Polymerase Chain Reaction Technique Research Articles

The Assessment of IL-4 and IgG1 Levels in Patients with Rheumatoid Arthritis and Periodontitis: A Serological Study in Hilla City, Iraq

Abstract Background: Periodontal disease, which has a complex etiology, is prevalent and influenced by several factors, including microorganism invasion, host health, and external environmental factors, contributing to its development. It is unknown whether rheumatoid arthritis (RA) and periodontitis are related. Objectives: The current study aims to assess the relationship between periodontitis and RA by measuring the concentrations of interleukin 4 (IL-4) and immunoglobulin G1 (IgG1) in patients’ serum and quantifying Porphyromonas gingivalis in subgingival plaque samples. Materials and Methods: The study comprised patients with periodontitis, RA, and healthy controls. The levels of IL-4 and IgG1 in the serum samples were measured using an enzyme-linked immunosorbent assay. The identification of subgingival plaque bacteria is made by using the polymerase chain reaction technique. Results: The results showed the levels of IgG1 were high in healthy people compared to those with periodontitis and those with periodontitis and arthritis, where the levels in healthy people were 2.3 ± 0.28, 2.06 ± 0.55, 2.05 ± 2.34, 1.82 ± 0.77 in the age groups 41:50, 20:30, 51:60, 31:40, respectively. It was found that there are significant differences between the values of IgG1 and IL-4 in patients with periodontitis at P ≤ 0.05 which is considered a differential sign. Conclusion: The concentrations of IL-4 and IgG1 are decreased in patients (periodontitis and periodontitis with rheumatoid arthritis) compared to their concentrations in healthy people. Thus, we indicate IL-4 was related to reducing the severity of periodontitis disease and RA.

Selectively Isolation and Molecular Identification by dnaJ Gene of Legionella pneumophila Found in Various Water Sources in Dhaka, Bangladesh

Abstract BACKGROUND: Legionella spp. are the etiological agents of Legionnaires’ disease, often severe pneumonia. After the first severe outbreak of Legionella spp., it was found widely dispersed in natural water sources and humid environments and has become a concern for these sources’ water quality and safety. The fecal indicators used for water quality assessment do not predict the presence of these pathogens; thus, in the absence of targeted analysis, the risk is unobserved until the emergence of infections. There is no study on the presence of Legionella in unattended water bodies in Bangladesh, such as swimming pools, air-conditioned cooling water, and stagnant circulatory water systems. METHODS: A total of 42 water samples were collected from 12 different sites in Dhaka, Bangladesh. Acid, heat, and chlorine treatment were applied to reduce the load of non-Legionellae in water samples. Chlorine treatment was used to kill unwanted organisms in water samples. After successful pretreatment, samples were cultured on buffered charcoal yeast extract agar (BCYE) media. Optimum pH, concentrations of chlorine, and temperature were set up by extensive trial and error culturing of Legionella on BCYE media to minimize other bacterial growth and maximize Legionella growth. Isolated colonies were confirmed primarily by morphological characteristics on BCYE media. Isolates were then confirmed by polymerase chain reaction and gel electrophoresis techniques. We detected the presence of 16S rDNA for Legionella spp. and the dnaJ gene for Legionella pneumophila species identification. Antibiotic sensitivity tests (minimum inhibitory concentration) were also conducted to see if it was alarming for our community’s health. RESULTS: Most of the Legionella spp. could grow at various temperatures, such as 45°C, 50°C, and 55°C, and they could also survive at the normal levels of chlorination, such as 0.25 mg/L and 0.5 mg/L. We have found that, out of 12 different sites, 4 out of 5 (80%) isolates from cooling tower water, 1 out of 4 (25%) from the swimming pool, and 1 out of 3 (33%) household stagnant water are contaminated with Legionella. However, 3 out of 5 isolates found are L. pneumophila from cooling water sites. Furthermore, strains of Legionella spp. were resistant to many antibiotics, such as ciprofloxacin, azithromycin, and tetracycline. CONCLUSIONS: Serious and fatal Legionella spp. (L. pneumophila) infections may be transmitted because of the widespread presence of these water sources and the large number of users in the community. This threat is worsened by the survival of Legionella under various commonly used antibiotic conditions and further exacerbated by their antibiotic resistance. Health-care professionals may face great challenges due to the lack of studies about its prevalence and treatment procedures.

Antibiotic Resistance and Inflammatory Response in Urology Infections Caused by E. coli: A Molecular and Clinical Investigation

Background: Urinary tract infections (UTIs) are a common condition resulting from bacterial invasion of the urinary system, which includes the kidneys, ureters, bladder, and urethra. These infections are particularly prevalent among females due to anatomical differences and hormonal factors. UTIs can range from uncomplicated cases, where there are no structural abnormalities, to more severe infections that may ascend to the kidneys, causing pyelonephritis. Methods: This study aimed to isolate and diagnose Escherichia coli (E. coli) bacteria in patients with UTIs, identify specific virulence genes, and assess the bacteria's resistance to various antibiotics. A total of 100 clinical samples were collected from patients with UTIs at the General Hospital of Homadiyah and Ahmed Maher from October 2021 to December 2022. Bacterial identification was performed using biochemical tests and polymerase chain reaction (PCR) techniques, while antibiotic sensitivity was assessed using the Kirby-Bauer disk diffusion method. Additionally, inflammatory markers (TNF-α, IL-6, and IL-8) were measured using ELISA. Results: E. coli was isolated in 56% of the samples, with the highest prevalence in patients aged 31-40 years. The antibiotic sensitivity test revealed varying levels of resistance, with the highest sensitivity observed to third-generation cephalosporins, particularly ceftriaxone (32.2% resistance). PCR analysis identified the presence of several virulence genes, including the Hly gene in 96.6% of isolates and the iha gene in 10% of isolates. Elevated levels of inflammatory markers were observed in patients with UTIs, indicating a strong immune response to bacterial infection. Conclusion: E. coli is a significant pathogen in UTIs, exhibiting considerable antibiotic resistance, particularly to first-generation cephalosporins. The presence of specific virulence genes correlates with the severity of infection. These findings underscore the need for targeted antibiotic therapy and the development of strategies to manage antibiotic resistance in UTIs.

Significance of p16 in Site-Specific Human Papillomavirus-Positive and Negative Head and Neck Squamous Cell Carcinoma (HNSCC).

Head and neck squamous cell carcinoma (HNSCC) represents a group of cancers characterized by diverse origins and changing epidemiological patterns. The significance of high-risk human papillomavirus (HPV) infection in certain HNSCC cases has gained attention for its impact on the disease's behavior. Our current research focused on exploring the importance of using p16 as an HNSCC biomarker, particularly in the context of HPV infection, assessing its value in prognosis, and examining its variation across different tumor locations. A retrospective analysis was carried out on 100 HNSCC patients from a tertiary care center, with particular attention paid to p16 expression, HPV status, clinic-pathological characteristics, and prognosis. HPV was detected using polymerase chain reaction (PCR) techniques, and p16 expression was evaluated by immunohistochemistry. According to the ethical guidelines outlined in the Declaration of Helsinki, multivariate analysis assessed the prognostic value of p16. Our analysis demonstrated a significant correlation between HPV status and p16 expression in HNSCC cases. A vast majority of 58 (96.7%) HPV-+cases exhibited p16 overexpression, contrasting sharply with only two (5%) in the HPV--group. Patients with tumors that were both p16+and HPV+exhibited more favorable overall survival rates. In contrast, those with p16-and HPV-tumors experienced the poorest survival outcomes. Notably, having a p16--status in HPV+cases emerged as an independent factor for reduced survival. Additionally, the study revealed distinct variations in p16 expression based on tumor location, particularly within the oropharyngeal area. The study established that p16 is a dependable indication for the existence of HPV in HNSCC and highlights its significant role as a prognostic factor, particularly in cases that are p16--yet HPV-+. These findings underscore the importance of adopting site-specific treatment approaches in HNSCC management and contribute to a deeper understanding of p16's role in the disease, thereby aiding in more effective risk assessment and treatment planning.

Genetic Basis of Biofilm Formation Genes Ebp and Bph (phos) among Multidrug Resistance Enterococcus faecalis Isolates, Iraq

Abstract Background: Biofilm produced by Enterococcus spp. increase their inherent and acquired antibiotic resistance, posing a significant challenge to infection treatment, particularly in virulent strains. Objective: This study aimed to investigate some genes responsible for biofilm formation (Bph (phos) and EbpB) by polymerase chain reaction (PCR) technique. Materials and Methods: A total of 130 clinical samples were collected during this study, which were obtained from patients who were admitted to Feminine and Children Teaching Hospital in Al-Diwaniyah City during a period extending from (November 2022 to March 2023). All samples were subjected to culturing on different media (blood agar, MacConkey, Bile Esculin agar, and M-El chromogenic ager), after which catalase and oxidase tests were conducted biochemically. After cultivation, the VITEK-2 compact system was used to identify the samples. The isolates were investigated genotypically for harboring biofilm formation genes, including EbpB and Bph (phos), by molecular method (PCR). Results: The VITEK-2 compact system revealed that only 12 isolates were identified as E. faecalis. The most effective antibiotics against E. faecalis were (vancomycin, teicoplanin, linezolide, tigecycline, and ampicillin), and the highest resistance was against erythromycin (100%) and tetracycline (91.6%). Enterococcus faecalis isolates were investigated genotypically for harboring biofilm formation genes that include EbpB and Bph (phos) by molecular methods, PCR. The results showed that EbpB and Bph (phos) genes were positive for all isolates with a percentage of 100%. Conclusions: It was found that the presence of biofilm formation genes in E. faecalis EbpB, and phos (Bph) increase the pathogenicity of this pathogen. These genes showed a high percentage among E. faecalis isolates. Also, all isolates had the potential to form a biofilm, which complicates their treatment with antibiotics, confirming the critical need to develop novel antimicrobial agents that control the infection associated with the development.

The Association of MMP-2, MMP-9, and MMP-13 Gene Polymorphisms With Knee Osteoarthritis in the Greek Population.

Introduction Primary knee osteoarthritis (OA) is a multifactorial degenerative joint disorder characterized by articular cartilage degradation. Matrix metalloproteinases (MMPs) have been reported to play a vital role in OA pathogenesis, significantly contributing to extracellular matrix (ECM) catabolism. The purpose of this study is to investigate the association of MMP-2 -1575G/A (rs243866), MMP-9 836A/G (rs17576), and MMP-13 -77A/G (rs2252070) gene polymorphisms with knee OA in the Greek population. Methods One hundred patients (24% males, mean age: 68.3 years) with primary knee OA were included in the study along with 100 controls (47% males, mean age: 65.2 years). Genotypes were identified through polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) technique. Allelic and genotypic frequencies were compared between patients and controls. Results The MMP-13 -77A/G polymorphism was significantly associated with knee OA in the crude analysis (P = 0.008). After binary logistic regression analysis, the dominant model of the MMP-13-77A/G (AG + GG versus AA) was found to be associated with increased risk for knee OA (odds ratio (OR) = 2.290, 95% confidence interval (95%CI) = 1.059-4.949, P= 0.035).Compared to the A allele, the G allele in the MMP-13rs2252070 locus was a predictive factor for knee OA (OR = 2.351, 95%CI = 1.134-4.874, P= 0.022). No significant associations were detected for the MMP-2 -1575G/A and MMP-9 836A/G polymorphisms (P > 0.05). Conclusions The present study shows that the MMP-2 -1575G/A and MMP-9 836A/G polymorphisms are not significantly associated with primary knee OA in the Greek population. The MMP-13 -77A/G was found to be a significant risk factor for knee OA in the Greek population. Additional research is needed to verify this association in larger and different populations, in different joints, to elucidate the role of this single nucleotide polymorphism (SNP) in OA pathogenesis.

Temporal trends in respiratory pathogens following the COVID-19 pandemic and climate variables: A unicentric retrospective evaluation of 24 pathogens in a temperate subtropical region.

Temperature and humidity are studied in the context of seasonal infections in temperate and tropical zones, but the relationship between viral trends and climate variables in temperate subtropical zones remains underexplored. Our retrospective study analyzes respiratory pathogen incidence and its correlation with climate data in a subtropical zone. Retrospective observational study at Moinhos de Vento Hospital, South Brazil, aiming to assess seasonal trends in respiratory pathogens, correlating them with climate data. The study included patients of all ages from various healthcare settings, with data collected between April 2022 and July 2023. Biological samples were analyzed for 24 pathogens using polymerase chain reaction and hybridization techniques; demographic variables were also collected. The data was analyzed descriptively and graphically. Spearman tests and Poisson regression were used as correlation tests. Tests were clustered according to all pathogens, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza viruses, rhinovirus, and respiratory syncytial virus (RSV). Between April 2022 and July 2023, 3329 tests showed a 71.6% positivity rate. Rhinovirus and RSV predominated, exhibiting seasonal patterns. Temperature was inversely correlated with the viruses, notably rhinovirus, but SARS-CoV-2 was positively correlated. Air humidity was positively correlated with all pathogens, RSV, rhinovirus, and atmospheric pressure with all pathogens and rhinovirus. Our results showed statistically significant correlations, with modest effect sizes. Our study did not evaluate causation effects. Despite the correlation between climate and respiratory pathogens, our work suggests additional factors influencing transmission dynamics. Our findings underscore the complex interplay between climate and respiratory infections in subtropical climates.

Subinhibitory Concentrations of Kojic Acid Reduce Enterotoxins A and α-hemolysin Production in Staphylococcus aureus Isolates

Abstract Background: Staphylococcus aureus (S. aureus) is one of the most significant human pathogens that causes various sorts of diseases and invasive infections. Recently, due to the resistance of this bacterium to several medications, it has become difficult to treat infections caused by it. The use of biological compounds to control infections might be a solution to this challenge. This study aimed to investigate the influence of kojic acid (KA) on the expression of some virulence factors of S. aureus. Methods: Clinical samples were collected from hospital sources. The standard diagnostic test and molecular techniques were used to identify S. aureus strains. Minimal inhibitory concentrations (MICs) of KA were determined according to guidelines of the Clinical and Laboratory Standards Institute. The expression levels of α-hemolysin (hla) and enterotoxin A (sea) genes were measured in the presence and absence of KA using a real-time polymerase chain reaction technique. The obtained data were analyzed using standard statistical methods in SPSS software (version 16) and Prism GraphPad software. P < 0.05 was considered statistically significant in the comparative data. Results: The results indicated that the values of minimum inhibitory concentration (MIC) of S. aureus after KA treatment were 512 μg/mL. As a result, the expression of the hla gene was decreased 9 fold and the expression levels of the sea gene were decreased 2.7 fold. Conclusions: In this study, the desirable effects of KA on S. aureus strains were observed. In conclusion, the application of KA potentially reduces the virulence factors of S. aureus. These promising results can breathe hope in the fight against pathogenic S. aureus strains by repressing their virulence factors.

Antimicrobial susceptibility and virulence gene analysis of Shigella species causing dysentery in Iranian children: Implications for fluroquinolone resistance

Shigella species significantly impact global health due to their role in diarrheal diseases. A 2019–2022 cross-sectional study on 432 stool samples from pediatric patients in Mashhad, Iran, identified Shigella spp. and tested their susceptibility to 12 antimicrobials by the disk diffusion method. The presence of virulence factors, namely ipaH, virA, stx1, and stx2, as well as plasmid-mediated quinolone resistance (PMQR) genes, including qnrA, qnrB, qnrC, qnrD, and qnrS, were ascertained through the utilization of polymerase chain reaction techniques. Sequencing of 15 isolates detected mutations within quinolone resistance-determining regions (QRDRs) at the gyrA and parC genes, indicating fluoroquinolone (FQ) resistance. 19.2 % (83/432) of stool samples contained Shigella, primarily S. sonnei (77.1 %), followed by S. flexneri (21.6 %) and S. boydii (1.2 %). Most isolates were from children under five (55.4 %). All strains had the ipaH gene, lacked stx1 and stx2, and 86.7 % had virA. High resistance was noted for ampicillin and tetracycline (84.3 % each), trimethoprim-sulfamethoxazole (81.9 %), and azithromycin (60.2 %). 87.1 % of isolates were multidrug-resistant (MDR). The most common PMQR genes were qnrA and qnrS (41 % each). The qnrD gene, prevalent in 36.1 % of cases, is reported in Iran for the first time. The most common PMQR profile was qnrADS (15.7 %). Resistance to nalidixic acid and ciprofloxacin was 45.8 % and 12 %, respectively. The Shigella isolates exhibited mutations in the gyrA (at codons 83, 87, and 211) and parC (at codons 80, 84, 93, 126, 128, 129, and 132) genes. The D87Y mutation in the gyrA gene was the most common in Shigella isolates, occurring in 73 % of cases. The F93S and L132T mutations in the parC gene were unique to this study. Empirical FQ therapy in patients infected with MDR Shigella, possessing PMQR determinants and/or mutations in the QRDRs of gyrA and parC, may escalate the risks of secondary diseases, extended treatment duration, therapeutic failure, and resistance spread. Consequently, the necessity for continuous surveillance and genetic testing to detect FQ-resistant Shigella strains is of paramount importance.

Assessment of Commercial DNA Extraction Kits for Porcine Gelatin Detection Using RT-PCR and ddPCR

Gelatin is extensively applied in various industries, including food, beverages, cosmetics, and pharmaceuticals. Although the determination of gelatin species is essential for religious, health, and consumer preference reasons, a standardized analysis method is absent. The challenge in identifying gelatin through DNA-based methods arises from the low DNA content and extensive DNA denaturation in the gelatin matrix. This study assessed the efficacy of two commercial DNA extraction kits, namely the Processed Food DNA Extraction (PF kit) and the DNeasy Mericon Food Kit (DM kit), for extracting DNA from porcine gelatin powder and commercial products derived from gelatin. Additionally, we evaluate the amplification of the extracted porcine DNA using real time polymerase chain reaction (RT-PCR) and droplet digital polymerase chain reaction (ddPCR) techniques. The PF extraction kit demonstrated successful DNA extraction from porcine gelatin powder and commercial samples of porcine gelatin-based candies with a higher concentration (32.24-286.07 ng/uL) and purity (A260/A280 ratio of 1.82-2.33) compared to the DM kit (3.95-7.30 ng/uL and an A260/A280 ratio of 1.29-2.45). RT-PCR and ddPCR analyses yielded positive results for porcine DNA from gelatin powder for both PF and DM kits, albeit with differing Cq values and copy numbers. The choice of DNA extraction kit significantly impacted the amplification results when analyzing commercial samples of porcine gelatin-based candies. Using RT-PCR, all samples yielded negative results with the DM kit, while the PF kit detected one positive result for porcine DNA. Improved outcomes were observed with more sensitive analysis methods such as ddPCR, where the DM kit identified one positive result for porcine DNA while the PF kit detected positive results for all tested candies.

Risk Factors Associated with an Influenza B Outbreak due to Inefficient Screening in a Prison in Thailand

On 3 Oct 2019, the Office of Disease Prevention and Control Region 12 Songkhla received notification of a suspected influenzacluster in a prison in Songkhla Province, Thailand. In cooperation with the local public health teams, we investigated the event to confirm the diagnosis, identify the source of infection, and implement control measures. A suspected influenza case was defined as an individual with fever and at least one of the following symptoms: cough, sore throat, runny nose, or dyspnea. Aconfirmed case was a suspected case who tested positive to influenza by the reverse transcription polymerase chain reaction technique. An environmental survey was conducted to assess risk behaviors and determine the occupancy rate. A case-control study was performed to identify associated factors of developing influenza. We identified 128 suspected cases, of which seven were confirmed to have type B influenza. All were male and their median (interquartile range) age was 29 years (24–35). The overall attack rate was5.2%. The first case developed symptoms in the new prisoner zone and moved to another zone without undergoing respiratory screening. The analytical study revealed that sleeping within one meter of a confirmed case (adjusted odds ratio (AOR) 1.77, 1.07–2.92), and sharing drinking glasses with others (AOR 1.83, 1.18–2.82)were significant risk factors. The overcrowded prison (165% occupancy) led to limited availability of vaccines, causing 83.5% of prisoners to be unvaccinated. Strict screening of new prisoners before moving to another area and early isolation of symptomatic prisoners are recommended.

Mycological detection of aflatoxin producing gene (Aflr) in Aspergillus Spp found in commercial raw groundnut and groundnut cake (Kuli kuli) in some selected market in Katsina state, Nigeria

The effect of aflatoxin intoxication from groundnut and groundnut products has lead to a large amount of deaths some of which are not yet accounted of. Kuli-kuli formally referred to as groundnut cake, is a byproduct of oil extraction from groundnut which have been further fried, sweetened or spiced. This research aimed at determining the mycological quality and molecular detection of Aflatoxin gene aflR in Aspergillus spp found in raw Groundnut and groundnut cake (Kuli-kuli). A total of 120 samples (60 groundnut samples and 60 groundnut cake samples) were randomly collected from four selected markets in Katsina state and subjected to mycological assessment based on colonial morphology on PDA and microscopy. Out of the total samples, 16 samples (8 raw groundnut samples and 8 groundnut samples) were selected for determination of total Aflatoxin concentration using ELISA technique. Isolates were also screened for aflR gene using polymerase chain reaction technique, and subsequently sequenced using Sanger method. Eleven (11) fungi isolate including Aspergillus niger, Scedosporium apiospermum, Cladophialophora bantiana, Aspergillus flavus, Aspergillus nidulans, Microsphaeropsis arundinis, Cladosporium, Cladophialophora carrionii, Rhizopus arrhizus, Aspergillus parasitus and Aspergillus fumigatus were identified to produce aflatoxin concentration ranging from 0.4–3.5 ppb from the samples of the raw groundnut and groundnut cake (Kuli-kuli). Therefore, the findings of this research has raised serious food safety concerns, especially in consumption of raw groundnut and groundnut cake (Kuli-kuli) sold in Katsina as such relevant regulatory agencies such as NAFDAC are advised to deploy drastic measure to curtail a possible aflatoxin intoxication outbreak

The Prevalence, Predisposing Factors, and Outcomes of Pneumocystis jirovecii Pneumonia among Pediatric Inpatients, Northeastern Iran

Abstract Objective Pneumocystis jirovecii pneumonia (PJP), caused by Pneumocystis jirovecii, is one of the opportunistic fungal infections that can cause life-threatening pneumonia in children with underlying diseases. Due to the similarity of the symptoms of PJP with other lung infections, such as tuberculosis, differential and accurate diagnosis is necessary. The current study investigated the molecular diagnosis of P. jirovecii, predisposing factors and the outcomes, among pediatric inpatients in Northeastern Iran. Methods In this study, 180 bronchoalveolar lavage specimens were obtained from hospitalized children with respiratory disorders. The specimens were examined using Giemsa stain, and the genomic DNA was extracted according to the protocol of the AmpliSens kit. A real-time polymerase chain reaction (PCR) technique was used to detect P. jirovecii by the AmpliSens Pneumocystis jirovecii (carinii)-FRT PCR kit. Results Among the patients studied, 34 (18.9%) were positive and 8 (4.4%) were suspicious of the presence of P. jirovecii. Among the 34 positive cases, 12 (35%) were diagnosed before, and 22 (65%) during the coronavirus 2019 (COVID-19) pandemic. Only two cases (5.88%) among the positive ones detected by the real-time PCR method were observed using Giemsa staining. Also, no correlation was observed between positive cases of infection and the sex, the outcomes, and underlying diseases. Conclusion The results showed that PJP has a relatively high prevalence among pediatric inpatients with respiratory disorders. Neutropenia is a significant predisposing factor in these patients. However, there is no correlation between PJP cases and outcomes and underlying diseases. Most of the patients with PJP were affected during the COVID-19 pandemic, probably due to treatment with corticosteroids.

GRHPR, Targeted by miR-138-5p, Inhibits the Proliferation and Metastasis of Hepatocellular Carcinoma Through PI3K/AKT Signaling Pathway.

Background: Hepatocellular carcinoma (HCC) is a highly aggressive cancer. This study aims to elucidate the role of Glyoxylate reductase/hydroxypyruvate reductase (GRHPR) in HCC proliferation and metastasis, along with its molecular mechanism, and to identify miRNAs targeting GRHPR. Materials and Methods: Expression levels of GRHPR and miR-138-5p were assessed using real-time fluorescent quantitative polymerase chain reaction and Western blot techniques. Bioinformatic analysis was employed to identify miRNAs targeting GRHPR, and the results were confirmed via dual-luciferase reporter assays. HCC cell lines overexpressing GRHPR were established to investigate its roles in cell proliferation, migration, and invasion. The biological function of miR-138-5p targeting GRHPR in HCC cells was also evaluated. Furthermore, a xenograft mouse model was utilized to examine the invivo functions of GRHPR. Results: GRHPR expression was downregulated in HCC, whereas miR-138-5p was upregulated. Overexpression of GRHPR suppressed HCC cell proliferation, migration, and invasion. Conversely, inhibition of GRHPR by miR-138-5p promoted HCC cell proliferation and invasive properties. MiR-138-5p was found to regulate Phosphoinositide 3-kinase (PI3K) and protein kinase B (AKT) phosphorylation levels by inhibiting GRHPR expression. Conclusion: This study highlights GRHPR's role as a tumor suppressor in HCC, with its function being regulated by miR-138-5p.

Ginsenoside Rb3 alleviates the formation of osteoclasts induced by periodontal ligament fibroblasts in the periodontitis microenvironment through the STAT3 pathway.

This study explores the potential role and mechanism of Ginsenoside Rb3 (Rb3) in modulating osteoclastogenesis induced by human periodontal ligament fibroblasts (hPLFs) within the periodontitis microenvironment. We investigated the anti-inflammatory effects of Rb3 on hPLFs stimulated with Porphyromonas gingivalis lipopolysaccharide (P.g-LPS) utilizing quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay techniques. Moreover, the functional role of Rb3 in hPLFs-induced osteoclast formation was assessed by treating human bone marrow-derived macrophages (hBMMs) with conditioned medium from hPLFs, followed by analyses through qPCR, western blot analysis, and staining for tartrate-resistant acid phosphatase (TRAP) and phalloidin. The impact of Rb3 on the activation of the STAT3 signaling pathway was determined via western blot analysis. Results indicated that Rb3 treatment significantly suppressed the upregulation of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, MCP-1, and IL-18) at both gene and protein levels in hPLFs induced by P.g-LPS. Furthermore, conditioned medium from Rb3 plus P.g-LPS treated hPLFs notably decreased the number of TRAP-positive cells, actin ring formations, and the expression of osteoclast marker genes (including CTSK, NFATC1, and ACP5). Rb3 also inhibited the P.g-LPS-induced activation of the STAT3 pathway, with the activation of STAT3 partially reversing the effects of Rb3 on inflammation and osteoclast differentiation. Collectively, Rb3 ameliorates inflammation in P.g-LPS-stimulated hPLFs and reduces hPLFs-induced osteoclastogenesis by inhibiting the STAT3 signaling pathway, suggesting its potential as a therapeutic agent for periodontitis.

POLYMORPHISM OF THE BETA-CASEIN GENE IN HOLSTEIN-ZEBU COWS IN THE NORTH OF MINAS GERAIS, BRAZIL

This study aimed to investigate A1 and A2 polymorphisms of the beta-casein gene in crossbred Holstein-Zebu (HZ) cows from four herds in Northern Minas Gerais, Brazil. Blood samples were collected from 112 randomly selected cows, of wich 66 were at ½ HZ, and 46 were at ¾ HZ. DNA extraction was conducted using the phenol-chloroform method, and polymorphisms were evaluated using the amplification refractory mutation system - polymerase chain reaction technique. The results demonstrated that 72/112 (64.3%) cows were heterozygous (A1A2), 33/112 (29.5%) were homozygous for A2A2, and 7/112 (6.2%) were A1A1 homozygous. The percentage of A2A2 cows ranged from 10.7 to 41.4% among the evaluated herds. The frequencies of the A1 and A2 alleles were 38.4 and 61.6%, respectively. The A2 allele was the most frequent in both breeds. The ½ HZ cows were 1.8 times more likely (Odds Ratio) to belong to the A1A1 genotype. The distribution of genotypes was in Hardy-Weinberg equilibrium in three of the herds; however, the total of samples and the stratified samples according to breeds were not in equilibrium considering α = 0.05.

Identification of KRT80 as a Novel Prognostic and Predictive Biomarker of Human Lung Adenocarcinoma via Bioinformatics Approaches.

According to the 2022 Global Cancer Statistics, lung cancer is the leading cause of cancer-related mortality worldwide. Lung adenocarcinoma (LUAD), which is a histological subtype of Non- Small Cell Lung Cancer (NSCLC), accounts for 40% of primary lung cancer. Therefore, there is an urgent need to identify new prognostic markers as clinical predictive markers for LUAD. This study aimed to investigate the role of Keratin 80 (KRT80) in the prognosis of LUAD and its underlying mechanisms. Bioinformatics analysis was conducted using data retrieved from The Cancer Genome Atlas (TCGA) databases. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were employed to predict the involved biological processes and signaling pathways, respectively. The LinkedOmics database was utilized to identify differentially expressed genes (DEGs) correlated with KRT80. Nomograms and Kaplan-Meier plots were constructed to evaluate the survival outcomes of patients diagnosed with LUAD. Moreover, TIMER was employed to conduct correlation analyses between KRT80 expression and immune cell infiltration, shedding light on the intricate interplay between KRT80 and the tumor microenvironment in LUAD. To ascertain the RNA and protein expression levels of KRT80 in LUAD and adjacent normal tissues, Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) and immunohistochemistry techniques were employed, respectively. Scrutiny of the TCGA dataset revealed KRT80 up-regulation across pan-cancer tissues, notably elevated in LUAD compared to healthy lung tissues. This finding was validated in our clinical samples, where Kaplan-Meier survival curves indicated poorer survival rates for high KRT80 expression in LUAD. A positive correlation was found between the transcription level of KRT80 in LUAD samples and clinical parameters, such as lymph node metastasis stage, distant metastasis, and pathological stage. Survival, logistic regression, and Cox regression analyses emphasized the clinical prognostic significance of high KRT80 expression in LUAD. Nomogram results underscored the robust predictive potential of KRT80 for the survival of LUAD patients. Gene functional enrichment analyses mainly associated KRT80 with cytokine-cytokine receptor interactions, cell cycle, apoptosis, and chemokine signaling pathways. Based on the results of the immune infiltration analysis, it can be found that the expression of KRT80 is related to the immune cell subsets and survival rate of patients with LUAD. Our research revealed a significant upregulation of KRT80 in LUAD, with heightened KRT80 expression correlating with unfavorable prognosis. This study represents a comprehensive and systematic evaluation of KRT80 expression in LUAD, encompassing its prognostic and diagnostic significance, as well as underlying mechanisms. Our findings suggest that KRT80 may emerge as a novel prognostic and predictive biomarker in LUAD.

Archaeal communities in natural and artificially restored mangrove sediments in Tieshan Bay, China

Mangrove forests are crucial wetland ecosystems located in tropical and subtropical intertidal zones, but they have become extensively degraded. As a viable ecological restoration strategy, the cultivation of native mangrove vegetation in these degraded areas has gained considerable attention. Mangroves’ unique environmental conditions make them suitable habitats for diverse microbial communities, including Archaea - one of the main microbial communities in mangrove sediments - which plays a pivotal role in biogeochemical cycles. However, little is known about the dynamics of archaeal communities during mangrove restoration through phytoremediation. In this study, we investigated the physicochemical properties of sediment profiles from natural and artificially restored mangrove ecosystems in Tieshan Bay. We utilized quantitative polymerase chain reaction (qPCR) and 16S rRNA gene sequencing techniques to explore differences in abundance, community structure, and composition of archaeal communities between sediment profiles in natural and artificially restored mangrove ecosystems. We also examined correlations between archaeal communities and environmental factors. Our results revealed that Crenarchaeota, Thermoplasmatota, Asgardarchaeota, Nanoarchaeota, and Euryarchaeota were the predominant archaeal phyla, with significant variation in sediment composition observed for Crenarchaeota and Thermoplasmatota in different depths. We also found significant differences in archaeal abundance and community composition between natural and restored mangrove sediments. Furthermore, C/N ratio and pH emerged as primary drivers of archaeal communities in wet and dry season sediments, respectively. Additionally, the study revealed seasonal disparities in seasonal differences in the relative abundance of certain archaeal groups (such as Asgardarchaeota, Nanoarchaeota). Network analysis demonstrated stronger interconnections among archaeal communities in sediments from natural mangroves than from artificially restored ones. These findings enhance our knowledge of archaeal community succession patterns in mangrove restoration, as well as provide fresh perspectives for the sustainable management of mangrove ecosystems.

Association of insertion/deletion (I/D) polymorphism of angiotensin-converting enzyme gene (ACE) with Parkinson’s disease and factors risk in eastern Algeria: case–control study

BackgroundThis study aimed to explore the relationship between Parkinson’s disease and insertion/deletion polymorphism (I/D) of the angiotensin-converting enzyme gene (ACE) and to highlight the related risk factors within Eastern Algerian population. A total of 262 individuals were recruited, including 100 PD patients and 162 controls. Polymerase chain reaction technique was employed to determine the ACE polymorphism genotype of each participant. Logistic regression analysis was conducted to examine factors potentially contributing to Parkinson’s disease.ResultsThe mean age of onset of Parkinson’s disease in the patient group was 56.18 ±12.99\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\pm 12.99$$\\end{document}. The results revealed that the D allele of the angiotensin-converting enzyme gene and the DD genotype were most prevalent in both patients and controls. However, there were no significant differences in the genotype and allele frequencies of the I/D polymorphism of the angiotensin-converting enzyme gene between patients and healthy subjects. Cardiovascular disease, diabetes and profession were identified as potential risk factors for Parkinson’s disease.ConclusionThe obtained data indicated no correlation between the angiotensin-converting enzyme I/D gene polymorphism and Parkinson’s disease in our research cohort. Multiple logistic regression analysis revealed that cardiovascular disease, diabetes and profession were possible risk factors for Parkinson’s disease.

Association of single nucleotide polymorphism in the loxl1 gene with pseudoexfoliation syndrome in northwestern Rajasthan

Purpose: To identify, evaluate and establish the relation of LOXL1 gene polymorphism among patients with pseudoexfoliation syndrome in North West Rajasthan. Methods: 50 patients diagnosed with pseudoexfoliation syndrome and 50 healthy subjects of age >40 years were enrolled in the study. All participants underwent routine ophthalmic examination. 2ml blood samples were taken of each participant which were sent to Multi Disciplinary Research Unit to be checked for specific gene mutation using Polymerase Chain Reaction technique. Results and conclusion: Study showed that rs10486(G) SNP and rs21652(T) SNP has been present in general population also but is more prevalent in those with pseudoexfoliation syndrome. While, rs38259(G) SNP was found not to be a significant factor in development of pseudoexfoliation syndrome. Understanding the genetic basis of pseudoexfoliation syndrome would further contribute in aiding the early diagnosis and management of this disease.