Adhesion Genes Research Articles

Changes of osteodifferentiation potential of MSC-AT during in vitro co-cultivation with heparin

The aim of this study was to investigate the effect of heparin at a concentration of 1 IU/mL on changes in the osteodifferentiation potential of MSC from human adipose tissue under in vitro cocultivation.Materials and methods. Assessment of the phenotypic profile of MSC from human adipose tissue during cultivation in the presence/absence of heparin was performed by the flow cytometry method using the appropriate dyes according to the manufacturer’s protocol on a MACS Quant flow cytometer after 14 days of cultivation. To evaluate the migration and proliferation potential of MSCs in the presence of heparin, we were using a continuous monitoring electrode system, xCELLigence ® RTCA DP. After cultivation MSCs with heparin for 14 days, the intracellular expression of osteodifferentiation genes was evaluated by real-time PCR. In addition, the differentiation profile of MSCs from human adipose tissue cultured with heparin was evaluated by cytological staining with alizarin red to detect islands of mineralization after 21 days of cultivation. In addition, the amount of growth factors, chemokines, molecules with pro- and anti-inflammatory activity was estimated in the supernatants of the 14-day cultures.Results. There was a significant decrease ( compared with the control group of the study) in the number of cells with stem markers (CD73, CD90, CD105) on the cell surface of the culture in the MSC + heparin model; increase in proliferative and decrease in migratory activity of MSCs during co-cultivation with heparin; increased levels of relative mRNA expression of genes for osteodifferentiation (ALPL, RUNX2, BMP2, BMP6) and cell adhesion (CD49d); increase in mineralization area in the study model in the presence of heparin after 21 days of cultivation. There was a tendency to increase secretion of growth factor VEGF and pro-inflammatory factor IL -6 in the MSC + heparin model.Conclusion. The obtained results may serve as a basis for the development of new therapeutic tactics for the treatment of surgical patients undergoing osteosynthesis operations with a high risk of thrombosis.

DDDR-18. SINGLE-CELL FUNCTIONAL GENOMIC ANALYSIS OF PHARMACOLOGIC RESPONSES IN NF1 MUTANT GLIOBLASTOMA REVEALS DISTINCT MECHANISMS OF ACTION FOR MEK AND SHP2 INHIBITION

Abstract BACKGROUND Glioblastomas harbor recurrent mutations in NF1 that activate Ras/Raf/MEK signaling, motivating pharmacologic approaches targeting downstream (MEK) or upstream (SHP2) of Ras signaling. However, the mechanisms underlying therapeutic response remain unclear. Here, we integrate genome wide CRISPRi screens and single cell sequencing in cell lines and mouse xenografts to identify functional genomic mechanisms underlying responses to the MEK inhibitor selumetinib or the SHP2 inhibitor RMC-4550. METHODS Human NF1 mutant GBM43 cells expressing CRISPRi machinery were used for CRISPRi screens with selumetinib or RMC-4550. Intracranial xenografts were established with GBM43 cells and treated with selumetinib or RMC-4550. Tumor growth kinetics were measured with bioluminescent imaging. RESULTS Genome-wide CRISPRi screens with selumetinib or RMC-4550 in NF1 mutant human GBM43 cells identified 39 genes (14 sensitivity, 25 resistance) mediating selumetinib response and 327 genes (51 sensitivity, 276 resistance) mediating RMC-4550 response. While both selumetinib and RMC-4550 hits were enriched for components of Ras/Raf/MEK, hits sensitizing RMC-4550 response were uniquely enriched for PI3K signaling and focal adhesion components while hits mediating resistance were enriched for cell cycle genes. In NF1 mutant GBM43, RMC-4550 significantly reduced tumor volume (p=0.02) while selumetinib did not. Single cell RNA-sequencing (scRNA-seq) analysis of 90,782 tumor cells from NF1 mutant GBM43 xenografts treated with selumetinib or RMC-4550 identified 12 clusters across all samples. RMC-4550, but not selumetinib, resulted in significant depletion of cells expressing an epithelial-mesenchymal transition (EMT) signature marked by expression of focal adhesion genes mediating RMC-4550 sensitivity, suggesting this signature comprises a distinct mechanism of action in response to RMC-4550 but not selumetinib. CONCLUSIONS In NF1 mutant glioblastoma in vitro and in vivo models, cellular response to both upstream (SHP2) and downstream (MEK) inhibition are mediated by Ras/Raf/MEK signaling while RMC-4550 alone uniquely regulates a focal adhesion network associated with EMT and demonstrates improved efficacy in vivo.

Single-cell transcriptomics uncover hub genes and cell–cell crosstalk in patients with hypertensive nephropathy

Hypertensive nephropathy (HTN) is one of the leading causes of end-stage renal disease, yet the molecular mechanisms are still unknown. To explore novel mechanisms and gene targets for HTN, the gene expression profiles of renal biopsy samples obtained from 2 healthy living donor controls and 5 HTN patients were determined by single-cell RNA sequencing. Key hub genes expression were validated by the Nephroseq v5 platform. The HTN endothelium upregulated cellular adhesion genes (ICAM2 and CEACAM1), inflammatory genes (ETS2 and IFI6) and apoptosis related genes (CNN3). Proximal tubules in HTN highly expressed hub genes including BBOX1, TPM1, TMSB10, SDC4, and NUP58, which might be potential novel targets for proximal tubular injury. The upregulated genes in tubules of HTN were mainly participating in inflammatory signatures including IFN-γ signature, NF-κB signaling, IL-12 signaling and Wnt signaling pathway. Receptor-ligand interaction analysis indicated potential cell–cell crosstalk between endothelial cells or mesangial cells with other renal resident cells in HTN. Together, our data identify a distinct cell-specific gene expression profile, pathogenic inflammatory signaling and potential cell–cell communications between endothelial cells or mesangial cells with other renal resident cells in HTN. These findings may provide a promising novel landscape for mechanisms and treatment of human HTN.

Comparison of phylogenetic and virulence factors between Escherichia coli isolated from biliary tract infections and uropathogenic Escherichia coli

Escherichia coli is a gram-negative intestinal commensal that can also cause various infections, including urinary tract infections, biliary tract infections, neonatal meningitis, and septicemia. Although the characteristics of uropathogenic E. coli and the mechanisms of urinary tract infection have been well studied, the genetic distinctions among E. coli isolates from different types of infections have not yet been determined. This study compared the phylogenetic and virulence factors of E. coli isolates from bacteremic biliary tract infections with those from bacteremic urinary tract infections. The phylogenetic B2 group was the most prevalent in both pathogenic groups (68 % in biliary pathogenic isolates and 85 % in uropathogenic isolates), but the frequency pattern of the phylogenetic group was different. Half of the uropathogenic isolates belonged to ST95 and ST131 (51 %). Among the biliary pathogenic isolates, ST131 was the most prevalent, while the remaining half belonged to other STs outside the four major STs. The frequency of some virulence factors, such as papC, papG2, hlyA, tcpC, fyuA, kpsMT2, sat, and traT, was lower in the biliary pathogenic isolates than in the uropathogenic isolates. The frequency of phylogenetic groups and STs in MLST differed between E. coli isolates from bacteremic biliary tract infections and urinary tract infections. Additionally, some virulence factors, including adhesion and toxin gene groups, showed lower frequencies in the biliary pathogenic group than in the uropathogenic group. Studying the differences in E. coli pathovars from different infection sites is important for developing pathovar-specific targeted therapies such as vaccine therapy.

Whole-genome sequencing reveals virulence and antibiotic resistance determinants in Enterococcus faecium strains isolated from the dairy industry in Mexico

Enterococcus faecium is a part of the native microbiota in fermented and dairy products. However, the acquisition of virulence determinants turns this bacterium into a pathogen. To evaluate the genetic relatedness and virulence profile, we performed whole-genome sequencing of four E. faecium strains isolated from the dairy industry and compared them with clinical and environmental isolates reported in Mexico. Our findings revealed close genetic relationships between certain dairy-associated and clinical isolates, with shared antibiotic-resistance determinants and prophages. Notably, the tetL, tetM, lnuA, and lsaE genes, associated with tetracycline and lincosamide resistance, were present in the two dairy-related strains. Clinical isolates exhibited a higher prevalence of adhesion and biofilm-related genes, including acm, ecbA, fss3, and sgrA. Additionally, the identification of prophages in dairy-related strains suggests their potential for genetic exchange. Thus, it is crucial to assess the risk posed by E. faecium as a potential pathogen in the dairy plant environment.

Investigation of Virulence Factors, Phylogenetic Group Features, and the Presence of ST131 Clone in Escherichia coli Isolates, a Urinary Tract Infection Agent in Children

Urinary tract infection (UTI) caused by Escherichia coli is a significant health issue in children. Today especially E.coli O25b/ST131, defined as a pandemic clone, is a serious public health problem due to its high virulence and antimicrobial resistance rates. In this study, a total of 200 (100 first and 100 recurrent UTI-causing) E.coli isolates from urine samples sent to the Ankara University School of Medicine Cebeci Training and Research Hospital Central Laboratory between January and September 2021 with the preliminary diagnosis of UTI in pediatric patients aged three to 18 years were analyzed for antimicrobial resistance rates, phylogenetic group distributions, virulence factor frequencies and whether they belong to the O25b/ST131 clone. It is aimed in this study that, the obtained data will shed light on new studies for diagnosis, treatment and prophylaxis options that can be developed for more effective UTI management by contributing to the surveillance studies in our country. Antimicrobial susceptibility of E.coli isolates identified by conventional methods was evaluated by Kirby-Bauer disc diffusion method and extended spectrum beta-lactamase (ESBL) production was evaluated by double disc synergy test. Polymerase chain reaction (PCR) was used for the investigation of phylogenetic grouping, the O25b/ST131 clone, virulence genes and the molecular level classification of the isolates detected as uropathogenic E.coli (UPEC). Pulsed-field gel electrophoresis (PFGE) was performed with the isolates collected at different times from the same patient. The highest antimicrobial resistance rates observed were against ampicillin (n= 100, 50%), cefazolin (n= 99, 49.5%), trimethoprim-sulfamethoxazole (n= 55, 27.5%), amoxicillin-clavulanic acid (n= 43, 21.5%) and cefotaxime (n= 43, 21.5%). In recurrent UTI agents, resistance rates were higher for cefotaxime (n= 29, 29%), trimethoprim-sulfamethoxazole (n= 35, 35%) and cefepime (n= 25, 25%) and in O25b/ST131 isolates (n= 67) the rates were higher for amikacin (n= 3, 4.5%), gentamicin (n= 10, 14.9%) and ciprofloxacin (n= 17, 25.4%) when compared to the first UTI agents and non-O25b/ ST131 isolates (p< 0.05). It was found that 29% (n = 58) of the isolates were multidrug resistant (MDR) and 19% (n = 38) produced ESBL.The rate of recurrent UTI agents was found to be higher among ESBL producing isolates and/or MDR isolates (n= 36, 62% and n= 27, 71%, respectively, p< 0.05). It was found that 45.5% (n= 91) of the isolates were in D, 37.5% (n= 75) in B2, 12.5% (n= 25) in A, and 4.5% (n= 9) in B1 phylogenetic groups and isolates belonging to B2 and D phylogenetic groups had higher antibiotic resistance rates and carried more virulence genes (p< 0.05). Of the isolates, 33.5% (n= 67) were found to belong to the O25b/ST131 clone, no significant difference was found between the O25b/ST131 rates among the first and recurrent UTI agents (p> 0.05). It was determined that the isolates most frequently carry virulence genes for adhesion [fimH 97% (n= 194), papA 57% (n= 114), yfcV 49.5% (n= 99)] and iron uptake systems [fyuA 85.5% (n= 171), chuA 78% (n= 156), iutA 73% (n= 146)]. All virulence factors were detected more frequently in isolates belonging to the O25b/ST131 clone (p< 0.05). Of the isolates, 97% (n= 65) belonging to the O25b/ST131 clone and 27.1% (n= 36) not belonging to this clone were defined as UPEC with molecular analysis (p< 0.0001). Thirty-three isolates belonging to 15 patients were evaluated with PFGE, and it was observed that the latter isolate and the first isolate of eight patients (53%) had the same band profile. Focusing on surveillance, diagnostic testing, treatment algorithms, and preventive measures for E.coli and especially for ST131 clone, which is frequently observed as causative agent in childhood UTIs, will help to manage challenging E.coli infections.

MrCreC, a carbon catabolite repression gene, is required for the growth, conidiation, stress tolerance and virulence of Metarhizium robertsii

As a key component of carbon source metabolism in fungi, CreC WD40 repeat protein is regulated by carbon catabolite repression (CCR). However, the understanding of the functions of CreC in entomopathogenic fungi is currently limited. Here, CreC in Metarhizium robertsii (MrCreC) was identified, and its roles in fungal development, conidiation, environmental stress response, and insecticidal virulence were explored. MrCreC is localized to cytoplasm, and MrCreC deletion affects fungal growth on various nutrients. Compared to the wild type, the sporulation of ΔMrCreC strain was significantly decreased by 60.3%. Further qPCR analysis found that deletion of MrCreC resulted in repression of sporulation-related genes such as AbaA, FlbA, Flbc, MedA, FlbD, FluG, and wetA. In addition, MrCreC loss did not alter heat stress tolerance but resulted in enhanced tolerance to UV-B. Interestingly, bioassays showed that the virulence following exposures to topical applications or injection of conidial suspensions of both infection and injection was impaired compared with that of the wild type. Further analysis showed that the adhesion and cuticle penetration genes in ΔMrCreC was down-regulated during infection, and the appressorial formation rate was significantly reduced. A deletion of MrCreC significantly also reduced immune escape and nutrient utilization genes in insect hemocoel. In conclusion, MrCreC is involved in the growth, development and virulence of M. robertsii. These findings advance our understanding of the function of CCR pathway-related genes.

Association between bacterial pathogenicity, endometrial histological changes and clinical prognosis in canine pyometra

Despite the high frequency and clinical relevance of canine pyometra, its pathogenesis remains poorly understood. In this study, the clinical data, histopathological alterations, and microbiological findings of 39 dogs with pyometra were analyzed to assess possible associations. The mean age of the affected animals was 9.6 ± 3.8 years; 76.3 % (29/38) had open cervix pyometra, 88 % (22/25) had tachypnea, 71 % (27/38) had anorexia, and 60.5 % (23/38) had leukocytosis. Histopathological analysis revealed that 66.5 % (26/39) of the uteri had a high degree of inflammation (score 4). Third-degree hyperplasia of the endometrial epithelium (72 %, 28/39) and intralesional or intrauterine bacteria (66.5 %, 26/39) were identified in most animals. Bacterial isolates were obtained from 82 % (32/39) of the uterine contents and five bacterial species were identified. Escherichia coli, classified in phylogroup B2, is associated with virulent adhesion genes (fimH, focG, and papC), and serum resistance (traT) was the most common isolate. There was an association between the detection of papC in E. coli isolates and higher necrosis scores. Additionally, the necrosis score was positively associated with the length of hospitalization, with each point increase in the necrosis score leading to two more days of hospitalization. These results suggest that papC-positive E. coli play an important role in the severity of pyometra in dogs. The present study revealed the possibility of using this virulence gene to better understand the prognosis of the disease in an affected animal.

Insight into the mechanism of CD34+ cell mobilisation impairment in multiple myeloma patients treated with anti-CD38 therapy.

Induction therapy followed by CD34+ cell mobilisation and autologous transplantation represents standard of care for multiple myeloma (MM). However, the anti-CD38 monoclonal antibodies daratumumab and isatuximab have been associated with mobilisation impairment, yet the mechanism remains unclear. In this study, we investigated the effect of three different regimens (dara-VCd, isa-KRd and VTd) on CD34+ cells using flow cytometry and transcriptomics. Decreased CD34+ cell peak concentration and yields, longer collection and delayed engraftment were reproduced after dara-VCd/isa-KRd versus VTd induction in 34 patients in total. Using flow cytometry, we detected major changes in the proportion of apheresis product and bone marrow CD34+ subsets in patients treated with regimens containing anti-CD38 therapy; however, without any decrease in CD38high B-lymphoid progenitors in both materials. RNA-seq of mobilised CD34+ cells from 21 patients showed that adhesion genes are overexpressed in CD34+ cells after dara-VCd/isa-KRd and JCAD, NRP2, MDK, ITGA3 and CLEC3B were identified as potential target genes. Finally, direct in vitro effect of isatuximab in upregulating JCAD and CLEC3B was confirmed by quantitative PCR. These findings suggest that upregulated adhesion-related interactions, rather than killing of CD34+ cells by effector mechanisms, could be leading causes of decreased mobilisation efficacy in MM patients treated with anti-CD38 therapy.

Transcriptomics reveals the mechanism of high-adhesive Bacillus subtilis inhibiting fresh fish spoilage

Aquatic products are highly susceptible to spoilage caused by adherence of spoilage bacteria during storage, so it is crucial to develop new microecological preservatives. The probiotic B. subtilis can achieve potential preservation effects by adhering to the fish gut to regulate the amount of probiotic colonization in the flora. Here, we demonstrated the surface adhesion characteristics of B. subtilis and the mechanism of adhesion-associated gene-mediated adhesion; the regulation of B. subtilis adhesion ability by nitrogen and carbon sources, and the application of tilapia preservation. Our results revealed 10 B. subtilis adhesion-related genes, among which nitrogen sources up-regulate the adhesion genes hag and motA, carbon sources up-regulate sinR and fliG. Based on flagellum-associated high adhesion strain C6 and high adhesion regulation of B. subtilis C6_H allowed the abundance of probiotic bacteria Bacillus, Rhodobacteraceae to increase, attenuate the content of spoilage flavor substances such as 1-Octen-3-ol, 15-Crown-5 and maintain the freshness of tilapia. The results helped to reveal the adhesion mechanism of B. subtilis and to develop and apply microecological preservatives with high quality adhesion ability.

Neutrophil Extracellular Traps: Potential Prothrombotic State Markers and Therapeutic Targets for Atrial Fibrillation.

Recently, the mechanism of thrombogenesis has taken a new direction with the involvement of neutrophil extracellular traps (NETs). However, little is known about the relationship between NETs and thrombogenesis in atrial fibrillation (AF). Our study aimed to evaluate NETs in AF patients and their potential association with thrombogenesis. In addition, we studied the effect of NETs on thrombogenesis in rat models. A total of 125 AF patients and 172 controls were studied. Spontaneous echo contrast (SEC) was examined using transesophageal echocardiography to assess the prothrombotic state. We used rapid atrial pacing (RAP) rat models to study NETs' formation and their effects on thrombogenesis. The levels of NETs were analyzed by flow cytometry. To deeply understand the regulatory mechanism of NET formation, the transcriptional characteristics of the left atrial appendage (LAA) tissue from RAP rats were analyzed. We found that NETs were increased significantly in AF patients and positively correlated with SEC grades. And inserting the NET level could significantly enhance the predictivity of CHA2DS2-VASc scores for the AF prothrombotic state. In the RAP models, we observed that NET levels increased significantly in the LAA and promoted thrombosis. Meanwhile, we found that these changes could be suppressed by the NET formation inhibitor. Transcriptomic analysis of the LAA tissue from RAP rats suggested that RAP might stimulate the NET formation by promoting the expression of inflammatory cytokine and adhesion genes. NETs may constitute useful thrombogenesis risk markers in AF patients and provide a potential therapeutic strategy for AF management.

Virulence Characterization of Faecal Escherichia coli Isolates from Wild Animals

The present study was undertaken to characterize virulence factors associated with fecal Escherichia coli (E.coli) isolates (n=22) from wild animals. Isolates were phenotypically characterized for haemolytic, haemagglutination and gelatinase activity whereas presence of adhesion genes viz. fimH and papG1 was detected genotypically by PCR. Further, whole blood survival assay of E. coli isolates (n=6) was performed using human and goat blood. Isolates exhibited heamolytic (45.45%) as well as gelatinase activity (27.27%) and led mannose sensitive agglutination of human (45.45%), dog (40.90%) and chicken (27.27%) RBC. fimH gene was detected in 27.27% of the isolates whereas none of the isolates was found positive for PapG1 gene. Isolates survived, got adapted and then grew successfully in human as well as goat blood. It can be concluded from present study that wild animals are potent reservoir of virulent E. coli that could adversely affects in-contact human and livestock population.

Virulence factors analysis and determination of the suitable chemical agent to inhibit Streptococcus mutans growth and biofilm formation

Streptococcus mutans is one of the most important causes of tooth decay and oral diseases, and it has received wide attention from researchers worldwide. The present work aimed to investigate, evaluation and comparisons effect of the antimicrobial and anti-biofilm property of some commercial compounds (chlorhexidine, hyaluronic acid, sodium fluoride and silver nanoparticles) against oral bacteria pathogens of S. mutans. In this work, S. mutans ATCC 25175 was used and its virulence factors were analyzed using Seed Viewer and The Comprehensive Antibiotic Resistance Database (CARD). The antibacterial susceptibility testing (AST) confirms effects of chlorhexidine mouthwash on S. mutans ATCC 25175 beggar than all components. The minimum inhibitory concentration (MIC) minimum bactericidal concentration (MBC) and Minimum biofilm inhibition concentration (MBIC) of the commercial compounds were determined. The statistical analysis was performed using One-Way ANOVA (Tukey test). The VITEK2system was used to confirm the biochemical features of S. mutans ATCC 25175 and susceptibility testing was done to compare the results with Seed Viewer and CARD analysis. The findings showed that The MICs, MBCs, and MBIC means of chlorhexidine against S. mutans ATCC 25175 was 0.0079, 0.0038, and 0.0039 μg/mL respectively, where its biological activity was significantly (P < 0.05) higher compared to other commercial compounds included in this study. The S. mutans strain used in this study showed sensitivity to all tested antibiotics and the Seed Viewer and CARD reported that it has several virulence factors such as adhesion genes (hsp33, Cpa, PrtF, PrtF2, MsmRL, Nra, SOF, LepAL, Spy0135, and RofA) and antibiotic-resistant genes (vanY, vanT, and mdeA). This study concluded that chlorhexidine was the best in inhibiting S. mutans growth as well as preventing biofilm formation, and the study recommends further investigation about antibiotic resistance genes and susceptibility profile.

Lymph Vessels Associate with Cancer Stem Cells from Initiation to Malignant Stages of Squamous Cell Carcinoma.

Tumor-associated lymph vessels and lymph node involvement are critical staging criteria in several cancers. In skin squamous cell carcinoma, lymph vessels play a role in cancer development and metastatic spread. However, their relationship with the cancer stem cell niche at early tumor stages remains unclear. To address this gap, we studied the lymph vessel localization at the cancer stem cell niche and observed an association from benign skin lesions to malignant stages of skin squamous cell carcinoma. By co-culturing lymphatic endothelial cells with cancer cell lines representing the initiation and promotion stages, and conducting RNA profiling, we observed a reciprocal induction of cell adhesion, immunity regulation, and vessel remodeling genes, suggesting dynamic interactions between lymphatic and cancer cells. Additionally, imaging analyses of the cultured cells revealed the establishment of heterotypic contacts between cancer cells and lymph endothelial cells, potentially contributing to the observed distribution and maintenance at the cancer stem cell niche, inducing downstream cellular responses. Our data provide evidence for an association of lymph vessels from the early stages of skin squamous cell carcinoma development, opening new avenues for better comprehending their involvement in cancer progression.

Abstract P307: What A Difference Hypertension Makes: The Adventitia Hands Off To PVAT For Extracellular Matrix Remodeling As Blood Pressure Rises In The High Fat Fed Dahl SS Rat.

Tunica media extracellular matrix remodeling is well understood to occur in response to elevation of blood pressure, considered a protective response. Remodeling of other tunicas is not well known. We have a long-term hypothesis that perivascular adipose tissue (PVAT) is responsive to hypertension and likely remodels to assist the blood vessel in protection against higher pressures. Here, we used the tunica adventitia and PVAT of the thoracic aorta in measuring extracellular matrix genes from 5 pairs of Dahl SS male rats on 8 or 24 weeks of feeding from weaning on a control (10% fat) or high fat (HF; 60%) diet. These times were picked given relative increases mean arterial blood pressure (mm Hg) in the HF vs control (mmHg increase; 8 week: 14±5; 24 week = 35±8; p&lt;0.05). The adventitia and PVAT were dissected apart, RNA isolated, cDNA synthesized and run on RT2 profiler array kits for ECM and Cell Adhesion genes. Differentially expressed genes (HF 2 fold &gt; Con) were determined in Cytoscape, followed by identification of Hub Genes and construction of a Gene Regulatory network. At 8 weeks on HF diet, the adventitia had 29 upregulated genes (vs Con diet) while the PVAT had none. By contrast, at 24 weeks on diet, the HF adventitia had 3 upregulated genes while PVAT had 47. The top two genes elevated at 8 weeks in adventitia where thrombospondin 1 and collagen 8a1. Surprisingly, at 24 weeks, thrombospondin 1 was one of the three top genes upregulated in PVAT. The following transcription factors were identified as regulators of those genes upregulated in the 8 week adventitia: Adarb1, Fos, Gm10323, Smad3 and Mapk1 . By contrast, the key transcription factors for DEGS in the 24 week PVAT were Stat1, Bcl6B, Tsnax, Pax8, and Jun. Adarb1 (Adenosine deaminase RNA specific B1) was also determined to be a key regulator in PVAT remodeling. These data support that the PVAT responds biologically once blood pressure is elevated.

Physiological and Transcriptomic Analyses of Escherichia coli Serotype O157:H7 in Response to Rhamnolipid Treatment.

Rhamnolipid (RL) can inhibit biofilm formation of Escherichia coli O157:H7, but the associated mechanism remains unknown. We here conducted comparative physiological and transcriptomic analyses of cultures treated with RL and untreated cultures to elucidate a potential mechanism by which RL may inhibit biofilm formation in E. coli O157:H7. Anti-biofilm assays showed that over 70% of the E. coli O157:H7 biofilm formation capacity was inhibited by treatment with 0.25-1 mg/mL of RL. Cellular-level physiological analysis revealed that a high concentration of RL significantly reduced outer membrane hydrophobicity. E. coli cell membrane integrity and permeability were also significantly affected by RL due to an increase in the release of lipopolysaccharide (LPS) from the cell membrane. Furthermore, transcriptomic profiling showed 2601 differentially expressed genes (1344 up-regulated and 1257 down-regulated) in cells treated with RL compared to untreated cells. Functional enrichment analysis indicated that RL treatment up-regulated biosynthetic genes responsible for LPS synthesis, outer membrane protein synthesis, and flagellar assembly, and down-regulated genes required for poly-N-acetyl-glucosamine biosynthesis and genes present in the locus of enterocyte effacement pathogenicity island. In summary, RL treatment inhibited E. coli O157:H7 biofilm formation by modifying key outer membrane surface properties and expression levels of adhesion genes.

Effect of thymosin β4 on lipopolysaccharide‑stimulated brain microvascular endothelial cell remodeling: A possible role in blood‑brain barrier injury.

War veterans, in particular, are more prone to mental illness as they are more likely to have encountered multiple traumatic brain injuries (TBIs) whilst serving on active duty in war zone areas. A TBI is known to cause mortality or serious neurological disabilities among survivors and elicits a number of pathological processes, including neuroinflammation and blood brain barrier (BBB) disruption, leading to secondary brain damage and subsequent impairment of the neurovascular unit. Although several drugs exhibit promising effects for TBI, the repertoire of currently available therapeutic strategies remains limited. Thymosin 4 (Tβ4) is a 43-amino acid G-acting sequestering peptide that confers neuroprotective potential in TBI models. However, its role in BBB function remains unclear. Further research into the mechanism of BBB disruption induced by TBI and its specific role in neurovascular pathophysiology is necessary. In the present study, the protective effects of Tβ4 in lipopolysaccharide (LPS)-stimulated gene expression of several tight junction proteins, inflammatory genes, apoptotic genes, and adhesion genes in human brain microvascular endothelial cells (hBMVECs), one of the pivotal cell types in the BBB, were reported. The results suggested that pretreatment with Tβ4 reversed the LPS-induced damage of BBB components in hBMVECs. Furthermore, these results identified neuregulin 1 as a possible target for Tβ4. Therefore, it is proposed that Tβ4-mediated cellular signaling in hBMVEC may be vital for understanding the association between the BBB and TBI pathophysiology, which warrants further investigation.

Prevalence and virulence genes of Staphylococcus aureus from food contact surfaces in Thai restaurants.

Staphylococcus aureus is one of the most common pathogens responsible for food poisoning due to its ability to produce staphylococcal enterotoxin (SE). S. aureus can form biofilms on the surfaces of food processing devices, enabling the distribution of SE on foods through cross-contamination events. Thailand is known for its exotic cuisine, but there is no data on the prevalence of SE-harboring S. aureus in restaurants in Thailand. In this study, we conducted surface swabs on surfaces of kitchen utensil that come into contact with food and on the hands of food handlers working in restaurants in the north part of Thailand. Isolated S. aureus was investigated for biofilm formation, virulence, and SE genes. Two hundred S. aureus were isolated from 650 samples. The highest prevalence of S. aureus contamination was detected on the hands of food handlers (78%), followed by chopping boards (26%), plates (23%), knives (16%), spoons (13%), and glasses (5%). All of them were methicillin-sensitive S. aureus (MSSA) and the mecA gene was not present in any strains. Biofilm formation was detected using the CRA method, and 49 (24.5%) were identified as biofilm-producing strains, with the hands of food handlers identified as the primary source of biofilm-producing strains. The prevelence of biofilm-related adhesion genes detected were: icaAD (13%), fnbA (14.5%), cna (6.5%), and bap (0.5%). Two classical enterotoxin genes, sec and sed, were also found in four and six of the S. aureus isolates, respectively, from hands and utensils. The highest prevelence of S. aureus was detected on the hands of food handlers. S. aureus strains with biofilm and enterotoxin production abilities were discovered on food contact surfaces and the hands of food handlers, implying significant risk of food contamination from these sources that could be harmful to consumers. To avoid cross-contamination of food with food contact items, the food handlers' hands should be properly washed, and all food preparation equipment should be thoroughly cleaned.

Anti-inflammatory probiotic Lactiplantibacillus plantarum HF05 screening from Qula: Genomic analysis and alleviating effect on intestinal inflammation

The use of probiotics for their anti-inflammatory potential is an area with limited research, and there is currently limited understanding of the connection between the genotype and phenotype of the majority of probiotics. In this study, Lactiplantibacillus plantarum HF05 with prominent anti-inflammatory effects and excellent probiotic characteristics was screened from Qula by combining RAW264.7 inflammatory cell model screening with in vitro comparison of probiotic properties. Through whole genome analysis, we found that the HF05 genome encoded a large number of environmental adaptation, adhesion, colonization, anti-inflammatory and antioxidant-related genes. The in vitro experiments demonstrated that HF05 inhibited pro-inflammatory cytokine release in RAW264.7 inflammatory cells. Furthermore, we found that prophylactic intake of HF05 alleviated intestinal inflammation and oxidative damage in mice with acute colitis and inhibited the value-added of enteropathogenic bacteria (Escherichia coli, Enterococcus faecalis, Staphylococcus xylosus) and reversed the depletion of Lactobacillus (Lactobacillus johnsonii, Lactobacillus intestinalis). Collectively, we demonstrated that HF05 can serve as a new probiotic for the relief of intestinal inflammation.

Trans-cinnamaldehyde loaded chitosan based nanocapsules display antibacterial and antibiofilm effects against cavity-causing Streptococcus mutans

ABSTRACT Background Dental caries is a multifactorial disease, and the bacteria such as Streptococcus mutans (S. mutans) is one of the risk factors. The poor effect of existing anti-bacterial is mainly related to drug resistance, the short time of drug action, and biofilm formation. Methods To address this concern, we report here on the cinnamaldehyde (CA) loaded chitosan (CS) nanocapsules (CA@CS NC) sustained release CA for antibacterial treatment. The size, ζ-potential, and morphology were characterized. The antibacterial activities in vitro were studied by growth curve assay, pH drop assay, biofilm assay, and qRT-PCR In addition, cytotoxicity assay, organ index, body weight, and histopathology results were analyzed to evaluate the safety and biocompatibility in a rat model. Results CA@CS NC can adsorb the bacterial membrane due to electronic interaction, releasing CA slowly for a long time. At the same time, it has reliable antibacterial activity against S. mutans and downregulated the expression levels of QS, virulence, biofilm, and adhesion genes. In addition, it greatly reduced the cytotoxicity of CA and significantly inhibited dental caries in rats without obvious toxicity. Conclusion Our results showed that CA@CS NC had antibacterial and antibiofilm effects on S. mutans and inhibit dental caries. Besides, it showed stronger efficacy and less toxicity, and was able to adsorb bacteria releasing CA slowly, providing a new nanomaterial solution for the treatment of dental caries.