Need help with questions 5) Given your review of the Gram Stain and the structural basis for how it works, why would a cell wall synthesis inhibitor be more effective for a Gram-positive cell type?6) Given your knowledge of molecular biological principles, why would a protein synthesis inhibitor work with both cell types? Why might one protein synthesis inhibitor be more effective than another? For the second part - Hint: Think of change....what might have changed?7) Given your knowledge of molecular biological principles, why are metabolic inhibitors not effective? Hint: Think of change....what might have changed?8) A Gram-negative infection has been determined in a patient, what antibiotic would you suggest for treatment and why? shown below.Gram positivebacteriaGram negativehacteriadreamstime.comThe representative data set you will work with would have been acquired after inoculating twoplates of medium (one for each organism) and placing disks permeated with varied antibiotics.The procedure for generation is as follows (source: www.slideshare.net):How to perform Kirby- Bauertesting• The basics are easy: The bacterium isswabbed on the agar and the antibioticdiscs are placed on top. The antibioticdiffuses from the disc into the agar indecreasing amounts the further it is awayfrom the disc. If the organism is killed orinhibited by the concentration of theantibiotic, there will be NO growth in theimmediate area around the disc: This iscalled the zone of inhibitionEach plate what have looked like the one below (source: www.researchgate.net). The Zone ofInhibition is what you would have measured in mm. The mechanism for the Zone of Inhibition iseither death or growth inhibition. This type of testing does not allow the distinction between a| 2bactericidal (killing) or bacteriostatic (not allowing population expansion) mechanism. What ismeasured is the clear area from the edge of the disk to the start of the growth. The greater theclearing, the greater the impact. The lack of clearing means no impact.Data:Staphylococcus aureus (G+)20 mmEscherichia coli (G-)22 mmAntibiotic UsedChloramphenicolErythromycinPenicillinStreptomycinSulfamethoxazoleTetracyclineTriple SulfaVancomycinProtein Synthesis Inhibitor; Cell Wall Synthesis Inhibitor (Peptidoglycan Impact);Metabolic Inhibitor12 mm5 mm1 mm8 mm2 mm18 mm6 mm3 mm19 mmO mm17 mm20 mmO mmO mmQuestions:1) Evaluate the data for each organism. Is each organism equally impacted by thech Antibiotic Sensitivity with the Gram Stain as Background InformationThe Gram Stain is the most important stain used in microbiology. Knowing whether a bacterialcell is Gram-positive or Gram-negative determines the type of antibiotic used to treat aninfection. There are antibiotics that target Gram-positive cells, there are antibiotics that targetGram-negative cells, and then there are those that target both Gram-positive and Gram-negativecells (e.g. broad-spectrum antibiotics).Danish bacteriologist Hans Christian Gram developed the Gram Stain in 1884. The Gram Stainis a staining technique that separates bacteria into the two groups mentioned above. It is based onthe ability of bacteria to retain the purple color of crystal violet during decolorization withalcohol. Gram-positive bacteria remain purple after decolorization and Gram-negative bacteriaare decolorized as they lose the purple color. To visualize the Gram-negative cells, the cells arecounterstained with safranin. After staining with safranin, the cells look pink. See below:RodGram Stain(gram-negative)Celer efGram-Posve Cell Graegative CelCeler oPrimary StainCrystal VieletPurplePurpleCoceiMordantPurplePurpleIgram pesitiveladineDecalarizing AgentANhelAcetonePurpleCaloressCounterstainPurpleRedPrkSatranin(b)Microbiology, An Introduction, 11 cd., Tortora et al., Pearson, 2013The two cell types differ structurally in the following way:Figure 4.130eBecterial cell walls.PeptidogycanWell teichoic acidCell wallLUpoteichoicacidPlasmamembraneProteinO polysaccharideCore pelyaccharide(b) Gram-positive cell wllCore polysaccharideLipopatycharide o polyaccharideLipid AParta of the LPSPorin proteinLipoproteinCell wa Outer membraneLPeptideglycanPlasmamebranePhospholipidPeriplasmProtein(e) Gramegtive cell wallarntThe video link below is a short presentation on the cellular differences between the two celltypes. Please access and watch for clarification on the figure above.https://www.bing.com/videos/search?q=gram+positive+vs+gram+negative+bacteria&ru=%2fvidcos%2fsearch%3fq%3dgram%2bpositive%2bvs%2bgram%2bnegative%2bbacteria%26FORM%%203dVDVVXX&view detail&mid-E52E108AA9D86F20114CE52E108AA9D86F20114C&&FORM=VDRVRVThe exercise will focus on the analysis of antibiotic impact on a Gram-positive cell type,Staphylococcus aureus, and Gram-negative cell type, Escherichia coli. These are not the onlyspecies that exhibit similar behavior in the presence of antibiotics. Other examples of bacteria areshown below.Gram positivebacteriaGram negativehacteria

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1. Most pathogenic bacteria have a cell wall that provides tensile strength and maintains…