Cross-linking receptors brings them close together and stimulates internal signaling, including p21.

Antibodies can function as enzymes (see https://en.wikipedia.org/wiki/Abzyme).
If you use the right monoclonal antibody, the variable end can be the right shape to bind to the site you mention.

This is incorrect. The antibodies in question are not abzymes, they're simply cross-linking antibodies that mimic the binding of a ligand to an extracellular receptor.


text:
What has a more negative impact: Having only a partial deletion of a specific gene or the having entire gene deleted?
I am looking through a genomics database to see whether patients who have intellectual disability have deletion of only a specific gene of interest and no other mutations (e.g. deletions or copy number variations of other genes).
I have seen that some patients have deletion of the entire gene whereas other patients have deletion of only part of the coding region of the gene. However, even if part of the gene is deleted, patients have intellectual disability and other abnormalities.
I was wondering, in general, is it more serious for a person to have a deletion of an entire gene in contrast to having part of the gene deleted? Can having part of the coding region of a gene deleted be just as serious as having the entire gene deleted?
Thanks.

Highly gene dependent I suspect - delete an important motif in one gene then yes, delete one repeat in a highly repeat rich gene (e.g. Ankyrin repeats) and it might not be.

I strongly agree with bob1, a particularly clear example is that some partial deletions will result in a dominant negative allele — these will have a stronger effect than a complete deletion. ——— Consequently, this is only answerable for specific deletions of specific loci. As written this isn't a question with a useful answer, so I've voted to close.

The answer depends as @bob1 says on the gene and the mutation probably.
For a simple analogy, think of a gene like a chainsaw or other piece of machinery.
To break it down a little:
Full gene deletion
We expect a full gene deletion to be associated with a loss of function. This is very bad if the gene is essential to life
If you lose your chainsaw, you can't cut anything, which is bad if you need to cut things e.g. for your job.
Partial gene deletions
Are more ambiguous. They might cause:

Full loss of function of the gene (equally bad to full deletion).
A crucial piece of your chainsaw is broken, so you can't start it.
Partial loss of function of the gene ("hypomorphs", less bad than full deletion).
There's something wrong the chain on the chainsaw, so it's slower than usual, but you can still cut some wood.
Gain of function of the gene (could be better or worse than full deletion!)
There is something wrong with the gas tank, and the chainsaw might explode and hurt you!

Gain of function is the most interesting but probably the rarest. Here is one example, where a partial deletion of the tumor suppressor gene p53 switches its function so it turns into an oncogene (a tumor promoter), effectively switching its function from protecting against cancer to making cancer more likely.
Here is a powerpoint from a class that goes over gain-of-function and loss-of-function in a little more detail.


text:
Can Western Blots be used to quantify the activity of a protein?
I am new to Western Blot analysis and I have recently done my first two. I am studying a phosphoprotein (a protein kinase) that can be both activated and inactivated via phosphorylation at a specific amino acid residue. I have labelled my membrane against the active and inactive forms of my protein of interest (using phosphospecific antibodies), as well as the total protein (using pan-specific antibodies).
I know that Western Blots are used to quantify the expression of a protein (i.e. how much of a specific protein there is in a sample). But I was wondering how they can be used to quantify the activity (e.g. how functional the protein is) of a protein, e.g. how can I measure/determine whether my protein is more active/inactive in my treatment samples vs. control?
For example, I have normalised the integrated density values for my active protein bands against the integrated density values for my total protein bands. Then I have calculated the fold change in protein level for my active protein in treatment samples relative to control.
If I have observed that the fold change for my active protein is reduced by 40% (as an example) in treatment samples relative to controls, would this be sufficient data to say that the activity of my protein of interest is reduced in treatment samples compared to control?

You may need to optimise your question here: 'How functional is the protein?'. Too broad, what does functional mean? Also, more importantly what are the differences between your control and treatment groups? Are you asking: Because there is more protein expression, is there more protein activity? If so, you do not have sufficient evidence yet. More protein does not prove more activity, especially since the activity is undefined.

The protein I am studying is constitutively active, so by functional I mean what proportion of the protein is in its active form compared to total protein (active + inactive forms of the protein) in my samples. The difference between my control and treatment groups is that I am comparing knockout and wild-type brain lysate samples. I am investigating whether knocking out a protein affects the activity of my protein of interest.

Thank you for your response, it adds clarity.

I see 3 parts to a complete answer:

If you define the phosphorylated protein as active, and non-phosphorylated protein as inactive, and you have a total protein blot (as you say, via pan-specific antibodies) as you describe, then you have done everything correctly.

It's simple math from there on based on radioactivity or fluorescence or mass or whatever you use to quantify your band intensity. The 3 components should match up with the inactive protein amount (active + inactive = total). If this is consistent, you have additional evidence that you have done it correctly.

However, here come the hidden assumptions: this is all assuming that (i) you are not experiencing significant protein loss during your preparation steps, that (ii) your antibodies are good (specific and yield the complete protein extract), that (iii) the blotting procedure is performed correctly, and most importantly, that (iv) your initial definition is valid, that the phosphorylated protein is truly the active form of the protein, and that the unphosphorylated counterpart is the inactive form. You see, this is not always the case, though with some (most?) kinases, it is. If it has not previously been demonstrated, one may raise the reasonable objection that your assumption is wrong. However, I take it that in your specific case, the kinase activation by phosphorylation is well understood.


text:
Are there any online resources that have a list of post-translational modifications and their molecular weight?
I am studying a protein and I am interested in learning about all the post-translational modifications that it has (as I am analysing it via Western blot). I have found a list of the post-translational modifications that it has, however, I want to know what are the molecular weights of the post-translational modifications (in daltons/kilodaltons). I have found this site but I am not sure what the difference is between 'Monoisotopic Mass Change' and 'Average Mass Change' and what units the values are in.
I was wondering if there are any reliable online resources that have a list of all the post-translational modifications that exist and their associated molecular weights? Any insights are appreciated.

To a man with a hammer, everything looks like a nail. If you want to know what modifications a protein might have you use the most appropriate method. Western blots of SDS gels isn’t it. Your intelligence is the resource you need to consult. Calculate the molecular weight of an acetyl group and then what increase N-acetylation will have on the molecular weight of your protein. Then determine whether SDS gel electrophoresis can resolve that difference. You have accepted an answer to your question, but it is not an answer to your — or anyone else’s biological problem.

You've found one. Since the differences between the "Monoisotopic" and "Average" columns in the Sigma-Aldrich link you found are extremely low, functionally we can say they are approximately equal. The masses are in atomic mass units, or Daltons.
However, these numbers are from actual mass changes seen in modified peptides analyzed by LC/MS/MS, typically from digested proteins. Mass spec is a highly precise platform, as you can see by the number of decimal places in the numbers in the table. When estimating protein size by Western blot, the margin of error is at least 1 kilodalton for smaller (10-50 kDa) proteins and 5-10 kDa or more for larger ones. Since a single phosphorylation event would only change the mass of a target protein by 0.08 kDa, determining that by Western blot is essentially impossible. However, evidence shows that proteins modified by a single phosphorylation event typically also have (sometimes many) other post-translational modifications (PTMs) as well, including other phospho sites, methylation, acetylation, glycosylation, etc., that can visibly affect a modified protein's mobility in an SDS-PAGE gel.
Another factor to think about is that in denaturing SDS-PAGE gels (which most Western blots are generated from), migration rates are not only influenced by total molecular mass, but also by charge, as the proteins are migrating in an electrical field. When these proteins are linearized by SDS, a charged detergent, their overall charge is roughly proportional to their size. However, charged PTMs like phosphorylation will affect that overall charge, and may modify the migration rate according to that charge in addition to their extra mass.
Combine all this together, and you have PTM-modified proteins migrating at somewhat different rates than one might expect based on the physical mass of the modification(s) alone. When you start combining multiple PTMs, especially different kinds of PTMs (phosphorylation and acetylation, for example), it can become very difficult or impossible to predict what the expected apparent molecular masses will be.


text:
Why is it good practice to prepare western blot samples in pairs when you have two experimental groups?
I have done some Western blots where I have analysed the expression levels of a certain protein in brain homogenates from wild-type and knockout mice. When preparing brain homogenates, I was advised to prepare them in pairs, so that on the same day two brain homogenates, one from a wild-type mouse and the other from a knockout mouse were prepared.
I know that paired analysis is useful for comparing differences visually on the blot between samples from two experimental groups. However, I was wondering whether there are any other benefits (e.g. for statistical analysis) to preparing samples in pairs for a Western blot analysis besides just ease of visual comparison? Any insights are appreciated.

This point is not unique to Western Blot, but to any case-control study: if there is variability inherent in the preparation procedure, this variability will be reduced when the two samples are prepared simultaneously. This is perhaps easier understood using an analogy: if a toaster fits two toasts, but there is variation in the toasting time - the toasts that were in the toaster together are likely to be cooked/burned to the same level.
Statistically, one takes the full advantage of such improvements when using the statistical techniques specifically designed for paired data, such as paired difference test (see also paired samples t-test). But even unpaired statistical methods are likely to show better performance (in terms of identifying significant results).


text:
What is meant by 'genes overlapping inherited structural variants'?
I am reading a journal paper about genes that are related to autism, and I have come across the following statement:

To assess commonality of biological function among rare risk alleles,
we compared functional knowledge of genes overlapping inherited
structural variants in idiopathic ASD subjects relative to healthy
controls

I am not sure what is meant by 'genes overlapping inherited structural variants'. I searched the definition for structural variants and according to Wikipedia, structural variation refers to the variation in structure of an organism's chromosome, such as deletions, duplications, copy-number variants, insertions, inversions and translocations.
However I am not sure what is meant by genes overlapping the inherited structural variants. Does this mean for structural variants that are common between ASD subjects, there are some common genes that are found in these regions (the inherited structural variants)? Any insights are appreciated.

If I understand correctly, what this paper is looking at is rare copy-number variation in ASD cases relative to controls. So in the context of the paper, the passage you quoted is basically saying we found genes that had copy-number variation between cases and controls, i.e. the cases had a pattern of more/fewer copies of a stretch of the genome that includes a gene than the controls. Then they look at the functions of the genes they found in a gene ontology database to see if they seem like they might relate to ASD.


text:
Receptor tyrosine kinases: Clarification about what is meant by stabilization of the receptor
I am reading a journal paper. This paper focuses on how neural cell adhesion molecule (NCAM) interacts with fibroblast growth factor receptor 1 (FGFR1) and promotes its stabilization.
When FGF binds to FGFR1, the receptor gets internalized and degraded by lysosomes.
In contrast, when the soluble ectodomain of NCAM binds to FGFR1 the receptor gets internalized and recycled back to the cell surface, resulting in sustained signaling.
I asked in another question, what is meant by the stabilization of a receptor, and the response stated that it means the receptor stays in the plasma membrane and doesn't get re-internalized.
However, in the case of this paper, it seems that stabilization means that the receptor doesn't get degraded after it is endocytosed.
What exactly does 'stabilization' mean in the context of a cell-surface receptor? Any advice is appreciated.

I would look at their methods: what are they measuring? That's what they mean by stabilization.


text:
What exactly is this plant resembling Conyza?
This plant grows on my balcony in Vienna, Austria. It gained most of it’s current height (30-35cm) during the last 2 weeks. Before I didn’t notice it.

It is covered with tiny white hairs. I conducted a couple of tests. They never stung.
The leaves are toothed (I believe “serrate” is the official terminus for this type of edge).
From a central leaf vein, more veins spread out in a manner that seems somewhat irregular to my untrained eye:

Edit: Now, in July, that is about 2 months later, it bears flowers.  Furthermore it is about 3 times taller. I believe there were seed heads, which have been blown and washed away. All of this would fit Conyza mentioned in the comments, but the shape and density leaves  appear to differ from C. boranensis or C. canadensis.

Edit: now, still July some seed heads have reappeared

It is similar to Conyza canadensis, but the shape of the leaves makes me uncertain

Conyza canadensis can have these serrated leaves, although usually not this strong. The amount of flowers is also a bit much. Hmm, dunno...


text:
Is this not a moth?
I apologise for the terrible photographs. I tried many times and can not shoot better ones.
I came back from vacation and moths (I assume) came creeping out of two packages containing oatmeal and rice respectively. I bought pheromonic traps for moths attacking food stuffs, but they have not caught a single one, although they are inches from it. So, I wonder, are these no moths:
Here slain:
[]
Here alive:

I notice they don't match the colour of any food stuffs moth mentioned here.
The slain moths wingspan (distorted as it is) equals the width of my thumb.

Well, they are definitely moths of some sort, but I wouldn't dare identifying them from those photos (maybe try in better lighting?). Since they came out of food packages, that makes me think it's likely that they live there. Have you checked for larvae? I'd recommend sealing the packages.

It is a moth.  Width of the thumb is big for a moth that attacks food, at least in the US.  Pheromones are specific to specific species of insects.  Otherwise you would show up ready to mate and find something of a different species waiting.  You could freeze the packages for a day or 2 to kill anything in them.

@Will thanks. Already threw away everything I thought they could reach, but I am still worried


text:
Explaining a branch with visibly different leaves on Citrus japonica
My father was so kind as to send me a tree of Citrus japonica, aka Fortunella japonica aka kumquat (all names mean the same according to Wikipedia). Though I have seen this species many times, it is only now that I noticed two very distinct kinds of leaves.
One kind of leaves that I call  'double leaf' (second image) is only found on one single branch. The branch is still green down to where it merges with the tree, but many of the leaves are among the biggest on the tree. Excepting one sickly not fully formed leaf, all leaves on this branch seem to be of this 'double' type. I checked carefully.

All other leaves are of the kind shown in the first image.

This is the tree as a whole:

How can this different type of leaves on just one branch be understood? Does that branch belong to a different kind of Citrus (meaning grafting occurred), or is this a stage in leaf development? I don't think the latter, because I found small single leaves in the top branches of the tree. But I also don't think the former, because father said no such thing. If it is a mutation, why does it solely affect one branch?
I am a particularly ignorant amateur, but if you instruct me what to observe or photograph, I shall gladly do so.
Edit:
I thought I should supply a picture of where the unique branch emerges from the tree:

The tree bears fruit and at its thickest point is marginally thinner than my thumb.
Edit:
It just occurred to me that the entire upper kumquat might be grafted onto a different kind of citrus, which would explain such behaviour in the lower branch. I will search for evidence of this in the morning.

You are interested in developmental biology.

@SANBIsamples I am a layman with very little knowledge about biology. I think you are saying it is a mutation. Why does it appear in this branch only?

Don't worry @Ludi, I'm not British. You don't have to say you're a layman at the onset or even play humble :) A mutation can appear whenever cell division goes wrong. Yes, it's probably a mutation.

It was probably a graft onto another citrus rootstock, so you might get shoots of either types of citrus

What you are seeing here is an example of a winged petiole. The petiole is the leaf stalk and these can have wings called stipules.
According to this source kumquat leaves have narrow petioles. I can't explain why you are seeing variation on a single plant, but I recall seeing similar variation on garden citrus in California, so I suspect it isn't unusual.


text:
Vultures in Germany in the past
I noticed that since the 1970s the bearded vulture , which had disappeared from the Alps due to hunting, has been successfully reintroduced. I have searched for the modern habitats of vultures, but it appears (the wording on the site is a bit imprecise) none occur in Germany beyond the very mountainous regions.
I want to know whether at any point in the past vultures of any species where common across large parts of Germany other than the Alps. I am especially curious about the Middle Ages.
Edit:
This site in German claims that vultures in Medieval Germany followed sheep herds, eating dead, stillborn and afterbirth of cattle. It mentions, beside the Alps, the upper Danube Valley and the Moselle–Rhine region:

Bis dahin hatte es vom Mittelalter ausgehend, auch in deutschen Ländern eine einträgliche Zusammenarbeit gegeben. Geier folgten den wandernden Schafherden, beseitigten verendete Alttiere, totgeborene Junge, die Nachgeburt oder die Knochenreste vom Essen der Schäfer. Geier nisteten im Mosel-Rheingebiet, den Alpen und im oberen Donautal.


text:
What did the ancestors of textile moths eat?
These days I had to wrestle a lot with textile moths. From the Internet I got the impression that these moths don’t feed on naturally available furs in the wild, but have specialised on man made textiles(?). From the Wikipedia entry on textiles I gather humans started wearing textiles (not furs) about 6500 years ago.
Did the ancestors of textile moths (from the time of textile adoption) feed on furs and such, or were they akin to today’s kitchen moths, which eat food stuffs?
In the latter case, is not 8500 years a very small time for such a fundamental change in animal biology? Or must I take into account the short lifespan of insects, equating each human year to a dozen generations and each human generation to about 300 moth generations, so that the change moths can undergo in 6500 years is more akin to that humans can undergo in 250.000 years (which perhaps has to be increased even further due to moth simplicity and large population making non lethal mutation more common) ? For comparison, it appears Homo  Sapiens emerged some 200.000 years ago.

Modern textile moths actually eat a wide variety of material, including fur, feathers, shed exoskeletons, bran, and grain. humans just pay more attention to them when they eat textiles. They prefer human handled material because we tend to create larger concentrations than occur in nature.
if you are interested in changes in biology in response to humans you may want to look at starch digestion in dogs, nylon eating bacteria, and body lice.


text:
Is this malus baccata?
Sunday 22 September 2024
Location: Vienna 1190, Austria
This tree (which my app can’t identify) reminds me of an apple tree (I am terrible at tree identification). Its fruit are the size of cherries, but taste like apples with a bitter aftertaste. Seeds and fruit stalks are also reminiscent of apples.
I am now thinking it might be malus baccata.

This is a very well put together question! Showing images of all parts of the plant and giving a location makes this a fun question to answer.
You are really close in thinking that this is Malus baccata. However this tree is actually more correctly identified as Malus prunifolia or the Pear Leaf Crabapple.
Let me explain why. Starting with the family, and working down to the species.
Family
We know that this is in the Family Rosaceae or the Rose family. Due to its fruiting nature and alternating leaf pattern. (Yes, apples are in the Rose family, as well as pears, plums, cherries, and most other exported edible fruits)
Genus
Let's keep narrowing down our possibilities.  This tree is in the Malus Genus which is better known as the Apples and Crabapples Genus. There are a two main reasons:

The leaves have a serrated margin as well as being alternate and simple.
The fruit pictured is a globose pome. Bearing distinctive Malus traits. (In more simple words: the fruit resembles an apple or crab-apple)

Apples versus Crab-apples
In order to correctly identify this tree the difference between a crab-apple tree and a regular apple tree must be understood. Britannica sums it up fairly well:

Crabapple trees are stiffer in form and spinier than the common apple. The plants are deciduous and often have attractive fall foliage. The simple ovate leaves are serrated along the margins and are borne alternately along the twigs. The fragrant white, pink, carmine, or purplish flowers appear early in showy masses—in some species and cultivars as five-petaled blossoms, in others as semidouble (having 6–10 petals) or double (having more than 10 petals) blossoms. The pome fruits often persist through the winter and are generally less than 5 cm (2 inches) in diameter. Source: https://www.britannica.com/plant/crabapple

For the purpose of identification, it only needs to be understood that the tree pictured is a crab-apple tree due to the smaller size of the fruit. For more information on why crab-apple trees are in the same genus as apple trees or what their differences are. Visit the following links:

The most helpful: https://arboretum.harvard.edu/wp-content/uploads/2020/06/2009-67-2-Arnoldia.pdf
An interesting forum: https://forum.inaturalist.org/t/apples-vs-crabapples/34888
https://link.springer.com/article/10.1007/s40502-023-00712-0
https://www.actahort.org/books/696/696_4.htm
https://link.springer.com/article/10.1007/s10722-020-01026-y

Species
One helpful process of getting from genus to species is using the location where the model was found. In this case: Vienna Austria. Using the geographic model of the genus malus on inaturalist.org and the list of all the species in the malus genus. (List Here). We can determine that there are four possible trees in the genus Malus that inhabit that region in Austria.  (with the help of this map). These possible species are:

The Domestic Apple Malus domestica
The European Wild Apple Malus sylvestris
The Pear-Leaf Crabapple Malus prunifolia
The Siberian Crabapple Malus baccata

We can knock out M. domestica and M. sylvestris because we already determined that this tree is a crab-apple tree.
That leaves us with M. baccata and M. prunifolia. From here, these two trees are remarkably similar. The genetic variations of the two trees make it even trickier to differentiate the two trees. This is illustrated here.
There are two primary ways to tell between the two trees. The blossom shape and color, and the fruit size and weight. Since we don't have a sample of the blossom, the best way is to look at the fruit size. M. baccata has a smaller, lighter fruit averaging at about 1-5 grams. While M. prunifolia has a larger, heavier fruit averaging anywhere from 10-25 grams. This confirms this species as Malus prunifolia or the Pear Leaf Crabapple:

Malus prunifolia is a species of crabapple tree known by the common names plumleaf crab apple, plum-leaved apple, pear-leaf crabapple, Chinese apple and Chinese crabapple. It is native to China, and is grown elsewhere for use as an ornamental tree or as rootstock. It reaches from between 3 and 8 meters tall and bears white flowers and yellow or red fruit.  Source: https://en.wikipedia.org/wiki/Malus_prunifolia

A diagram showing the differences in fruit between M. baccata and M. prunifolia

Journal of the American Society for Horticultural Science J. Amer. Soc. Hort. Sci. 142, 2; 10.21273/JASHS03968-16, https://journals.ashs.org/jashs/view/journals/jashs/142/2/article-p126.xml
A picture of M. baccata. Note the smaller fruit and cleaner leaves

Photo 30691961, (c) Александр Скачко, some rights reserved (CC BY-NC), uploaded by Александр Скачко, https://www.inaturalist.org/photos/30691961
A picture of M. prunifolia, note the alternate leaf placement

Photo 148881668, (c) Magazova Lubov, some rights reserved (CC BY-NC), uploaded by Magazova Lubov


text:
When transmembrane proteins destined for the plasma membrane are in the ER membrane are they in their final folded form?
I am studying how transmembrane proteins are made and I have read that proteins that are destined for the plasma membrane are initially in the ER membrane and do not get translocated into the ER lumen unlike other proteins.
I was wondering when a transmembrane protein destined for the cell membrane is in the ER membrane, is in its native form (e.g. final, folded form?). I am asking this because I read that the ribosomes are on the ER membrane, and that protein translocation through the ER membrane occurs co-translationally, meaning that it's occurring as the mRNA is being translated. Would the protein get folded as it's moving through the ER membrane?
Any insights are appreciated.

This topic isn't fully understood, but proteins seem to be considered to become folded within the ER membrane, and the transmembrane integration system is compared to chaperone functionality. I found a nice review-paper to this topic. Check its sources for further reading to specific interests. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2814870/

The speed of folding/refolding is much faster than translation speed. So I expect much folding to happen during translation. Of course, proteins that become exposed to the different extracellular environment may refold then.


text:
Which mosquito species is this?
Today, when I just woke up and saw this. I need to identify the species of this mosquito. I don't think its dangerous like Female Anopheles. I'm afraid if it is a carrier of any dangerous diseases such as Filaria, Dengue, etc.

Location: Bokaro, Jharkhand, India
Temprature: $ 29°C $

It is a female. From Wikipedia: "Anopheles mosquitoes can be distinguished from other mosquitoes by the palps, which are as long as the proboscis,and by the presence of discrete blocks of black and white scales on the wings." I can't see the palps for sure, but they look short (if what I am seeing are the palps).

@Karl Is it Anopheles?

I don't know because I cannot clearly see the palps. Mosquitos are not my specialty though, and I am sure that an expert will be better able to tell.


text:
How does the first organism infected by a disease get infected?
How does a micro-organism causing some communicable disease infect the first organism it infected?
I was reading about HIV, when I found that HIV has jumped from chimpanzees to human beings. But, how did the first chimpanzee affected by HIV get affected?
In other words, is it possible (without artificial means) for a new infection to get started? How?
NOTE:
I am not aksing about the origin of pathogens, rather, about how they infect the first organism infected from it, and thus, become a disease causing microorganism.

Does this answer your question? How did viruses come to be?

Another related question. Please note that you are expected to do prior research (including, but not limited to searching on this site) before posting. In addition, micro-organisms includes many pathogens that are not viruses, so your question is unfocused and poorly worded as well as under researched. Please take the [tour] and consult the [help] starting with [ask] for details about what is expected.

@tyersome sorry, but I didn't mean only viruses, its just while reading about HIV, this question came in my mind

and @tyersome, I did not want to know how viruses or other microorganisms are "formed", instead, my focus was on how it "infects" the first organism it infected. For example, Rhinovirus (causes common cold) may have formed from dna fragmemts, or whatever. I was asking that how did the first rhinovirus-infected organism get infected?

The key term to find information pertinent to this question is Zoonosis (link to the Wikipedia article).
However, one should not generalize zoonotic origin to all the diseases affecting humans - in fact, the relationship between humans, diseases, and other organisms carrying these diseases is rather complex.

Some diseases are uniquely human: their origin is unknown - they might have evolved together with humans or were transmitted to humans or their predecessors millions or billions years ago. Smallpox is one example - it became eradicated as soon as it was eradicated in human population.
Other diseases have evolved to be transmitted between different types of hosts, e.g., such as humans and mosquitoes. Flaviviruses, such as Zika, Yellow fever, dengue, west Nile virus are examples of such pathogens, adapted to two different types of hosts. In fact, some of these are adapted to mammals other than humans, but capable of infecting humans due to sufficient similarity. Some of these might not be able to effetcively replicate in humans, even though being a serious health hazard.
For some diseases insects (or other organisms) serve as mechanical vectors, merely carrying disease from one human to another or from a human-like animal to humans, while not infecting the vector.
There are animals that can be infected by both animal and human viruses. E.g., the types of influenza affecting many birds are not dangerous for humans, and these birds do not get infected with human influenza. However, pigs and paultry may get infected by both. The influenza viruses coexisting in a pig then undergo gene reshuffling, producing new viral strains infectious to humans, that humans get from pigs.
Finally, what the OP probably had in mind is the transmission of viruses such as Ebola, SARS/MERS, and HIV which exist in nature as animal viruses and suddenly transform to become human viruses. This is an evolutionary process, which is rarely successful.

HIV
E.g., the bushmeat consumption has probably on many occasions introduced SIV to human population - perhaps thousands or millions times during the history of this virus. However, not being adapted to humans, the virus didn't cause pandemics - until the moment when this transmission as accompanied by a fortunate mutation (or mutations) that allowed it successfully replicated and be transmitted in human population. It is quite possible that it took some time for this virus to further adapt to human population before it became as lethal as it is.
Ebola
Ebola is an example of virus that seems to be poorly adapted to human population - although successful in replicating itself, it fails to keep enough hosts alive to assure its permanent existence in the population. Last century has seen multiple Ebola epidemics, all successfully contained by quarantine measures. There have been probably many more spillovers over the course of human history, but they all ended with virus becoming extinct in the human population by eliminating available hosts.

Can you tell me how: 1) Uniquely human diseases got into the human population? 2) Multi-host disease causing pathogens infect the first host it infected? 3) Pathogens which use insects as vectors infect the first human it infected? 4) Pig influenza virus get into the first pig it infected? 5) HIV/ Ebola viruses (before mutation) infect the first animal they infected?

The two threads linked (here and here) have some information, but no one knows for sure, since it happened long time ago. HIV, Ebola, Zika, SARS/MERS are well studied, since they happened recently. Some other disiseases are merely mentioned in historical records from a thousand or two thousand years ago (see also this thread), but we cannot look back any further than that. Unlike dinosaurs, viruses and bacteria have not left fossil records.

okay.. thanks:)

Viruses evolve like all other biological species. The family of SIVs that gave rise to HIV goes back millions of years across many hosts.
Just like you can build a "family tree" out of related plant or insect or mammal or bacterial species, you can do the same with viruses. The family tree of HIV and SIV goes back to a broader classification of lentiviruses that includes some affecting non-primate mammals.
Viruses affect all types of life: bacteria and animal, plant and protist. They've likely existed as long or nearly as long as cellular life.

Oversimplified, your answer means new infectious diseases are caused by evolution of already existing micro-organisms?

If that's true, then I got it, thanks.

@AsmitKarmakar Yes, that's accurate.


text:
What species of flying insect is this? Resembles a fly with the wings of a dragonfly
Locus: Vienna, Austria
Local Time : 12:24 today, 6th of January
Size: length approximately a large thumb. Width much smaller (pic).
Behaviour: possibly attracted by honey in my medicine, along with flies.

Definitely in Syrphidae.

Might be Episyrphus. In any case, the common name for the family is "Hoverfly".

@KarlKjer After consulting Wikipedia, I tend to agree. Furthermore, I am amazed to see that this kind of eyes apparently makes it a female! https://en.m.wikipedia.org/wiki/Episyrphus_balteatus

Insect ID from photos is difficult, but this is definitely in Syrphidae. The alternating thick and thin stripes on the abdomen resemble very much those in a picture I found here: https://alchetron.com/Episyrphus-balteatus
I would say that it is likely in the genus, Episyrphus, which is present in Austria.

Syrphidae are usually easily identified from photo. This is without a doubt E. balteatus. There is no other species that resembles it in Europe.

Could you provide further support using Comstock-Needham system out of curiosity?


text:
Can any species be bred selectively/engineered to become as diverse looking as dogs?
I've done some research and it appears that dogs are the most diverse looking single species of mammals. The questions that interest me is - are dogs special in respect to genes/gene activation mechanisms related to appearance? Or does this dramatic difference in appearance have something to do with dog anatomy and how they give birth?
If dogs are not special, this makes me interested if other species of mammals can also be bred selectively (or genetically engineered) to produce such dramatic variation?

good question.. no idea, as of now, about what are the genes in dog that give morphological diversity.. i think another species that has been selected for morphological features is pigeon.. you can check this paper out.. I shall also go through it..

Dogs have a genomic structure that allows breeding with high variation in size, shape, coat quality, color and other qualities particular to each breed as well.
Other domesticated animals can be bred for as many qualities, but dogs in particular show a wide level of morphological traits - varying in size from just over a pound to the size of a wolf, from which dogs are derived and genetically are still compatible.  But more interesting than just size or coat color/texture and even their intelligence and personalities, the proportions of their bodies, of their skull length and breadth, are remarkable.
There are over 160 registered breeds of dogs, but this is only a measure of how much time people have put into them.  I think its possible to get nearly anything you want with animals, if you are patient enough - its not clear what is and is not possible with enough genetic manipulating. For instance, horses can be bred over nearly as great a size range for instance (the miniature horse the size of a large dog, the Shire is 3,300 pounds), but it would not be as easy to get both the size and muscularity and shape of a bulldog in a horse.  Breeding a mouse of various colors can be done, and so can interesting behaviors, but body shape seems to be harder: a Weimaraner mouse could take a tremendous amount of time and animals.

Great answer, from your first link, it appears that dogs have smaller number of genes that strongly morphology, while other organisms, like humans have larger number of genes that have only mild to moderate effect

thanks for saying that outright - i should have said that in the answer... :)  This is the reason that there is such an interest in dog genetics.

Good answer, and there are other animals bred for weird stuff. Pigeons and rabbits, etc. Pigeons in particular we've pretty thoroughly reshaped.

Also chickens have been bred into many different varieties.  The size range doesn't seem to be as great as with dogs, but feathers give a larger range of color & form.

Its true nearly any animal can produce many breeds with behavioral, colorful and fascinating variations.  dogs seem to have a remarkable variation in body shape and size in particular though.

its really more about the kinds of genes and how sensitive they are to mutation - i'm sure dogs have a similar number of genes that control development, but with other animals (such as cats) they are not as sensitive to breeding.

Rabbits have been bred domestically since at least ancient egypt.
Rabbits vary in size from > 15 kg (35lb) for a Continental Giant to .5kg(1lb) for A small Netherland Dwarf.
The American Rabbit Breeders Association recognizes 47 different breeds of rabbit for shows and there are a number of breeds that are either not recognized or are being developed for acceptance.
Breeders break the rabbits down by type of body, size, fur, and ear type.  Each breed is then broken down into Groups and Varieties.  It may be hard to believe but each of these rabbits share the same basic DNA.  Though interestingly some breeds will express the genes in manners that are often not the same visually.

Interesting, I didn't know there were so many rabbits. It would be interesting to know if the genes for "floppy ears" are similar between dogs and rabbits

There is not actually a single gene for floppy ears.  It is a number of genes that go into the expression of their ears.  One part that is especially important to the display of their ears is the crown, which is the muscle and tendons that provide ear support.  In french lops you want a very strong and visible crown.  In English lops you want no visible crown at all.

Not a mammal. But goldfish have also been modified both in shape and colour
http://40.media.tumblr.com/ef8838366e4b7a51646b74b5330238e3/tumblr_mi60i8S52c1rmp9qqo1_1280.jpg


text:
Composition of EZ-Vision® DNA Dye?
I'm looking for the composition of the EZ-Vision DNA Dye for electrophoresis visualization. I could not find anything helpful on the web.
I would appreciate a lot your help :)

Presumably you're asking for safety reasons. Their safety data sheet is missing for that product on their website. You'd be justified in directly contacting the manufacturer on this topic.

The MSDS does not specify which compound is actually present in the solution. A quick google search does not show what is composition of the solution, except for DMSO.

@AlwaysConfused I found that info sheet as well, but sadly it doesn't provide the components. Thanks, anyway :)

Contacted technical support and received the following response:
"Unfortunately, the information you requested is proprietary and we will not be able to share the images of excitation-emission spectra of EZ-Vision® Bluelight DNA Dye and the chemical structure of the dye. "


text:
What is Modulation Power Spectrum?
I'm reading this paper https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4562283/ , but I cannot make sense of what MPS nor how should I read the pictures that make use of this method.
The authors say

The MPS, beyond classical representations such as the waveform and spectrogram (Figure 1A and B, upper and middle panels), displays the time-frequency power in modulation across both spectral and temporal dimensions (Figure 1A and B, lower panels). The MPS has become a particularly useful tool in auditory neuroscience because it provides a neurally and ecologically relevant parameterization of sounds

I'm clueless about what the meaning of time-frequency power in modulation across both spectral and temporal dimensions. Can someone provide a brief explanation about this and MPS?

A time-frequency graph also know as a TF distribution, is a spectrogram, a spectrograph, a sonogram, a waterfall display of sound. They are all different names for the same thing. They look like this:
https://www.google.fr/search?q=dolphin+spectrogram&num=100&source=lnms&tbm=isch&sa=X&ved=0ahUKEwiD3vv-9cvdAhUCzhoKHf4jDM0Q_AUIDigB&biw=1429&bih=734
Just as white light contains many wavelengths of colors, sound energy contains bass, mids and high all in the same moment in time, a bit like a water wave has multiple wave sizes, sound energy is the same with sound pressure levels.
Modulation refers to the variation of a simple signal, like tarzan modulates his voice using a kind of tremolo effect, so does a car alarm, and a bird has very varied squeak effects with tremolo effects and also changes from squeaks into clicks into hisses, spectral changes in the timbral quality of the sound. Modulation is a scientific word for the variations of the amplitudes and frequencies of a communications signal.
They write about temporal modulation and spectral(pitch) modulation, it's just hte changes on the time-frequency graphs.
Modulation power spectrum is a relatively obscure field of bioacoustics, it attempts to generalize the fluctuations and patterns of loudness and squeak spectrums into another graph, which is a bit like a secondary time and freqency graph derived from the first sound time and frequency graph, else a 2D graph as in the illustrations, which show sound fluctuation prevalence as an average of the TF graph(a averaged sound fluctuation prevalence of the spectrograph). so, if tarzan's voice oscillates every 1 second, he has a major modulation at 1Hz spectrally, and a minimal amplitude modulation, and they will graph that as a peak as 1Hz on the MPS graph.

We can see on this image that The human speech spectrogram on the top left is mostly at 500HZ, with some S and CH sounds at 2000hz.
For the MPS of the human voice, they have correlated Spectral modulations against Temporal modulations. It is easy to see that the human voice mosly changes at arond 1HZ, same as tarzan's voice, because syllables and pauses take about .5 to 1 second (average words per minute of a human is about 50, average syllables per minute is about 120). So it is easy to see a temporal modulation peak at 1Hz.
The less clear thing is that it is plotted against spectral modulation, which is 2-3 octaves of change, an octave being a doubling of wave speed and frequency. So we can say that the main spectral modulations of the human voice occur over 2-3 octaves at a speed of about 1Hz.
There is a secondary peak, which suggest that there is a change of about 1 octave that occurs at about 200-300 HZ. That is the gruff buzz that you hear in your own man voice, it is about 150HZ and it has about .5-1octave, the base frequency of your voice being 150Hz and various 3rds adn 5ths of an octave, and another octave at 350Hz, seems to be visible from that graph.
tarzan voice: https://www.researchgate.net/profile/Alexandre_Hyafil/publication/283260815/figure/fig2/AS:293940512215079@1447092287682/A-Phase-Frequency-coupling-can-be-mistaken-for-Phase-Amplitude-Coupling-Top-curves-An.png
MPS seems to have been used to study neuron responses in psycho-acoustics.
Neurons interpret sounds using some kinds of complex time and freqency variation(modulation) recognition(resolution) and the researchers use MPS as they say it fits well with the neuron psychoacoustics study.
I would suggest that there are many other ways of measuring psycho acoustic nerve responses and bioacoustics than only MPS, it is an over simplification of sound analysis, and the Digital-Signal-Processing forum on this website will inform you that there are also 100ds of other maths and analysis algorythms that you can run on sound to study Time-Frequency Graphs, and to define sound in many different ways than MPS.
very pure sounds like sine waves make tight lines on a time frequency graph, noise makes very scattered signals at all frequencies, and sounds with many harmonic modes make nice ordered lines of smaller subharmonics that reflect the base frequency of a signal, which resemble echoes of the original sound in parralel with it, and that are on a different frequency.
Time frequency graphs are quite interesting to study and there are probably some studies of bird calls on YT: https://www.youtube.com/watch?v=_FatxGN3vAM

Still not sure if I'm getting it. (1) What (I think) I understand is that MPS graph shows the variation of pitch (spectral modulation) vs variation of time (temporal modulation). So if Tarzan's voice jumps from a tree using his voice with a tremolo effect, and his voice oscillates from 10 to 20 hz every second, should I expect to see at the 1hz a distribution of red to blue from 10 to 20hz corresponding to the change in pitch? (2) Consider the "fall" in the MPS for human speech, nearer to 0, do I see more red because within invervals close to zero voice shows a greater change in pitch?

that's about right. (2)The human voice spectrogram is illustrated as a correlation of spectral versus temporal modulations. We can see at .5Hz the giant peak at the start corresponds to the slow change in words and syllables, which varies by 2-3 octaves of pitch. There is also another peak at 150Hz which corresponds to the gruff tone of a male voice, which varies by 1 octave, and which has 3rd and 5th harmonics and at 300hz, an octave above 150hz. I'll add a bit just under the image with another image of tarzan's vice. given a moment to read the peacks, spectrograms are easy to read.

MPS is the two-dimensional Fourier transform of the spectrogram. Assuming that you know what a Fourier transform and a spectrogram is, the MPS method consists in treating the spectrogram as an image on which to apply the Fourier transform. Thus, one can extract an estimate of the joint modulation on the frequency and temporal domain of a signal. If I attempt to provide an intuitive explanation of how to interpret an MPS, I would say that it allows the researcher to quantify the irregularity of a signal across time and frequency bands.

Here a clear explanation, cheers!
https://journals.plos.org/ploscompbiol/article/figure?id=10.1371/journal.pcbi.1000302.g001

Link-only answers are generally not acceptable on Stack Exchange sites. The link may change or become unreachable in the future, and without a summary of what the link contains this answer would be useless. Please summarize what is in the link (don't just copy and paste) and use the link solely for reference. If you remove the link and the answer cannot stand on its own, it is not a good answer.


text:
Horizontal gene transfer and Botulinum toxin
Is it theoretically possible that any aerobic bacterium becomes able to produce Botulinum toxin as a result of horizontal gene transfer from Clostridium botulinum to that bacterium?


text:
How do we call a region of low carrying capacity?
I am looking for a convenient word to define a region of the landscape where the  local carrying capacity (population size) for a given species is low but different from zero. The species is still present in this zone.
How would you call such area?

I hesitated to ask my question here or on ELL.SE. I asked my question here as I am hoping that there is a term in used in the literature.

If you ask about quantity/numbers only, I would suggest using "the zone of low population density" or "intermittent zone" which in turn borders the "buffer zone" and "the zone of intermediate/high population density".
The "buffer zone" is the zone between two different populations where the number of the animals in question is zero.
J.Potts, M.Lewis "How do animal territories form and change? Lessons from 20 years of mechanistic modelling"
There are other classifications based on the "main usage" of the territories.

Thank you for your answer. Reading your answer I reazized my question was unclear. I meant a zone of low carrying capacity surrounded by zones of high carrying capacity. I edited my post. Your answer seemed very correct as one possible interpretation of my previously answered question. Sorry, my mistake.

@Remi.b IMHO, you should roll back to your original formulation and use ADD word (or UPD) before a new part of the question. I answered the original question, now you ask another question (which is obscure as well), so the answer looks not relevant...

I rolled back to my previously unclear question. I'll open a new post (or just ask my colleagues) for the question I really wanted to ask. Thank you. +1 Btw, what does "ADD" and "UPD" mean?


text:
Eukaryotic cell lysate-based protein expression efficiency
What is average efficiency of eukaryotic cell lysate-based protein expression systems in terms of (mg of protein expressed) / (mg of lysate) ?

This is nothing what you can determine that easy. The question is from what cells your lysate is prepared, how does your protein sequence look like (long or short and so on) and other parameters. You can have a look into commercial kits, which may give you a yield estimate, but this might not be true. Also: Functional protein or just total protein?

Total protein .


text:
Sprawling locomotion, are knee/elbow joints perpendicular to erect locomotion?
The first terrestrial animals used sprawling locomotion. Is the knee and elbow jointed perpendicular to the joints in erect locomotion? It is easy to assume so, based on how the limbs move in sprawling locomotion.

There was a question on exactly what is meant with jointed perpendicularly. It means the joint angle is perpendicular. Here is a rough ugly sketch, does not have to be a joint like that, question is if the movement in the joint is in any way in that direction.

Here, Baier seem to support that the knee and elbow joint in sprawling locomotion supports movement perpendicular to the simple hinge joint in erect locomotion, which is exactly what question is asking.

"The elbow was not a simple hinge; motion entailed substantial
abduction/adduction"

Baier, D. B., & Gatesy, S. M. (2013). Three-dimensional skeletal kinematics of the shoulder girdle and forelimb in walkingAlligator. Journal of Anatomy, n/a–n/a. doi:10.1111/joa.12102
Edit: the "diagram" caused some controversy in comment section. It was a copy+paste, simplest fastest possible I could make, in response to comment "What do you mean by "jointed perpendicular"?", because I considered that comment to be asking something that is clear. It is meant to show the perpendicular movement direction, not the joint design. It is not needed for the question, at all.

They don't appear to be in the images, why do you think that they would be?

It is easy to assume so, based on how the limbs move in sprawling locomotion. In that particular image, they are jointed perpendicular, but it is a rough schematic, could be wrong.

What do you mean by "jointed perpendicular"? They appear to be jointed in the same orientation in both pictures.

What do these diagrams have to do with the comparison in the original images?

They were a response to your comment. But nevermind. If you could falsify that abduction/adduction in the elbow joint of alligators is not the perpendicular movement I ask about (since besides flexion/extension, and pronation/supination, it is what is left..), then you prove my question to be wrong. I would guess it is correct though. If it is correct, why are you critiquing it so much?

A question is not correct or wrong, only an answer can be. I am critiquing because it is unclear. Probably it is just an issue of writing it in English, but that's still important because you won't get good answers if the question is unclear. In the diagram you drew, it seems like you are calling "perpendicular" motion to be the hinge-like motion you indicate in green, but when you write "perpendicular to the simple hinge" it sounds like you mean motion not in the direction indicated in green.

But it isn't unclear. Forget the diagram. You were asking questions, "What do you mean by "jointed perpendicular"?" it was a response to that, nevermind it. The question is clear. I suggest the answer by Baier, that there is significant adduction/abduction movement, means that there is some perpendicular movement. This means my assumption was right.

Probably the simplest source of lack of clarity is that you wrote "are knee/elbow joints perpendicular to erect locomotion?" when it seems you meant to ask "can knee/elbow joints move in a direction that is not the primary hinge direction?" These are not interchangeable questions.

This question could be better explained, perpendicular to a previous orientation? what the migration of the joint and bone orientation is through evolution? perperdicular is a very specific word which requires more specific references to it's surroundings than you give. Are you saying that the knee could fit image 4 rather than image 3?

This article by Baier seem to support that the knee and elbow joint in sprawling locomotion supports movement perpendicular to the simple hinge joint in erect locomotion, which is exactly what question is asking. The joints in sprawlers also supports flexion/extension axis though, and the question could have been a bit clearer on that. If I misinterpret Baier feel free to point that out (but since besides flexion/extension, and pronation/supination, it is what is left, I think I do not), and if not, question answered.

"The elbow was not a simple hinge; motion entailed substantial abduction/adduction"

References
Baier, D. B., & Gatesy, S. M. (2013). Three-dimensional skeletal kinematics of the shoulder girdle and forelimb in walkingAlligator. Journal of Anatomy, n/a–n/a. doi:10.1111/joa.12102


text:
Does tardigrade eat mould?
There is a problem of devastating mold in ventilation of old buildings in Europe. I guess tardigrades might be better solution because chemicals is not a good idea for ventilation because people are breathing with this air.

can you add some links to the terms - especially "tardigrade" - in your question? it's kind of hard to follow - it would definitely benefit from some more background information.

Majority of more then nine hundred of known species of tardigrades are populates mossy and lichen pillows on the earth, the trees, the cliffs and the rock walls. I seen huge black tardigrades about 1.5mm at wall with mold.

Mould in buildings can be serious.  It is treated by removing and destroying (fire is good) the things the mould is growing on, and lots of cleaning.
The mould grows on wet cellulosic material like fiber board, gypsum board, paper, or wood.  It can be prevented by fixing the leaks so stuff doesn't get wet, and more cleaning.
You could pour bleach on the contaminated material, kill some of the mould, and what survived would grow back.  Neither chemicals, nor tartigrades are any help here.  Just more cleaning.
Details:  http://www.cdc.gov/mold/stachy.htm


text:
What winged insect is in this photo? Is it Axymyia furcata?
Posting on behalf of a neighbor who asked:

Can anyone identify this insect? It's about 3/8" long. There are, it seems, hundreds flitting low over our lawn. There were none 2 days ago.

This is the Northern California Bay Area Peninsula.

$\frac{3}{8}$'' is a weridly accurate measure so just to confirm, it was $\frac{3}{8}$ of an inch, that is 0.95 cm, is that right?

Yes, " indicates inches. So about 1 cm.

One concern is that they are swarming termites.

My neighbor submitted the photo to http://bugguide.net and it was identified as a sugar cane soldier fly,
"Inopus rubriceps"- Female
http://bugguide.net/node/view/1299782
They are from Australia and were accidentally introduced to San Francisco 50 years ago.  It feeds on grasses.

Great pictures.  First, fly or wasp?
In the second picture, under the head you don't see jaws of a wasp, but do see a bit of a tongue-like organ, like a fly.  Further down, you can see that there are only 2 wings.  Just below where the left wing meets the thorax, you can see what looks like a tiny wing or paddle.  That's a haltere, a reduced wing that vibrates to provide stability, like a gyroscope.  Flys have halteres.  Finally, in the first picture at the very end of the abdomen are 2 points.  That ain't a stinger, it's an ovipositor.  It's a fly, female, and from the size of the abdomen, gravid.
The closest I've gotten to identifying this one is Axymyia furcata http://bugguide.net/node/view/1216479.  Everything matches, as well as we can see from a photo, except the coloration.  Also, A furcata is associated with streams and is found in the north-east and north-central US, while, your bug was found on a lawn in CA.  For more information about A. furcata than you need, there is a master's thesis available:  http://lib.dr.iastate.edu/cgi/viewcontent.cgi?article=1882&context=etd.
I've looked at hundreds of fly pics to find the one I linked to above.  That's the only fly I found that matches the shape of yours.  Maybe it's a different fly within that genus, maybe it's just a color variation.
Their maggots live in rotten wood and decaying plant material, and the adults are mostly interested in more maggots, so this isn't a pest, just a neighbor.
Edit: furicata --> furcata, oops.

Can you fix furicata to furcata?  (I can't -- the edit isn't large enough.)


text:
How does bath application of KCl depolarize a neuron?
Increasing extracellular KCl is often a way to depolarize neurons in experiments.
My understanding is that increasing K+ extracellular concentration changes K+ reversal potential to more positive values and hence depolarises the neuron as the open leaky potassium channels will lead to influx of K+ into the neuron. Is this correct?
However, this explanation does not take into account Cl- ions.
How can this be explained in terms of Goldman equation?
Also, how does intracellular injection of KCl affect neuron membrane potential as compared to extracellular KCl application? In patch clamp experiments, often the pipette is loaded with KCl and injected into neurons that leads to depolarization. How can this be explained?

Cl does not play an important role in membrane depolarization and AP. Chloride channels are not very common in neurons (unlike K⁺, Na⁺ and Ca²⁺ channels) and therefore play a minor role. However there are reports of chloride channels that can regulate excitability of neurons

Yes. But how can you explain this in terms of changes in electrochemical driving force and derive from Nernst/Goldman equations?

Well. It essentially means membrane has a low permeability for chloride and therefore it (Cl0 does not diffuse much. Therefore its contribution in membrane potential (described by GHK equations) would be less.

Membrane potential (measured electrophysiologically, or calculated using the Goldman equation) depends on two things: concentrations and permeabilities. In the Goldman equation you see this directly: concentrations multiplied by permeability, in both the numerator and denominator.
If the membrane is not permeable to an ion (PIon=0), that ion will not affect the reversal potential. In a typical resting neuron, in the absence of any synaptic activity, Chloride conductance is near zero compared to ions like potassium.
I'm not as clear what you are asking about patch clamp experiments, I will try to just describe the process and hopefully that helps.
In a whole-cell patch clamp, the inside of the cell is continuous with the patch pipette solution, which usually contains high concentrations of potassium, like the normal intracellular environment. This is done on purpose, to try to record from cells in the most natural condition that can be replicated simply.
During the process of making a whole-cell recording, however, the experimenter has to first place a patch pipette down near the cell. While doing this, some solution is leaking out of the pipette and increasing extracellular potassium concentrations, which can depolarize surrounding cells. So, patch quickly my friends, you are making it uncomfortable for everybody before you make a good seal.
(the process would be similar for inside-in patches; for on-cell or inside-out patches, one would use different ionic concentrations in the pipette to mimic the extracellular space, or these would vary depending on the exact experiment performed)
edit: Just wanted to note that a statement in your question is mostly wrong; "the pipette is loaded with KCl and injected into neurons that leads to depolarization" - KCl inside a cell would not depolarize that cell, unless the concentration of K+ was lower than the typical K+ concentration of the cell, in which case you would probably have other issues and a dead cell from osmolarity problems.


text:
What kind of salamander is this?
This guy was found in central Virginia, under a flower pot. He's about 3 inches long, because part of his tail is either missing or it's just short. He looked dried out, so of course I wet my hand, scooped him into a small container, and moved him to our birdbath (which is on the ground and had been recently rinsed so the ground was plenty moist). I ran my hand (still wet) over his back and he seemed happier and started moving around.
However, I can't seem to find on Google just what kind of salamander he is. The VA Herpetological Society has an entry for a Jefferson Salamander, but those are only found in North/West VA. Attached is a picture of him (blurry, sorry) when I found him. I have a video of him coming out of the container and going under the leaves next to the birdbath, but it's too big of a file to put on here. Instead, here is a link to the file in my Google Drive: Salamander
Edit: I checked some pictures I had of him, and he has five toes on his hind legs. He is a dark gray-brown with faint mottling and a semi-light underbelly. Sorry that I didn't say this earlier. Hope this helps!
Can anyone identify him?

be careful of tapwater with amphibians, you probably know, because of the chlorine content.

yea i used spring water for my hand, but I used garden hose water for the birdbath. That should be okay, because chlorine evaporates out after 24 hours, and it's been about 48+

@PvParkour thanks for the updated clearer photo. I think you should keep it, but I still want to recommend that you [edit] to also add a gif of your video. EZgif is a very easy website to use.

Maybe it's a Mabee's Salamander, Ambystoma mabeei.  Unfortunately, I can't really see the underside in your video, but I have the sense that it is a lighter color.  The clubbed tail, with an apparent break, seems like it may have been autotomized or otherwise injured, making it harder to compare.  I had the impression your example had dark spots dorsally and lightening ventrally, as can be seen in the image above.  Some other images are a bit different.  However, Mabee's Salamander is only found in "six localities in the coastal plain in extreme southeastern Virginia: the cities of Hampton and Suffolk and the counties of York, Southampton, Gloucester, and Isle of Wight. It is also found in Newport News." and is considered threatened, so unless your location matches what you read there it is still not likely to be the answer.

i updated it for a better picture i you want to look at it again

The appearance seems similar to my eye, the frequently confused species suggested at the first site don't look like they could be options, but the range and rarity of Mabee's Salamander concerns me.  If you're not in the area they describe, I'm very likely to be mistaken.

no, Im close to richmond (central VA). its ok though, you're still helpful

ok since the bounty expires in one day, i dont want 50 rep. to disappear, so ill give it to you