u/Ornery_Restaurant375

https://wynnbuilder.github.io/builder/#CN0Q01-CWi3sQGoDIkHUG00Wg689G11uzvBA6h2s

Can someone give me an Idol build? I have like max 2m budget, can someone give me a wynnbuilder link to a build?

I'm thinking of Performing Ghost Garden, Joy - adam tan or Melody in Repose, can someone provide their feedback on these pieces?

I'm thinking of Performing Ghost Garden, Joy - adam tan or Melody in Repose, can someone provide their feedback on these pieces?

Can someone make me a lootrunning build for a warrior? Anything is fine

Hi, can someone give mea good, cheap warrior build? Lowkey broke. Levle 98 rn, but dont mind any build requirinjg higher level

Hi, I'm trying to get my friend to run a good Assassin Archtype for this party: Level 60 Paladin+Monk mix. Level 63 rift, Level 48 Acolyte and level 45 Assassin. I'm thinking of trickobat, but not sure

Hi, I've been trying to find 4 mallet pieces to play at my school's tallent show, the hardest 4 mallet piece I played is Raindance or yellow after the Rain. I could only find Melody In Repose for any piece In Repose by Michael Charles Smith for free. I'm looking for ANY 4 mallet piece with similar vibes that you can provide for free! Thank you for helping.

How would cross resistance affect bacteria once It had developed tolerance against a chemical?

Introduction

This is an experiment to test the rate that bacteria develop resistance against a substance that would kill the bacteria.  We are choosing this question as the investigation question to explore how quickly bacteria would develop tolerance and eventual resistance against a chemical that has proven to damage the bacteria.

Preparation

Pathogen choice      

We decided to use bacteria for their ability of Horizontal gene transfer, causing them to be able to share resistance once a bacterium had evolved, its natural resilience due to its cellular structure unlike virus, adaptability at different environments unlike fungi and not requiring a host unlike parasitical organisms. We decided to use bacillus subtilis for the bacteria choice, as it can turn into an endospore, an almost immune version of the bacterium, hoping to increase the chances of survival when we attempt to kill the bacterium, and also to observe would a colony of bacteria prefer the longevity of the species by developing resistance or focus on its current survival by turning itself into an endospore, as it can’t reproduce in that form. Safety wise, it is classified as a gram positive due to the lack of an outer membrane. Also, the bacteria have a safety level of BSL-1 and doesn’t infect humans unless they have an open wound and has cancer, going through chemotherapy, or has been infected by human immunodeficiency virus (HIV).

Agar choice

We have decided to use the normal nutrient agar with a drop of tea tree oil to be antifungal. We had chosen this agar type to prevent the growth of dangerous bacteria like, tetanus, staph, and more, and nutrients agar has sufficient nutrients to encourage the growth of the bacteria without inducing dangerous microorganisms.

Chemical choice

Nothing

We are using a plate with nothing, to see how bacteria would a bacteria change without any external factors affecting it, just food for rapid reproduction. This is to prove that survival of the fittest is a real concern, and to prove that without necessity, an organism wouldn’t evolve significantly.

Honey

We intended to use honey, specifically manuka honey to see how bacteria would create tolerance in such conditions, because Honey can kill bacteria using osmotic pressure, a phenomenon when waters at different concentration of nutrients meet, creating pressure which in this case, destroys the cell membrane of the bacteria, causing plasmolysis, the contraction of a protoplast (a bacteria/fungus/plant cell that lost their cell wall), that causes the water to leave, killing with extreme dehydration. Next, it can kill bacteria through hydrogen peroxide, as when diluted, an enzyme of it creates hydrogen peroxide (h2o2), a bond with 2 hydrogen atoms and 2 oxygen atoms. It works like a neutrophil, also being a ROS, (reactive oxygen species). Next, we have a rather simple way of killing bacteria, acidity. Bacteria need a PH level of 6-8 to grow, while honey has a PH level of 3-5, significantly more acidic than preferred. Next, for some types of honey, specifically manuka honey (the type we’re using) can produce methylglyoxal, which in recent research, has showed antibacterial properties, specifically killing bacteria by destroying the structure of bacterium by interacting with the amino acids arginine and lysine, which appears on the surface on bacteria. We decided to use honey to see how a substance with multiple methods of killing a bacteria affect its speed of evolution and to see against a such a powerful substance, would the bacteria focus on survival through endospores or attempt to preserve the species by creating tolerance?

Vinegar

We chose vinegar as an inferior option to honey, as it relies on acidity to kill, (its acidity level is 2.0-3.5) and only acidity. This is to see would one singular variable ease the pressure on the pathogen, and to see would developing tolerance against only one part of the opposing chemical boost or lower its chance of survival?

Variables

The time for bacteria to cultivate	The temperature we will keep it at	The agar type used
The time for bacteria to die	Observing the change in the percentage of dead bacteria relative to alive bacteria.	Changing the fluid used
The ratio from fluid to water	the dye used	the bacteria type used

 

 

Risk evaluation

Risk	Solution
Growing dangerous bacteria and mold (E.g Staphylococcus)	Using a fume hood to prevent other bacteria, mold spores or chemicals from getting in, and preventing headaches caused by tea tree oil.
Developing a new strain of bacteria	Making sure that all areas it is exposed in is decontaminated, avoid letting it contact other bacteria and always protect your eyes, mouth and nose when opening the dish. After the experiment, we autoclave or disinfect the dish before disposing it, just to make sure the gene doesn’t get any contact with the outside world.

 

 

 

Thesis

I believe that there would be a steep logarithmic pattern in the amount of death caused by honey, because at the start, the bacteria had no reason to become endospores or have significant tolerance against honey, but once we drop in the honey,  most bacteria alive would have tolerance against honey or become endospores, significantly lowering the death count, but as the dosage increases, the resistant becomes less useful, as the bacteria is developing resistance against 4 factors instead of one at the same time. For vinegar, I believe there would be a steep increase of deaths, than exponentially down, as acidity doesn’t focus much on the killing aspect of a antibacterial agent, but instead, stops reproduction, so even though most survivors would stop reproducing, they wouldn’t die to slight increase in dosage, and because of the lack of new generations, the kill count would likely be lower.

Process

1.      Disinfect the area that we’re going to be opening the agar plates

2.      Slightly open the agar plates

3.      swab the b. subtills sample on the agar

4.      close the plate

5.      leave the plate at a place with temperature of 25-30C for 24hours

6.      equip safety gear

7.      open the plate slightly

8.      put in 10%  substance to 90% water, 0.5ml worth total of liquid.

9.      Close and tape the petri dish

10. Leave it for 72 hours

11. Re-equip safety gear

12. Reopen the petri dish

13. Drop 1 part to 3-part erythrosine dye on the plate

14. Disinfect the cotton swab and dispose it

15. Observe with a microscope

16.  Swab the survival of the survivor of the experiment, put it onto a petri dish and let it grow for another 72 hours

17. We repeat step 8-16 2 times, but every time, we increase the % of the substance to water ratio by 2%

18.  We put a 3:17 ratio of the chemical to see did the bacteria evolve.

19. We swab out the survivor of step 16, and put it onto another dish

20. We use the opposite chemical to an almost lethal rate that has been scientifically proven for a normal version of the bacterium, and see, would the survival cause any defects for its ability against other chemicals.

21. Disinfect through autoclaving or cleaning alcohol

Note: all times when opening a petri dish, it will and MUST be under supervision by an educator inside a fume hood with eye protection, a mask and lab coat. You also must disinfect them right after.

 

Observation

Written during experiment

First test with honey

	Dead bacteria	Alive bacteria	Endospores	% of dead bacteria
Spot 1
Spot 2
Spot 3
Spot 4
Spot 5
Average

Second test with honey

	Dead bacteria	Alive bacteria	Endospores	% of dead bacteria
Spot 1
Spot 2
Spot 3
Spot 4
Spot 5
Average

Third test with honey

	Dead bacteria	Alive bacteria	Endospores	% of dead bacteria
Spot 1
Spot 2
Spot 3
Spot 4
Spot 5
Average

 

Vinegar with honey bacteria

	Dead bacteria	Alive bacteria	Endospores	% of dead bacteria
Spot 1
Spot 2
Spot 3
Spot 4
Spot 5
Average

 

Test 1 with Vinegar  

	Dead bacteria	Alive bacteria	Endospores	% of dead bacteria
Spot 1
Spot 2
Spot 3
Spot 4
Spot 5
Average

Test 2 with Vinegar   

	Dead bacteria	Alive bacteria	Endospores	% of dead bacteria
Spot 1
Spot 2
Spot 3
Spot 4

the main problem is the bacteira just keeps dying, even when I reduce the doses, any ideas?