Abstract: INTRODUCTION: The bacterium “ Mycobacterium tuberculosis (MTB)” causes tuberculosis, which is currently regarded as the most common cause of bacterial infection-related death globally. In addition to eukaryotes, MTB also encodes 11 “serine/threonine protein kinases (STPKs),” including PknA, B, C, D, E, F, G, H, J, K, and L that control cell division, cell development, metabolism, and dormancy. MATERIALS AND METHODS: The antisense and sense-orientated PknL have been cloned utilizing the shuttle mycobacterial vector-pAZI9018b, which is IPTG inducible. Utilizing the appropriate primers, the PknL (Rv2176) full-length gene from MTB has been amplified. The product which was amplified was then inoculated inside the vector at the “5’BamH1 and 3’Nde1” regions to produce PknL in the sense orientation (PknL-S) resembling PknL-S, and likewise, amplified substance was introduced into the vector at the 3’BamHI and 5’NdeI sites for producing PknL in the antisense orientation (PknL-AS). RESULTS: Regardless of the absence or presence of an inducer, PknL-AS had lower transcription levels than Wild type and PknL-S, but PknL-AS revealed a significant decrease in PknL expression during the end of 2 nd week with 1 mM of IPTG and a further decrease on day 10 with 10 mM IPTG. In the presence of 10 mM IPTG, PknL expression has been close to, 1.8 log in Wild type, 1.4 log in PknL-AS, and 2.4 log in PknL-S. CONCLUSION: It has been clear that PknL may read environmental cues and respond appropriately, assisting bacteria to adapt to stressful situations in the host by lowering bacterial development, leading to a dormant state, and allowing bacteria to stay inside the host.

Background and Aim: Rheumatoid arthritis (RA) is a prevalent and debilitating disease that affects the joints. The study was intended to evaluate the anti-arthritic activity of aerial parts of Heliotropium zeylanicum (EEHZ). EEHZ constituting phytoconstituents to treat adjuvant induced arthritic rats to minimize the side effects. Materials and Methods: The anti-arthritic activity study was carried out by using adjuvant induced model of Wistar albino strain rats. EEHZ was injected at different doses such as 100 and 300 mg/kg/i.p., and the study was compared with standard drug dexamethazone (5 g/kg). The results obtained from the above methods were subjected to statistical analysis. Results: The study was conducted various hematological parameters and histopathological sections were carried out. The plant has various phytoconstituents such as reducing sugars, saponins, starch, steroids and terpenoids. Significantly one among those phytoconstituent lessens to oedema and arthritis of the rodents. The EEHZ showed the maximum inhibitory activity at (300 mg/kg/i.p.) by dose dependent manner. These inhibitions were statistically significant (P < 0.01-0.001). Conclusion: From these results indicate that EEHZ is a bioactive agent and having significant results in anti-inflammatory and anti-arthritic action by inhibition of the exudation, and leukocytes recruitment into the inflamed tissues and bone regeneration calcium deposition.

The COVID-19 pandemic has forced the public health authorities to impose a lockdown as an epidemiological containment strategy. Due to COVID-19, the world is facing an unprecedented challenge that has changed people's lifestyles. This study aimed to evaluate how the COVID-19 pandemic lockdown affected students in Tamilnadu in terms of changes to their eating habits, sleeping patterns, and level of physical activity. A cross-sectional study was conducted using a random representative sample of 263 undergraduate students (mean age 19.63+1.86 years, males 33.46%) from Tamilnadu voluntarily filling out an online questionnaire. The Pittsburgh Sleep Quality Index (PSQI) and The International Physical Activity Questionnaires, Eating Attitude Test (EAT-26) were used to assess sleep quality, Physical Activity, and Eating Attitude disorder, respectively. The prevalence of poor sleep quality among participants was 43.3% (males: 44.32% and females: 42.86%). The mean score of the participants' EAT-26 score was found to be 10.92 ± 8.63. Students' high body weight and BMI risk factors for eating disorders (2=9.68 p=0.02, 2=9.59 p=0.02). 46% of students did not do any physical activity. There is no significant correlation between sleep and Eating attitude scores (p=0.21). 66.16% of participants experienced Psychological stress due to lock down. Findings from this study indicate that a lockdown period due to COVID-19 negatively impacted young undergraduate adults' physical activity levels and sleep quality. Finally, body weight, BMI, monthly family income, and poor physical activity with excess eating during the COVID 19 lockdown were the common exposures that are significantly associated with a higher risk of developing sleep disturbances and eating disorders. Several efficient strategies, programs, and coordinated efforts must be rigorously executed to encourage a seamless transition between school and daily life.

The main aims in the development of a novel drug delivery vehicle is to efficiently carry therapeutic drugs in the body’s circulatory system and successfully deliver them to the targeted site as needed to safely achieve the desired therapeutic effect. In the present study, a passive targeted functionalised nanocarrier was fabricated or wrapped the hollow mesoporous silica nanoparticles with 3-aminopropyl triethoxysilane (APTES) to prepare APTES-coated hollow mesoporous silica nanoparticles (HMSNAP). A nitrogen sorption analysis confirmed that the shape of hysteresis loops is altered, and subsequently the pore volume and pore diameters of GaC-HMSNAP was reduced by around 56 and 37%, respectively, when compared with HMSNAP. The physico-chemical characterisation studies of fabricated HMSNAP, Ga-HMSNAP and GaC-HMSNAP have confirmed their stability. The drug release capacity of the fabricated Ga-HMSNAP and GaC-HMSNAP for delivery of gallium and curcumin was evaluated in the phosphate buffered saline (pH 3.0, 6.0 and 7.4). In an in silico molecular docking study of the gallium-curcumin complex in PDI, calnexin, HSP60, PDK, caspase 9, Akt1 and PTEN were found to be strong binding. In vitro antitumor activity of both Ga-HMSNAP and GaC-HMSNAP treated MCF-7 cells was investigated in a dose and time-dependent manner. The IC50 values of GaC-HMSNAP (25 µM) were significantly reduced when compared with free gallium concentration (40 µM). The mechanism of gallium-mediated apoptosis was analyzed through western blotting and GaC-HMSNAP has increased caspases 9, 6, cleaved caspase 6, PARP, and GSK 3β(S9) in MCF-7 cells. Similarly, GaC-HMSNAP is reduced mitochondrial proteins such as prohibitin1, HSP60, and SOD1. The phosphorylation of oncogenic proteins such as Akt (S473), c-Raf (S249) PDK1 (S241) and induced cell death in MCF-7 cells. Furthermore, the findings revealed that Ga-HMSNAP and GaC-HMSNAP provide a controlled release of loaded gallium, curcumin and their complex. Altogether, our results depicted that GaC-HMNSAP induced cell death through the mitochondrial intrinsic cell death pathway, which could lead to novel therapeutic strategies for breast adenocarcinoma therapy.

Clinical pathogens, especially Gram-negative bacteria developing resistance to third-generation cephalosporins, are making clinical outcomes more complicated and serious. This study was undertaken to evaluate the distribution of CTX-M-type extended-spectrum β-lactamases (ESBLs) in Tamil Nadu, India. For this study, clinical samples were collected from five different hospitals located in Tamil Nadu and the ESBL-producing Gram-negative isolates were characterized. MIC was performed using cefotaxime and ceftazidime. The bla ESBL-producing genes were screened using multiplex PCR for the genes, CTX-M group-1, -2, -8, -9, -26. The conjugation studies were performed using Escherichia coli AB1157 as a recipient for the isolates harbouring plasmid-borne resistance following broth-mating experiment. In total, 1500 samples were collected and 599 Gram-negative bacteria were isolated that included E. coli (n=233), Klebsiella pneumoniae (n=182), Pseudomonas aeruginosa (n=79), Citrobacter spp. (n=30), Proteus mirabilis (n=28), Salmonella spp. (n=21), Acinetobacter baumannii (n=12), Serratia spp. (n=6), Shigella spp. (n=4), Morganella morganii (n=3) and Providencia spp. (n=1). MIC results showed that 358 isolates were resistant to cefotaxime and ceftazidime. Further, ESBL gene-amplification results showed that 19 isolates had CTX-M group-1 gene including E. coli (n=16), K. pneumoniae (n=2) and P. aeruginosa (n=1) whereas one M. morganii isolate had CTX-M group-9, which was plasmid-borne. Through conjugation studies, 12/20 isolates were found to be involved in the transformation of its plasmid-borne resistance gene. Our study highlighted the importance of horizontal gene transfer in the dissemination of plasmid-borne bla CTX-M-type resistance genes among the clinical isolates.

Curcumin delivery to cancer cells is challenging due to its hydrophobic nature, low bio distribution and low availability. Many nano vehicles suffer from low stability and toxicity, and hence the prerequisite of a non-toxic nano vehicle with effective drug delivery is still being delved. The present study investigates the delivery efficiency of curcumin with non-spherical mesoporous silica nanoparticles (MSNAs). Their mechanism of drug delivery and signalling proteins activated to induce apoptosis was further explored in MCF-7 cells. A non-spherical MSN was synthesised, functionalised with PEI (MSNAP) and analysed its intracellular behaviour. Our result indicates that MSNAP was non-toxic until 20 µg/mL and likely localizes in cytoplasmic vesicles. On contrast, well-known MCM-41P induced autophagosome formation, indicating cellular toxicity. Curcumin was loaded on MSNAP and its effectiveness in inducing cell death was studied in MCF-7 and in MCF-7R cells. Curcumin loading on MSNAP induces better cell death with 30 µM curcumin, better than unbounded curcumin. Western blot analysis suggest, curcumin induce apoptosis through the activation of caspase 9, 6, 12, PARP, CHOP and PTEN. The cell survival protein Akt1 was downregulated by curcumin with and without the nanostructure. Interestingly, cleaved caspase 9 was activated in higher amount in nano-conjugated curcumin compared to the free curcumin. But other ER resident protein like IRE1α, PERK and GRP78 were downregulated indicating curcumin disturbs ER homeostasis. Further, electron microscopic analysis reveled that nanocurcumin induced apoptosis by disrupting mitochondria and nucleus. Our results with doxorubicin resistant MCF-7 cell lines confirm nanodelivery of doxorubicin and curcumin sensitised cells effectively at lesser concentration. Further docking studies of curcumin indicate it interacts with the apoptotic proteins through hydrogen bonding formation and with higher binding energy.

Endoplasmic reticulum (ER) stress-mediated apoptosis is a well-known factor in the pathogenesis of age-related macular degeneration (AMD). ER stress leads to accumulation of misfolded proteins, which in turn activates unfolded protein response (UPR) of the cell for its survival. The prolonged UPR of ER stress promotes cell death; however, the transition between adaptation and ER stress-induced apoptosis has not been clearly understood. Hence, the present study investigates the regulatory effect of (–)-epigallocatechin gallate (EGCG) on ER stress-induced by hydrogen peroxide (H2O2) and disturbance of calcium homeostasis by thapsigargin (TG) in mouse retinal pigment epithelial (MRPE) cells. The oxidant molecules influenced MRPE cells showed an increased level of intracellular calcium [Ca2+]i in ER and transferred to mitochondria through ER-mitochondrial tether site then increased ROS production. EGCG restores [Ca2+]i homeostasis by decreasing ROS production through inhibition of prohibitin1 which regulate ER-mitochondrial tether site and inhibit apoptosis. Effect of EGCG on ER stress-mediated apoptosis was elucidated by exploring the UPR signalling pathways. EGCG downregulated GRP78, CHOP, PERK, ERO1α, IRE1α, cleaved PARP, cleaved caspase 3, caspase 12 and upregulated expression of calnexinin MRPE cells. In addition to this, inhibition of apoptosis by EGCG was also confirmed with expression of proteins Akt, PTEN and GSK3β. MRPE cells with EGCG upregulates phosphorylation of Akt at ser473 and phospho ser380 of PTEN, but phosphorylation at ser9 of GSK3β was inhibited. Further, constitutively active (myristoylated) CA-Akt transfected in MRPE cells had an increased Akt activity in EGCG influenced cells. These findings strongly suggest that antioxidant molecules inhibit cell death through the proper balancing of [Ca2+]i and ROS production in order to maintain UPR of ER in MRPE cells. Thus, modulation of UPR signalling may provide a potential target for the therapeutic approaches of AMD.

Objective: To isolate. Screen and characterize an effective phage for MDR and ESBL producing pathogenic bacterial strains. Methods: Bacteriophages were isolated from hospital effluent samples by double layer agar method. Isolated phages were propagated by liquid enrichment technique and its host range was analyzed by double layer agar method. Morphology of the isolated phages was identified by Transmission Electron Microscope (TEM). Genomic and proteomic analysis was confirmed by electrophoresis technique. Results & Discussion: 46 bacteriophages were isolated against 20 different MDR and ESBL strains of those 7 phages (Mm81, Ec84, Ps85, En833, Sal836, Ec8ATCC and Ec8PMG) were selected for further studies. According to the host range analysis result the 7 phage has been shown narrow host range. The phage genomic DNA and structural proteins were analyzed. In addition to based on the TEM analysis two phages viz., Mm81 and Ec84 were belongs to Siphoviridae and Podoviridae family respectively. Present study evaluates the extensive occurrence of phages in the hospital effluent. In addition, this is first report of isolation and characterization of Morganella morganii lytic phage in Tamil Nadu, India. Conclusion: The study highlights the distribution of bacteriophages in the hospital effluent and it gives the therapeutic potential of isolated phages for the treatment of MDR and ESBL Producing Pathogens. Keywords: Phage therapy, Transmission electron microscope, Extended-spectrum of β-lactamase, Multi-drug resistant.

Several enzymes are known to accumulate in the cornea in unusually high concentrations. Based on the analogy with lens crystallins, these enzymes are called corneal crystallins, which are diverse and species-specific. Examining crystallins in lens and cornea in multiple species provides great insight into their evolution. We report data on major proteins present in the crocodile cornea, an evolutionarily distant taxon. We demonstrate that tau-crystallin/alpha-enolase and triose phosphate isomerase (TIM) are among the major proteins expressed in the crocodile cornea as resolved by 2D gel electrophoresis and identified by MALDI-TOF. These proteins might be classified as putative corneal crystallins. tau-Crystallin, known to be present in turtle and crocodile lens, has earlier been identified in chicken and bovine cornea, whereas TIM has not been identified in the cornea of any species. Immunostaining showed that tau-crystallin and TIM are concentrated largely in the corneal epithelium. Using western blot, immunofluorescence and enzymatic activity, we demonstrate that high accumulation of tau-crystallin and TIM starts in the late embryonic development (after the 24th stage of embryonic development) with maximum expression in a two-week posthatched animal. The crocodile corneal extract exhibits significant alpha-enolase and TIM activities, which increases in the corneal extract with development. Our results establishing the presence of tau-crystallin in crocodile, in conjunction with similar reports for other species, suggest that it is a widely prevalent corneal crystallin. Identification of TIM in the crocodile cornea reported here adds to the growing list of corneal crystallins.

Several enzymes are known to accumulate in the cornea in unusually high concentrations. Based on the analogy with lens crystallins, these enzymes are called corneal crystallins, which are diverse and species‐specific. Examining crystallins in lens and cornea in multiple species provides great insight into their evolution. We report data on major proteins present in the crocodile cornea, an evolutionarily distant taxon. We demonstrate that τ‐crystallin/α‐enolase and triose phosphate isomerase (TIM) are among the major proteins expressed in the crocodile cornea as resolved by 2D gel electrophoresis and identified by MALDI‐TOF. These proteins might be classified as putative corneal crystallins. τ‐Crystallin, known to be present in turtle and crocodile lens, has earlier been identified in chicken and bovine cornea, whereas TIM has not been identified in the cornea of any species. Immunostaining showed that τ‐crystallin and TIM are concentrated largely in the corneal epithelium. Using western blot, immunofluorescence and enzymatic activity, we demonstrate that high accumulation of τ‐crystallin and TIM starts in the late embryonic development (after the 24th stage of embryonic development) with maximum expression in a two‐week posthatched animal. The crocodile corneal extract exhibits significant α‐enolase and TIM activities, which increases in the corneal extract with development. Our results establishing the presence of τ‐crystallin in crocodile, in conjunction with similar reports for other species, suggest that it is a widely prevalent corneal crystallin. Identification of TIM in the crocodile cornea reported here adds to the growing list of corneal crystallins.