91 replies to this topic

[Mind]

Interesting to see the challenges of raising a large population of mice. Thanks for keeping us up to date.

[AgeVivo]

Thank you Mind! - it is! Now that I have 30 males and 30 females of known age, it is insufficient to do a well powered lifespan test with N=60 per group.
I wonder if I can handle 120 mice. Not sure for now. It takes quite time, energy, and even money (food)
If someone was doing the same as me, we would both use N=15 per group, perfect. Anyone?

 

The other challenge is that I will start experiments in more than a year, after all these efforts. This, because I had to make babies to know their age and then wait. So how can I prepare a new cohort while the previous cohort has reached its mid-life, to wait much less next times, without handling 120 mice?
The solution would be to have someone doing the same as me living nearby: I would give him my males (for example; to test viagra females are better to permit some blind study) and take his females (for each next cohort we can jointly decide who has males and who has females; in fact, exchanges can be done a month earlier of starting an experiment - estimated time for animals to adapt well to their new environment and new human contact)
 

The other things is how to get reagents (though one can find its ways since one have months to find the solution - I now have sildenafil at home). And how to organize placebos. Anyone to know for the placebos?

Cheers
 

Edited by AgeVivo, 19 January 2023 - 07:41 PM.

[AgeVivo]

Changed cages, and found one of the 10 females of the first offspring dead, under the wheel, hidden in the hay carpet where they like to hide. Not sure what happened, first time there is a death that is not obviously due to a fight; so sign of injury, this type of carpet is thin so I don't think asphyxia. Hit by another mouse with the spinning wheel? Doesn't seem very probably. 5 month old. Not sure what happened.
While I am still preparing the colony and establishing things, I am thinking of trying next week-end sand + hay: sand instead of litter. Better do all kinds of adjustments now than later, to be well established as soon as possible. I did read many website in the past months but didn't see anyone will a clear knowledge on what's best. If you are a mouse lover don't hesitate to provide your knowledge. Cheers

[AgeVivo]

Changed cages, all is perfect. The mice seem well and happy. The lastest 8 newborns from January 15 have mixed black&white shapes and are very cute. In an nutshell, now that I have a stable installation, I feel that N=60 mice is perfectly OK.

 

I wonder though now how I will have enough mice of one gender. I feel that having more mice is a challenge. i) I don't have much room and ii) it takes quite much time per cage to change cages so the most practical for me would be to "pack" cages with mice in order not to further increase the number of cages. If I do that, I need to have various mouse-houses per cage for them to find their collective equilibirum, otherwise they will fight, there will be deaths and injuries, especially in the few days when merging mice who don't know each other. Also, iii) I have to make sure that if I mix animals of different ages in the same cage, I am able to distinguish the mice from the two ages! (for the longevity experiment).

Also, iv) I am afraid it will smell much if I have 120 mice, not 60, branched on the vaccum of the kitchen. Though, I could always add a pump if needed, plugged on an eletrical plug, to facilitate the air extraction to the general vaccum of the kitchen. So, even if I am a bit afraid of not handling the smell well, somehow I know this aspect should be fine.

 

If I could go back in time, I would have kept the first male newborns with females-and-mothers until age 1.5 month, then move them away, in order to have a huge number of newborns at once in a cage, and split the males from females one month later. This would have given me crowded-but-happy cages with one age and one gender per cage. Exactly what I would need now. But I didn't know in advance how mice would feel when staying together since they are born (very well, even in large groups) versus when mixing groups (not so well, except if cages are "messy" with many toys and homes for mice to find their own room), I didn't know how much time it would take to change cages once regular, I didn't know how much it would smell when using the air extractor, etc etc etc.

 

 

This is about the N. Now, about the treatment. I will definitely test viagra, if it works this post will make the buzz and it will be good for longevity - for people. I was able to get viagra, now I need to organize the treatment versus placebo aspect.

I prepared for how to make a placebo versus treatment and obtained good results: I crushed pellets into power, mixed with a bit of water, rolled the mixture in paper and put one minute in the microwave: it is like pellets again:

So if I do this both with water+nothing and water+something and I don't know what-is-what, I do a blind experiment. Perfect
I can ask one of my children or a neighbour, who are interested in what I do, to prepare the water and water+nothing without me knowing it.

It can also be in the water they drink, directly -- it doesn't have to be via food. But it is good to know I can do it with food. Sildenafil (viagra) dissolves well in water it is probably best to put it directly in the drink.

Edited by AgeVivo, 29 January 2023 - 12:14 PM.

[AgeVivo]

Theoretical way to go with my little acquired experience:

 

t=0

buy 6 white initial males with red eyes (albinos)

buy 6 colored initial females with black eyes (normal)

Put them in 2 cages (3 males and 3 females per cage)

 

t=2 weeks

Put the males back together with many homes in a new large cage (to limit fight; not obvious) and same for females (a very large cage if possible)

t=3 weeks, 4 weeks

Limit the number of homes in the male cage so that they live together as a group that appreciates each other (ok, sometimes one or a few will kick the ass of others, but it gets accepted by the group), reduce slightly the size of the cage to spare room.

 

Let us take an assumption to understand what we are doing: at this stage, say 3 initial females were pregnant and then make about 30 newborns total. This is the "first offspring". Write their date of birth carefully. Let us say 16 males and 14+6=20 females (including the initially bought females).

 

t= 2 months and later. Make sure you distinguish the initial females from other females as the daughters will grow and size will not be a distinction but you will need to know the age of every mouse of the lifespan experiment.

t=about 3 months (precisely, when newborns are 8 week old and have had time to mate with mothers and sisters) = time to stop preparing new children: Put the males of the first offspring away.

You can put them together with the initial males, with many homes in a new large cage (to limit fight). You can not put them together if you wish, but it means one more cages so more work for the next years every week-end or other week-end to come..

Again, make sure you recognize the initial females

 

Now, let's continue with the assumption: out of the 20 females, 10 are pregnant, they lead to 100 babies. They were not all born in one day, so you have splitted the cage with many many mice 10 days after the first newborns (of the "second offspring") in order to know the age of newborns up to one week uncertainty, approximately. When you did the split, you also moved the mothers so that the babies have their mothers. So you have
CAGE A - 1 cage of 6 initially bought males and 16 males from the 1st offspring -- probably less sadly due to males fights

CAGE B - 1 cage of 50 babies of the 2nd offspring (0-10 days after the first newborn of this "offspring") and 3 females initially bought and 7 females from the 1st offspring -- perhaps less sadly due ot fights

CAGE C - 1 cage of 50 babies of the 3rd offspring (>10 days) and 3 females initially bought and 7 females from the 1st offspring -- perhaps less sadly due to fights

So, you have approximately 130 animals of known ages, in 3 cages

 

When the babies are 3-4 week old, move the adult females away in order to avoid new offsprings. Put them together in a large cage with many homes.

When they are 4-5 week old, move the young females away from the young males. So now, you have:

CAGE A1 (previously named A): 10-20 males of known age + a few initially bought males

CAGE A2 10-20 females of known age + a few initially bought females

CAGE B1 about 25 males of known age

CAGE B2 about 25 females of known age

CAGE C1 about 25 males of known age

CAGE C2 about 25 females of known age

Now, the goal is
1. to maintain this as happilly as possible, while mastering the vaccum and the efficiency to changes cages every week-end or every other week-end : important to be able to do this well for when you take vacations, this comes before the beginning of experiments so it is the first priority

2. to carefully prepare the experiment: get reagents, learn to prepare them, study how someone can help you be blind on "placebo" versus control". There is more time but it can be complex, so prepare much in advance

Edited by AgeVivo, 29 January 2023 - 01:40 PM.

[AgeVivo]

The above is quite theoretical as it very much depends on how many females are pregnant at the very beginning. So it is hard to land right, and additional cycles might be needed, a bit like what I did.

 

To move probabilities on the side of caution, it might be better to buy more animals at the beginning, to have a larger initial first offspring, then to move the males from the first offspring one week earlier than described here above in order to have a much reduced third offspring.

 

Still, I do not have the experience on that so things might be wrong by a week and it might be needed in all cases to complement with a fourth offspring much after. This leads to more complexity, two more months, more cages (with less animals), but the fourth/fifth offspring becomes the main adjustment and it is much less dependent on how many initially bought females are initially pregnant. I did this, or with a few more cycles. It leads to 10 cages, not 6 : it is not possible to merge cages of similarly-looking animals born at different ages, otherwise one doesn't know what animal is of what age.

 

With that approach it takes 7 months to have the right cohorts. They are then aged 0, 2 months, 2.5 months, and some 4.5 month and 5 months months, and a few initially bought of unknown exact age (8-12 months, probably)

Edited by AgeVivo, 29 January 2023 - 02:09 PM.

[AgeVivo]

re-reading above, there was a typo here. I meant N=30 per group (control or treatment, for a given gender). So 60 females for example. This typo is probably obvious to people with some knowledge in survival stats but probably not to others and it can affray

Thank you Mind! - it is! Now that I have 30 males and 30 females of known age, it is insufficient to do a well powered lifespan test with N=60 per group.
I wonder if I can handle 120 mice.

 

Edited by AgeVivo, 29 January 2023 - 05:27 PM.

[AgeVivo]

I have changed the cages every other week without giving information here. In short the mice do well and I am in a routine mode, I take care of them until they are 18 month old to start an experiment. I will need to investigate if it is sildenafil or something else but I have time.

 

cage 1a: 10 females (3rd offspring, born 2022-12-23; all kinds of colors)

cage 2a: 6 black males (1st offspring, born 2022-09-01) + 1 bought male (mixed colors, born approx 2022-10-15)

cage 3a: 4 females (3rd offspring, born 2022-12-23) + 1 female (born 2022-05-16, initial sister numbered 8) + 1 bought female (bought as male but it was a female, strange but it was not obvious; born approx 2022-10-15)

cage 4a: 9-or-10 black females [incl 3 gray] (each time I moved cages while not counting; 1st offspring, born 2022-09-01) + 4 females (multicolor but always which white so distinguishage from the born 2022-05-16); 

cage 5a: 12 males (3rd offspring, born 2022-12-23; all kinds of colors)

cage 6a: 12 recently born 2023-02-07; all kinds of colors; separation of genders to be done soon

[AgeVivo]

changed cages yesterday

 

cage 1a: same as before: 10 females (3rd offspring, born 2022-12-23; all kinds of colors)

cage 2a: same as before + I added males from cage 6a: 6 black males (1st offspring, born 2022-09-01) + 1 bought male (mixed colors but no white on the head, born approx 2022-10-15) + 4 males (mixed colors and white on the head, born 2023-02-07)

cage 3a: same as before: 4 females (3rd offspring, born 2022-12-23) + 1 female (born 2022-05-16, initial sister numbered 8) + 1 bought female (bought as male but it was a female, strange but it was not obvious; born approx 2022-10-15)

cage 4a: same as before: 9 black females [incl 3 gray] (each time I moved cages while not counting; 1st offspring, born 2022-09-01) + 4 females (multicolor but always which white so distinguishage from the born 2022-05-16); 

cage 5a: same as before: 12 males (3rd offspring, born 2022-12-23; all kinds of colors)

cage 6a: kept females only: 9 females (born 2023-02-07; all kinds of colors)

 

Things to note:

- for 2 months, a female in cage 4a has been having a big bump. Tumor or infection - she doesn't smell like she has an infection; it seems to have appeared within a week, it is about 2 cm long and 1 cm wide and tall, it is now stable though over the weeks it seems it very slowly grows. She doesn't show strong signs of suffering from it, she is active and among the most playful mice of the cage, it seems it reduces a bit her balance when she plays equilibrium games with the bars of the weel

- when merging the males in cage 2, based on my past experience to try to reduce fights, I first put animals in a house in two different cages then I grouped the two carpets and two houses in the same cage, such that the two groups have their natural territory and can smell if other and get closer when they want but return to their house if they feel afraid. I hope and guess I will avoid fights and deaths this way.

- in cage 6a, some females are pregnant. I have the room for 2 more cages so for now I will stop with these forthcoming young mice. For now I have 10+6+13+9 = 38 females and 13+12 = 25 males. So a current total of 63 animals.

Edited by AgeVivo, 26 March 2023 - 08:39 AM.

[AgeVivo]

In cage 4a it is very very likely a tumor (http://www.rescue-fo...ne-2-ans-16969/)

 

I take pictures from cage 2a to later help distinguish mice: when some mouse dies I must be sure of its age:

Corresponding comments in the preceeding post

[AgeVivo]

cage 1a: 10 females as before

cage 2a: 10 males: following the mixing of two populations of males, fights happened and one of the 4 males with white on the head died (as sadly expected in the last post; but I would say "not too bad"; at my beginnigs this was usually much worse). The 6 black males are still there as well as the bought male (who seems to be dominant)

cage 3a: 6 females as before but only on one side of the cage; on the other side I have put yesterday the mouse with a tumor from cage 4a: I am trying some special keto diet with dark green leaves to see if it potentially resues her as she would likely die within a few days otherwise (the tumor is on the back/side/shoulder and is overall about the size of the head, it becomes heavy and probably hurts which makes her fall on the side from time to time). The two sides of the cage are separated by a metal grid inside the cage - this permit the isolated mouse not to feel too isolated.

cage 4a: 12 females as the one with the tumor was moved to cage 3a. 

cage 5a: same as before: 12 males (3rd offspring, born 2022-12-23; all kinds of colors)

cage 6a: 9 females + 25 newborns, born around 2023-04-1 (half of them born 3 days before, half born 3 days after)

Edited by AgeVivo, 08 April 2023 - 06:05 PM.

[AgeVivo]

So if the half of newborns are males, I should soon have about 85 mice in 8 cages:

• 37+12=49 females (+1 but out of the lifespan test due do its cancer at a relatively young age and the special treatment I try) in 5 cages
• 24+12=36 males in 3 cages

This is quite clearly the maximum I can do in my current setup.

It is strongly limited in terms of statistical power for females and inadequate for males (except for very large life extensions of course)

[AgeVivo]

an event happened: the top of cage 5a apparently fell as the males discovered that by jumping it made it move (I wasn't there, my son saw it) and five males where able to enlarge a whole to enter in the neighboring cage of females (cage 1a). I put them back to their cage a few hours later -- perhaps there will be many babies in cage 1a...

[AgeVivo]

cage 1a: 10 females as before

cage 2a: 9 males. 1 black male died, obviously from a fight as it happens across males

cage 3a: 6 females as before; the mouse with a tumor died a few days ago; despite the keto+green leaf died the tumor was increasing and she stopped eating over the last two days. I did a necrospy, didn't find any metastasis but the tumor was larger than the head of the animal.

cage 4a: 12 females as before 

cage 5a: 12 males as before

cage 6a: 9 females as before and... I accidentally killed one as it tried to escape when I was adjusting the place of the cage while the top was removed. I feel extremely bad about that. It won't happen again now that I see that this can happen but.. there are many things to know in my installation to make sure they have food, drink and don't die from an accident...

cage 6a: the 25 newborns, who are beautiful, with black and white patterns

So about 50 females and 34 males. It starts to be enough for the experiments. Perfect.

Edited by AgeVivo, 22 April 2023 - 09:45 PM.

[AgeVivo]

an event happened: the top of cage 5a apparently fell as the males discovered that by jumping it made it move (I wasn't there, my son saw it) and five males where able to enlarge a whole to enter in the neighboring cage of females (cage 1a). I put them back to their cage a few hours later -- perhaps there will be many babies in cage 1a...

There were 10 newborns this last week following the event ...that I killed immediately (after moving the adult females to another cage) because I absolutely do not have the space nor time to handle them - no choice.

 

Other than that, no death, and in cage 5 the sister of the one who had a tumor and recently died also has a tumor. This time, not around its right shoulder but on its right side next at the belly/chest. It is roughly about 1 cm of diameter (I observed the tumor by handling the mouse, it is not very visible otherwise).

=> I will try a ketotarian diet (plants, proteins and fat but no sugar) + antiglutamine treatment (fenbendazole, 2 mg per day in water or food, I will see), as there is strong science suggesting that tumor cells make their energy from glucose and glutamine

[AgeVivo]

I splitted the 25 young mice into males (14 I think, I didn't specifically count; cage 6a) and females (11 I think; cage 7a)

 

In cage 4a, a dark gray female mouse (born 2022-09-01) also had a tumor, I put it together in cage 8a with the white-brown female (that must have been in cage 4a before, not 5a males: typo in my previous post obviousy)

the white-brown female has been treated with fenbendazole dissolved in water since Friday (so 3 days before this gray mouse)

is it a bit strange to have 3 tumors in the same cage...

 

[AgeVivo]

On May 7, I used the same hemp litter as long time ago (when I had quite many deaths, you will see why I mention this is a second), and I have 3 deaths quite deep in the litter. I am going to through this type of litter (that is sold everywhere and used like I do in petshops, and about which I receive ads, which is why I thought let's try again...). I had read in forums that it is not good for them but I had read the contrary as well, it is a bit like tabacco or fertilizers, a big lie...

cage 1a: 9 females (3rd offspring, born 2022-12-23; 2 brown (agouti), 1 black with very little white, 1 black & white, 1 white, 1 light gray, 3 brown & white [one a bit yellow, two quite dark])+ 1 death this week deep in the litter (black)

cage 2a: 8 males (4 black males, 1st offspring, born 2022-09-01 all quite bited with little hair as they constantly bite each other in this cage; 1 bought male mixed colors but no white on the head, born approx 2022-10-15; 3 males mixed colors and white on the head, born 2023-02-07) //4 black not 6 because 1 died from a fight very likely as indicated above (April 22) and 1 death this week deep in the litter (black)

cage 3a: 6 females (1 large hairy white & light gray bought as "male" born approx 2022-10-15; 1 agouti, 1 black, 1 black & white (incl one that looks like initial sister 8...), 2 brown & white: all 4 born born 2022-12-23)

cage 4a: 9 females (5 black & 2 agouti 1st offspring born 2022-09-01; 1 black & white and 1 brown & white born 2022-05-16); //7 black/agouti instead of 9 because one is aside due to a tumor and 1 death this week deep in the litter; 2 colored mice instead of 3 due to a tumor (and instead of 4 previously due to another tumor) -- I put in bold the tumors because it is strange to have the 3 tumors in the same cage. However it is the oldest cage and perhaps (I don't feel so but it is still possible) for some reason I saw the tumors in this cage and other deaths are linked to tumors -- next time I change cages I will pay attention to tumors everywhere (it takes some time to scrutinize all mice though while handling them)

cage 5a: 12 males (3rd offspring, born 2022-12-23; all kinds of colors)

cage 6a: 8 females (born 2023-02-07; all kinds of colors)

cage 7a: 12 females born 2023-04-01 (3 white with red eyes, 1 brown with a little white, 4 brown and white, 4 black & white)

cage 8a: 13 males born 2023-04-01 (4 white, 1 brown and white, 8 black & white)

 

The 2 living mice with a tumor (one agouti with a tumor on the top of the back and one brown/yellow&white on the right side) are in a separated cage 9a with the good hemp and a great and fun environment (I didn't use the bad hemp in that cage because the good one allows to make lawyers, wholes etc and I made a fun environment), they eat green leaves, salad, avocado, tomato sides, pepper sides, a bit of their normal (quite keto) food and they have mebendazole in their drink (to reduce glutamine). Their tumors were growing fast before this treatment and seem to be stabilized but I might be biased and it might be that since they are larger it is difficult to see the growth as much. I guess I have to wait two weeks to see if it continues growing. In any case, it is not reducing, so I can't shout victory at all and I am wondering what I should do to avoid the scenario where I am unclear if I should have tested something more severe. Any thought is welcome

[Mind]

Are you wondering if you should have tried a "stronger" treatment? (You wrote "Severe")

 

 

 

so I can't shout victory at all and I am wondering what I should do to avoid the scenario where I am unclear if I should have tested something more severe

 

[AgeVivo]

Are you wondering if you should have tried a "stronger" treatment? (You wrote "Severe")

 

thank you Mind, yes.

actually the tumors were & are still growing - I needed a few more days to see it. Yesterday evening I multiplied the dosage by 5 (now their water looks like milk given the white treatment), they seem to do well and be happy so no side effect, I continue with such a high dosage in their water.

 

Tonight instead of normal food I mixed the content of a green tea bag with a bit of olive oil. This is the keto - against-cancer solution without taking time to read, but I will google this week-end. actually, I guess that I may well find what I need on Longecity !

Edited by AgeVivo, 24 May 2023 - 10:29 PM.

[QuestforLife]

thank you Mind, yes.

actually the tumors were & are still growing - I needed a few more days to see it. Yesterday evening I multiplied the dosage by 5 (now their water looks like milk given the white treatment), they seem to do well and be happy so no side effect, I continue with such a high dosage in their water.

 

Tonight instead of normal food I mixed the content of a green tea bag with a bit of olive oil. This is the keto - against-cancer solution without taking time to read, but I will google this week-end. actually, I guess that I may well find what I need on Longecity !

 

From my understanding keto is relatively effective in humans but less so in mice. 

 

This is a good paper. 

 

https://www.nature.c...2003-019-0455-x

[AgeVivo]

From my understanding keto is relatively effective in humans but less so in mice. 

 

This is a good paper. 

 

https://www.nature.c...2003-019-0455-x

Thank you QuestforLife for the paper;
I am essentially testing the same in my 2 mice but what they do is a (really great) lab-world study and what I do is more like a real-worl study:

- their conditions of tumor and treatment schedule are optimized to have a higher chance of success (they inject some tumor and two days after they start a diet at a specific dose; they find that half of the dose works less). They measure many things, they inject. In my case I wait for naturally occuring tumor and only when the tumor starts to be big I try to do something with oral intakes that one can easily find. I do this because I am not in a laboratory.

 

- To deplete the body from glutamine, I use oral mebendazole ; they use DON injections (6-diazo-5-oxo-L-norleucine; both aim at depleting the body of glutamine, an amino-acid that tumors crave for). Medendazole was showed to help mice fight cancers, so I guess that both should have the same type of effet and mebendazole doesn't need to be injected. In the discussion, they are not sure that the therapeutical effect is really from depleting glutamine or something else (it is great that they keep their understanding wide like this)

 

- To lower sugar blood levels I did my own invented ketotarian diet (keto = where fat/oil replaces sugar). I do a ketotarian diet (plant-based keto) because both keto and plant-based diets are known to help fight cancers : dark green leaves (roquette, spinach), green tea (content of a bag), brocoli, olive oil (mixed with everything green), strawberries. If you would like to suggest me some simple adjustment, don't hesitate. 

They buy/are given a commercial diet, KetoGEN (Medica Nutrition, Canada). Table 1 in the supplement file says it has little carbohydrates (3% vs 62%), much fat (72% vs 3%), a slightly low protein content (15% vs 27%), and contains almost the double calories per gram. It made the mice loose about 15% of their weight.
 

- They inject two types of (glioblastomal) tumors; with the first one the mice fully recover from the tumor; with the second one the mice die later but still die fast. They detect things in both cases. In my case, if what I do slightly increases the survival I won't probably notice any effect: I would clearly notice things in their first case but probably not the second one.

So... we will see what happens with what I do but it might be that I need to get one step more precise. Perhaps I need to inject DON instead of giving mebendazole in water, but I am not sure where to buy DON without a research lab -- would you/anyone know how to organize this? Coincidentally, would you happen to know these researchers? Because perhaps they could guide me: what I do is a real-life translation of lab work. With my very basic model and my 76 mice, I will see many tumors and might gradually optimize the technique towards a non-invasive cancer solution available to anyone (that would be quite immense).

---

Regarding my many mice,

 

I changed cages yesterday, putting the correct litter everywhere (never the desastrous litter anymore!). I checked for visible tumors and did not find any. Nothing particular except i) death in cage 1 (females) with unknown cause (consequence of the desastrous littler? I didn't have the time to do a necrospy) and ii) an ill mouse in cage 3a (it was ill in the previous weeks with an eye slightly bleeding on the side, but now the eye is pink/livid and the belly is particularly large)

 

Now that I have quite solid conditions (the last issue with the desastrous litter, but this being fixed I think that now all is OK), I consider that now the lifespan test starts, with for now all mice as a non-treated non-placebo lifespan control group (once the treatment starts, some mice will be the treated group, others the placebo group, other will remain the nothing control until they have the age to start)

 

I don't know if I shoud include or exclude the 2 mice with a tumor and the one already quite ill. I guess I can do a mouse "healthy lifespan test" where I exclude mice with strong illness. In that case their exclusion are before the start of the lifespan test. In any case, it doesn't change conclusions because I will start treatments the age of approx 18 months and for now mice are younger than that.

 

cage 1a: 9 females ; cage 2a: 8 males ; cage 3a: 6 females (-1 ill) ; cage 4a: 9 females ; cage 5a: 12 males ; cage 6a: 8 females ; cage 7a: 12 females ; cage 8a: 13 males. N = 76 mice : 43 females and 33 males. I wished I had more females or that someone would do the same as me to complement (now that I start to master things I can teach), but this number of animals is the max I can handle (it takes me about 5 hours of care per week; financially, it costs me between 30 and 50$ per month).

[AgeVivo]

PS: this is the article following which I am testing mebendazole (MBZ): https://www.ncbi.nlm...les/PMC4096024/

There, notably,

- it seems that 1 mg MBZ orally every day reduces cancer growth by 80%: "In untreated controls, approximately 300 metastatic colonies appeared in the lungs by 21 days. Mice treated with 1 mg of MBZ every other day showed a mean colony count 80% lower than controls."

- and that the dose can be increased significantly for more effects: "Mice treated with MBZ showed no side effects" and "Treated animals showed a dose-dependent arrest in tumour growth"

 

And I estimate that my 2 large-tumor-bearing mice take about 5-10 mg of MBZ now. My fault until a few days was perhaps to give only about 2 mg and not to make them eat less than they want. I will see within the coming week if the tumors shrink. For now, since I started the high dose MBZ and the low food content, I am not able to judge. I will probably be able to appreciate in a few days but based on my first observations I am not very optimistic - my guess so far is that at best the tumors are stable while the mice lose weight due to the limited diet and appetite due to MBZ, and I don't know how safe this situation is over time.

[AgeVivo]

I was talking here above... and coming back from a 3 day vacations I see that the tumor of the side of the light gray tumor is enormous and just after I see that the other mice with a tumor is dead (the dark gray mouse). So the treatment, the green leaves etc: it absolutely does not work           

 

And about 20 minutes later I hear a huge fight, I was on something else so I come back a few minutes later, and I look at the cage (males), there is blood on the glass and on the tail of one of the mice, blood that is not from the tail, so I guess they just killed one of their counterparts. This is a cage with old males (born september 2022) who have regularly had fights and deaths in their cage. Perhaps this fight comes from the stress of having us come back after 3 days of absence ? In any case, I still have male fights. 

Edited by AgeVivo, 29 May 2023 - 06:43 PM.

[AgeVivo]

I investigated inside the cage where there is blood, actually no mouse died; one got beaten on the side and a mouse continues to fight with it; I had to separate them outside of the cage for their tempers to go down. Clearly, I don't know what to do about male fights.

[AgeVivo]

Changed the cages,
- a black & white mouse is missing in cage 8.. was there 13 before ? did it go to another cage ? I clearly did not find any death and given the room an escape is quite impossible ; I will count cages when I can, perhaps next week-end (busy times currently)
- no death over the last 3 weeks

- no new tumor (I did not count each cage... ; but I checked every animal individually)

Frail mice:

- the one with a tumor (cage 4a) is healthy despite the tumor becoming gigantic (perhaps it increases the weight of the mouse by 50%..); she starts to scratch itslelf where the tumor is, while she is healthy I guess that this scratching will end up kill her.

- the mouse who is blind from an eye (cage 3a) is slower getting better - this cage has a particularly good atmosphere of respect, it is obvious it plays a big role.

- in cage 4a, there is a black and white mouse that is slightly frail
- cage 8a: only 12, not 13 mice ?? what happened ??

I should probably put the tumor mouse back in a separate cage with a mouse without any tumor (puting it alone may make her die/feel bad/not eat..) in order to test the treatment against tumors but
- it means one less mouse for the lifespan test ("sadly" no other mouse has a tumor)
- I have reached the maximum number of cages I can handle
- I currently don't have the time toprepare special meals for them several times a week 

[AgeVivo]

the mouse who was fail in cage 3a died on Tuesday. Not unexpected.

 

Other mice are fine, including the one with a tumor on the side,

who since Tuesday is back on a ketotarian diet with many green leaves and butter (no sugar) and mebendazole (to remove glutamine)

I put her together with the mouse from cage 3a whom is different from others (long hair, I bought her apart) as she would not count like others in the lifespan experiment anyway, they go well along

the tumor is particularly visible on this picture as the animal has started to move (movement) when I take the picture and the tumor is just starting (virtually no movement)

[AgeVivo]

rapidly changed cages

- no death

- the frail white and black mouse if cage 4a is still as such (nothing changed)

- the cage where males fight gradually fight even more than in the past, they are all bitten everywhere with loss of hair on the back/side.. I don't know what to do about it.

- the mouse with the tumor is still alive though after scratching much on the tumor (as indicated in an previous post) it is now eating its tumor where there is whole in the skin -- probably it does not hurt her and she is hungry (I am trying to starve the tumor with mebendazole and no sugar and to be on the side of a diet) or the tumor is scratching with medenzaleo? or just inconfortable with the tumor so trying to get rid of it. I am not sure what to do about it/if I should do about it. the best is probably to continue and clearly putting her back to a cage with normal food and many mice would be bad for her.

 

cage 1a: 9 mice //like on May 28

cage 2a: 8 mice //like on May 28

cage 3a: 4 mice //was 6 on May 28 but there was a frail mouse who recently died and one strange mouse was moved as a companion with the mouse having a tumor

cage 4a: 9 mice //like on May 28

cage 5a: 12 mice //like on May 28

cage 6a: 8 mice //like on May 28

cage 7a: 12 mice //like on May 28

cage 8a: 12 mice //was 13 on May 28 but one mouse disappeared as indicated in a previous post (I don't think I will ever understand this)

cage 9a: mouse with a tumor and companion mouse (they go very well along together)

 

This is a total of 74+2 mice

I should soon start viagra vs placebo - when I find time

Edited by AgeVivo, 24 June 2023 - 12:23 PM.

[AgeVivo]

The one with a tumor died a few days after my last post, it was eating its tumor and the tumor has started to get infected (as I could tell by the smell). So perhaps the treatment would work in humans, but here the mice eats the tumor which (the causality is only a guess) infects it and kills it. The companion went back to cage 3a; it was not in a great shape (not self-washed fur): perhaps this is because it felt alone once the tumor-bearing mouse died or perhaps this is because the ketotarian food I was preparing was quite bad for them actually or perhaps this is the medenbadole. Now, one week later, it is good looking. So, if another mouse has a tumor again, it will be a companion again but I will try to keep the normal food and the sole treatment will be medendazole only, to limit confounders (see if it is medendazole that is bad over time) both for the control and ill mouse.

cage 1a: 9 females - unchanged

cage 2a: 8 males - unchanged

cage 3a: 5 females - was 4, as the companion went back when the tumor-bearing mouse died. 

cage 4a: 8 females - was 9, one died this week, we knew it was frail as indicated above

cage 5a: 12 males - unchanged

cage 6a: 8 females - unchanged

cage 7a: 12 females - unchanged ; one of them is frail (it has a large bully as if it was pregnant, and is more precautionous than others: the belly must hurt)

cage 8a: 11 males - was 12, one died; unknown cause (too busy to investigate) but it might well be some age-related cause.

 

Going away for 10 days, setting the automated on/off light, full of water and food for all. Based on the last week it should do well but fingers crossed - it is always a bit of stress for me to do that. Last time it was during summer vacations, it went well (there were newborns) but any mistake in one cage and the whole cage may go wrong. Now animals start to be old, I will try to start the treatment-placebo very soon (lacking time no my side)

[AgeVivo]

Went to vacations 10 days, put 3 water bottle in all cages to ensure they had water (and enough food and day/night automated light). As a result, I was short in water bottle which are clearly functional so in the male cages I put the water bottles for which I wasn't sure (sometimes the tip is blocked). And.. all cages had largely enough water ...except one, cage 8, where appearently only one bottle was ok when I left (but I didn't know), and only 2 males in that cage were still alive when I arrived, extremely thirsty. I feel bad - in short I absolutely wanted to protect the females for the lifespan test to start soon and it went at the cost of a cage of males (due to bad luck also).

Edited by AgeVivo, 20 July 2023 - 03:46 PM.

[AgeVivo]

6 August - nothing new