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Gokshura tablet is an ayurvedic formulation with gokhru (Tribulus terrestris) as best fixing recommended for building vitality levels. It enhances life, sexual want and drive. Pushyanug churna is an ayurvedic polyherbal formulation, hence this seems essential to explain the material institutionalization, various bioactive markers, blends exhibit in the polyherbal ayurveda compositions such as pushyanug churna. Point of the exhibit effort has been to create what’s more, approve a high-performance thin layer chromatography (HPTLC) strategy for assurance of diosgenin present in gokshura tablet. Mangiferin and chlorogenic acid are present in pushyanug churna. Diosgenin, a biomarker chemical found in gokshura tablets, and mangiferin, a biomarker compound found in pushyanug churna, were standardized using recently developed easy and accurate HPTLC procedures. Pre-coated silica gel 60-F254 was employed at the stationary phase and a mixture of toluene, ethyl acetate, and formic acid (in the proportions 5:4:1) was employed for the mobile phase in the development methodology for diosgenin. In the mobile phase, mangiferin, ethyl acetate and methanol were added in a ratio of 40:60 v/v were utilized. In the chlorogenic acid mobile phase, ethyl acetate:formic acid:acetic acid:water (10:1.1:1.1:2.6 v/v). It was determined that the Rf value of the marker chemical was 0.77 (diosgenin) in gokhsura tablet and 0.23 mangiferin, 0.75 chlorogenic acid in pushyanug churna. For bioactive marker chemicals found in in-house and commercially available formulations, the developed HPTLC approach has shown to be straightforward, sensitive, specific, and dependable.

Bacteriophages and phage-derived proteins are a promising class of antibacterial agents that experience a growing worldwide interest. To map ongoing phage research in Singapore and neighboring countries, Lee Kong Chian School of Medicine, Nanyang Technological University Singapore (NTU) and Yong Loo Lin School of Medicine, National University of Singapore (NUS) recently co-organized a virtual symposium on Bacteriophage and Bacteriophage-Derived Technologies, which was attended by more than 80 participants. Topics were discussed relating to phage life cycles, diversity, the roles of phages in biofilms and the human gut microbiome, engineered phage lysins to combat polymicrobial infections in wounds, and the challenges and prospects of clinical phage therapy. This perspective summarizes major points discussed during the symposium and new perceptions that emerged after the panel discussion. Agency for Science, Technology and Research (A*STAR) Ministry of Health (MOH) National Medical Research Council (NMRC) National Research Foundation (NRF) Following supports are acknowledged: N.K.V. to LeeKong Chian School of Medicine Strategic Academic In-itiative Grant (L0494003); H.C. to A*STAR Grant(C210812044); P.H., B.C.G., and W.M. to Singapore Na-tional Research Foundation (NRF)’s Intra-Campus for Re-search Excellence and Technological Enterprise (CREATE)Seed Collaboration Grant; P.H. and B.C.G. to Singapore-MIT Alliance for Research and Technology (SMART) An-timicrobial Resistance Interdisciplinary Group funded by theNRF under its CREATE program; T.A.A. to NUSMedpostdoctoral fellowship (NUHSRO/2019/046/PDF/19); J.P.B.to National University Hospital System Seed Fund (NUHSRO/2019/050/T1/Seed-Mar/03) and SMART (ING-001014 BIO);W.M. to Singapore Ministry of Health’s National MedicalResearch Council (OFIRG21jun-0038); M.A.M.V.S. to Na-tional Health Innovation Centre Singapore Innovation to De-velop Grant (NHIC-I2D-2104625).

Purpose: This study aims to determine the siege and related factors of nurses working in psychiatric clinics. Method: This study was conducted as a related description design. The sample of the study consisted of 204 nurses working in psychiatric clinics. The data was collected using questionnaires generated by researchers from literature and workplace psychological abuse scales. Descriptive statistics (frequency and percentage) and chi-square independence test are used to determine whether there is a significant relationship between variables. Results: The conditions for more frequent siege behaviors are that nurses have graduate education, work at night, are dissatisfied with work methods and institutions, have been besieged before, and have an understanding of the number of legal persons and consulting services related to Go. Conclusion: The nursing service management department of the hospital should effectively organize the types of employment and develop strategies that can improve nurse satisfaction. It is believed that raising nurses’ awareness of siege will effectively reduce psychological violence in high-risk wards.

Novel therapeutic options are needed to halt the unabated spread of multiple-drug resistance. Phage-derived proteins, especially enzymes that can degrade the bacterial cell wall and cause cell-lysis, offer an attractive new paradigm for treating serious drug-resistant infections. Recent research and clinical data on this class of anti-infective agents have revealed their novel properties. This chapter describes the development path for phage-derived lysins, starting with discovery and leading to the strategy into the clinic, with P128, a novel chimeric antistaphylococcal lysin, as example. Standard regulatory pathways that apply to chemical antibiotics need to be modified to accommodate and accelerate the development of this class. The similarities and differences between these macromolecular bactericidal agents, and the traditional small molecule antibiotics are highlighted throughout this chapter. The data-intensive exemplification of research and development of lysin P128 in this write-up is aimed at influencing all the stakeholders, research community, clinicians, patients, biopharma industry, and regulators, into capitalizing the benefits that lysins offer.

We demonstrate that the prophage status of bacteria plays a critical role in achieving homogenous population of a phage preparation. When a lytic Staphylococcus bacteriophage 44AHJD was propagated in a Staphylococcus clinical isolate, the enriched phage showed 44AHJD phage virions along with the released prophages from the baiting host. The released prophage was identified as a siphophage by transmission electron microscopy. To obtain a phage preparation free of prophages, when we carried out multiplication of the 44AHJD phage in a prophage free Staphyloccoccus aureus host namely RN4220, we were surprised not to see any phage plaques in spite of the phage exhibiting >99.9% adsorption to such cells. Since RN4220 host is devoid of restriction modification system and prophages, we hypothesized that in spite of successful infection and multiplication, the phage virions might have failed to show plaques due to its insignificant release from the cell possibly due to insufficient endolysin expressed from phage virions during phage development and assembly. Our hypothesis was confirmed when we observed plaques of 44AHJD phage in RN4220 cells where additional phage endolysin protein was supplemented via a plasmid. Endolysin protein from various types of Staphylococcus phages showed plaques of 44AHJD in RN4220 cells confirming our hypothesis. Also, we demonstrate for the first time that propagation of 44AHJD phage with endolysin supplementation in prophage free RN4220 host yields pure phage preparation.

The emergence of multi-drug resistant (MDR) microbial pathogens threatens the very foundation upon which standard antibacterial chemotherapy is based. We must consider non-antibiotic solutions to manage invasive bacterial infections. Transition from antibiotics to non-traditional treatments poses real clinical challenges that will not be easy to solve. Antibiotics will continue to reliably treat some infections (e.g., group A streptococci and Treponema pallidum) but will likely need adjuvant therapies or will need to be replaced for many bacterial infections in the future.

Today the world has suffered to the Zika virus is a flavivirous related to yellow fever, dengue, West Nile and Japanese encephalis virus. Zika virus causes acute, serious illness which is often fatal if it is not treated. Its name comes from the Zika forest of Uganda. Zika is a mosquito born disease. Zika virus disease is caused by a virus transmitted primarily by Aedes mosquitoes. People with Zika virus disease can have symptoms including mild fever, skin rash, conjunctivitis, muscle and joint pain, malaise or headache. These symptoms normally last for 2-7 days. There is scientific consensus that Zika virus is a cause of microcephaly and Guillain-Barre syndrome. Links to other neurological complications are also being investigated. Neither an effective treatment nor a vaccine is available for Zika virus; therefore, the public health response primarily focuses on preventing infection, particularly in pregnant women. This article also covers from where the zika was firstly introduced or its history, its transmissions, causes, diagnosis, treatment and some current affairs..

P128 is a chimeric anti-staphylococcal protein having a catalytic domain from a Staphylococcus bacteriophage K tail associated structural protein and a cell wall targeting domain from the Staphylococcus bacteriocin-lysostaphin. In this study, we disclose additional properties of P128 and compared the same with lysostaphin. While lysostaphin was found to get inactivated by heat and was inactive on its parent strain S. simulans biovar staphylolyticus, P128 was thermostable and was lytic towards S. simulans biovar staphylolyticus demonstrating a difference in their mechanism of action. Selected mutation studies of the catalytic domain of P128 showed that arginine and cysteine, at 40th and 76th positions respectively, are critical for the staphylolytic activity of P128, although these amino acids are not conserved residues. In comparison to native P128, only the R40S mutant (P301) was catalytically active on zymogram gel and had a similar secondary structure, as assessed by circular dichroism analysis and in silico modeling with similar cell binding properties. Mutation of the arginine residue at 40th position of the P128 molecule caused dramatic reduction in the Vmax (∆OD600 [mg/min]) value (nearly 270 fold) and the recombinant lysostaphin also showed lesser Vmax value (nearly 1.5 fold) in comparison to the unmodified P128 protein. The kinetic parameters such as apparent Km (Km APP) and apparent Kcat (KcatAPP) of the native P128 protein also showed significant differences in comparison to the values observed for P301 and lysostaphin.

Background Bacterial drug resistance is one of the most significant challenges to human health today. In particular, effective antibacterial agents against methicillin-resistant Staphylococcus aureus (MRSA) are urgently needed. A causal relationship between nasal commensal S. aureus and infection has been reported. Accordingly, elimination of nasal S. aureus reduces the risk of infection. Enzymes that degrade bacterial cell walls show promise as antibacterial agents. Bacteriophage-encoded bacterial cell wall-degrading enzymes exhibit intrinsic bactericidal activity. P128 is a chimeric protein that combines the lethal activity of the phage tail-associated muralytic enzyme of Phage K and the staphylococcal cell wall targeting-domain (SH3b) of lysostaphin. Here we report results of in vitro studies evaluating the susceptibility of staphylococcal strains to this novel protein. Results Using the broth microdilution method adapted for lysostaphin, we found that P128 is effective against S. aureus clinical strains including MRSA, methicillin-sensitive S. aureus (MSSA), and a mupirocin-resistant S. aureus. Minimum bactericidal concentrations and minimum inhibitory concentrations of P128 (1-64 μg/mL) were similar across the 32 S. aureus strains tested, demonstrating its bactericidal nature. In time-kill assays, P128 reduced colony-forming units by 99.99% within 1 h and inhibited growth up to 24 h. In an assay simulating topical application of P128 to skin or other biological surfaces, P128 hydrogel was efficacious when layered on cells seeded on solid media. P128 hydrogel was lethal to Staphylococci recovered from nares of healthy people and treated without any processing or culturing steps, indicating its in situ efficacy. This methodology used for in vitro assessment of P128 as an agent for eradicating nasal carriage is unique. Conclusions The novel chimeric protein P128 is a staphylococcal cell wall-degrading enzyme under development for clearance of S. aureus nasal colonization and MRSA infection. The protein is active against globally prevalent antibiotic-resistant clinical isolates and other clinically significant staphylococcal species including S. epidermidis. The P128 hydrogel formulation was bactericidal against Staphylococci including S. aureus recovered from the nares of 31 healthy people, demonstrating its in situ efficacy.

Background Interest in phage therapy has grown over the past decade due to the rapid emergence of antibiotic resistance in bacterial pathogens. However, the use of bacteriophages for therapeutic purposes has raised concerns over the potential for immune response, rapid toxin release by the lytic action of phages, and difficulty in dose determination in clinical situations. A phage that kills the target cell but is incapable of host cell lysis would alleviate these concerns without compromising efficacy. Results We developed a recombinant lysis-deficient Staphylococcus aureus phage P954, in which the endolysin gene was rendered nonfunctional by insertional inactivation. P954, a temperate phage, was lysogenized in S. aureus strain RN4220. The native endolysin gene on the prophage was replaced with an endolysin gene disrupted by the chloramphenicol acetyl transferase (cat) gene through homologous recombination using a plasmid construct. Lysogens carrying the recombinant phage were detected by growth in presence of chloramphenicol. Induction of the recombinant prophage did not result in host cell lysis, and the phage progeny were released by cell lysis with glass beads. The recombinant phage retained the endolysin-deficient genotype and formed plaques only when endolysin was supplemented. The host range of the recombinant phage was the same as that of the parent phage. To test the in vivo efficacy of the recombinant endolysin-deficient phage, immunocompromised mice were challenged with pathogenic S. aureus at a dose that results in 80% mortality (LD80). Treatment with the endolysin-deficient phage rescued mice from the fatal S. aureus infection. Conclusions A recombinant endolysin-deficient staphylococcal phage has been developed that is lethal to methicillin-resistant S. aureus without causing bacterial cell lysis. The phage was able to multiply in lytic mode utilizing a heterologous endolysin expressed from a plasmid in the propagation host. The recombinant phage effectively rescued mice from fatal S. aureus infection. To our knowledge this is the first report of a lysis-deficient staphylococcal phage.

A variety of bacterial pathogenicity determinants, including the type VI secretion system and the virulence cassettes from Photorhabdus and Serratia , share an evolutionary origin with contractile-tailed myophages. The well-characterized Escherichia coli phage P2 provides an excellent system for studies related to these systems, as its protein composition appears to represent the “minimal” myophage tail. In this study, we used nuclear magnetic resonance (NMR) spectroscopy to determine the solution structure of gpX, a 68-residue tail baseplate protein. Although the sequence and structure of gpX are similar to those of LysM domains, which are a large family associated with peptidoglycan binding, we did not detect a peptidoglycan-binding activity for gpX. However, bioinformatic analysis revealed that half of all myophages, including all that possess phage T4-like baseplates, encode a tail protein with a LysM-like domain, emphasizing a widespread role for this domain in baseplate function. While phage P2 gpX comprises only a single LysM domain, many myophages display LysM domain fusions with other tail proteins, such as the DNA circulation protein found in Mu-like phages and gp53 of T4-like phages. Electron microscopy of P2 phage particles with an incorporated gpX-maltose binding protein fusion revealed that gpX is located at the top of the baseplate, near the junction of the baseplate and tail tube. gpW, the orthologue of phage T4 gp25, was also found to localize to this region. A general colocalization of LysM-like domains and gpW homologues in diverse phages is supported by our bioinformatic analysis.