Multitarget action of Benzothiazole-piperazine small hybrid molecule against Alzheimer's disease: In silico, In vitro, and In vivo investigation.

A novel small molecule based on benzothiazole-piperazine has been identified as an effective multi-target-directed ligand (MTDL) against Alzheimer's disease (AD). Employing a medicinal chemistry approach, combined with molecular docking, MD simulation, and binding free energy estimation, compound 1 emerged as a potent MTDL against AD. Notably, compound 1 demonstrated efficient binding to both AChE and Aß1-42, involving crucial molecular interactions within their active sites. It displayed a binding free energy (ΔGbind) -18.64± 0.16 and -16.10 ± 0.18 kcal/mol against AChE and Aß1-42, respectively. In-silico findings were substantiated through rigorous in vitro and in vivo studies. In vitro analysis confirmed compound 1 (IC50=0.42 µM) as an effective, mixed-type, and selective AChE inhibitor, binding at both the enzyme's catalytic and peripheral anionic sites. Furthermore, compound 1 demonstrated a remarkable ability to reduce the aggregation propensity of Aß, as evidenced by Confocal laser scanning microscopy and TEM studies. Remarkably, in vivo studies exhibited the promising therapeutic potential of compound 1. In a scopolamine-induced memory deficit mouse model of AD, compound 1 showed significantly improved spatial memory and cognition. These findings collectively underscore the potential of compound 1 as a promising therapeutic candidate for the treatment of AD.

Use of inferior vena cava guided fluid therapy in the treatment of septic shock: A randomised controlled trial.

INTRODUCTION: By administering inferior vena cava (IVC) directed fluid, it is possible to avoid the use of additional fluid and fluid overload in patients with septic shock (SS) and sepsis-induced hypoperfusion (SIH). METHODOLOGY: In patients with SIH and SS, we conducted prospective observational research on fluid therapy. A time-motion trace of the IVC diameter was created using M-mode imaging. The ability to predict fluid responsiveness was based on the IVC collapsibility index (cIVC) > 40%. Participants were randomised into 2 groups using a permuted block-of-four randomization list, with the investigators being blinded prior to patient allocation. They were split equally between the usual-care (UC) group, which received sepsis-guided fluid treatment, and the interventional ultrasound-guided fluid therapy (UGFT) group. RESULTS: The average age of the participants was 63.2 years (62.8 years for the UGFT group and 63.7 years for the UC group). Co-morbid health conditions were practically the same in both arms at baseline. Prior to enrolment, both groups received the same quantity of fluid as part of resuscitation (UGFT arm received 2.4 0.6 L, UC group received 2.2 0.7 L). The UGFT group outperformed the UC group with a P value of 0.02 due to a significantly lower positive fluid balance after 72 hours of ICU discharge (-1.37 L), which rendered the UGFT group superior to the UC group. Even after accounting for the fluids consumed before enrolment, there was still a sizable difference in the fluids infused. When the pre-enrolment fluids were counted at 72 hours, UGFT participants still displayed a decreased positive fluid balance. However, there was no discernible difference in the 30-day mortality rate overall (6.3% difference, UGFT: 15.7%, and UC: 22.0%). CONCLUSIONS: In contrast to the UC group, the UGFT arm of our study demonstrated a statistically significant benefit of Point of Care USG (POCUS) guided fluid therapy during resuscitation in sepsis in reducing the positive fluid balance in 72 hours, preventing fluid overload, and reducing the need for dialysis and invasive ventilation. However, there was no statistically significant variation in the 30-day mortality rate.

Kolaviron suppresses dysfunctional reproductive axis associated with multi-walled carbon nanotubes exposure in male rats.

Reproductive toxicity associated with excessive exposure to multi-walled carbon nanotubes (MWCNTs), which are commonly used in medicine as valuable drug delivery systems, is well documented. Kolaviron, a bioflavonoid isolated from Garcinia kola seeds, elicits numerous health beneficial effects related to its anti-inflammatory, anti-genotoxic activities, anti-apoptotic, and antioxidant properties. However, information on the role of kolaviron in MWCNTs-induced reproductive toxicity is not available in the literature. Herein, we assessed the protective effects of kolaviron on MWCNTs-induced dysfunctional reproductive axis in rats following exposure to MWCNTs (1 mg/kg) and concurrent treatment with kolaviron (50 or 100 mg/kg body weight) for 15 successive days. Results showed that MWCNTs-induced dysfunctional reproductive axis as evidenced by deficits in pituitary and testicular hormones, marker enzymes of testicular function, and sperm functional characteristics were abrogated in rats co-administered with kolaviron. Moreover, co-administration of kolaviron-abated MWCNTs-induced inhibition of antioxidant enzyme activities increases in oxidative stress and inflammatory indices. This is evidenced by diminished levels of tumor necrosis factor-alpha, nitric oxide, lipid peroxidation, reactive oxygen, and nitrogen species as well as reduced activity of myeloperoxidase in testes, epididymis, and hypothalamus of the rats. Biochemical data on the chemoprotection of MWCNTs-induced reproductive toxicity were corroborated by histological findings. Taken together, kolaviron suppressed dysfunctional reproductive axis associated with MWCNTs exposure via abrogation of oxidative stress and inflammation in male rats.

Development of novel carbazole derivatives with effective multifunctional action against Alzheimer's diseases: Design, synthesis, in silico, in vitro and in vivo investigation.

Carbazole based novel multifunctional agents has been rationally designed and synthesized as potential anti-Alzheimer agents. Multi-functional activity of these derivatives have been assessed by performing various in-vitro assays and these compounds appeared to be potent AChE inhibitors, Aß aggregation inhibitors, anti-oxidant and neuroprotective agents. Among the entire series, MT-1 and MT-6 were most potent multifunctional agents which displayed effective and selective AChE inhibition, Aß disaggregation, anti-oxidant and metal chelation action. Neuroprotective activity of MT-6 has been examined against H2O2 induced toxicity in SHSY-5Y cells and they have shown effective neuroprotection. Additionally, MT-6 did not display any significant toxicity in SHSY-5Y cells, indicating its non-toxic nature. Molecular docking and MD simulation studies have been also performed to explore molecular level interaction with AChE and Aß. Finally, MT-6 was evaluated against scopolamine induced dementia model of mice and this compound actively improved memory deficit and cognition impairment in scopolamine treated mice. Thus, novel carbazole derivative MT-6 has been explored as an effective and safe multifunctional agent against AD and this molecule may be used as a suitable lead for development of effective anti-Alzheimer agents in future.

Development of novel N-(6-methanesulfonyl-benzothiazol-2-yl)-3-(4-substituted-piperazin-1-yl)-propionamides with cholinesterase inhibition, anti-ß-amyloid aggregation, neuroprotection and cognition enhancing properties for the therapy of Alzheimer's disease.

A novel series of benzothiazole-piperazine hybrids were rationally designed, synthesized, and evaluated as multifunctional ligands against Alzheimer's disease (AD). The synthesized hybrid molecules illustrated modest to strong inhibition of acetylcholinesterase (AChE) and Aß1-42 aggregation. Compound 12 emerged as the most potent hybrid molecule exhibiting balanced functions with effective, uncompetitive and selective inhibition against AChE (IC50 = 2.31 µM), good copper chelation, Aß1-42 aggregation inhibition (53.30%) and disaggregation activities. Confocal laser scanning microscopy and TEM analysis also validate the Aß fibril inhibition ability of this compound. Furthermore, this compound has also shown low toxicity and is capable of impeding loss of cell viability elicited by H2O2 neurotoxicity in SHSY-5Y cells. Notably, compound 12 significantly improved cognition and spatial memory against scopolamine-induced memory deficit in a mouse model. Hence, our results corroborate the multifunctional nature of novel hybrid molecule 12 against AD and it may be a suitable lead for further development as an effective therapeutic agent for therapy in the future.

Effect of periodontal therapy on disease activity in patients of rheumatoid arthritis with chronic periodontitis.

BACKGROUND: Evidence have been proposed a positive association between severity of Periodontitis and Rheumatoid arthritis (RA) activity, individuals with advanced RA are more likely to develop periodontal problems compared to their non-RA counterparts, and vice versa. Studies have been suggested that RA manifest as a result of an inflammatory imbalance and autoimmunity. In this perspective, treatment modalities that lead to inhibition of proinflammatory mediators, may prove beneficial for reducing the severity of RA. This study examined the effects of non surgical periodontal therapy (NSPT) on disease activity of RA. METHODS: Diagnosed patients of active rheumatoid arthritis with chronic periodontitis were recruited in this study and divided in to treatment and controls groups, both groups were similar in all demographics assessed. Treatment group (n = 20) and controls group (n = 20) underwent assessment for periodontal clinical parameters (plaque index, gingival index, probing pocket depth, clinical attachment level), Rheumatologic clinical (simplified disease activity index) and biochemical parameters(C-reactive protein, Rheumatoid factor, Anti-cyclic citrullinated protein) at baseline and 8 weeks. Serum levels of biochemical parameters were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The statistically significant (p < 0.001) reduction observed in mean values of PI, GI, PPD, CAL, SDAI in treatment group at 8weeks after NSPT as compare to control group. However serum level of ACCPA, CRP and RF did not show statistically significant (p > 0.05) changes from baseline to reassessment (8 weeks) in both groups. CONCLUSIONS: The improvement in RA disease activity may occurs after non surgical periodontal therapy.

Bovicins: The Bacteriocins of Streptococci and Their Potential in Methane Mitigation.

Bovicin is a type AII lantibiotic, possessing two ß-methyllanthionine and a disulfide bridge encoded by bovA gene hitherto unknown a couple of decades ago. Bacteriocins can be useful in directly inhibiting methanogens and/or redirecting H2 to other reductive microorganisms, in particular, propionate producers or reductive acetogens. So far, the role of nisin and bovicin to suppress greenhouse gas (GHG) production under in vitro conditions has been documented. GHG emissions from ruminants are a threat to the environment, because of their role in global warming as well as in climate change. Methane (CH4) produced from livestock farming practices is a potent GHG, comprising 18% of total GHG emissions in the world. Therefore, minimizing enteric CH4 production is quite essential from both the economical livestock production as well as environment perspectives. Strategies for the abatement of CH4 have provided two-way opportunities, viz., improved livestock productivity and reduced GHG emissions. In the past, different strategies have been proposed and tested to mitigate CH4, such as the dietary composition of feeds, ionophores, antibiotics, vaccines, analogues, probiotics, and secondary metabolites of plants and fungi. However, quite a few of these strategies have been adopted at farm level due to their varied effect on animal health and/or residues on animal products. The use of bacteriocins might have potential in inhibiting methanogens in the rumen. A bacteriocin produced by Streptococcus bovis (an isolate from rumen) named bovicin HC5 has been exhibited to decrease CH4 production to an extent of 50%. In this review, authors intend to discuss the sources, structure, biochemical properties, and antimicrobial spectra of bovicins, besides the potential applications with special reference to CH4 mitigation.

Bacteriocins: Classification, synthesis, mechanism of action and resistance development in food spoilage causing bacteria.

Huge demand of safe and natural preservatives has opened new area for intensive research on bacteriocins to unravel the novel range of antimicrobial compounds that could efficiently fight off the food-borne pathogens. Since food safety has become an increasingly important international concern, the application of bacteriocins from lactic acid bacteria that target food spoilage/pathogenic bacteria without major adverse effects has received great attention. Different modes of actions of these bacteriocins have been suggested and identified, like pore-forming, inhibition of cell-wall/nucleic acid/protein synthesis. However, development of resistance in the food spoilage and pathogenic bacteria against these bacteriocins is a rising concern. Emergence and spread of mutant strains resistant to bacteriocins is hampering food safety. It has spurred an interest to understand the bacteriocin resistance phenomenon displayed by the food pathogens, which will be helpful in mitigating the resistance problem. Therefore, present review is focused on the different resistance mechanisms adopted by food pathogens to overcome bacteriocin.

Evaluation of combined B cell specific N-terminal immunogenic domains of LipL21 for diagnosis of leptospirosis.

Leptospiral outer membrane protein LipL21 and its truncated N-terminal immunogenic region (I-LipL21) were evaluated for diagnosis of leptospirosis. The complete coding sequence of LipL21 nucleotide sequence was subjected to BCPred and VaxiJen analysis for determination of B cell specific immunogenic epitopes. Epitope1 ACS STD TGQ KDA TTV GDG (1.8837), Epitope2 WGG PPE QRN DGK TPR DTN (0.9483), Epitope3 VKG VGV YEC KAT GSG SDP (1.4077) and Epitope4 NEW ECQ CVI YAK FPG GKD (0.4462) were predicted. LipL21 and N-terminal fragment having B-cell specific epitopes with higher VaxiJen score >0.9 as truncated I-LipL21 were cloned independently in pET15b and expressed in Escherichia coli. IgM ELISA and dot blot assay was performed for sera samples collected from Delhi-NCR for leptospiral whole cell lysate (WCL), recombinant LipL21 and I-LipL21. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were found to be 92.5%, 92.8%, 83.3%, and 97% respectively for recombinant I-LipL21 by IgM-ELISA. 11-14.8% increased sensitivity was observed over LipL21 and WCL. The I-LipL21 dot blot assay showed a further increased sensitivity of 3.8% over the IgM-ELISA. Therefore I-LipL21 may be the ideal candidate protein for diagnosis of leptospirosis.

Contribution of Cell Surface Hydrophobicity in the Resistance of Staphylococcus aureus against Antimicrobial Agents.

Staphylococcus aureus is found in a wide variety of habitats, including human skin, where many strains are commensals that may be clinically significant or contaminants of food. To determine the physiological characteristics of resistant strain of Staphylococcus aureus against pediocin, a class IIa bacteriocin, a resistant strain was compared with wild type in order to investigate the contribution of hydrophobicity to this resistance. Additional clumping of resistant strain relative to wild type in light microscopy was considered as an elementary evidence of resistance attainment. A delay in log phase attainment was observed in resistant strain compared to the wild type strain. A significant increase in cell surface hydrophobicity was detected for resistant strain in both hexadecane and xylene indicating the contribution of cell surface hydrophobicity as adaptive reaction against antimicrobial agents.

Bacteriocin production and different strategies for their recovery and purification.

Bacteriocins from lactic acid bacteria (LAB) are a diverse group of antimicrobial proteins/peptides, offering potential as biopreservatives, and exhibit a broad spectrum of antimicrobial activity at low concentrations along with thermal as well as pH stability in foods. High bacteriocin production usually occurs in complex media. However, such media are expensive for an economical production process. For effective use of bacteriocins as food biopreservatives, there is a need to have heat-stable wide spectrum bacteriocins produced with high-specific activity in food-grade medium. The main hurdles concerning the application of bacteriocins as food biopreservatives is their low yield in food-grade medium and time-consuming, expensive purification processes, which are suitable at laboratory scale but not at industrial scale. So, the present review focuses on the bacteriocins production using complex and food-grade media, which mainly emphasizes on the bacteriocin producer strains, media used, different production systems used and effect of different fermentation conditions on the bacteriocin production. In addition, this review emphasizes the purification processes designed for efficient recovery of bacteriocins at small and large scale.

Highly Specific Culture-Independent Detection of YGNGV Motif-Containing Pediocin-Producing Strains.

A novel method based on (1) initial microbiological screening and (2) a highly specific PCR is described for selection of strains expressing YGNGV motif-containing pediocin. Initial screening is carried out using spot on the lawn assay for selection of acid-free, hydrogen peroxide (H2O2)-free and secreted heat-stable inhibitory activity producing strains. This is followed by highly specific PCR for amplification of 406-bp fragment using forward primer: 5'-tggccaatatcattggtggt-3' targeting signal peptide sequence of pediocin structural gene and reverse primer: 5'-ctactaacgcttggctggca-3' encoding N-terminus of immunity gene. The assay was validated with Pediococcus pentosaceus NCDC273 and Pediococcus acidilactici NCDC252 using (1) digestion of amplified 406-bp fragment with HindIII restriction enzyme-producing two restriction fragments of expected sizes (227 and 179 bp), (2) nucleotide sequencing of 406-bp fragment from both strains found these pediocins identical to pediocin PA-1/AcH and (3) identification of both pediocins as pediocin PA-1 at protein level using RP-HPLC. The assay was used for screening six strains (3 pediococci, 2 lactobacilli and an Enterococcus faecium) producing acid-free, hydrogen peroxide (H2O2)-free and secreted heat-stable inhibitory activity. This resulted in the detection of three new strains (P. pentosaceus NCDC35, E. faecium NCDC124 and Lactobacillus plantarum NCDC20) producing YGNGV motif-containing pediocins.

Simple and rapid purification of pediocin PA-1 from Pediococcus pentosaceous NCDC 273 suitable for industrial application.

The use of pediocins as food additives or drugs requires a simple and rapid method by which large quantities of homogeneous pediocin are produced at industrial level. Two centrifugation steps required during initial stages of purification i.e. separation of cells from fermentation broth and collection of precipitates after ammonium sulphate precipitation are the major bottlenecks for their large scale purification. In the present work, pediocin production by a new a dairy strain, Pediococcus pentosaceous NCDC 273 (identical to pediocin PA-1 at nucleotide sequence level), was found to be optimum at initial pH of 6.0 and 7.0 of basal MRS supplemented with 20 g/l of glucose or lactose at 20 and 24 h, respectively. Immobilization of cells through entrapment in alginate-xanthan gum gel beads with chitosan coating resulted in negligible cell release during fermentation. Thus, the cell free extract was directly collected through decantation, avoiding the need of centrifugation step at this stage. Subsequent ammonium sulphate precipitation at isoelectric point of pediocin PA-1 (8.85), using magnetic stirrer at high speed (approx. 1200 rpm), resulted in forceful deposition of precipitates on the wall of precipitation beaker allowing their collection using a spatula, avoiding centrifugation step at this stage also. Further purification using cation-exchange chromatography resulted in yield of 134.4% with more than 320 fold purification with the specific activity of 19×105 AU/mg. The collection of single peak of pediocin at 41.9min in RP-HPLC, overlapping with standard pediocin PA-1, resulted in yield of 1.15 µg from 20 µl of sample applied. The overlapping of RP-HPLC peak and SDS-PAGE band corresponding to 4.6 kDa, confirmed the purity and identity of pediocin 273 as pediocin PA-1.

Morinda revisited: changes in nutritional well-being and gender differences after 30 years of rapid economic growth in rural Punjab, India.

A follow-up study of malnutrition and its determinants among children 6 to 24 months of age was carried out in rural areas of Punjab State in India 30 years after the original study, and following a period of rapid economic growth. The original 1971 study had found a high prevalence of mortality and malnutrition and the worst gender difference in nutritional status ever recorded in an Indian study. The 2001 follow-up study found dramatic reductions in child mortality, child malnutrition, gender-based imbalances in child well-being and care, and family size, the result of participatory economic growth coupled with broad-based educational, health, and family-planning services. Despite overall improvements in caloric intake, however, 40% of lower-class children in 2001 were still consuming less than 50% of their caloric allowance. With minimal gender-based abortion and significantly reduced neglect and mortality offemale children, gender balance among children in this area of rural Punjab improved markedly over the 30-year period.