Part 2:Paper 1: The Brucella abortus S19 vjbR live vaccine candidate is safer than S19 and confers protection against wild-type challenges in BALB/c mice when delivered in a sustained-release vehicle.Describe the hypothesis of the paper.The hypothesis of this article is that the amount of infections will be reduced by the sustained-release encapsulated alginate microspheres.What background information was important in forming this hypothesis?To understand the effects the S19 vaccine, the scientists looked at how this was specifically used to prevent bovine crucellosis. –vjbR, encoding the luxR-like quorum sensing-related transcription regulator is required for VirB expression, virulence in mice and survival in macrophagesWhen vjbR is deleted in S19 diminished inflammation and reduced persistence occurThis helps with safety and reduces side effectsWhat data do the scientists need to have to support their hypothesis?Mice that have received the sustained-release encapsulated alginate microspheres. Expose mice to the bacteria and see how well they fight off the bacteria. Conduct a comparison of vaccinated and unvaccinated mice and show that the vaccinated mice become infected less often. Describe the experiments that produce the data needed in question 3. Separate the mice into groups of 5. Each group was vaccinated with different strengths Seven weeks post vaccinated, the mice were then given the B. abortus (challenged).The mice are killed at weeks 1, 3, 5, 7, 9 after being infected Spleens, lungs, livers, kidneys, and the hearts of the mice are dissected for observation. The evidence is shown what strain is the most effective and helps with the inflammatory response. Describe the 3 most important conclusions and the evidence that support these conclusionsThe mutated strand vaccine compared to the normal S19 strand, there was a 1.47 log different which indicated the mutated strain works more efficiently. The spleens showed that the mutant type spleen was significantly smaller when compared to the regular vaccinated mice and the unvaccinated mice. Immunization with encapsulated S19 ?vjbR::Kan resulted with a more noticeable IgG1 and IgG2a responses when compared to the nonencapsulated mutant2nd paperDescribe the hypothesis of the paper.The hypothesis of this article is that a dual-targeting approach will inhibit Brucella abortus replication in human cells. What background information was important in forming this hypothesis?Intracellular pathogenEvasion of host immune response is possible because phagocytes are taken over by Brucella spp. This causes a weakened effect of antimicrobial compounds for the cell.Knowing what ICCB compounds inhibit B. abortus metabolismHow to find the Tanimoto similarity score to determine if the compound was statistically significant.What data do the scientists need to have to support their hypothesis?480 bioactive molecules inhibited B. abortus replication in the intracellular niche of a human cell line.Identified an additional 26 pathogen-targeting that inhibited B. abortus Two of the five, A-3 and W-7, host-specific compounds that target host kinase signaling pathways inhibiting infection. AA-861, HA14-1, and Cape inhibited replication of intracellular B. abortus Describe the experiments that produce the data needed in question 3. Metabolism- Using the streaking method, B. abortus was put on an Schaedler blood agar and cultivated at 37 degrees Celsius for 48 hours. This process was repeated a second time to be cultivated for 48 hours again. Next, the cell concentration was diluted in an inoculating solution and transferred to well plates and incubated at 37 degrees Celsius for two days. Intracellular growth- Macrophage-like cells were grown to max density at 37 degrees Celsius. Pre-infection the THP-1 cells were transferred to well plates for differentiation. Next, the cultures were centrifuged to attempt higher rate of infection then incubated for 37 degrees Celcius. The Describe the 3 most important conclusions and the evidence that support these conclusions1. Discovered certain compounds that target the inhibition of B. abortus infection. -When applied post infection the screening yielded 45 compounds that inhibited the intracellular growth of B. abortus -The single compound was only effective when it was applied prior to the infection. The result of this suggested that it may prevent Brucella entry in the host cell.2. Identify compounds that directly inhibited B abortus metabolism in axenic culture. -Measuring the reduction of the dye spectrophotometrically. By doing this they could see the tetrazolium being reduced by the metabolic active cells which result in color change. 3. Compounds with activities against host and pathogen potently inhibit intracellular replication of B. abortus. -Used two approaches in order to avoid false-positive results. -Of 44 compounds, 13 were toxic to human cells. -These 13 compounds were described as inhibitors of B. abortus metabolism in axenic culture. -They took another step by testing 18 non-cytotoxic host-specific compounds that did not have an effect on the metabolism which gave them measurements of compounds that inhibit replication of intracellular Brucella in the initial bulk fluorescence assay. Out of 18 non-cytotoxic compounds, 9 were shown in the bulk fluorescence screen inhibiting replication of intracellular bacteria.Finally, give a synopsis of the 3 papers. 1. How did they connect to one another?The first paper connects to the original paper by the fact that both were mutating strains of B. abortus. Both experiments had positive results demonstrating that when the bacterium is mutated it had better effects. 2. Describe any major contradictions, if there were any.There was no big contradictions within the first paper. 3. What major overall significant conclusions can be drawn from the 3 papers.B. abortus greatly affects animals but humans can also become infected but it has been shown different ways to vaccinate or inhibit this bacterium.