Staining Reaction Research Articles

The expression of repulsive guidance molecule a in the rat brain and the diffusion tensor imaging evaluation for crossed cerebellar diaschisis.

Cerebral infarction is one of the most common diseases. Diffusion tensor imaging (DTI) has been used to evaluate for crossed cerebellar diaschisis (CCD) to observe the expression of repulsive guidance molecule a (RGMa), the axonal regeneration as well as the effect on neural functional recovery in the middle cerebral artery occlusion (MCAO) rat model. To certify the expression pattern of RGMa in cerebral infarction and the mechanism of CCD to provide a new target for clinical therapy. Building the MCAO rat model, every 16 rats were randomly divided into one of six groups. The brain was scanned over the time points above and the rats were sacrificed for immunohistochemistry staining and reverse transcription polymerase chain reaction (RT-PCR) to assay the RGMa expression. The apparent diffusion coefficient (ADC) and fractional anisotropy (FA) value of MCAO rats declined and peaked at 12 h. The contralateral cerebellum had a lower parameter than the other side. The expression of RGMa kept climbing and achieved the maximum at 48 h (P < 0.05). The value of the protein in the cerebellum was higher (P < 0.05) compared with controls, especially the right cerebellum. The expression of RGMa was negative compared to the parameter of magnetic resonance imaging (MRI) and the axonal regeneration. The MRI and pathology parameters after MACO had significant differences compared to the controls, as well as the bilateral cerebellum, which provided evidence of CCD. RGMa was related to the axonal regeneration in the injured brain.

Prevalence of Extended Spectrum Beta-Lactamase Producing Bacteria in Patients with Wound Infections Attending Tertiary Hospitals in Enugu, Nigeria

Background: The problem of antibiotics usage against bacterial infection is the modifications of such antibiotics by the bacteria thereby rendering them ineffective. Extended spectrum β-lactamase (ESBL) producing bacteria invading wound infections may lead to long term hospitalization, financial burden and limited antibiotics for therapy. The goals of this study were to determine the prevalence of ESBL-producing bacteria colonization of wound infections among individuals with non-healing wounds and the antibiotic susceptibility pattern of the ESBL isolates. Methodology: The study adopted a cross-sectional research design. A total of 266 samples were collected from different wound infections which included diabetic foot ulcers, burn wounds, post-surgical wounds, non-diabetic foot ulcers, pressure ulcers, accident and open cancer wounds that met the inclusion criteria. The patients were consecutively selected. That is, any individual that has a wound and was willing to participate was selected. A structured questionnaire was administered to the patients to obtain information on demographic characteristics, antibiotic usage, and duration of infection, herbal medication, type and site of wound. Identification of bacterial isolates was done using colony/ microscopic morphology, gram stain reaction and standard biochemical tests. Antibiotic susceptibility testing was done using modified Kirby - Bauer disc diffusion method. ESBL detection was done following the recommendations by the Clinical and Laboratory Standard Institute (CLSI) which involves a 2-step approach of initially screening for ESBL producers and phenotypic confirmatory test using a combination disc test method. Molecular screening of the genes encoding for ESBLs was done at the Central Science Laboratory, University of Nigeria, Nsukka. Results: A total of 196 isolates were recovered from the wound swabs. Pseudomonas aeruginosa 56 (28.6%) was the leading organism causing wound infection followed by Staphylococcus aureus 24 (12.2%). The bacteria isolates showed that 157 (80.1%) were gram negatives as against 39 (19.9%) that were gram positive bacteria. Among the gram-negative bacteria isolates, 21.7% (34/157) were confirmed as ESBL producers. The ESBL- producers were Pseudomonas aeruginosa, Esherichia coli, Proteus vulgaris, Proteus mirabilis, Klebsiella oxytoca, Klebsiella pneumonia and Klebsiella granulomatis at frequencies of 41.2%, 20.6%,14.7%,11.8%,5.9%,2.9% and 2.9% respectively. This showed that these isolates have the ability to resist penicillins and cephalosporins of the first, second and third generations. The ESBL-producing bacteria isolated exhibited high degree of multidrug resistance especially to tetracyclines, cefuroxime, ceftriaxone, cefotaxime and ceftazidime. The antibiotics amikacin was sensitive to most of the ESBL-producing bacteria isolated. The assessment of the risk factors showed that none of the variables was statistically significant, though those risk factors were still important in evaluating wound infections. Both hospital and community acquired infections showed no statistical significance (P= 0.072) which means that both had the same degree of pathogenicity. Among the 10 samples screened for ESBLs genes namely bla SHV, bla TEM, bla CTX-M, bla GES, and bla OXA-50, only bla OXA- 50 was detected in 8 out of the ten samples. Thus, the persistent gene circulating in this region is bla OXA-50, which confers high rate of infection and persistence. Conclusion: The presence of ESBL-producing bacteria in wounds remains a challenging issue, as the majority of the patients may suffer from long term infected wounds due to treatment failure.

Gonokokkenartritis bij een 69-jarige man

Gonococcal arthritis in a 69-year-old male A 69-year-old man presented with fever and a painful and swollen left wrist and right ankle joint. Aspiration of the left wrist showed purulent fluid and culture was positive for Neisseria gonorrhoeae. The diagnosis of gonococcal arthritis, a manifestation of disseminated gonococcal infection, was made. History revealed he had unprotected sexual contacts with men. Treatment with intravenous ceftriaxon for 7 days and surgical lavage of the joints led to clinical improvement. Gonorrhea has an increasing incidence in Belgium and is now the most diagnosed sexually transmitted disease (STDs) in men. If untreated, infection with N. gonorrhoeae can lead to a disseminated gonococcal infection, sometimes resulting in serious complications such as arthritis, osteomyelitis, and meningitis. A thorough sexual history is essential in cases of acute monoarthritis or oligoarthritis. Joint aspiration with gram stain, culture and/or polymerase chain reaction (PCR) is diagnostic. Treatment of disseminated gonococcal infection consists of intravenous ceftriaxon. Surgical drainage may be necessary in case of purulent arthritis. Notification and testing of sexual partners is crucial to prevent further spread of N. gonorrhoeae. This case illustrates the importance of considering N. gonorrhoeae as a cause of septic arthritis in sexually active individuals. Early recognition and adequate treatment can prevent complications and further transmission of infections. Physicians should be alert to the increasing incidence of gonorrhoea and its potential complications.

Nocardiosis in domestic ferrets (Mustela putorius furo).

Nocardia spp are ubiquitous, gram-positive, variably acid-fast, branching and beaded filamentous, facultative intracellular bacteria that are resistant to phagocytosis and can cause localized or systemic disease in a variety of mammals, including humans, as well as in birds, fish and reptiles. Seventeen pet domestic ferrets (Mustela putorius furo) were diagnosed with nocardiosis by several methods including cytological evaluation, histopathology, Ziehl-Neelsen staining and polymerase chain reaction (PCR). All except two ferrets were 2 years old or older at the time of clinical presentation. Clinical findings included anorexia, weight loss, lymphadenomegaly, splenomegaly, masses/nodules in internal organs, hindlimb weakness or paresis, vomiting, dyspnoea, central nervous system signs, diarrhoea, abdominal pain and coughing. Anaemia, leucocytosis, neutrophilia, hypoalbuminaemia and hyperglobulinaemia were frequent. All ferrets had granulomatous or pyogranulomatous inflammation involving most commonly the lymph nodes, spleen, lungs, liver and adipose tissue. Intralesional acid-fast, branching filamentous bacilli were detected in 15 of 17 ferrets. PCR for the 16S rRNA gene of Nocardia spp was positive in 13 of 15 ferrets, including the two that were negative by acid-fast staining; of these two ferrets, one had intralesional coronavirus antigen. BLASTn analysis of eight sequences revealed six different Nocardia spp, including N.globerula, N.seriolae and N.donostiensis, that have rarely been reported to cause disease in humans and terrestrial animals. Antibiotic treatment (most commonly trimethoprim-sulfamethoxazole alone or in combination with clarithromycin and marbofloxacin) was followed by a marked improvement in several patients, although relapses were frequent. Six ferrets had concurrent neoplasia as a potential predisposing factor. Nocardiosis should be included in the differential diagnosis of granulomatous/pyogranulomatous inflammation in ferrets, particularly in those over 2 years of age.

Biodegradable PHBVHHx-PEG/Collagen Hydrogel Scaffolds for Cartilage Repair.

Recently, there has been increased attention on the treatment of cartilage repair. Overall, we constructed PHBVHHx-COL, a composite hydrogel of PHBVHHx-co-PEG and collagen, and evaluated its cartilage repair efficacy through in vitro and invivo studies using hydrogel loaded with peripheral blood-derived mesenchymal stem cells (PBMSCs). Rheological properties and compressive mechanical properties of the hydrogels were systematically evaluated. The cytocompatibility of the hydrogels was evaluated using the Cell Counting Kit-8 test, live/dead staining, scratch test, and transwell test. The effect of chondrogenic differentiation of PBMSCs on hydrogels was evaluated using immunofluorescence staining and reverse transcription-polymerase chain reaction. Furthermore, the invivo cartilage repair ability of the hydrogels was confirmed following in situ injections in rabbit chondral defect models. Finally, the induced polarization of the hydrogel scaffold on macrophages was explored by the expression of CD86 and CD206. In vitro experimental results confirmed that PHBVHHx-COL-gel led to better cell migration, proliferation, and chondrogenic differentiation than PHBVHHx-PEG and COL hydrogels. Hematoxylin and eosin staining indicated that the tissue of the repaired area in the PHBVHHx-COL group was nearly in fusion with the surrounding normal tissue and the reconstruction of subchondral bone was good. Safranin-O staining and COL-2 immunohistochemistry indicated that the tissue of the repaired area in the PHBVHHx-COL group had more cartilage-specific matrix secretion. The PHBVHHx-COL group exhibited more M2 macrophage infiltration and less M1 macrophage presentation than the other groups. This study demonstrated that PHBVHHx-COL scaffolds loaded with PBMSCs significantly promoted the repair of cartilage injury through immune regulation by M2 polarization and could be potential candidates for cartilage tissue engineering.

Low molecular weight galactomannan alleviates diarrhea induced by senna leaf in mice via intestinal barrier improvement and gut microbiota modulation.

Low molecular weight galactomannan (LMGM), a soluble dietary fibre derived from guar gum, is recognized for its prebiotic functions, including promoting the growth of beneficial intestinal bacteria and the production of short-chain fatty acids, but the mechanism of alleviating diarrhea is not fully understood. This study established an acute diarrhea mouse model using senna leaf decoction and evaluated the therapeutic effects of LMGM by monitoring diarrhea scores, loose stool prevalence, intestinal tissue pathology and gene expression, and gut microbiota composition and metabolisms. The results indicated that LMGM significantly reduced diarrhea scores and loose stool prevalence within two hours post-treatment. Hematoxylin and eosin staining and quantitative real-time polymerase chain reaction analysis revealed that LMGM improved intestinal epithelial structure and up-regulated the expression of zonula occludens 1, occludin, mucin 2, aquaporin 3, and aquaporin 4 in ileum, jejunum, and colon tissues. Moreover, LMGM increased the abundance of beneficial bacteria such as Lactobacillaceae and Lachnospiraceae, and decreased Prevotellaceae in the cecum. Furthermore, LMGM promoted short-chain fatty acid production and reduced ammonia nitrogen and skatole concentrations in the intestinal content. The study suggests that LMGM could serve as a functional prebiotic for diarrhea alleviation, potentially by enhancing the intestinal barrier, modulating water transportation, and regulating the microbiota composition.

M2-like macrophage-derived exosomes inhibit osteoclastogenesis via releasing miR-1227-5p.

Macrophages play a pivotal role in regulating inflammatory response in periodontitis, a condition characterized by excessive osteoclast differentiation. This study aimed to investigate whether exosomes derived from M2 macrophages regulate osteoclast differentiation and to identify the underlying molecular mechanisms. Exosomes were isolated from M2 macrophages and used to treat osteoclasts. Osteoclastogenesis was assessed using tartrate-resistant acid phosphatase staining and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The molecular mechanism was evaluated using microarray analysis, RT-qPCR, dual-luciferase reporter analysis, and RNA pull-down assay. The results showed that exosomes from M2 macrophages inhibited receptor activator of nuclear factor κ-B ligand (RANKL)-induced osteoclast differentiation. Additionally, miR-1227-5p expression in osteoclasts was increased after treatment with exosomes, and inhibition of miR-1227-5p counteracted the suppressive effects of exosomes on osteoclastogenesis. Moreover, OSCAR is a target of miR-1227-5p. In conclusion, exosomal miR-1227-5p suppresses osteoclast differentiation, potentially via targeting OSCAR. These findings provide new insights into the pathogenesis of periodontitis.

Enhancement of Differentiation and Mineralization of Human Dental Pulp Stem Cells via TGF-β Signaling in Low-Level Laser Therapy Using Er:YAG Lasers.

Low-level laser therapy (LLLT) using an erbium-doped yttrium aluminum garnet (Er:YAG) laser provides a non-invasive approach applicable to various dental treatments. Here, we investigated the effects of Er:YAG laser irradiation on human dental pulp stem cells (hDPSCs) in an in vitro experiment. The hDPSCs were categorized into four groups: laser-irradiated with activators (VLT: activated vitamin D3, bone morphogenetic protein receptor inhibitor, and transforming growth factor-beta (TGF-β)) (LLLT(+)VLT), laser-irradiated without activators (LLLT(+)-only), non-irradiated with activators (LLLT(-)VLT), and non-irradiated without activators (control). Cell proliferation, hard tissue differentiation, TGF-β signaling pathway activity, mineralization induction, and gene expression levels were assessed using several approaches, including cell proliferation assays, ALP assays, western blotting, Alizarin Red S staining, X-ray diffraction, and quantitative polymerase chain reaction. Cell proliferation was similar between the LLLT(+)-only and control groups. The ALP activity was significantly higher in LLLT(+)VLT group than in LLLT(-)VLT group (p < 0.05); however, it was suppressed by TGF-β signaling inhibitors. Western blotting showed enhanced SMAD3 phosphorylation in the LLLT(+)VLT group. The mineralization nodules and mRNA levels of matrix vesicle marker genes were significantly higher in LLLT(+)VLT group, and the nodules were partially composed of hydroxyapatite. The hard tissue formation marker gene expression in LLLT(+)VLT group was significantly higher (p < 0.05) than that in the LLLT(+)-only and control groups; however, it was unchanged or suppressed compared with that in LLLT(-)VLT group. LLLT using an Er:YAG laser, combined with VLT, may promote the differentiation of hDPSCs into hard tissue-forming cells and enhance mineralization.

KLF10 Induced by Hypothermic Machine Perfusion Alleviates Renal Inflammation Through BIRC2/Noncanonical NF-κB Pathway

Background. Hypothermic machine perfusion (HMP) is becoming the main preservation method for donation after circulatory death (DCD) kidneys. It can provide continuous flow and form shear stress (SS) upon endothelial cells (ECs), thereby regulating EC injury. Krüppel-like factor 10 (KLF10) has been shown to lessen vascular damage. However, how SS and KLF10 impact HMP-regulated injury is unclear. Methods. In this study, we investigated the influences of KLF10 on HMP in animal models and human renal biopsy and explored how SS affected KLF10 expression in a parallel-plate flow chamber system. Chromatin Immunoprecipitation sequencing and luciferase assay were performed to seek the target genes of KLF10. The influences of KLF10 on HMP-regulated injury were investigated by transfecting si-KLF10 adeno-associated virus serotype 9 into rat kidneys. The molecular expression was examined using immunofluorescence staining, Western blotting, and quantitative polymerase chain reaction. Results. Our results show KLF10 expression was augmented in human, rabbit, and rat DCD kidneys after HMP. HMP improved ECs and tubule injury and attenuated inflammation; however, the knockdown of KLF10 reversed this effect. SS regulated KLF10 expression in ECs by affecting F-actin, and KLF10 could maintain ECs homeostasis. Chromatin Immunoprecipitation sequencing and luciferase assay revealed that baculoviral inhibitor of apoptosis protein repeat-containing 2 (BIRC2 ) is a target gene of KLF10. Furthermore, BIRC2 linked to nuclear factor kappa B (NF-κB)-inducing kinase, induced NF-κB)-inducing kinase ubiquitination, and resulted in inhibiting the noncanonical NF-κB pathway. Conclusions. SS can mediate KLF10 expression, whereas HMP can protect against warm ischemic injury by reducing inflammation via KLF10/BIRC2/noncanonical NF-κB pathway. Therefore, KLF10 might be a novel target for improving DCD kidney quality.

Impaired Wnt/Planar Cell Polarity Signaling in Yellow Nail Syndrome.

Yellow nail syndrome (YNS) is a rare disorder characterized by a triad of yellow dystrophic nails, lymphedema, and chronic lung disease. Most patients present in adulthood, with only a few congenital or familial cases described. The cause of YNS remains largely unknown, although defects in lymphatic vessel development are suggested to play a significant role. To elucidate the genetic mechanisms underlying YNS. Analysis of genetic sequencing data and gene and protein expression studies. A tertiary care academic medical center. 6 patients with congenital YNS (cYNS) and 5 with sporadic YNS (sYNS). Exome and genome sequencing were used to detect disease-causing variants, complemented by RNA analyses for intronic variants. Protein and gene expressions were studied by immunofluorescence staining and real-time reverse transcriptase quantitative polymerase chain reaction analyses. Biallelic variants in CELSR1 (n = 5) or likely FZD6 (n = 1), both core molecules in the Wnt/planar cell polarity (PCP) pathway, were identified in all patients with cYNS; none of the patients with sYNS had candidate genetic variants. Immunofluorescence staining showed that CELSR1 colocalizes with lymphatic vessels in the skin but not in the lungs or the intestine. Moreover, levels of CELSR1 and FZD6 proteins were negligible to zero in patient tissues (n = 2) compared with control tissues. Gene expression of Wnt/PCP-related genes was reduced in patients with cYNS (n = 3), and patients with sYNS (n = 4) showed milder gene expression impairments. Small cohort size and limited sample availability. Defects in PCP organization may play a major role in the pathogenesis of YNS. To the authors' knowledge, this is the first demonstration of a mechanism explaining YNS development, mainly in its congenital form but also in patients with sporadic disease. The Prof. Baum Research Fund of Israel Lung Association.

Withdrawal Effects on Antioxidative and Histochemical Deficits in Acetaminophen Induced Neurotoxicity in Cerebellar Cortex of Adult Wistar Rats

Introduction: Neurotoxicity refers to the harmful effects of chemical, biological, or physical agents on the nervous system. Acetaminophen, a common pain reliever whose abuse has been linked to neurotoxicity at high doses, causing oxidative stress and neuronal damage. Aim: The present study aimed to assess the withdrawal effects of Acetaminophen use on the Cerebellar cortex of adult wistar rats. Methodology: Thirty (30) Adult wistar rats weighing 200±50g were assigned into three groups (n=10) of Control (C), Treatment 1(T1) and Treatment 2 (T2). The control group C received distilled water, while Treatment groups T1 and T2 received 200mg/kg of acetaminophen for 4 weeks. However, treatment group T2 were allowed a 2-week acetaminophen withdrawal period. At the end of exposure, all animals were sacrificed via cervical dislocation and eventual removal of cerebellar specimens which were processed for some Neurohistochemical staining reactions as well as Biochemical Quantifications of LDH enzyme and Antioxidative stress markers (SOD &amp; CAT). Quantitative analyses of all data obtained including Body weight, brain weight and Cerebellum weight were analysed using GraphPad® (version 8) and plotted using ANOVA followed by Tukey's multiple comparisons test. Significance was set at p&lt;0.05* (95% confidence interval). Result: Results showed a significant increase in body weight (P=.05) and a decrease in cerebellum weight (P=.01) in both acetaminophen groups. SOD and CAT activity decreased significantly (P=.01) in the T1 groups while T2 groups shows a significant increase, LDH activities decreased significantly (P=.01) in both groups. The Neurohistochemical findings showed severe degeneration, loss of the Purkinje neurons and poorly differentiated DNA on the T1 group compared to control group C while the withdrawal group showed onset of regenerative changes in neurons improved differentiation in DNA stained. Conclusion: The study's findings suggest that long-term acetaminophen use can lead to neurotoxic effects on the cerebellum, but partial recovery is possible upon withdrawal.

Masticatory Stress Maintains Mucosal Homeostasis via M2 Polarization.

In addition to breaking down food, mastication plays regulatory roles in tissue homeostasis. During mastication, the oral mucosa is subjected to masticatory stress, and the maintenance between mucosal damage and repair is a key process. Despite rapid healing in the oral mucosa during chewing, the molecular mechanisms underlying repair remain unclear. In this study, we investigated the impact of masticatory stress on masticatory mucosal wound healing. Our data showed that reduced masticatory stress on the oral mucosa in mice fed a soft food diet resulted in decelerated hard palate mucosal wound healing and decreased numbers of Ki67-positive cells as compared with the hard food diet group. An RNA sequencing analysis revealed lower expression levels of the mechanosensitive gene Piezo1 in the hard palate mucosa, as well as lower levels of transforming growth factor β1 (Tgf-β1) and Tgf-β receptor 2 (Tgf-βr2), in the soft food diet group than the hard food diet group. Immunofluorescence staining, flow cytometry, and polymerase chain reaction analyses demonstrated that masticatory stress induced M2 polarization of macrophages surrounding the wound in the hard food diet group, leading to increased Tgf-β1 secretion. The specific deletion of Piezo1 in macrophages (Piezo1DLysm) attenuated masticatory stress-induced accelerated healing in mice. These findings reveal the crucial role of masticatory stress-induced Piezo1 expression in tissue repair, potentially influencing M2 polarization of mucosal macrophages and Tgf-β1 secretion. These findings underscore the pivotal role of physical stimulation in the immune response and tissue repair and may provide important insights into therapeutic interventions for tissue repair.

Prenatal Histomorphological and Histochemical Study of Stomach in Indigenous Rabbit (Oryctolagus cuniculus)

The present study was designed to investigate the normal histological features of the stomach at the prenatal stage from ten healthy pregnant rabbits at the third period of pregnancy. The stomach appeared as J-shaped situated at the left part of the abdominal cavity. It found in the front part entirely within the rib. The non-glandular region act as a reservoir, the septa prevented ingesta reflux into the esophagus. Histologically the stomach in both periods composed of four tunics', which were mucosa, tunica submucosa, tunica muscularis and tunica serosa. The stomach also has different regions cardiac, fundic and pyloric regions, glands in the cardiac region were coiled branched tubular gland and almost present of the mucous cells with a few chief cells. In the fundic region the glands in this part were long and simple tubular. There are four secretory cell types: mucous cells, chief cells, parietal cells and endocrine cells, glands in the pyloric region were branched coiled tubular glands which opened at the pits of the pyloric part. The mucous-secreting cell always persists, the pyloric sphincter muscle found as thickened muscle in the pyloric duodenal junction. With PAS stain reaction gave a positive reaction for Cardiac and pyloric glands secretion which indicates the present of neutral mucopolysaccharides. In conclusion, it was found that the histomorphological features of the stomach in prenatal period embryos at the third stage of pregnancy of rabbit it was changed according to the age and food intake after birth.

Identification and verification of feature biomarkers associated with choline metabolism in sepsis-induced cardiomyopathy.

Background: Sepsis-induced cardiomyopathy (SIC), one of the most common complications of sepsis, seriously affects the prognosis of critically ill patients. Choline metabolism is an important biological process in the organism, and the mechanism of its interaction with SIC is unclear. The aim of this study was to reveal the choline metabolism genes (CMGs) associated with SIC and to provide effective targets for the treatment of SIC.Methods: Through a comprehensive analysis of the microarray dataset GSE79962 (comprising 20 SIC patients and 11 healthy controls) from the GEO database, suspected co-expression modules and differentially expressed genes (DEGs) in SIC were identified. Hub CMGs were obtained by intersecting choline metabolism database with DEGs and key model genes. Afterward, hub CMGs most significantly involved in prognosis were further analyzed for the verification of major pathways of enrichment analysis. Finally, the expression of hub CMGs in in vivo and in vitro SIC model was verified by immunohistochemistry staining and quantitative real-time polymerase chain reaction analysis (qPCR).Results: WGCNA identified 1 hub gene panel and 3867 hub genes, which were intersected with DEGs and CMGs to obtain the same 3 hub CMGs:HIF-1α, DGKD and PIK3R1. Only HIF-1α shows significant association with mortality (P = 0.009). Subsequent differential analysis based on the high and low HIF-1α expression yielded 63 DEGs and then they were uploaded into Cytoscape software to construct a protein-protein interaction (PPI) network and 6 hub genes with the highest priority were obtained (CISH, THBS1, IMP1, MYC, SOCS3 and VCAN). Finally, a multifactorial COX analysis revealed a significant correlation between HIF-1α and survival in SIC patients, which was further validated by in vitro and in vivo experiments.Conclusion: Our findings will provide new insights into the pathogenesis of SIC, and HIF-1α may have important applications as a potential biomarker for early detection and therapeutic intervention in SIC.

PTPMT1 inhibition induces apoptosis and growth arrest of human SCLC cells by disrupting mitochondrial metabolism.

Many cancer cells exhibit aberrant metabolic reprogramming through abnormal mitochondrial respiration. Protein tyrosine phosphatase mitochondrial 1 (PTPMT1) is a protein tyrosine phosphatase localized to the mitochondria and linked to mitochondrial respiration. However, the expression and role of PTPMT1 in regulating the biological characteristics of small cell lung cancer (SCLC) has not yet been explored. The aim of this study was to evaluate the role of PTPMT1 on SCLC cell survival and mitochondrial function. SCLC and adjacent normal tissues were obtained from surgery. The expression level of PTPMT1 in the SCLC tissues and cell lines was determined by immunohistochemical staining, western blot, and quantitative real-time polymerase chain reaction (qRT-PCR). PTPMT1 knockdown was induced by lentivirus-mediated short-hairpin RNA (shRNA) transduction and PTPMT1 inhibition (alexidine dihydrochloride). The biological characteristics of the cells were measured by cell counting kit 8 (CCK-8), colony formation assay, and cell migration assay. The mitochondrial function of the cells was measured by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) staining. The H69 cells were treated with alexidine dihydrochloride, after which transcriptome sequencing and an untargeted metabolomic analysis were performed. The transcriptome differentially expressed genes were measured by qRT-PCR. PTPMT1 was upregulated in the SCLC tissues compared to the adjacent normal tissues. PTPMT1 inhibition by lentiviral shRNA transduction or specific inhibition resulted in significant growth arrest and apoptosis. The transcriptome sequencing analysis revealed that pathways related to the respiration chain and mitochondrial member protein were disrupted. Several mitochondrial metabolism-related genes, such as FGF21, GDF-15, APLN, and MT-DN6, were dysregulated. Further, PTPMT1 inhibition was found to downregulate Glut expression and disturb mitochondrial function. PTPMT1 was shown to play a critical role in the survival and growth of SCLC cells, and may become a potential therapeutic target.

Preparation and Characterization of Essential oil from Lavandula spica Plant and its Antimicrobial Activity against Pseudomonas aeruginosa and Staphylococcus aureus

The biological properties of herbs and essential oils (EOs), such as their antibacterial, analgesic, anti-inflammatory, antioxidant, and anticancer characteristics, make them widely used in a variety of fields. This research aims to assess the antibacterial efficacy of lavender oil against Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa). The essential oil from Lavandula spica was extracted via water distillation and characterized by using Gas Chromatography-Mass Spectrometry (GC-MS) and Fourier Transform Infrared Spectroscopy (FTIR). Bacterial strains were isolated from burn wounds, confirmed by polymerase chain reaction technique, and were tested using disc diffusion method and Minimum Inhibitory Concentration (MIC) calculations. The study identified 28 components in the EO, with monoterpenes being the predominant. Out of 150 samples, 56 (45.52%) were positive for P. aeruginosa and 67 (54.47%) for S. aureus by using standard microbiological techniques including Gram staining, biochemical tests and Polymerase chain reaction technique. P. aeruginosa showed high resistance to cefazolin (100%) and levofloxacin (83.3%), while S. aureus was highly resistant to cefoxitin, piperacillin, amoxicillin/clavulanic acid, and ampicillin/sulbactam. The zone of inhibition and MIC for EO against P. aeruginosa were 9.910±0.866 and 2.376±0.352 while for S. aureus were 10.597±0.818 and 0.894±0.073 respectively with significance levels of p>0.05 and p<0.01. The study concluded that L. spica EO shows promising antimicrobial activity, particularly against Gram-positive bacteria suggesting its potential for further research and antimicrobial use.

Escherichia coli in betel leaves: prevalence, virulence characterization and antibiogram

Escherichia coli is a significant foodborne pathogen, frequently linked to hemorrhagic diarrhea, especially in developing nations, where it presents considerable public health concerns. This study was conducted to examine the virulence gene profiles and antibiotic resistance patterns of E. coli strains isolated from piper betel leaves. A total of 100 betel leaf samples, including fresh (n = 60) and ready-to-eat (n = 40) specimens, were collected and tested for the presence of E. coli using standard diagnostic techniques, such as selective culture methods, staining, latex agglutination, and polymerase chain reaction (PCR) assays. Further, the identified E. coli isolates underwent PCR-based testing for virulence genes and disk diffusion assays to assess antibiotic susceptibility. Among the 100 samples screened, 4% (n = 4/100; 95% CI: 1.57–9.84; P = 0.1126) were identified as E. coli O157, and 33% (n = 33/100; 95% CI: 24.56–42.69; P = 0.4011) were classified as non-O157 isolates. The virulence gene stx1 was found in 10.81% of isolates, while stx2, eaeA, and hlyA genes were not detected in any samples. Antibiotic resistance analysis showed that all isolates (100%, 37/37) were resistant to amoxicillin and erythromycin, with 75.68% (28/37) demonstrating resistance to tetracycline. Notably, all isolates were fully susceptible to ceftriaxone and ciprofloxacin. A majority (72.97%, 27/37) of isolates were sensitive to streptomycin, and 67.57% (25/37) were sensitive to gentamicin. Additionally, 86.48% of the E. coli isolates exhibited multidrug resistance (MDR), showing 10 resistance patterns, including 8 MDR patterns. The most common MDR pattern was AMX-TE-E, observed in 56.76% (21/37) of isolates. One isolate demonstrated resistance to six of the eight tested antibiotics across four distinct classes, with the resistance pattern AMX-TE-GEN-S-E-AZM. The MAR indices for E. coli isolates ranged between 0.25 and 0.75. These findings highlight the significant threat posed to global public health by multidrug-resistant shiga toxin-producing E. coli found on piper betel leaves in urban environments. Asian Australas. J. Biosci. Biotechnol. 2024, 9(3), 33-44

Clinical characteristics, progression patterns and treatment outcomes in microsporidial keratoconjunctivitis: a prospective study in Thailand.

To assess the clinical characteristics, progression patterns, and treatment outcomes of microbiologically confirmed microsporidial keratoconjunctivitis (MKC). This prospective cross-sectional study included patients with superficial punctate epithelial keratitis clinically suspected of MKC. Comprehensive slit-lamp examinations were conducted, and corneal scraping was performed for Gram-chromotrope staining and polymerase chain reaction (PCR) analysis. A standardized questionnaire gathered demographic data, clinical features, and risk behaviors. Treatment regimens and corneal findings, including medication and frequency, were documented at each visit. PCR confirmed the diagnosis of MKC in 96 out of 117 eyes (82.1%), identifying Vittaforma corneae in 93.7% of cases, Microsporidium sp. in 4.2%, and Encephalitozoon hellem in 2.1%. All cases exhibited similar characteristics and pattern of progression, including elevated epithelial lesions in diffused distribution (34.1%), typical target lesions (31.3%), and subepithelial infiltrations (41.7%). Treatment with topical moxifloxacin, with or without oral albendazole, followed by topical steroids for subepithelial infiltrates, led to clinical improvement within approximately two weeks, with 52% of patients achieving complete recovery. This study identifies key clinical features and progression patterns in MKC. Topical fluoroquinolone monotherapy, or its combination with topical steroids or oral albendazole, results in favorable visual outcomes without corneal scarring. These insights may inform and enhance clinical management of MKC.

Histopathological characterization of mandibular condyles in four temporomandibular joint osteoarthritis mouse models

ObjectiveTemporomandibular joint osteoarthritis (TMJOA) has been modeled in different ways with a lack of uniformity. We aimed to investigate four TMJOA mouse models and assess histopathological changes in condyles, which could assist in the selection of animal models in further TMJOA-related studies. DesignFour TMJOA mouse models were established, including unilateral hyperocclusion, discectomy, monosodium iodoacetate injection and aged model. Bilateral condyles were collected at different time points. The condylar alterations were analyzed by Micro-CT, Hematoxylin and eosin staining, Toluidine blue staining, Safranin O staining, Trap staining, immunofluorescence staining, immunohistochemistry staining and quantitative polymerase chain reaction. ResultsRadiographic and histopathological analysis indicated that all four methods could cause condylar degeneration successfully. Differences in morphologic and histologic changes were found among four models. The hyperocclusion model was time-dependent and the lesions got worse over time. Discectomy model presented obvious damage of cartilage and subchondral bone. Injected model showed severe inflammation and chondrocyte hypertrophy. The aged model was characterized by decreased of proteoglycan and osteolysis. ConclusionsThe four methods had different characteristics and applicability. The harvest time affected the degree of cartilage degradation. Hyperocclusion was suited to explore the early-stage of TMJOA. Discectomy present advantages in investigating the long-term restoration of cartilage and subchondral bone. Monosodium iodoacetate-injection was appropriate for screening the agents for inflammatory relief. The aged model more naturally facilitated discovering underlying mechanisms in primary TMJOA. Unilateral modeling methods could initiate contralateral condylar alterations. The TMJOA models should be selected based on experimental requirements and applicability of each model.

Use of the immunohistochemical marker HBME-1 to optimize the diagnosis of follicular carcinomas

Background. In clinical practice, there are often patients with a diagnosis of benign follicular adenoma (FA) and follicular tumor with an uncertain malignant potential, whose diagnoses must be changed to malignant processes due to relapse or metastasis observed over time. The purpose of the study was to optimize the differential diagnosis of FA and follicular thyroid carcinoma (FTC) using the immunohistochemical (IHC) marker HBME-1 on the histological material of patients who were operated for nodular goiter with a cytological conclusion corresponding to the Bethesda categories III–V. Materials and methods. One hundred and twenty-four patients underwent surgery for follicular nodular neoplasms with a cytological conclusion according to Bethesda Gray zone and had a histological diagnosis of FTC in 23 ca­ses (18.55 %) and FA in 101 cases (81.45 %). IHC analysis was performed using mouse monoclonal antibodies against human HBME-1 (Bio SB, USA). Evaluation criteria corresponded to the strength of staining (from 0 to 3). Results. According to the intensity of IHC staining, there was an increase in the diagnosis of FTC (p &lt; 0.05): 0 — 0 %, 1 — 12.7 %, 2 — 25 %, 3 — 36.4 %. Histopathological findings were reexamined for FA that had the strongest staining reaction: in 3 (2.97 %) cases, the diagnosis was changed to FTC. Accor­dingly, the total number of detected FTC increased to 26 (20.96 %). These cases showed a good positive result regarding IHC study with HMBE-1 on FA preparations for a differential diagnosis between FA and FTC. Conclusions. HBME-1 can serve as an IHC marker for the differential diagnosis of FA and FTC in cases considered as FA, as after the study, the diagnosis was changed to FTC in 2.97 % of patients.