Staining Test Research Articles

P1223 Lactobacillus rhamnosus GG/p40 encapsuled cationic host defense peptide play protective roles on immune-related colitis by inducing differentiation and immune suppression of regulatory T cells

Abstract Background The immune-related colitis (IRC) is the most prominent in immune-related adverse events (irAE). The association between gut microbiota and IRC development has been revealed. Our study aimed to explore the protective effect and mechanism of the probiotic Lactobacillus rhamnosus GG (LGG) and its secretory protein p40 on IRC. The self-assembled cationic host defense peptide (CHDP) hydrogel was applied to protect oral p40 protein from digestive damage and target enriched the site of colitis. Methods The IRC model mice were intervened with LGG by gavage (i.g.), p40 by enema (en.), or p40 encapsuled by CHDP (i.g.). The T cell phenotype of colonic lamina propria mononuclear cells (LPMC) was measured by flow cytometry. The indocyanine green (ICG) encapsuled by CHDP was gavage in IRC model mice, the distribution of CHDP was monitored by fluorescence signals imaging via IVIS, to assess the adhesive effect of CHDP in mice colitis. In vitro, the proportion of regulatory T cells (Treg) differentiated was measured by LGG supernatant co-cultured with CD4+ naïve T cells, and the division index of CD4+T cells was measured by LGG supernatant co-cultured with CD4+T cells and Treg (CD4+T cells: Treg=1:1) by carboxyfluorescein succinimidyl ester (CFSE) staining test. Results Compared with DSS+anti-PD-1+anti-CTLA-4 IRC model group, LGG (i.g.) group had lower weight loss, DAI score, histopathological score, relative IL-6 and TNF-α mRNA level, IL-6 and TNF-α concentration and higher colon length (P<0.05) (Fig.1A-L). P40 (en.) group had lower weight loss, DAI score, histopathological score and higher colon length (P<0.05) (Fig.1M-T). CHDP+p40 (i.g.) group had lower weight loss, DAI score, histopathological score, relative IL-6 and TNF-α mRNA level, IL-6 concentration and higher colon length (P<0.05) (Fig.1M-W). The percentages of CD25+FoxP3+Treg/CD4+T cells of colonic LPMC in LGG, p40 (en.), and CHDP+p40 (i.g.) group were significantly higher than IRC model group (P<0.05) (Fig.1X-Z, α). The average 28hr-residual rate of fluorescence imaging radiance in IRC-CHDP+ICG group was higher than both NC-CHDP+ICG and IRC-ICG group (Fig.2β-δ), and the average radiance of colon in IRC-CHDP+ICG group was also the highest (P<0.05) (Fig.2ε-ζ). In vitro, LGG 107 CFU/mL, 108 CFU/mL and 5 × 108 CFU/mL (P<0.05) could significantly promote Treg differentiation with concentration dependent manner (Fig.2η-ι), and the suppression of Treg on CD4+T cells with lower division index compared with control group (Fig.2κ). Conclusion LGG/p40 played protective role by promoting the differentiation and immune suppression of Treg cells in IRC. CHDP hydrogel could protect p40 protein from the digestive damage, and target enriched the inflamed lesion to improve the efficacy.

A sustainable strategy for generating highly stable human skin equivalents based on fish collagen

Tissue engineered skin equivalents are increasingly recognized as potential alternatives to traditional skin models such as human ex vivo skin or animal skin models. However, most of the currently investigated human skin equivalents (HSEs) are constructed using mammalian collagen which can be expensive and difficult to extract. Fish skin is a waste product produced by fish processing industries and identified as a cost-efficient and sustainable source of type I collagen. In this work, we describe a method for generating highly stable HSEs based on fibrin fortified tilapia fish collagen. The fortified fish collagen (FFC) formulation is optimized to enable reproducible fabrication of full-thickness HSEs that undergo limited contraction, facilitating the incorporation of human donor-derived skin cells and formation of biomimetic dermal and epidermal layers. The morphology and barrier function of the FFC HSEs are compared with a commercial skin model and validated with immunohistochemical staining and transepithelial electrical resistance testing. Finally, the potential of a high throughput screening platform with FFC HSE is explored by scaling down its fabrication to 96-well format.

Isolation, Identification and Comparative Analysis of Oral Microbial Communities in Smokers and Non-Smokers: A Scientific Investigation

Smoking tobacco considerably changes the Gram-positive bacteria in saliva, including pathogens that may be involved in the development of tobacco-related illnesses such as periodontitis and peri-implantitis. A poor diet, poor oral hygiene, and other health problems can change the balance between these bacteria, allowing the dangerous bacteria to gain control. Pathogens can spread to distant places in the body if they overgrow in the mouth and blood vessels. Some bacteria have adapted to survive in the oral cavity by surviving within our bodies' defense mechanisms. Smokers are more likely to experience issues following gum and oral surgery, gum disease, tooth loss, decay on the roots of teeth, and oral cancer of the mouth. It has been reported that smoking has an impact on the oral microbiota, which has been connected to a number of human disorders. This study was designed for the isolation of bacteria from smokers and non-smokers. Oral health can be impacted by smoking. Smokers are more likely to experience issues following gum and oral surgery, gum disease, tooth loss, decay on the roots of teeth, and oral cancer of the mouth. The bacterial were isolated from the samples collected from non- smokers and smokers. The culture technique was used to grow bacteria. For further identification Gram staining, microscopy and biochemical tests were performed. Gram positive and gram negative bacteria were isolated from both smokers and non-smokers. The prevalence of gram positive bacteria was high in smokers and the prevalence of gram negative bacteria among smokers was high. Total 33% of gram positive bacteria were isolated from non-smokers and 67% from smokers. The gram positive species isolated were S. aureus and S. pneumoniae and 33% of gram negative bacteria were isolated from non-smokers and 67% from smokers. The gram negative species isolated were E.coli and K. pneumoniae.

Integrated histopathology and transcriptome metabolome profiling reveal the toxicity mechanism of phenazine-1-carboxylic acid in zebrafish

Phenazine-1-carboxylic acid (PCA) is a new type of agrochemical used to prevent plant diseases, but its effects on aquatic organisms are unclear. To comprehensively assess the impacts of PCA for aquatic organisms and its associated environmental risks, this study investigated, taking zebrafish as the research object, the toxicological mechanism of PCA by means of optical microscopy, hematoxylin and eosin (HE) staining, ultrastructural observation, physiological and biochemical testing, transcriptome sequencing, metabolome analysis, fluorescence quantitative PCR and molecular simulation. The results indicated that PCA was detrimental to zebrafish embryos, larvae and adults, with LC50 values at 96 h of 3.9093 mg/L, 8.5075 mg/L, and 13.6388 mg/L, respectively. PCA caused abnormal spontaneous movement, slowed the heart rate, delayed hatching, shortened the body length, slowed growth, and caused malformations. PCA mainly affected the brain, liver, heart, and ovaries. PCA distorted cell morphology, damaged mitochondrial membranes, disintegrated mitochondrial ridges, and dissociated nuclear membranes. PCA inhibited the enzyme activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX), decreased the malondialdehyde (MDA) content and disrupted antioxidant effects. The results of omics studies confirmed that PCA interfered with the transcriptional and metabolic network of zebrafish, downregulating most genes and metabolites. PCA mainly affected functions related to mitochondrial steroids, lipids, sterols, oxidoreductase activity and pathways involving cofactors, steroids, porphyrin, cytochromes, which specifically bound to targets such as panx3, agmat, and ace2. PCA was moderately toxic to zebrafish, and its usage should be strictly controlled to reduce toxic effects on aquatic organisms. The results of this study provide a new insights for ecotoxicology research.

Isolation and Identification of Biosurfactant-Producing Bacteria in An-aerobic Palm Oil Waste Pools at PT. Aek Loba Plantation

Biosurfactant-producing bacteria are bacteria that have alternative energy sources, namely surfactants which contain hydrophilic and hydrophobic groups based on their molecular structure so that they can survive between different polar and hydrogen bonding fluids. Palm oil mill liquid waste is waste in the form of water or liquid, oil and organic solids produced from the processing of palm oil. The purpose of this study was to determine the genera of biosurfactant-producing bacteria found in palm oil wastewater in the an-aerobic pond of PT. Aek Loba plantation and to find out the bacterial species with the highest emulsification index. This study used descriptive methods and identification of bacteria through morphological characterization, gram staining tests and biochemical tests. Bacteria were isolated from palm oil wastewater by dilution method 10-4, 10-5 and 10-6. Bacterial emulsification index is calculated by emulsification test. In this study, 8 biosurfactant-producing bacterial isolates were found, consisting of 2 Proteus genus isolates, 2 Enterobacter genus isolates, 2 Bacillus genus bacterial isolates and 2 Pluralibacter genus isolates. The bacteria that had the highest emulsification index was Pluralibacter gergoviae with an emulsion index of 43% which was identified by molecular testing.

Identification of Methicillin-resistant Staphylococcus aureus Isolated from Dairy Cow’s Milk in Tulungagung District, Indonesia

Background: Mastitis is one of the factors contributing to the health-related decreased milk production and quality for dairy cows. Mastitis in ruminants is a serious bacterial disease caused by Staphylococcus aureus. Staphylococcal bacteria are becoming increasingly resistant to many classes of antibiotics, particularly β-lactam families like the MRSA strain. Laboratory tests are required to determine the level of bacterial resistance and to identify MRSA isolates sourced from dairy cows in Tulungagung District. Methods: 110 milk samples were isolated on MSA media followed by Gram’s staining and biochemical tests. Kirby-bauer diffusion test-based assessment of antibiotic sensitivity. The S. aureus isolates that underwent the MRSA identification test were S. aureus isolates that had developed a resistance to β-lactam antibiotics. Result: A total 81 samples of the 110 isolated milk samples were determined to be positive for S. aureus. Out of total isolates, 25 isolates of S. aureus had the highest level of oxacillin resistance. As many as 4 isolates were confirmed to be MultiDrug Resistance (MDR) and 17 MRSA isolates were discovered from 100 samples of dairy cows. Early diagnosis of MRSA infection is crucial since it can be challenging to treat because this type of bacteria is known to be resistant to several drugs and spreads readily.

Antagonistic Test of Bacteria Producing Siderophore and Protease Enzymes from The Rhizosfer of Peanut Plants on The Growth of Pathogenic Fungus Colletotrichum gloeosporioides

This study aims to determine the inhibitory effect of bacterial isolates producing siderophores and protease enzymes on the growth of the pathogenic fungus Colletotrichum gloeosporioides. The initial stage of research begins with the isolation of pathogenic bacteria and fungi, and is followed by testing the production of siderophores and protease enzymes. Bacteria were isolated from the rhizosphere of peanut plants, while pathogenic fungi were isolated from large chili fruit infected with anthracnose disease taken from Jelantik Village, Central Lombok Regency. Characterization of isolates for siderophore production used the Arnow method, while the protease enzyme production test used SKIM Milk Agar media. Next, the inhibition test of bacterial isolates against pathogenic fungi was carried out using the dual culture method. Characterization of potential isolates was carried out by observing bacterial colony morphology, gram staining and biochemical tests. The results of the siderophore production test showed that there was one isolate capable of producing siderophores with the isolate code RKT2. Meanwhile, the protease enzyme production test showed that all bacterial isolates produced protease enzymes, where isolate RKT9 had the highest Proteolytic Index, namely 1.57. The two isolates showed different inhibitory test results, namely isolate RKT2 had high inhibition, while RKT9 had low inhibition. The results of the research showed that a bacterial isolate (RKT2) from the rhizosphere of peanut plants was able to inhibit the growth of the pathogenic fungus Colletotrichum gloeosporioides in the high category.

Transcriptomics-based analysis reveals the nephrotoxic effects of triphenyltin (TPT) on SD rats by affecting RAS, AQPs and lipid metabolism

Triphenyltin (TPT) is a class of organotin compounds that are extensively used in industry and agriculture. They have endocrine-disrupting effects and cause severe environmental contamination. Pollutants may accumulate in the kidneys and cause pathological complications. However, the mechanism of TPT's toxicological effects on the kidney remains unclear. This study aimed to investigate the toxic effects and mechanism of action of TPT exposure on renal impairment in rats. Male SD rats were divided into four groups: the Ctrl group (control group), TPT-L group (0.5 mg/kg/d), TPT-M group (1 mg/kg/d), and TPT-H group (2 mg/kg/d). After 28 days of exposure to TPT, we observed the morphology and structure of kidney tissue using HE, PASM, and Masson staining. We also detected serum biochemical indexes, performed transcriptome sequencing of rat kidney tissue using RNA-seq. Furthermore, protein expression levels were measured through immunohistochemistry and gene expression levels were determined using RT-qPCR. The study results indicated a decrease in kidney weight and relative kidney weight after 28 days of exposure to TPT. Additionally, TPT caused damage to kidney structure and function, as evidenced by HE staining, PASM staining, and serum biochemical tests. Transcriptomics identified 352 DEGs, and enrichment analyses revealed that TPT exposure primarily impacted the renin-angiotensin system (RAS). The expression levels of water channel proteins were reduced, and the expression levels of RAS and lipid metabolism-related genes (Mme, Ace, Fasn, Cyp4a8, Cpt1b and Ppard) were significantly decreased in the TPT-treated group. In summary, exposure to TPT may impair renal structure and function in rats by affecting RAS, AQPs, and lipid metabolism.

Short chain fatty acids inhibit corneal inflammatory responses to TLR ligands via the ocular G-protein coupled receptor 43

PurposeShort chain fatty acids (SCFAs) produced by gut microbiota are known to play primary roles in gut homeostasis by immunomodulation partially through G-protein coupled receptors (GPR) 43. Using mouse models of TLR ligand induced keratitis, we investigated whether SCFAs and GPR43 play any regulatory roles in the pathogenesis of inflammatory responses in the eye. MethodsBoth human and mouse eyes were labeled with a specific antibody for GPR43 and imaged by a laser scanning confocal microscope. Corneal cups from naïve C57BL/6J (B6) and GPR43 knockout (KO) mice were stimulated with TLR ligands in the presence or absence of sodium butyrate overnight and then processed for RT-PCR assay for expression of GPR43 and cytokines. Keratitis was induced by Poly I:C in wild type (WT) B6, GPR43KO and chimeric mice and the disease severity was evaluated by the corneal fluorescein staining test, and infiltrating cell staining and calculating in corneal whole mount. ResultsGPR43 is expressed in both human and mouse eyes and the expression is bidirectionally regulated by TLR ligands and butyrate. Butyrate significantly inhibited inflammation caused by several TLR ligands such as Poly I:C, Flagellin, and CpG-ODN (TLR-3, 5 and 9 agonists, respectively) in WT, but not GPR43KO, mice. Butyrate inhibition of TLR-induced keratitis is mediated by the GPR43 expressed in tissue but not hematopoietic, cells. ConclusionsThis is the first report to demonstrate of the protective effect of SCFAs on microbial keratitis, and the dynamic expression and anti-inflammatory function of GPR43 in the eye. SCFAs can modulate inflammation and immunity in the eye through GPR43.

Accuracy of gastric nodule combined with rapid urease test prediction in diagnosing Helicobacter pylori infection in children.

The diagnosis of Helicobacter pylori (H. pylori) infection in children remains challenging with the lack of a rapid, cost-effective, and highly accurate diagnostic method. Consequently, this study aimed to investigate the accuracy of the combination of gastric nodule and rapid urease test (RUT) as a diagnostic method for H. pylori infection in children. The study included participants who underwent a thorough examination, including gastroscopy, a 13C breath test, RUT, and pathological methylene blue staining, with the gold standard for diagnosing of H. pylori infection being a positive result from both pathological methylene blue staining and 13C breath test. The sensitivity, specificity, positive and negative predictive values, and accuracy of the diagnostic methods were calculated. The accuracy of the different tests for H. pylori infection was evaluated in 2202 participants. A total of 730 (33.2%) children were diagnosed with H. pylori infection (pathological methylene blue staining and 13C breath test, both positive). Gastric nodule had a sensitivity of 87.1% and a specificity of 93.1%, whereas combining gastric nodule and RUT in parallel had the higher accuracy of 95.4%. The accuracy of gastric nodule diagnosis was higher in younger age groups and increased after excluding patients with a history of anti-H. pylori treatment. The findings of this study suggest that gastric nodules, particularly when combined with RUT, can be a valuable predictor of H. pylori infection in children, offering a simple and feasible alternative to other invasive methods.

Survey of Cholera Outbreak and its Resistant Pattern in Baluchistan, Pakistan

Objective: The present study was conducted to identify the isolates of V. cholerae as causative agent for the current outbreak in Baluchistan and evaluate the antibiotic sensitivity profile of the isolates Methodology; This cross-sectional study was conducted at Bolan Medical Complex Hospital Quetta from March to October 2022. Suspected stool samples were received with a history of untreated severe diarrhea within 12-hour duration from different affected districts of Baluchistan Province, Suspected colonies were subjected to gram staining, biochemical analysis and antibiotic susceptibility testing was performed by the Kirby-Bauer disk diffusion method. Results; Total 483 diarrheal samples were collected out of which 43 (8.90 %) were positive and 440 (91.09%) were negative. The all isolates 100 % were resistant to common antibiotics were whereas Ciprofloxacin100%, Levofloxacin 100% and Azithromycin 100% were sensitive to all isolates. Conclusion: Culture methods and biochemical identification of V. cholerae from suspected cholera may be helpful. There needs to be systematic surveillance of commonly used antibiotics and encourage the design and implementation of the cholera control policy advised by the WHO

Antimicrobial Activity of Azithromycin and Erythromycin against Streptococcus Pyogenes Isolated from Sore Throat Patients in Shendi, Sudan

Background: Streptococci is considered one of the predominant flora colonizing the respiratory tract of humans. The group A Streptococci (GAS) causes the broadest range of diseases that can lead to the asymptomatic carriage, superficial infection of the upper respiratory tract mainly throat infection. Objectives: The study was carried out to assess the antimicrobial activity of azithromycin and erythromycin against Streptococcus pyogenes (Group A) isolated from sore throat patients. Methods: Sixty-one throat swab samples from both sexes were collected randomly from different clinics in Shendi, Sudan from patients with clinical findings suggestive of throat infection between August to November 2021. Streptococcus pyogenes were isolated by standard cultural techniques and identified by using Gram stain and biochemical tests. Also, the antimicrobial activity of Azithromycin and Erythromycin were assessed using the disc diffusion method. Result: 19 throat swab samples (31%) out of 61 had S. pyogenes growth, whereas 42 (69%) did not. Of the patients, 12 (63.2% of them) were men, and 7, 36.8%, were women. The ages of the infected patients ranged from 1 to 10 years old in 2 (5.3%) cases, 11 to 20 years old in 2 (10.5%), 21 to 30 years old in 15 (78.9%), and 41 to 50 years old in 2 (5.3%) cases. In contrast to the other 5 (26.3%), 14 of them (73.7%) had recurring throat infections. Out of the 19 S. pyogenes isolates that tested positive, only 12 (63.6%) were susceptible to azithromycin and just 7 (36.8%) were resistant. 13 (68.4%) of the 19 S. pyogenes positive isolates were erythromycin sensitive, whereas 6 (31.6%) were resistant. Conclusion: Azithromycin and erythromycin are more sensitive to S. pyogenes, which indicates less excessive usage of these antibiotics in Shendi. Streptococcal infections in the respiratory tract are challenging to treat, and selecting an antibiotic treatment involves numerous considerations. Any isolated strain's susceptibility to antibiotics should be assessed because this is the only way to ensure quick and successful treatment. In order to improve public health, antibiotic therapy should be accompanied by adequate preventive measures, such as training nursing staff to prevent as many nosocomial infections as possible, educating the general public about the importance of hygiene and encouraging them to stop self-medicating and fostering closer scientific collaboration between clinicians and microbiologists.

CALCIUM SULFATE/HYDROXYAPATITE-MEDIATED CONTROLLED CO-DELIVERY OF BMP-2 AND ZOLEDRONIC ACID ENHANCES SPINAL FUSION

Although bone morphogenetic protein 2 (BMP-2) has been FDA-approved for spinal fusion for decades, its disadvantages of promoting osteoclast-based bone resorption and suboptimal carrier (absorbable collagen sponge) leading to premature release of the protein limit its clinical applications. Our recent study showed an excellent effect on bone regeneration when BMP-2 and zoledronic acid (ZA) were co-delivered based on a calcium sulphate/hydroxyapatite (CaS/HA) scaffold in a rat critical-size femoral defect model. Therefore, the aim of this study was to evaluate whether local application of BMP-2 and ZA released from a CaS/HA scaffold is favorable for spinal fusion. We hypothesized that CaS/HA mediated controlled co-delivery of rhBMP-2 and ZA could show an improved effect in spinal fusion over BMP-2 alone. 120, 8-week-old male Wistar rats (protocol no. 25-5131/474/38) were randomly divided into six groups in this study (CaS/HA, CaS/HA + BMP-2, CaS/HA + systemic ZA, CaS/HA + local ZA, CaS/HA + BMP-2 + systemic ZA, CaS/HA + BMP-2 + local ZA). A posterolateral spinal fusion at L4 to L5 was performed bilaterally by implanting group-dependent scaffolds. At 3 weeks and 6 weeks, 10 animals per group were euthanized for µCT, histological staining, or mechanical testing. µCT and histological results showed that the CaS/HA + BMP-2 + local ZA group significantly promoted bone regeneration than other treated groups. Biomechanical testing showed breaking force in CaS/HA + BMP + local ZA group was significantly higher than other groups at 6 weeks. In conclusion, the CaS/HA-based biomaterial functionalized with bioactive molecules rhBMP-2 and ZA enhanced bone formation and concomitant spinal fusion outcomeAcknowledgements: Many thanks to Ulrike Heide, Anna-Maria Placht (assistance with surgeries) as well as Suzanne Manthey & Annett Wenke (histology).

Avaliação da viabilidade do pólen em algumas variedades de crisântemo em spray no período de armazenamento

Abstract In producing chrysanthemum hybrids, more seed sets per fruit are preferred. Only successful pollination and fertilization allow seeds to de-velop. High pollen viability and pollen germination rate are intimately correlated with successful fertilization. Chrysanthemum pollen and their storage duration have only been the subject of a relatively few investigations. The study aimed to determine pollen's viability and germination during the 4 days, which were kept at 24 ºC. In the present study, two different Chrysanthemum species (Chrysanthemum coronarium L. and Chrysanthemum segetum L.) and two commercial Chrysanthemum varieties (Chic and Haydar) that belong to Chrysanthemum morifolium Ramat. species as spray chrysanthemum pollens were used for experimental material. The grains pollen were stored in an incubator which was 24 ºC and 60% humidity. The pollen viability was tested with TTC (2,3,5 Triphenyl Tetrazolium Chloride) staining test and pollen germination was evaluated hanging drop method with modified ME3-m medium, daily for 4 days including day 0. The results showed that the viability and germination of all pollens used in this study decreased day by day. Depending on the species/varieties, the viability rates ranged from 12.83% to 32.04% on the first day and between 0.57-2.33% on the last day. Pollen germination rates differed between 16.76% - 3.45% on the 0th day and 0.0-0.17% on the 4th day.

Health Risk Assesment of Air Emitting from Different Abattoirs in Akure Metropolis of Ondo State, Nigeria.

The burden of diseases can be greatly reduced by reducing air pollution which is one of the greatest risks to health in our environments. The study assessed the different types of micro-organisms present in a major and minor abattoir in Akure metropolis of Ondo state, Nigeria and their sensitivity to Antibiotics. Pre-tested questionnaires were used to obtain socio-demographic information, how the environment of the abattoir is sanitized and diseases likely to be contacted from the workers working in the abattoirs. Different levels of occurrence of isolates were observed in the major and minor abattoirs and the total count was done with the total bacteria counts ranging from lowest to highest. The total count for Escherichia coli range from 3.6×10 cfu/ml to 6.5 ×10 cfu/ml, while the total bacteria count ranged from 1.34×10 cfu/ml to 2.55×10 cfu/ml, also total coliform ranged from 2.8×10 cfu/ml to 4.1×10 cfu/ml at the major abattoir. The total count for Escherichia coli range from 5.7×10 cfu/ml to 7.2×10 cfu/ml, while the total bacteria count range from 5.2×10 cfu/ml to 1.56×10 cfu/ml, also total coliform ranged from 3.4×10 cfu/ml to 4.3×10 cfu/ml at the minor abattoir. During gram staining test it was observed that three out of the eight organisms namely, Micrococcus luteus, bacillus spp, Staphylococcus aureus, were gram positive, while Escherichia coli, Proteus mirabilis, Proteus vulgaris, Enterobacter aerogenes and Aeromonas sppwere gram negative. Antibiotic sensitivity was carried out on each of the isolates. Ciprofloxacin antibiotic had inhibitory effect on both grampositive bacteria with the highest effect on Staphylococcus aureus at 24mm and gram negative bacteria with highest result on Escherichia coli at 21mm. Therefore, there is presence of pathogenic micro-organisms especially Escherichia coli in the air environment of the abattoirs. Conclusively, there is need for routine environment sanitation of these slaughter houses because inhalation of air from these abattoirs can constitute a great environmental menace to the workers and residents around the area where they are situated, resulting in health complications to those with existing health challenges.

Genomic characterization and probiotic potential of lactic acid bacteria isolated from feces of guinea pig (Cavia porcellus).

Presently, there exists a growing interest in mitigating the utilization of antibiotics in response to the challenges emanating from their usage in livestock. A viable alternative strategy encompasses the introduction of live microorganisms recognized as probiotics, exerting advantageous impacts on the immune system and nutritional aspects of the host animals. Native lactic acid bacteria, inherently possessing specific properties and adaptive capabilities tailored to each animal, are deemed optimal contenders for probiotic advancement. In the current investigation, microorganisms exhibiting probiotic potential were isolated, characterized, and identified from the fecal samples of guinea pigs (Cavia porcellus) belonging to the Peruvian breed. The lactic acid bacteria isolated on Man, Rogosa, and Sharpe agar underwent Gram staining, catalase testing, proteolytic, amylolytic, and cellulolytic activity assays, low pH tolerance assessment, hemolytic evaluation, antagonism against Salmonella sp., determination of autoaggregation and coaggregation capacity, and genotypic characterization through sequencing of the 16S rRNA gene. A total of 33 lactic acid bacteria were isolated from the feces of 30 guinea pigs, also 10 isolates were selected based on Gram staining and catalase testing. All strains exhibited proteolytic activity, while only one demonstrated amylolytic capability, and none displayed cellulase activity. These bacteria showed higher tolerance to pH 5.0 and, to a lesser extent, to pH 4.0. Furthermore, they exhibited antagonistic activity against Salmonella sp. Only two bacteria demonstrated hemolytic activity, and were subsequently excluded from further evaluations. Subsequent assessments revealed autoaggregation capacities ranging from 4.55% to 23.19%, with a lesser degree of coaggregation with Salmonella sp. ranging from 3.53% to 8.94% for the remaining eight bacterial isolates. Based on these comprehensive tests, five bacteria with notable probiotic potential were identified by molecular assays as Leuconostoc citreum, Enterococcus gallinarum, Exiguobacterium sp., and Lactococcus lactis. The identified bacteria stand out as promising probiotic candidates, deserving further assessment in Peruvian breed guinea pigs. This exploration aims to enhance production outcomes while mitigating the adverse effects induced by pathogenic microorganisms.

Unlocking potential of oxytocin: improving intracranial lymphatic drainage for Alzheimer's disease treatment.

Background: The impediment to β-amyloid (Aβ) clearance caused by the invalid intracranial lymphatic drainage in Alzheimer's disease is pivotal to its pathogenesis, and finding reliable clinical available solutions to address this challenge remains elusive. Methods: The potential role and underlying mechanisms of intranasal oxytocin administration, an approved clinical intervention, in improving intracranial lymphatic drainage in middle-old-aged APP/PS1 mice were investigated by live mouse imaging, ASL/CEST-MRI scanning, in vivo two-photon imaging, immunofluorescence staining, ELISA, RT-qPCR, Western blotting, RNA-seq analysis, and cognitive behavioral tests. Results: Benefiting from multifaceted modulation of cerebral hemodynamics, aquaporin-4 polarization, meningeal lymphangiogenesis and transcriptional profiles, oxytocin administration normalized the structure and function of both the glymphatic and meningeal lymphatic systems severely impaired in middle-old-aged APP/PS1 mice. Consequently, this intervention facilitated the efficient drainage of Aβ from the brain parenchyma to the cerebrospinal fluid and then to the deep cervical lymph nodes for efficient clearance, as well as improvements in cognitive deficits. Conclusion: This work broadens the underlying neuroprotective mechanisms and clinical applications of oxytocin medication, showcasing its promising therapeutic prospects in central nervous system diseases with intracranial lymphatic dysfunction.

Integration of network pharmacology, lipidomics, and transcriptomics analysis to reveal the mechanisms underlying the amelioration of AKT-induced nonalcoholic fatty liver disease by total flavonoids in vine tea.

Nonalcoholic fatty liver disease (NAFLD) is the main reason for chronic liver diseases and malignancies. Currently, there is a lack of approved drugs for the prevention or treatment of NAFLD. Vine tea (Ampelopsis grossedentata) has been used as a traditional Chinese beverage for centuries. Vine tea carries out several biological activities including the regulation of plasma lipids and blood glucose, hepato-protective function, and anti-tumor activity and contains the highest content of flavonoids. However, the underlying mechanisms of total flavonoids from vine tea (TF) in the attenuation of NAFLD remain unclear. Therefore, we investigated the interventions and mechanisms of TF in mice with NAFLD using an integrated analysis of network pharmacology, lipidomics, and transcriptomics. Staining and biochemical tests revealed a significant increase in AKT-overexpression-induced (abbreviated as AKT-induced) NAFLD in mice. Lipid accumulation in hepatic intracellular vacuoles was alleviated after TF treatment. In addition, TF reduced the hepatic and serum triglyceride levels in mice with AKT-induced NAFLD. Lipidomics results showed 32 differential lipids in the liver, mainly including triglycerides (TG), diglycerides (DG), phosphatidylcholine (PC), and phosphatidylethanolamine (PE). Transcriptomic analysis revealed that 314 differentially expressed genes were commonly upregulated in the AKT group and downregulated in the TF group. The differential regulation of lipids by the genes Pparg, Scd1, Chpt1, Dgkz, and Pla2g12b was further revealed by network enrichment analysis and confirmed by RT-qPCR. Furthermore, we used immunohistochemistry (IHC) to detect changes in the protein levels of the key proteins PPARγ and SCD1. In summary, TF can improve hepatic steatosis by targeting the PPAR signaling pathway, thereby reducing de novo fatty acid synthesis and modulating the glycerophospholipid metabolism.

Vertical Transmission of Group B Streptococcus, Prevalence, Associated Factors, and Antimicrobial Susceptibility Profile among Newborns Delivered at Health Facilities in Jigjiga City, Ethiopia.

Group B Streptococcus (GBS) colonizes the rectovaginal area of women and vertically transmitted to neonates. This bacterium has been linked to severe neonatal complications including pneumonia, septicemia, and meningitis. GBS shows an increased resistance to commonly used antibiotics. Thus, we assessed the vertical transmission, contributing factors, and antimicrobial resistance patterns of GBS among newborns delivered at selected Hospitals in Jigjiga City. Hospital-based cross-sectional study was conducted from 1st June 2022 to 30th April 2023. A total of 849 pregnant women admitted to delivery wards from two hospitals were screened for GBS colonization. Subsequently, 162 GBS-colonized pregnant women and their newborn babies were included. A semistructured questionnaire and a review of medical records were used to collect the sociodemographic and clinical characteristics of the study participants. Trained nurses collected swab samples from the vaginal-rectal area of pregnant women and the nasal, ear canal, and umbilical areas of newborn babies. Samples were inoculated on Todd Hewitt broth media supplemented with gentamycin and nalidixic acid and then subcultured on blood agar. Colony characteristics, Gram stain, and catalase test were used for identification. All gram-positive cocci, B-hemolytic, and catalase-negative isolates were further identified using Christie-Atkins-Munch-Petersen and a bacitracin test. The modified Kirby-Bauer disk diffusion method was used for antimicrobial susceptibility testing. Data were analyzed using SPSS version 26. Logistic regression analysis was used to determine the factors associated with vertical transmission of GBS, and statistical significance was set at p values <0.05. The overall vertical transmission rate was 41.4% (67/162). History of preterm labor (Adjusted odds ratio (AOR) = 2.25; 95% CI: 1.11, 4.59), history of urinary tract infection (UTI) at current pregnancy (AOR = 2.25; 95% CI: 1.11, 4.59), and prolonged rupture of membranes greater than 18 hours (AOR = 2.23; 95% CI: 1.13, 4.4) were significantly associated with vertical transmission of GBS from previously colonized mothers to their newborn babies. Regarding GBS antibiotic susceptibility profile, a significant degree of resistance was observed to penicillin (29.9%), tetracycline (22.4%), ampicillin (20.9%), and clindamycin (19.4%). Our study documented a high prevalence of vertical transmission rate of GBS from pregnant women to their babies, with an overall transmission rate of 41.4%. The study identified the presence of antimicrobial-resistant GBS to penicillin, ampicillin, clindamycin, ciprofloxacin, and chloramphenicol. The organism was susceptible to vancomycin, followed by azithromycin, ceftriaxone, and erythromycin. Our study also reported MDR at 13.4%. Based on our findings, there is a need for antenatal culture-based GBS screening, maternal vaccination, and large-scale epidemiological and serotype identification studies to be put into practice in the study area.

Bacillus Calmette-Guérin Adenitis: An Intriguing Series of Four Cases

Bacille Calmette-Guérin (BCG) vaccine is part of the immunisation schedule in India. It is a live-attenuated vaccine with very few adverse reactions, one of which is BCG adenitis. The intriguing fact is that there has been a sudden upsurge in the number of BCG adenitis cases (two males and two female children), with four reported cases within three months (March 2023-June 2023). There were two male and two female children, aged between 2-8 months. All of them presented with left axillary swelling, and all but one had low grade fever only in the initial days. They all had a healed BCG vaccination scar on their left arm. Fine Needle Aspiration Cytology (FNAC) showed multiple epithelioid granulomas and, in some cases, frank caseation necrosis with the demonstration of tubercle bacilli using either Ziehl-Neelsen (Z-N) stain or Cartridge-based Nucleic Acid Amplification Test (CBNAAT). These cases mostly benefited from pus aspiration from the swelling site, with no definite role for oral antibiotics. Proper training of nursing or paramedical staff regarding vaccination technique and dosage, proper maternal and child healthcare, early detection of these cases with a high index of suspicion, as well as adequate knowledge of the treating doctors and parents/caregivers, may help in prompt management of these cases in future occurrences.