Inhibitors Of Bacterial Protein Synthesis Research Articles

Strategies for the Discovery of Oxazolidinone Antibacterial Agents: Development and Future Perspectives.

Oxazolidinones represent a significant class of synthetic bacterial protein synthesis inhibitors that are primarily effective against Gram-positive bacteria. The commercial success of linezolid, the first FDA-approved oxazolidinone antibiotic, has motivated researchers to develop more potent oxazolidinones by employing various drug development strategies to fight against antimicrobial resistance, some of which have shown promising results. Thus, this Perspective aims to discuss the strategies employed in constructing oxazolidinone-based antibacterial agents and summarize recent advances in discovering oxazolidinone antibiotics to provide valuable insights for potentially developing next-generation oxazolidinone antibacterial agents or other pharmaceuticals.

Chlamydia trachomatis induces the transcriptional activity of host YAP in a Hippo-independent fashion.

The obligate intracellular pathogen Chlamydia trachomatis is the causative agent of the most common bacterial sexually transmitted disease worldwide. While the host response to infection by this pathogen has been well characterized, it remains unclear to what extent host gene expression during infection is the product of Chlamydia-directed modulation of host transcription factors. To identify transcription factors potentially modulated by Chlamydia during infection, we infected immortalized endocervical epithelial cells (End1/E6E7) with the anogenital C. trachomatis serovar L2, harvesting polyadenylated RNA for bulk RNA-sequencing. Subsequent experiments elucidating the mechanism of infection-mediated YAP activation assayed YAP target gene expression via qRT-PCR, YAP nuclear translocation via quantitative immunofluorescence, and YAP phosphorylation via Western blotting. RNA sequencing of Chlamydia-infected endocervical epithelial cells revealed gene expression consistent with activity of YAP, a transcriptional coactivator implicated in cell proliferation, wound healing, and fibrosis. After confirming induction of YAP target genes during infection, we observed an infection-dependent increase in YAP nuclear translocation sensitive to inhibition of bacterial protein synthesis. While Hippo-mediated phosphoinhibition of YAP at S127 was unaffected by C. trachomatis infection, Hippo-independent phosphorylation at Y357 was increased. Infection did not enhance nuclear translocation of Y357F mutant YAP, illustrating a requirement for phosphorylation at this residue. Pharmacological inhibition of host Src-family kinase activity attenuated YAP Y357 phosphorylation, but not nuclear translocation - which was instead sensitive to inhibition of Abl. Our results define a transcriptome-altering mechanism of pathogen-directed YAP activation that bypasses canonical inhibition by the Hippo kinase cascade, with a potential link to chlamydial fibrosis and other advanced disease sequelae. Additional study is required to determine the specific role of infection-associated Y357 phosphorylation and Abl activity in chlamydial induction of YAP.

Thermorubin Biosynthesis Initiated by a Salicylate Synthase Suggests an Unusual Conversion of Phenols to Pyrones.

Thermorubin is a tetracyclic naphthoisocoumarin natural product that demands investigation due to its novel mechanism of bacterial protein synthesis inhibition and its unusual structural features. In this work, we describe the identification of the biosynthetic cluster responsible for thermorubin from the sequenced Laceyella sacchari producer species and its confirmation via heterologous production in Escherichia coli. Based on an in-depth annotation of the cluster, we propose a biosynthetic pathway that accounts for the formation of the unique, nonterminal pyrone. Additionally, the expression and use of salicylate synthase TheO enabled testing of the stability properties of this extremophile-derived enzyme. TheO displayed rapid kinetics and a remarkably robust secondary structure, converting chorismate to salicylate with a KM of 109 ± 12 μM, kcat of 9.17 ± 0.36 min-1, and catalytic efficiency (kcat/KM) of 84 ± 9 nM-1 min-1, and retained significant activity up to 50 °C. These studies serve as the basis for continued biosynthetic investigations and bioinspired synthetic approaches.

Antibacterial Mechanism of Action of Two Types of Honey against Escherichia coli through Interfering with Bacterial Membrane Permeability, Inhibiting Proteins, and Inducing Bacterial DNA Damage.

Honey is a sweet natural food produced by bees from flower nectar or some part of plant secretions that exhibit antimicrobial activity against many microorganisms. It has been used as traditional therapy for skin infections. Antibiotics play an essential role in managing wound infection; however, some pathogenic bacteria have begun to possess resistance against them, which may cause chronic infections and severe adverse effects. This study investigates the antibacterial activities and mechanism of action of Yemeni Sidr honey (SH) and Manuka honey (MH) against Escherichia coli. The inhibitory effects of SH and MH using the disk diffusion method on bacterial growth were remarkable at 700 mg/disk. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were similar for both kinds of honey. However, MH showed a better bactericidal effect (30%) than SH (50%). The antimicrobial mechanism of action showed that SH substantially impacted the bacterial membrane’s permeability and increased the potassium and protein leakage rate. On the contrary, MH demonstrated remarkable inhibition of bacterial protein synthesis, while both kinds of honey caused bacterial DNA damage. These data reveal that SH and MH could be used as a remedy for skin infections and might be further developed as a promising dressing for bacterial wound infections.

Synthesis of Pleuromutilin.

Synthesis of a potent inhibitor of bacterial protein synthesis, pleuromutilin, is described. Assembly of the critical cyclooctane fragment relies on an oxidative ring-expansion, and complete stereochemical relay in the synthetic sequence is enabled by the judicious choice of tactics. The requisite connectivity pattern of the perhydroindanone motif is rapidly established in a sequence of cycloaddition and radical cyclization events. Application of this strategy allows for preparation of the target natural product in 16 steps from commercially available material.

A Novel Proline-Rich Cathelicidin from the Alpaca Vicugna pacos with Potency to Combat Antibiotic-Resistant Bacteria: Mechanism of Action and the Functional Role of the C-Terminal Region.

Over recent years, a growing number of bacterial species have become resistant to clinically relevant antibiotics. Proline-rich antimicrobial peptides (PrAMPs) having a potent antimicrobial activity and a negligible toxicity toward mammalian cells attract attention as new templates for the development of antibiotic drugs. Here, we mined genomes of all living Camelidae species and found a novel family of Bac7-like proline-rich cathelicidins which inhibited bacterial protein synthesis. The N-terminal region of a novel peptide from the alpaca Vicugna pacos named VicBac is responsible for inhibition of bacterial protein synthesis with an IC50 value of 0.5 µM in the E. coli cell-free system whereas the C-terminal region allows the peptide to penetrate bacterial membranes effectively. We also found that the full-length VicBac did not induce bacterial resistance after a two-week selection experiment, unlike the N-terminal truncated analog, which depended on the SbmA transport system. Both pro- and anti-inflammatory action of VicBac and its N-terminal truncated variant on various human cell types was found by multiplex immunoassay. The presence of the C-terminal tail in the natural VicBac does not provide for specific immune-modulatory effects in vitro but enhances the observed impact compared with the truncated analog. The pronounced antibacterial activity of VicBac, along with its moderate adverse effects on mammalian cells, make this molecule a promising scaffold for the development of peptide antibiotics.

Mechanism of action, resistance, synergism, and clinical implications of azithromycin.

BackgroundAzithromycin (AZM), sold under the name Zithromax, is classified as a macrolide. It has many benefits due to its immunomodulatory, anti‐inflammatory, and antibacterial effects. This review aims to study different clinical and biochemisterial aspects and properties of this drug which has a priority based on literature published worldwide.MethodsSeveral databases including Web of Science, Google Scholar, PubMed, and Scopus were searched to obtain the relevant studies.ResultsAZM mechanism of action including the inhibition of bacterial protein synthesis, inhibition of proinflammatory cytokine production, inhibition of neutrophil infestation, and macrophage polarization alteration, gives it the ability to act against a wide range of microorganisms. Resistant organisms are spreading and being developed because of the irrational use of the drug in the case of dose and duration. AZM shows synergistic effects with other drugs against a variety of organisms. This macrolide is considered a valuable antimicrobial agent because of its use as a treatment for a vast range of diseases such as asthma, bronchiolitis, COPD, cystic fibrosis, enteric infections, STIs, and periodontal infections.ConclusionsOur study shows an increasing global prevalence of AZM resistance. Thus, synergistic combinations are recommended to treat different pathogens. Moreover, continuous monitoring of AZM resistance by registry centers and the development of more rapid diagnostic assays are urgently needed.

Local antibiotic therapy options for treating acute rhinosinusitis in children

The article presents various types of acute rhinosinusitis progression in pediatric practice depending on the duration of the disease, based on the available modern consensus papers and clinical guidelines. The main clinical terminology of the nosological forms of rhinosinusitis was determined, and aspects of their differential diagnosis and stages of the treatment were studied. The authors provided insight into disease pathogenesis, taking into account literature review data on the spectrum of the most common bacterial agents of the nose and paranasal sinuses pathologies. The available data on the treatment of rhinosinusitis are summarized from the perspective of consensus papers and modern clinical guidelines of the Ministry of Health of the Russian Federation, the use of which for the prescription of treatment is the most important quality criterion for medical care provided. A clinical case of the treatment of acute bacterial uncomplicated rhinosinusitis in a 12-year-old child is described. In this case, an aerosolized form of topical antibiotic therapy combined with a mucolytic component was used as a drug of choice alongside with the standard irrigation-elimination therapy and nasal decongestants, which gives an advantage in achieving the maximum effective concentration of the active substance at the site of inflammation. A drug with a mucoactive effect helps to decrease mucus viscosity and discharge, and at the same time facilitates the entry of an active substance into the tissues and inhibits bacterial adhesion to the epithelium of the respiratory tract. Antibacterial component – thiamphenicol, a derivative of chloramphenicol with a mechanism of action associated with inhibition of bacterial protein synthesis has a broad spectrum of action and a maximum degree of accumulation and action in the site of inflammation due to the method of delivery to the epithelium of the respiratory tract.

1370. Role of Clindamycin Versus Linezolid for Serious Group A Streptococcal Infections

Background Streptococcus pyogenes can cause severe illnesses such as toxic-shock syndrome and necrotizing fasciitis due to pyrogenic exotoxins. Clindamycin is added to penicillin for treatment of severe S. pyogenes infections as it is a bacterial protein synthesis inhibitor which reduces toxin production. However, clindamycin is associated with several adverse effects including C. difficile infection. Linezolid is a bacterial protein synthesis inhibitor that has been shown to provide excellent coverage of S. pyogenes including toxin inhibition in vitro, but clinical evidence is lacking. We compared outcomes of patients treated with linezolid versus clindamycin for serious S. pyogenes infections. MethodsThis was a retrospective study of patients with necrotizing fasciitis or toxic shock syndrome caused by S. pyogenes admitted to the Shock Trauma Center at University of Maryland Medical Center treated with at least 48 hours of either clindamycin or linezolid. Data collected included Sequential Organ Failure Assessment (SOFA) and Laboratory Risk Indicator for Necrotizing Fasciitis (LRINEC) severity scores, time to resolution of infection, number of surgeries, C. difficile infection, other antibiotic associated adverse effects, and mortality. Associations between patient characteristics, antibiotic groups, and outcomes were analyzed using the chi-square test, Fisher’s exact test and t-test or Wilcoxon rank-sum test as appropriate (SAS v 9.4). Results52 patients were included, 26 treated with clindamycin and 26 with linezolid. Most patients (85% clindamycin and 96.2% linezolid) were treated for necrotizing fasciitis. Baseline characteristics, including SOFA and LRINEC scores, were similar between the groups. There was no difference in mortality between patients treated with clindamycin versus linezolid (11.5% vs. 7.7%, p = 0.22), and resolution of infection was similar between the groups (92.3% vs. 88.5%, p = 1.0). There was no difference in adverse effects between the clindamycin and linezolid groups, including C. difficile infection (3.9% vs. 0% p = 1.0) and thrombocytopenia (30.8% vs. 42.3%, p = 0.4).ConclusionLinezolid could be an alternate to clindamycin for the treatment of serious toxin producing S. pyogenes infections. Further prospective studies are needed. Disclosures Emily Heil, PharmD, MS, BCIDP, Nothing to disclose

In vitro synergistic effect of minocycline combined with antifungals against Cryptococcus neoformans

BackgroundCryptococcus neoformans infections occur in immunocompromised patients, especially those with HIV infection, chemoradiotherapy after cancer, and organ transplantation. Infection can cause pneumonia and meningoencephalitis in severe cases with a high mortality rate if not treated. Although fluconazole and amphotericin B are the first-line treatments for cryptococcosis, the rate of fluconazole resistance has increased significantly due to long-term use. Minocycline is a derivative of tetracycline that exerts its antibacterial effect through inhibition of bacterial protein synthesis. It is also able to pass the blood-brain barrier to act on the central nervous system. The present study investigates the effects of minocycline in combination with antifungals in treating C. neoformans. ObjectiveTo determine in vitro interactions of minocycline combined with itraconazole, voriconazole, posaconazole, fluconazole and amphotericin B against C. neoformans. MethodsThe minimum inhibitory concentrations (MIC) of the antifungals were determined by the CLSI Clinical and Laboratory Standards Institute M27-A3 microdilution method. The in vitro synergistic effects of minocycline combined with itraconazole, voriconazole, posaconazole, fluconazole, and amphotericin B on C. neoformans were detected by the broth microdilution checkerboard technique and disk diffusion testing. Results and ConclusionThe working concentration ranges were 0.125–4 µg/mL for itraconazole, 0.03–0.125 µg/ml for voriconazole, 0.03–1 µg/ml for posaconazole, 0.25–16 µg/ml for fluconazole, and 0.125–2 µg/ml for amphotericin B. The synergistic rates of minocycline combinations against C. neoformans were 55% with itraconazole, 10% with voriconazole, 85% with posaconazole, 20% with fluconazole, and 70% with amphotericin B. The effective MIC value of minocycline in the synergistic combination decreased to 2–32 µg/ml, while the MIC of itraconazole decreased to 0.03–0.125 µg/ml, voriconazole 0.03–0.125 µg/ml, posaconazole 0.03–0.125 µg/ml, 0.125–4 µg/ml fluconazole, and 0.06–0.50 µg/ml amphotericin B. The disk diffusion assay showed that the plates containing minocycline and antifungal drugs produced inhibition zones with diameters larger than the single drug plates. Minocycline showed no antagonistic effect in the combinations. In conclusion, the combination of minocycline and azoles or amphotericin B has synergistic effects against C. neoformans in vitro.

RISE OF RIFAXIMIN IN PAEDIATRIC INFECTIOUS DIARRHOEA

The incidence and risk of morality is high under the age of five years is due to diarrhoea. In India twenty percentages of paediatric deaths is due to diarrhoeal disease. It is the second cause of leading mortality in children after acute respiratory infections. Following on from the GEMS study when we looked at the data in the management of bacterial diarrhoea the primary treatments included oral rehydration solution, oral zinc probiotics and antibiotics. Anecdotally we noted that a lot of children were treated with oral antibiotics and the majority of them were treated with fluoroquinolones like Ofloxacin and drugs like Metronidazole there are multiple side-effects associated with the usage of fluoroquinolones and a major side-effect of that is the risk to the growing cartilage. We also noticed that irrational combinations such as imidazole like metronidazole have shown more problems than solutions .usage of Ofloxacin in children should be discouraged for all the side-effects plus the infectivity in the management of bacterial diarrhoea. The duration of treatment have paved the way for usage of drugs such as new molecules in paediatric i.e. rifaximin comes under the group of oral non-absorbable GI antibiotics broad-spectrum agents with antibacterial activity against gram positive and gram negative aerobes and Anaerobes makes Rifaximin a bactericidal drug in nature. Rifaximin is particularly recommended in children above the age of two years in acute infectious diarrhoea the mechanism of action of rifaximin is by inhibition of bacterial protein synthesis by reversibly binding to bacterial DNA dependent RNA polymerase. In this review we discussed the “phrma” behind rifaximin, clinical experience of rifaximin in children and indications and dosage of rifaximin in paediatric.

Aminoglycosides Antagonize Bacteriophage Proliferation, Attenuating Phage Suppression of Bacterial Growth, Biofilm Formation, and Antibiotic Resistance.

The common cooccurrence of antibiotics and phages in both natural and engineered environments underscores the need to understand their interactions and implications for bacterial control and antibiotic resistance propagation. Here, aminoglycoside antibiotics that inhibit protein synthesis (e.g., kanamycin and neomycin) impeded the replication of coliphage T3 and Bacillus phage BSP, reducing their infection efficiency and mitigating their hindrance of bacterial growth, biofilm formation, and tolerance to antibiotics. For example, treatment with phage T3 reduced subsequent biofilm formation by Escherichia coli liquid cultures to 53% ± 5% of that of the no-phage control, but a smaller reduction of biofilm formation (89% ± 10%) was observed for combined exposure to phage T3 and kanamycin. Despite sharing a similar mode of action with aminoglycosides (i.e., inhibiting protein synthesis) and antagonizing phage replication, albeit to a lesser degree, tetracyclines did not inhibit bacterial control by phages. Phage T3 combined with tetracycline showed higher suppression of biofilm formation than when combined with aminoglycosides (25% ± 6% of the no-phage control). The addition of phage T3 to E. coli suspensions with tetracycline also suppressed the development of tolerance to tetracycline. However, this suppression of antibiotic tolerance development disappeared when tetracycline was replaced with 3 mg/liter kanamycin, corroborating the greater antagonism with aminoglycosides. Overall, this study highlights this overlooked antagonistic effect on phage proliferation, which may attenuate phage suppression of bacterial growth, biofilm formation, antibiotic tolerance, and maintenance of antibiotic resistance genes. IMPORTANCE The coexistence of residual antibiotics and phages is common in many environments, which underscores the need to understand their interactive effects on bacteria and the implications for antibiotic resistance propagation. Here, aminoglycosides acting as bacterial protein synthesis inhibitors impeded the replication of various phages. This alleviated the suppressive effects of phages against bacterial growth and biofilm formation and diminished bacterial fitness costs that suppress the emergence of tolerance to antibiotics. We show that changes in bacteria caused by environmentally relevant concentrations of sublethal antibiotics can affect phage-host dynamics that are commonly overlooked in vitro but can result in unexpected environmental consequences.

Peptide Inhibitors of Bacterial Protein Synthesis with Broad Spectrum and SbmA-Independent Bactericidal Activity against Clinical Pathogens.

Proline-rich antimicrobial peptides (PrAMPs) are promising lead compounds for developing new antimicrobials; however, their narrow spectrum of action is limiting. PrAMPs kill bacteria binding to their ribosomes and inhibiting protein synthesis. In this study, 133 derivatives of the PrAMP Bac7(1-16) were synthesized to identify the crucial residues for ribosome inactivation and antimicrobial activity. Then, five new Bac7(1-16) derivatives were conceived and characterized by antibacterial and membrane permeabilization assays, X-ray crystallography, and molecular dynamics simulations. Some derivatives displayed broad spectrum activity, encompassing Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Staphylococcus aureus. Two peptides out of five acquired a weak membrane-perturbing activity while maintaining the ability to inhibit protein synthesis. These derivatives became independent of the SbmA transporter, commonly used by native PrAMPs, suggesting that they obtained a novel route to enter bacterial cells. PrAMP-derived compounds could become new-generation antimicrobials to combat antibiotic-resistant pathogens.

Clinical Utility of Lefamulin: If Not Now, When?

Purpose of ReviewThe looming threat of antimicrobial resistance requires robust stewardship and new developments in infectious diseases pharmacotherapy. This review discusses the pertinent spectrum and clinical data of lefamulin (Xenleta®), with a focus on potential real-world use.Recent FindingsLefamulin is a novel pleuromutilin antibiotic that obtained Food and Drug Administration labeling for community-acquired bacterial pneumonia (CABP) in 2019. Lefamulin is available in both intravenous and oral formulations, and it inhibits bacterial protein synthesis inhibition through interactive binding to unique sites of the peptidyl transferase center of the 50s bacterial ribosome subunit. Resistance, including cross-resistance with other antibiotics, is infrequent. Lefamulin demonstrates activity against most Gram-positive pathogens and other organisms commonly associated with CABP, i.e., Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae, Legionella pneumophila, and Chlamydophila pneumoniae. Lefamulin may also be an option for serious public health threats like methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecium, and multi-drug-resistant organisms associated with sexually transmitted infections, e.g., Neisseria gonorrhoeae, Mycoplasma genitalium. Lefamulin lacks activity against Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacterales, most anaerobes, and E. faecalis. In Phase III trials, lefamulin monotherapy was non-inferior to moxifloxacin with or without linezolid for CABP.SummaryLefamulin is a well-tolerated agent with a unique mechanism, availability in both IV and PO formulations, and it has been rigorously studied for safety and efficacy for CABP.

Histidine-Triad Hydrolases Provide Resistance to Peptide-Nucleotide Antibiotics.

The Escherichia coli microcin C (McC) and related compounds are potent Trojan horse peptide-nucleotide antibiotics. The peptide part facilitates transport into sensitive cells. Inside the cell, the peptide part is degraded by nonspecific peptidases releasing an aspartamide-adenylate containing a phosphoramide bond. This nonhydrolyzable compound inhibits aspartyl-tRNA synthetase. In addition to the efficient export of McC outside the producing cells, special mechanisms have evolved to avoid self-toxicity caused by the degradation of the peptide part inside the producers. Here, we report that histidine-triad (HIT) hydrolases encoded in biosynthetic clusters of some McC homologs or by standalone genes confer resistance to McC-like compounds by hydrolyzing the phosphoramide bond in toxic aspartamide-adenosine, rendering them inactive.IMPORTANCE Uncovering the mechanisms of resistance is a required step for countering the looming antibiotic resistance crisis. In this communication, we show how universally conserved histidine-triad hydrolases provide resistance to microcin C, a potent inhibitor of bacterial protein synthesis.

Effect of Phosphatase Activity of the Control of Virulence Sensor (CovS) on Clindamycin-Mediated Streptolysin O Production in Group A Streptococcus.

Severe manifestations of group A Streptococcus (GAS) infections are associated with massive tissue destruction and high mortality. Clindamycin (CLI), a bacterial protein synthesis inhibitor, is recommended for treating patients with severe invasive GAS infection. Nonetheless, the subinhibitory concentration of CLI induces the production of GAS virulent exoproteins, such as streptolysin O (SLO) and NADase, which would enhance bacterial virulence and invasiveness. A better understanding of the molecular mechanism of how CLI triggers GAS virulence factor expression will be critical to develop appropriate therapeutic approaches. The present study shows that CLI activates SLO and NADase expressions in the emm1-type CLI-susceptible wild-type strain but not in covS or control of virulence sensor (CovS) phosphatase-inactivated mutants. Supplementation with Mg2+, which is a CovS phosphatase inhibitor, inhibits the CLI-mediated SLO upregulation in a dose-dependent manner in CLI-susceptible and CLI-resistant strains. These results not only reveal that the phosphorylation of response regulator CovR is essential for responding to CLI stimuli, but also suggest that inhibiting the phosphatase activity of CovS could be a potential strategy for the treatment of invasive GAS infection with CLI.

Aminomethyl spectinomycins: a novel antibacterial chemotype for biothreat pathogens

New antibiotics that are active against multi-drug-resistant strains and difficult-to-treat bacterial infections are needed. Synthetic modification of spectinomycin, a bacterial protein synthesis inhibitor, has been shown to increase antibacterial activity compared with spectinomycin. Aminomethyl spectinomycins are active against Gram-negative and Gram-positive bacterial pathogens. In this study, the ability of aminomethyl spectinomycins to treat biothreat pathogens is examined by MIC profiling, synergy testing, and in vivo efficacy experiments. Compound 1950 exhibited potent antibacterial activity against Gram-negative pathogens Brucella spp., Burkholderia mallei, and Francisella tularensis, but showed little to no growth inhibition against Burkholderia pseudomallei, Bacillus anthracis, and Yersinia pestis. Combination testing in checkerboard assays revealed that aminomethyl spectinomycin-antibiotic combinations had mainly an additive effect against the susceptible biodefense pathogens. The in vivo efficacy of compound 1950 was also demonstrated in mice infected with B. mallei (FMH) or F. tularensis (SchuS4). These results suggest that aminomethyl spectinomycins are promising new candidates for development of therapeutics against biodefense bacterial agents.

INVESTIGATIONS ON IN VITRO METABOLITES OF LINEZOLID IN VARIOUS SPECIES

Linezolid is a potent synthetic oxazolidinone used for the treatment of bacterial infections with a new mechanism of action that involves early inhibition of bacterial protein synthesis. In humans, linezolid circulates mainly as parent drug and is excreted primarily as parent drug and a major inactive, morpholine ring-opened carboxylic acid metabolite. In vitro studies were conducted to identify the hepatic enzymes responsible for the oxidative metabolism of linezolid using human liver microsomes. However the specific enzyme responsible for the oxidation of linezolid was not identified. The present study is to check with and identify the probable metabolic pathways in various species like monkey, mouse, rat, dog and human livermicrosomes and put up the best model for the contingency studies using modern analytical techniques. The current investigation on the metabolites obtained after 60 minutes incubation revealed three additional metabolites, namely, M10, M11 and M12, adding on to the list of already reported metabolites.>

Aminoglycoside Revival: Review of a Historically Important Class of Antimicrobials Undergoing Rejuvenation.

Aminoglycosides are cidal inhibitors of bacterial protein synthesis that have been utilized for the treatment of serious bacterial infections for almost 80 years. There have been approximately 15 members of this class approved worldwide for the treatment of a variety of infections, many serious and life threatening. While aminoglycoside use declined due to the introduction of other antibiotic classes such as cephalosporins, fluoroquinolones, and carbapenems, there has been a resurgence of interest in the class as multidrug-resistant pathogens have spread globally. Furthermore, aminoglycosides are recommended as part of combination therapy for empiric treatment of certain difficult-to-treat infections. The development of semisynthetic aminoglycosides designed to overcome common aminoglycoside resistance mechanisms, and the shift to once-daily dosing, has spurred renewed interest in the class. Plazomicin is the first new aminoglycoside to be approved by the FDA in nearly 40 years, marking the successful start of a new campaign to rejuvenate the class.

Chlamydia trachomatis fails to protect its growth niche against pro-apoptotic insults

Chlamydia trachomatis is an obligate intracellular bacterial agent responsible for ocular infections and sexually transmitted diseases. It has been postulated that Chlamydia inhibits apoptosis in host cells to maintain an intact replicative niche until sufficient infectious progeny can be generated. Here we report that, while cells infected with C. trachomatis are protected from apoptosis at early and mid-stages of infection, they remain susceptible to the induction of other cell death modalities. By monitoring the fate of infected cells by time-lapse video microscopy and by analyzing host plasma membrane integrity and the activity of caspases, we determined that C. trachomatis-infected cells exposed to pro-apoptotic stimuli predominately died by a mechanism resembling necrosis. This necrotic death of infected cells occurred with kinetics similar to the induction of apoptosis in uninfected cells, indicating that C. trachomatis fails to considerably prolong the lifespan of its host cell when exposed to pro-apoptotic insults. Inhibitors of bacterial protein synthesis partially blocked necrotic death of infected cells, suggesting that the switch from apoptosis to necrosis relies on an active contribution of the bacteria. Tumor necrosis factor alpha (TNF-α)-mediated induction of necrosis in cells infected with C. trachomatis was not dependent on canonical regulators of necroptosis, such as RIPK1, RIPK3, or MLKL, yet was blocked by inhibition or depletion of CASP8. These results suggest that alternative signaling pathways regulate necrotic death in the context of C. trachomatis infections. Finally, consistent with the inability of C. trachomatis to preserve host cell viability, necrosis resulting from pro-apoptotic conditions significantly impaired production of infectious progeny. Taken together, our findings suggest that Chlamydia’s anti-apoptotic activities are not sufficient to protect the pathogen’s replicative niche.