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Objective: In current study Molecular analysis of the recombinant clone ABA392 was carried out Methodology. In this study, detailed molecular analysis of the recombinant clone ABA392 were carried out by examining plasmid stability, plasmid profiling, determination of the plasmid molecular weight, and restriction endonuclease analysis. Results: From the experiments above, the plasmid size of ABA392 was estimated as 3.5 kb, while the size of the insert gene of ABA392 was estimated as 0.8 kb (double digested with PstI and SmaI restriction enzymes). A Pathogenicity study of recombinant clone ABA392 was carried out via a mouse virulence test and histopathological analysis, where it was found that mice, which were challenged with ABA392 and the parent strain of Pasteurella multocida serotype B strain PMB202, were lethal within 24 to 72 hours upon inoculation. Based on microscopic examination, the virulence effect was observed by marked haemorrhage in lung, liver, spleen, and kidney tissues. Conclusion: This study has also proven that the plasmid of recombinant clone ABA392 was stably maintained in the Escherichia coli JM101 host system as reported previously. A preliminary protein analysis via SDS-PAGE was performed and a distinct protein band was most likely to be detected, which was approximately 16kD. Nonetheless, further characterization of the recombinant clone ABA392 proteins needs to be done to determine the nature of these proteins, which are involved in the virulence mechanism of hemorrhagic septicemia. Keywords: Plasmid profiling; Pathogenicity; recombinant clone; Pasteurella multocida

Objective: Streptococcus equi has been identified to contain the hyaluronan synthase gene (HAS), which could have a role in the cell proliferation activity. The objective of this research was to determine anticancer properties of HAS113 recombinant clone. Methodology: The intracellular and extracellular portions of the expressed gene products of pQE-HAS113 were harvested and evaluate for anticancer properties against prostate cancer cell (PC3) and breast cancer cell (MCF-7). To evaluate anti-proliferative properties of pQE-HAS113, two assays were carried out: MTT assay, which was used for assessing percent cell inhibition and apoptosis assay, through the AO/EB stain of living, apoptotic, as well as necrotic cells. For evaluation of the immunomodulatory properties of pQE-HAS113 on RAW264.7 in terms of IL-6, IFN-γ & IL-8 expression, flow cytometry examination was done. The harvested intracellular and extracellular portions of pQE-HAS113 were tested. Results: The results indicated that the percentage proliferation of both the prostate cancer and breast cancer cells were 2.1% and 7.2% respectively against the intracellular portions of pQE-HAS113. While in case of the extracellular portions of pQE-HAS113 the proliferation rate of both the prostate cancer and breast cancer cells were 6.5% and 6.7% respectively. The PQE-HAS113 intracellular clone gene has shown proinflammatory activity by stimulating the expression of IL-6, INF-γ, & IL-8 in the RAW264.7 macrophage cells. Conclusion: Thus, the results from the present study indicated that the expressed product of recombinant clone of HAS 113, pQE-HAS113 has potential to be used as anti-proliferative and apoptosis inducing substance for prostate and breast cancer. Keywords: Streptococcus equi; breast cancer; cell proliferation; apoptosis ass

Introduction: Bacteria in the genus Pasteurella live as commensal parasites on the mucous membranes of vertebrates, particularly mammals and birds. Pasteurella species like Pasteurella multocida and Pasteurella haemolytica cause significant economic losses as a result of diseases caused by these two microorganisms. Objective: This study compares protein isolation and analysis methods for effective one and two-dimensional gel electrophoresis of Pasteurella multocidaproteins and a preliminary protein mapping and analysis of P. multocida serotype B. Methodology: Protein samples were obtained by two isolation methods: homogenisation/sonication and detergent lysis. 1D SDS-PAGE methodology was optimized for protein loading, separating gel percentages and gel size. Detergent lysis was the preferred isolation method of total proteins. Results: The optimum running condition for SDS-PAGE was determined to be 15 µg of protein loading run on a 12.5% separating gel with a gel size of 10cm x 10cm. Preliminary 1D SDS-PAGE analysis revealed 3 distinct protein bands (33 kDa, 39 kDa and >200 kDa), which could only be found specifically in serotype B samples. The samples were then separated by 2-DGE after optimization. The optimum running condition was 500µg of protein sample loaded using rehydration loading with immobilized pH gradient (IPG) strips pH 3–10 for the first-dimension isoelectric focusing (IEF) and 12.5% single percentage gel for the seconddimension SDS-PAGE. The protein map, which produced the most, spots were compared to in silico 2-DGE protein patterns of Pm70 in GELBANK database. Conclusion: A total of 23 proteins were identified and protein of 33 kDA was identified in both 1D SDS-PAGE and 2-DGE. This is the first protein analysis and the first protein map of P. multocida serotype B ever produced.

Introduction: Chitinases are enzymes that hydrolyze the internal β-1,4 glycosidic linkages of chitin, a significant structural component of arthropod exoskeletons and fungus cell walls. Objective: Main objective of current study was to determine the protein and to check the antimicrobial, antifungal activity of chitinase in Bacillus salmalaya strain 139SI. Methodology: Purified and estimation enzymes were quantified by the method of Lowry method. Antimicrobial action of chitinase was done using standard Disc Diffusion method while the hyphal extension inhibition assay was used to test the antifungal activity of pure chitinase strains 139SI, 140SI, and 141SI. Results: In this study, Bacillus salmalaya 139SIexhibited strong hemolytic activity and their protein concentration was measured as 56.43 mg/mL. In addition, strain 139SI had strong antifungal activity against phytopathogenic fungus including Fusarium sp., R. solani and Phytophthora sp. Strain 139SI had the capability in degrading the peptidoglycan component of cell walls of gram-negative bacteria such as Escherichia coli but not against the gram-positive, Staphylococcus aureus. Chitinase activity was observed when 200μl crude extract of 139SI able to degrade 0.09 g chitin of shrimp shell by breaking down shrimp shell structure and bonds of chitin effectively as early as 2 days or up to 7 days. Conclusion: Hence, based on the results, B. salmalaya 139SI has potential to be a novel biofunctional chitinase that could use as a biological agent in degrading the chitin component of fungal cell walls and shells waste of many kind of insect and crustaceans for solving future problem in the agricultural sector and fishery industry.

Introduction: Chitin degradation by chitinase enzyme can be used on a large scale for bioremediation of seafood waste and is environmentally friendly. Objective: The main aim of this study was to screen the potential of the strain as a chitin degrading agent. Methods: In this present study, the cell-free supernatant of Bacillus salmalaya was determined for its protein concentration. Bacillus salmalaya139SI was isolated from agricultural soil and it was identified by staining technique and colony morphology. The production of chitinase by Bacillus salmalaya139SI was optimized under different concentrations of substrate, pH and temperature. Results: Strain 139SI exhibited strong hemolytic activity and the crude protein concentration of Bacillus salmalaya was 84.09mg/ mL with OD value 0.462. Strain 139SI were also screened on colloidal chitin agar medium supplemented with mineral salt. Chikinase production was determined by clear zones of hydrolysis produced after 7 days of incubation at 37°C. The maximum chitinase production was observed in Brain Heart Infusion broth supplemented with 1.0% colloidal chitin at pH 7 and temperature 35°C after four days of incubation. Chitinase activity was observed when high concentrations of crude extract of 139SI able to degrade shrimp shell by showing the degradation zone at day 4. Conclusion: From the results, we concluded that the Bacillus salmalayahas potential to be a biofunctional chitinase that could degrade complex polysaccharides present in the organic wastes and applicable in cleaning the environment.

Newly emerging variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are continuously posing high global public health concerns and panic resulting in waves of coronavirus disease 2019 (COVID-19) pandemic. Depending on the extent of genomic variations, mutations and adaptation, few of the variants gain the ability to spread quickly across many countries, acquire higher virulency and ability to cause severe disease, morbidity and mortality. These variants have been implicated in lessening the efficacy of the current COVID-19 vaccines and immunotherapies resulting in break-through viral infections in vaccinated individuals and recovered patients. Altogether, these could hinder the protective herd immunity to be achieved through the ongoing progressive COVID-19 vaccination. Currently, the only variant of interest of SARS-CoV-2 is Omicron that was first identified in South Africa. In this review, we present the overview on the emerging SARS-CoV-2 variants with a special focus on the Omicron variant, its lineages and hybrid variants. We discuss the hypotheses of the origin, genetic change and underlying molecular mechanism behind higher transmissibility and immune escape of Omicron variant. Major concerns related to Omicron including the efficacy of the current available immunotherapeutics and vaccines, transmissibility, disease severity, and mortality are discussed. In the last part, challenges and strategies to counter Omicron variant, its lineages and hybrid variants amid the ongoing COVID-19 pandemic are presented.

Human DNA contains several variations, which can affect the structure and normal functioning of a protein. These variations could be single nucleotide polymorphisms (SNPs) or insertion-deletions (InDels). SNPs, as opposed to InDels, are more commonly present in DNA and may cause genetic disorders. In the current study, several bioinformatic tools were used to prioritize the pathogenic variants in the SLITRK1 gene. Out of all of the variants, 16 were commonly predicted to be pathogenic by these tools. All the variants had very low frequency, i.e.

The L-2-hydroxyglutarate dehydrogenase (L2HGDH) gene encodes an important mitochondrial enzyme. However, its altered activity results in excessive levels of L-2-hydroxyglutarate, which results in diverse psychiatric features of intellectual disability. In the current study, we executed an in-silico analysis of all reported L2HGDH missense and nonsense variants in order to investigate their biological significance. Among the superimposed 3D models, the highest similarity index for a wild-type structure was shown by the mutant Glu336Lys (87.26%), while the lowest similarity index value was shown by Arg70* (10.00%). Three large active site pockets were determined using protein active site prediction, in which the 2nd largest pocket was shown to encompass the substrate L-2-hydroxyglutarate (L2HG) binding residues, i.e., 89Gln, 195Tyr, 402Ala, 403Gly and 404Val. Moreover, interactions of wild-type and mutant L2HGDH variants with the close functional interactor D2HGDH protein resulted in alterations in the position, number and nature of networking residues. We observed that the binding of L2HG with the L2HGDH enzyme is affected by the nature of the amino acid substitution, as well as the number and nature of bonds between the substrate and protein molecule, which are able to affect its biological activity.

Garlic and its number of preparations are known to be effective for treatment of dyslipidemia, but the data about the specific active constituents of the garlic on the possible therapeutic value is scarce. Therefore, the aim of this research was to evaluate the role of garlic oil (GO) and its active element, diallyl disulphide (DADS) for obviating dyslipidemia in animal model.High fat diet (HFD) was given to animals to induce dyslipidemia. Animals of HFD groups were fed with atherogenic diet for 15 days prior to treatment. Animals in their respective groups received vehicle, GO (50 and 100 mg/kg), and DADS (4.47 and 8.94 mg/kg) for five consecutive days. Lipid profiles were estimated in serum, oxidant/antioxidant and liver profile were measured in liver tissue homogenate (LTH).Animals fed on HFD developed significant increase in the serum levels of triglycerides (TG), total cholesterol (TC), lactate dehydrogenase (LDL), malondialdehyde (MDA), glutathione peroxidase (GSHPx), glutathione (GSH), and glutathione disulfide (GSSG) that reduced significantly in groups that received GO and DADS treatments. Additionally, significant elevation in serum high density lipoprotein (HDL) level was observed in animals that received GO and DADS. Moreover, hepatic markers such as alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine transferase (ALT), that were abnormally altered by high fat diet, were significantly restored to almost normal values with GO and DADS treatments. Also, antioxidants such as superoxide dismutase (SOD), catalase (CAT), ferric reducing antioxidant power (FRAP), and total thiol (SH) levels in LTH were increased significantly in GO and DADS treated groups. When compared to DADS, GO showed better therapeutic effectiveness in terms of antihyperlipidemic and antioxidant properties.In hyperlipidemic rats, garlic and its principal active component, diallyl disulphide, were effective in avoiding dyslipidemia and neutralizing reactive free radicals induced by a high fat diet. It's an intriguing observation that GO has a larger therapeutic influence than its active constituent, DADS. These findings suggest that other constituents, in addition to GO's DADS, are involved in the compound's synergistic antihyperlipidemic and antioxidant activities.

Background and objective Although, the anti-depressant like effects of apigenin (APG) are documented in the literature, the underlying mechanism for exerting such an effect is still not clear. In this research, an attempt was made to determine the possible role of APG for antidepressant activity through serotonergic and catecholaminergic systems using standardized animal models. Materials and methods The antidepressant property of APG was determine by involving tail suspension (TST) and modified forced swimming tests (MFST). The effect of APG was evaluated at 25 and 50 mg/kg. In mechanistic models, animals were pretreated with catecholaminergic and serotonergic antagonists prior to administration of APG. The results obtained were statistically analyzed to determine the level of significance. Results The period of immobility in both models (TST and MFST) was significantly reduced by APG (25 and 50 mg/kg). The best therapetuic dose of APG (50 mg/kg) was selected for the mechanistic study. The anti-immobility effect of APG declined to a significant extent upon pretreatment with catecholaminergic antagonists (α-methyl-para-tyrosine methyl ester; SCH 23390; sulpiride; phentolamine) and serotonergic inhibitors (p-clorophenylalanine-methyl-ester; ondansetron) in both TST and MFST models. The antidepressant benefits of apigenin were only modestly reversed when rats were given propranolol. Conclusions The findings suggest that APG's antidepressant effect is mediated by the α-adrenergic, dopaminergic and 5-HT3 serotonergic receptors.

The SARS-CoV-2 outbreak is the COVID-19 disease, which has caused massive health devastation, prompting the World Health Organization to declare a worldwide health emergency. The corona virus infected millions of people worldwide, and many died as a result of a lack of particular medications. The current emergency necessitates extensive therapy in order to stop the spread of the coronavirus. There are various vaccinations available, but no validated COVID-19 treatments. Since its outbreak, many therapeutics have been tested, including the use of repurposed medications, nucleoside inhibitors, protease inhibitors, broad spectrum antivirals, convalescence plasma therapies, immune-modulators, and monoclonal antibodies. However, these approaches have not yielded any outcomes and are mostly used to alleviate symptoms associated with potentially fatal adverse drug reactions. Nanoparticles, on the other hand, may prove to be an effective treatment for COVID-19. They can be designed to boost the efficacy of currently available antiviral medications or to trigger a rapid immune response against COVID-19. In the last decade, there has been significant progress in nanotechnology. This review focuses on the virus’s basic structure, pathogenesis, and current treatment options for COVID-19. This study addresses nanotechnology and its applications in diagnosis, prevention, treatment, and targeted vaccine delivery, laying the groundwork for a successful pandemic fight.

Objectives: To find out the effect of mild heat shock on osteoblast proliferation and mineralisation following treatment with bisphosphonates. Materials and Methods: Cell culture of MC3T3 cells was performed in alpha minimum essential medium supplemented with 10% Foetal Calf Serum (FCS), 2 mmol L-glutamine and 100U/ml each of penicillin/streptomycin and were incubated at 37°C in humidified 5% CO2. The selected bisphosphonate drugs were administered, following which heat therapy was given by placing the flasks in a temperature-regulated water bath. The proliferation of osteoblast progenitors was measured using the methylene blue staining technique. Results: The results showed that mild heat-shock for 2 min at 42°C can stimulate proliferation of osteoblast progenitors, as well as bone nodule formation. In addition, mild heat-shock can reverse the action of bisphosphonate by stimulation of osteoblast proliferation and mineralisation. Conclusion: Mild heat-shock enhances cell differentiation and osteoblasts mineralisation following bisphosphonate treatment. Key words: Bisphosphonates, Heat-shock, Osteoblast, Osteoclast.

Since its outbreak, the coronavirus (COVID-19) pandemic has caused havoc on people’s lives. All activities were paused due to the virus’s spread across the continents. Researchers have been working hard to find new medication treatments for the COVID-19 pandemic. The World Health Organization (WHO) recommends that safety and self-measures play a major role in preventing the virus from spreading from one person to another. Wireless technology is playing a critical role in avoiding viral propagation. This technology mainly comprises of portable devices that assist self-isolated patients in adhering to safe precautionary measures. Government officials are currently using wireless technologies to identify infected people at large gatherings. In this research, we gave an overview of wireless technologies that assisted the general public and healthcare professionals in maintaining effective healthcare services during COVID-19. We also discussed the possible challenges faced by them for effective implementation in day-to-day life. In conclusion, wireless technologies are one of the best techniques in today’s age to effectively combat the pandemic.

Trastuzumab is a target-based recombinant humanized IgG1 monoclonal antibody (mAbs), extensively employed for treatment of metastatic breast cancer with human epidermal growth receptor 2 (HER2) overexpression. Studies around the world have reported that mAbs have substantial inter-patient unpredictable absorption, distribution, metabolism, and excretion (ADME-pharmacokinetics) because of multiple elements manipulating the concentration of mAbs in plasma. Herein, we have established a bioanalytical technique using UPLC-MS/MS with an easy sample workup method and in-solution digestion protocol to assay the trastuzumab plasma samples from breast cancer patients in clinical studies. Surrogated proteolytic peptides were used for accurate quantification of trastuzumab (CanMab) with a trastuzumab signature peptide with [13C6, 15N4]-arginine and [13C6, 15N2]-lysine stable isotope-labeled (SIL) peptide. Experiments to validate the method were accurately carried out according to the guidelines mentioned in the bioanalytical method validation protocol. The evaluation established excellent linearity over a wide range of 5–500 µg/mL. The experimental procedure was efficaciously performed in a pilot study of five breast cancer patients and residual concentrations of drugs from responding and non-responding subjects were compared. The receiver operating characteristic (ROC) examination displayed that 52.25 µg/mL was the Cmin threshold predictive response with a satisfactory sensitivity of 88.58% and specificity of 79.25%.

The objective of this study was to assess the prevalence of multi-drug resistance isolates of Klebsiella pneumoniae among the patients at Northern Area Armed Forces Hospital (NAAFH) of the Eastern region of Saudi Arabia. This retrospective, chart review observational study was conducted from February 1, 2016 to February 29, 2016 at Northern Area Armed Forces Hospital (NAAFH), a 330-bed community general hospital located in the Eastern region of Saudi Arabia. A total of 870 isolates were identified in 298 patients (3 isolates per patient involving multiple sites). It was observed that the risk of females being infected or colonized with K. pneumoniae was higher than males; the K. pneumoniae was more frequently encountered in hospitalized patient compared to outpatients; a greater number of isolates were obtained from medical service; and the K. pneumoniae was more likely to be isolated from the genitourinary system. Prevalence of multi-drug resistance of K. pneumoniae to antibacterial agents showed that 41% isolates were resistant to third generation cephalosporin primarily due to production of ESBLs, but this was considered statistically insignificant (RR 1.18; 95% CI, 0.96 to 1.47; p = 0.131). The K. pneumoniae producing carbapenemase (KPC) were significantly prominent in this series (RR 0.32; 95%CI, 0.23 to 0.43; p < 0.001). The rapid growth of the K. pneumoniae species that are resistant to carbapenem, a class of drugs considered the last-line of defense, is a matter of concern. Kew Words: Prevalence, multi-drug resistance isolates, Klebsiella pneumoniae, Saudi Arabia.

Purpose: To prepare some 2-morpholinomethylamino-4-(7 unsubstituted/substitutedcoumarin-3-yl)-6- chlorosubstitutedphenyl pyrimidines as antimicrobial agents. Methods: Some 2-morpholinomethylamino-4-(7 unsubstituted/substitutedcoumarin-3-yl)-6-chlorosubstitutedphenyl pyrimidines were prepared by reacting 2-amino-4-(7-substituted/unsubstituted coumarin-3 yl)-6-(chlorosubstitutedphenyl) pyrimidines with morpholine and formaldehyde. The chemical structures of the synthesized compounds were elucidated by their Fourier Transform infra-red (FTIR), 1H-nuclear magnetic resonance (1H-NMR) and mass spectra, as well as by elemental analysis. These compounds were investigated for their antimicrobial activity against ten bacteria and five fungi by serial plate dilution method using the standard drugs, ofloxacin and ketoconazole, respectively, and their minimum inhibitory concentrations (MICs) were determined. Results: A total of eighteen new compounds (1a-18a) were synthesized. Compound 3a (MIC = 75 μg/mL; p < 0.0001) and 15a (MIC = 125 μg/mL; p < 0.001) produced stronger antifungal activity than the standard drug, ketoconazole (MIC = 25 μg/mL; p < 0.0001) against P. citrinum . Compound 4a displayed higher but moderate activity against Gram-positive bacterium, S. aureus (MIC = 100 μg/mL; p < 0.05) than the standard drug, ofloxacin (MIC = 25 μg/mL; p < 0.0001). Compound 4a also displayed higher but moderate activity against the Gram-negative bacterium, E. coli (MIC = 75 μg/mL; p < 0.0001) than the standard drug, ofloxacin (MIC = 12.5 μg/mL; p < 0.0001). The structure activity relationship analysis revealed that the chloro- substitution at position 2 of the phenyl ring along with a chlorobromosubstituted coumarin moiety of the synthesized compounds is critical for activity against Gram-positive bacteria, Gram negative bacteria and fungi. Conclusion: The synthesized compounds are relatively active antifungal agents but are weak antibacterial agents. However, they require further evaluation of their antifungal activity against other fungal strains to ascertain their broad spectrum activity. Keywords: Pyrimidine, Coumarin, Morpholine, Antibacterial, Antifungal, Structure-activity relationship