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Biosynthesis of chitosan encapsulated silver- nanoparticles using Probiotic-Lactobacillus plantarum strain and it's in vitro anticancer assessment on HeLa cells

Cervical cancer remains the deadliest cancer among women worldwide. Investigating the molecular mechanisms of tumor progression by targeting signaling pathways can provide insights into novel therapeutic strategies to overcome the limitations of conventional treatments. Green synthesis of nanoparticles addresses conventional treatment drawbacks like chemotherapy and radiation. This study aims to green synthesize, characterize, and evaluate chitosan-encapsulated silver nanoparticles (AgNPs) using Lactobacillus plantarum probiotics against cervical cancer HeLa cells, targeting the EMT mechanism. The green synthesized chitosan encapsulated Silver- Nanoparticles using probiotic -Lactobacillus plantarum (CS-LP-AgNPs) were characterized using UV–vis spectroscopy, which showed a peak at 420 nm confirming the reduction of AgNPs. Zeta and DLS analysis revealed the particle surface charge and stability. TEM analysis demonstrated that CS-LP-AgNPs are spherically shaped, with a size of approximately 15.3 nm. XRD patterns confirmed the crystalline nature of CS-LP-AgNPs. FTIR spectroscopy confirmed that CS-LP-AgNPs were functionalized with biomolecules. DAPI and double staining were employed to examine characteristic nuclear and morphological changes during apoptosis. Gene expression profiles of EMT in HeLa cell lines were performed to evaluate the anticancer potency of CS-LP-AgNPs. MTT assay demonstrated cytotoxic activity, whereas DAPI/(AO/EB) double staining images showed the induced apoptosis in HeLa cells by CS-LP-AgNPs treatment. CS-LP-AgNPs treated HeLa cells showed decreased SNAIL/EMT via enhancing apoptotic cascade mechanism. Green synthesized CS-LP-AgNPs may be considered an effective anti-cancer drug delivery system for the treatment of CC in the future.

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Diagnosis of tuberculous lymphadenitis by molecular and immunological tools

IntroductionTuberculous lymphadenitis (TBL) represents the prevailing presentation of extrapulmonary tuberculosis (EPTB) that comprises ∼35 % of EPTB cases, respectively and mainly occurs at cervical lymph nodes. Diagnostic challenge in TBL is primarily due to paucibacillary nature of specimens, and most common laboratory tests produced inconclusive findings. Areas coveredWe evaluated the literature on current diagnostic methods for TBL. Smear microscopy, culture, tuberculin skin test, interferon-γ release assay, biochemical assessments, imaging, histopathological, and cytological examination, etc. are various conventional methods used to diagnose TBL but these are insufficient. Further, nucleic acid amplification tests (NAATs) such as loop-mediated isothermal amplification (LAMP), PCR/multiplex-PCR, nested-PCR, real-time PCR, and GeneXpert®MTB/RIF utilized for TBL diagnosis but they have their own merits and demerits. Presently, several tools have been employed for detection of circulating Mtb cell-free DNA (cfDNA) through NAATs, aptamer-linked immobilized sorbent assay, and immuno-PCR (I-PCR), etc. ConclusionCurrently, there is no single accessible test available for effective diagnosis of TBL. In this review, we summarized all detailed conventional methodologies along with additional tools such as ALISA, I-PCR, and cfDNA for detection of Mtb biomarkers that have been utilized for diagnosis of pulmonary TB (PTB) and various forms of EPTB that may also be investigated for diagnosis of TBL. Early diagnosis and treatment would help in reducing the severe complications associated with TBL such as fistula, ulceration, or abscess formation in lymph nodes.

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Open Access
Antibacterial activity of Ocimum sanctum L. essential oil against multidrug resistance bacteria vaginosis

The antibiotic resistance of Bacteria Vaginosis (BV) isolates intensifies vaginal morbidity and genital infections facilitating poor treatment outcome and severe vaginal pathology. The phytochemicals in Ocimum sanctum essential oil (OsEO) were investigated for their antibacterial activity against bacteria vaginosis and major metabolites on multidrug resistance (MDR) strains. Bacteria pathogens isolated from vaginal samples (n = 40) obtained from patients with confirmed BV were analysed for hemolytic activity, biofilm production and profiled for antibiotic resistance. Extracted OsEO was profiled with GC-MS and analysed for antibacterial activity. Of the recovered bacteria pathogens (n = 241) associated with vaginosis including Streptococcus pyogenes (34 %), Staphylococcus aureus (31 %) and Escherichia coli (10 %) and less than 10 % Klebsiella oxytoca, Enterobacter cloaca, Pseudomonas aeruginosa and Citrobacter freundii were identified. Significant rates of 21.6 %, 4.6 % and 2.3 % were weak, mild and strong biofilm producers respectively and overall 26.6 % were hemolytic strains (p < 0.05). More than 60 % resistance to ceftriaxone sulbactam, ampiclox, cefuroxime, cefotaxime, nalidixic acid and cefexime was observed in BV with significant proportion showing MARI>0.2 (p < 0.05). Hierarchical clustering of MDR BV strains provided related clustered bacteria pathogens having a very low susceptibility to iminepem, cefuroxime, and amoxycillin/clavulanate. More than 1.2 % saponin, alkaloids and flavonoids levels in OsEO gave significant inhibitory activities at IC50 (25.0 μg/mL) and IC90 (50.0 μg/mL) and significant inhibitory association with phytochemical compounds (eta = 0.457, p = 0.015). OsEO cyclohexene and methanoazulene metabolites showed significant antibacterial association with BV strains (p < 0.05). The OsEO phytochemical metabolites showed antibacterial activity against multidrug resistance BV and identified cyclohexene and methanoazulene are promising candidates for developing formulations as topical antimicrobial agents for BV treatment.

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Open Access
Exploring the Promise of Psychobiotics: Bridging Gut Microbiota and Mental Health for a Flourishing Society

Mental health problems have become one of the major issues worldwide. People of every age group and gender are facing psychological issues. Conventional medicines are not reliable due to their adverse effects like altered sleeping pattern, addiction and health problems throughout the entire body. Psychobiotics is a new class of probiotics that is serving a wide range of applications in psychological health. Psychobiotic refers to the biological formulation which when consumed in right amount, confers psychological benefits. A lot of studies have supported the function of gut microbiota in mood cognition and controlling anxiety. The mechanism of action of psychobiotics has not been completely investigated. However, it may confer benefits by modulating hypothalamic-pituitary-adrenal (HPA) axis, by directly influencing immune system and through production of various neurotransmitters and neurohormones like proteins and short fatty acids chains. This review highlights the potential of different bacterial strains in human and animal trials. It latter also covers various psychobiotics formulations marketed by different companies. In addition to this, we also tried to cover the various hurdles in psychobiotic research that need to be addressed in the future to build a prosperous society.

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Bacterial etiology and antimicrobial resistance pattern of community-acquired urinary tract infection in older adults

BackgroundUrinary tract infections (UTIs) are a significant cause of morbidity in elderly individuals and remain a persistent challenge for medical professionals. This study aimed to identify the bacteria causing community-acquired (CA) UTIs in older patients, determine their antimicrobial resistance patterns, assess the prevalence of polymicrobial infections, and identify the risk factors. MethodsUrine samples were obtained from patients with symptomatic UTIs and then cultured on blood and MacConkey agar. Positive cultures were identified and tested for antimicrobial susceptibility using the VITEK 2 system. ResultsPolymicrobial infections were found in 69/427 (16.16 %) of older patients with CA-UTIs and associated with diabetes (p = 0.007), previous antimicrobial use (p = 0.025), and recurrent urinary infections (p = 0.043). Escherichia coli was the leading pathogen (57.26 %), and Klebsiella pneumoniae was identified in 15.32 % of CA-UTIs. Escherichia coli was more common in non-diabetic patients (60.81 %) than diabetes (43.69 %). However, the rates of Klebsiella species were higher in diabetes (20.39 %) than non-diabetes (14.50 %). Gram-negative uropathogens showed 49.89 % resistance to amoxicillin-clavulanic acid, while imipenem is the least resistant (7.19 %). The gram-positive uropathogens were resistant to 9.80 % of linezolid and highly resistant to erythromycin (74.51 %), tetracycline (72.55 %), and gentamicin (70.59 %). ConclusionsEscherichia coli isolates were the predominant bacteria in the elderly and highly resistant to amoxicillin-clavulanic. The most effective drug against gram-negative bacteria was imipenem, while linezolid proved potently effective against gram-positive bacteria. Diabetes, previous antimicrobial use, and recurrent urinary infections are risk factors for polymicrobial UTIs.

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Open Access
Adherence and cytotoxicity of Acinetobacter baumannii on human cervical carcinoma epithelial cells: Exploring the role of anti-OmpA antibodies

We investigated the invasion of Acinetobacter baumannii strains to epithelial cells and elucidated the role of antibodies against outer membrane protein A (OmpA). A. baumannii ATCC 19606 and clinical isolate 58ST were utilized. OmpA was expressed, purified, and administered to BALB/c mice, inducing anti-OmpA antibodies. OmpA cytotoxicity was evaluated. Two A. baumannii strains were selected to infect human cervical HeLa cells. Serum resistance was determined at sera dilutions. Adhesion, internalization, and proliferation of live and killed A. baumannii in HeLa cells were examined with and without anti-OmpA sera. HeLa cell viability was assessed with and without exposure of live A. baumannii strains to anti-OmpA sera. Cytoskeleton inhibitor experiments were conducted on epithelial cells to probe microfilament and microtubule involvement in A. baumannii invasion. OmpA prompted antibody production without toxicity in mice. A. baumannii strains displayed varying cell invasion abilities, notably the clinical strain exhibiting the highest invasion. A. baumannii cells localized within vacuoles during internalization, migrating towards the nucleus, using a zipper-like invasion process. Bacterial proliferation within host cells led to HeLa cell death. Pre-treatment with anti-OmpA antibodies significantly curbed adhesion and invasion of A. baumannii in HeLa cells. Microscopic imaging provided proof of the intracellular presence of A. baumannii in HeLa cells. In conclusion, the OmpA plays a crucial part in A. baumannii - epithelial cell interactions. The results add to our knowledge of pathogenesis during the initial stages of infection by A. baumannii.

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Open Access
Microbiome and metabolomic profiling in humans with functional anorectal pain: Identifying key players in disease pathogenesis

Functional Anorectal Pain (FARP) is a chronic, functional, nonorganic disorder of the anorectal region, often misdiagnosed as various types of anorectal organic diseases in clinical practice. However, the pathogenesis and diagnostic markers of FARP remain poorly understood. In this study, we recruited 45 FARP patients and 35 healthy subjects to investigate the microbial and metabolomic profiles associated with FARP progression. Anorectal manometry was utilized to determine changes in anal pressure, revealing a significant decrease in FARP patients' anal resting pressure and maximum anal squeeze pressure. Through 16S rRNA gene sequencing and metabolomics analysis, we identified specific microbial genera, including Propionibacterium, Ruminococcus2, and Bilophila, and specific metabolite profiles, including Gentisic acid, 2-Naphthalenethiol, 2-Furancarboxaldehyde, Benzoic acid, Allose, and (S)-3-Amino-5-methylhexanoic acid, that were closely correlated with FARP. In-depth analysis revealed that the Phospholipase D (PLD) signaling pathway exhibited significant differences in FARP. Additionally, serum cortisol levels were found to be significantly elevated in FARP patients. These findings provide new insights into the anorectal manometry features, gut microbiota, and serum metabolome associated with FARP, potentially contributing to improved clinical diagnosis and therapeutic strategies for FARP.

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Open Access
Epsilon-poly-l-lysine inhibits biofilm formation and aids dispersion in Acinetobacter baumannii

Acinetobacter baumannii is a prominent hospital-associated bacterium whose eradication is increasingly challenging due to its remarkable ability to resist antibiotics. The most common illnesses caused by antibiotic-resistant A. baumannii are biofilm-associated. Therefore, novel methods to combat A. baumannii are urgently needed. Application of antimicrobial peptides (AMPs) is one of the avenues to be explored. Epsilon poly-l-lysine (ε-PL) is an antimicrobial peptide with low mammalian toxicity. It is commonly used as a food preservative and has advantages such as biodegradability, good water solubility, and thermal stability.Therefore, the antimicrobial activity of ε-PL against clinical isolates of A. baumannii was investigated. The effect of ε-PL on antimicrobial sensitivity was determined by broth dilution assay. The effect of ε-PL on biofilm formation and dispersion was studied using a crystal violet assay. The changes in the expression of quorum sensing related and virulence genes (abaI, csuE, pilT, bap, and luxI) were analyzed using qPCR by the Δ Δ CT method.All the A. baumannii clinical isolates (n = 28) tested, were resistant to multiple drugs. The treatment with ε-PL resulted in a significant (p < 0.05) reduction in the biofilm formation abilities of all the clinical isolates of A. baumannii. Also, ε-PL caused a significant (p < 0.05) decrease in the dispersion of preformed biofilms. The reduction in the biofilm formation could be attributed to the inhibition of autoinducer synthase (abaI) which is required for biofilm development in A. baumannii. Also, it could be due to altering of expression of biofilm-related genes like csuE, pilT, bap, and luxI. These results suggest that ε-PL could be effective in the elimination of A. baumannii biofilms and decreasing its virulence.

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Open Access
Allicin and Probiotics: Double-edged sword for the management of Striae distensae

Striae distensae (SD), commonly known as Stretch marks or striae, are one of the most common benign dermal lesions frequently seen in females that often cause a significant physical and psychological impact. A number of treatment modalities ranging from topicals to invasive approaches are commercially available, however, none of the available options is capable of complete eradication of SD. As effectiveness of most of the available topical formulations for SD is attributed to the combined effects of their antioxidant, anti-inflammatory and proliferative effects, allicin and probiotic based topical formulations are hypothesized to be effective in treatment and prevention of SD. Both allicin and probiotics are able to reduce the inflammatory response via suppression of transcription factor i.e., nuclear factor (NF)-κB, and pro-inflammatory cytokines and chemokines levels. Moreover, the antioxidant effect of allicin and probiotics is considered to decrease the reactive oxygen species induced fragmentation of collagen. Also, the effects of allicin on the collagen and elastin tissue as well as beneficial effects of probiotics and their metabolites on skin elasticity and skin hydration are expected to provide multiple target approach for the management of SD. Altogether, a combination formulation containing both allicin and probiotics is considered to be novel approach for the prevention and management of SD.

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Open Access