Polymerase Chain Reaction Technique Research Articles

Portable, and ultrasensitive HR-HPV tests based on nucleic acid biosensors.

Cervical cancer is the third most common cancer threatening women's health globally, and high-risk human papillomavirus (HR-HPV) infection is the main cause of cervical cancer worldwide. Given the recurrent nature of HR-HPV infection, accurate screening is essential for its control. Since the commonly used polymerase chain reaction (PCR) technique is limited by professional equipment and personnel, convenient and ultrasensitive detection methods for HR-HPV are still highly needed. As new molecular detection methods, nucleic acid amplification-based biosensors have the advantages of high sensitivity, rapid operation, and portability, which are helpful for point-of-care testing in rural and remote areas. This review summarized nucleic acid biosensors for HR-HPV screening based on a variety of nucleic acid amplification strategies involved in improved PCR, loop-mediated isothermal amplification, recombinase polymerase amplification, hybridization chain reaction, catalyzed hairpin assembly, and CRISPR/Cas systems. In combination with microfluidic technology, lateral flow assays, electrochemical analysis and other sensing technologies, HR-HPV nucleic acid biosensors have the advantages of high throughput, short response time, high sensitivity and easy operation in the field. Although there are still shortcomings, such as high cost and poor reproducibility, this approach will be suitable for on-site screening of HR-HPV infection or cervical cancer and for auxiliary clinical diagnosis in complex environments and poor areas in the future.

Identification ofantibiotic resistance genes inEscherichia coli from subclinical mastitis milk indairy cows and goats, East Java Province.

Antibiotics are still used totreat mastitis indairy cows inIndonesia. This study aimed toanalyse antibiotic resistance genes inEscherichia coli (E.coli) from subclinical mastitis milk inEast Java Province, Indonesia. Thesamples consisted ofsubclinical mastitis milk from cows and goats. Atotal of592-quarter cow's milk and 71goat's milk samples from both halves ofthe udder were collected from 67farms inLumajang, Banyuwangi, Malang, Sidoarjo, Jember, Pasuruan, Probolinggo, and Mojokerto. Subclinical mastitis samples were screened using the California mastitis test (CMT). E.coli was identified byphenotypic and genotypic methods. E.coli wasconfirmed with aprimer specific tothe polymerase chain reaction (PCR) technique. Gene resistance ofE.coli was tested using the multiplex-PCR (mPCR) technique with primers encoding the genes temoneira enzyme (TEM), oxacillinase (OXA), sulfhydryl variable (SHV), and cefotaximase-munich IV (CTX-M IV). These genes were chosen because mastitis treatment generally uses oxacilline and β-lactam antibiotics. All data obtained were analysed descriptively. Theresults show that six isolates ofE.coli (46.15%) carried asingle resistance gene (TEM orSHV) and two isolates (33.33%) were confirmed asmultiple drug-resistant organisms (MDROs) (TEM and SHV). Theresistance genes were found insamples originating from Blitar, Banyuwangi, Lumajang, and Pasuruan Regencies. This research implies that antibiotic-resistance genes found inE.coli oncertain farms are dangerous and may allow gene transmission toother bacteria that make treatment for mastitis orother bacterial infections ineffective.

Candida Variety in the Oral Cavity of Mexican Subjects with Type 2 Diabetes Mellitus and TLR2 Gene Expression.

The aim was to diagnose Candida in the oral cavity of subjects with type 2 diabetes mellitus (T2DM) using a genotyping technique and compare the results with those from conventional diagnosis by Papanicolaou (Pap) staining. Palatal mucosa smears were performed on 18 dental care patients diagnosed with T2DM and grade I, II, and III prosthetic stomatitis who met the inclusion criteria; 18 healthy control subjects were also included in the study. Hemoglobin A1c (HbA1c) levels were determined from total blood. Using exfoliative cytology, the Pap staining technique was used to diagnose candidiasis. Exfoliative cytology was also used for molecular diagnosis; DNA was obtained for Candida genotyping, and RNA was used for gene expression studies. Clinical patterns indicated that all subjects were positive for Candida; however, Pap analysis revealed only three positive subjects, whereas end-point polymerase chain reaction (PCR) analysis revealed 15 subjects with some type of Candida. The most common Candida species found were Candida guilliermondii (38.8%), Candida krusei (33.3%), Candida tropicalis, and Candida lusitaniae (22.2%). Interestingly, the coexpression of different species of Candida was found in various patients. In all patients, HbA1c levels were increased. Gene expression analysis showed a significant decrease (p ≤ 0.05) in TLR2 expression in positive subjects, whereas TLR4 expression did not differ significantly among patients. The end-point PCR technique showed better sensitivity for the diagnosis of Candida when compared with the diagnosis by Pap staining. T2DM subjects showed an increased presence of C. guilliermondii that was correlated with decreased TLR2 expression.

The Impact of Genetic Polymorphisms in (STIM1 and ORAI1) on Erythropoietin Resistance in Patients with Chronic Renal Failure on Hemodialysis in Iraq

Objective: This study aims to investigate the correlation between genetic polymorphisms in store-operated calcium channels (SOCs) signaling and resistance to erythropoietin treatment in patients with chronic renal failure (CRF). Anemia is a significant complication of CRF, and understanding the genetic factors contributing to resistance could provide insights into potential mechanisms influencing treatment outcomes. Specifically, the focus is on exploring the relationship between SOCs, a pathway activated by erythropoietin, and resistance to address the challenges associated with anemia in CRF patients.
 Methods: In this study, a single nucleotide polymorphism in each STIM1 and ORAI1 gene was selected. Genotyping was done by using allele specific polymerase chain reaction technique and the data was analyzed through the Statistical Package for the Social Sciences. 112 patients with CRF on hemodialysis were enrolled 58.9% of them were men and the mean age was 50.94±13.42.
 Results: We found that CC genotype of ORAI1 gene has a higher hemoglobin level, AA genotype of STIM1 has a higher EPO level, the interaction of the two genes showed that CCGG group has a higher Hb level and is considered good responder but represents only 4.5% of the patients
 Conclusion: After all the genetic polymorphisms of SOCs genes ORAI1 and STIM1 have no significant impact on erythropoietin resistance in patients with CRF on hemodialysis in Iraq.

CircCPA4 induces ASCT2 expression to promote tumor property of non-small cell lung cancer cells in a miR-145-5p-dependent manner.

Non-small cell lung cancer (NSCLC) is a type of lung cancer that occurs in the cells of the respiratory tract, and its development is influenced by the regulation of circular RNAs (circRNAs). However, the role of circRNA carboxypeptidase A4 (circCPA4) in the progression of NSCLC and the underlying mechanism remain relatively clear. The study utilized both real-time quantitative polymerase chain reaction (RT-qPCR) and western blot techniques to evaluate the levels of circCPA4, microRNA-145-5p (miR-145-5p), alanine, serine, or cysteine-preferring transporter 2 (ASCT2). To assess cell proliferation, cell counting kit-8 (CCK8) and 5-ethynyl-2'-deoxyuridine (EdU) assays were performed. Apoptosis was determined using flow cytometry, while cell migration and invasive capacity were evaluated through transwell and wound-healing assays. Intracellular levels of glutamine, glutamate, and α-KG were measured using specific kits. The relationship between miR-145-5p and circCPA4 or ASCT2 was confirmed using dual-luciferase reporter assay and RNA immunoprecipitation assay. CircCPA4 and ASCT2 RNA levels were elevated, while miR-145-5p was downregulated in both NSCLC tissues and cells. Depletion of circCPA4 significantly inhibited NSCLC cell proliferation, migration, invasion, and intracellular levels of glutamine, glutamate, and α-KG, and promoted apoptosis. Moreover, circCPA4 knockdown delayed tumor growth in vivo. Furthermore, circCPA4 was found to bind to miR-145-5p, thereby regulating the progression of NSCLC in vitro. ASCT2 was also identified as a downstream target of miR-145-5p, and its upregulation rescued the effects of miR-145-5p overexpression on NSCLC cell processes. CircCPA4 knockdown inhibited tumor property of NSCLC cells by modulating the miR-145-5p/ASCT2 axis.

Meningitis and Tuberculosis: A Correlation along with Diagnosis and Management of Tuberculosis Meningitis

Tuberculosis, caused by Mycobacterium tuberculosis (M. tb), is a communicable disease and a leading cause of death globally, especially in developing countries. It exists in active and latent forms, affecting both pulmonary and extrapulmonary sites. The pathophysiology of TB involves interactions with various immune cells, particularly T cells, in defense against the infection. On the other hand, meningitis is the inflammation of the membranes surrounding the brain and spinal cord, and it can be caused by bacteria or viruses. Bacterial meningitis, associated with high mortality and risk of disability. The article highlights the promulgation of TB into the brain and the immune responses within the brain, shedding light on the migration of M. tb into the central nervous system (CNS) and the innate immune responses involving microglia and cytokine secretion. The diagnostic methods for TB meningitis encompass neuroimaging, adenosine deaminase measurement in cerebrospinal fluid (CSF), microscopy, polymerase chain reaction (PCR) techniques, and enzyme-linked immunosorbent assay (ELISA). Moreover, it discusses the treatment of TB meningitis, emphasizing traditional antimicrobial therapies, systemic corticosteroid therapy, adjunctive therapy, fluid management, surgical intervention for hydrocephalus, and considerations for patients co-infected with HIV. The treatment approaches stress the importance of timely and empirical treatment, the use of newer generation fluoroquinolones, systemic corticosteroids as adjunctive therapy, and surgical interventions for hydrocephalus. The article also addresses the potential challenges and considerations for patients co-infected with HIV, such as the risk of immune reconstitution inflammatory syndrome (IRIS), drug interactions, and the efficacy of adjunctive corticosteroids. Overall, the document provides a comprehensive overview of the correlation between TB and meningitis, covering their pathophysiology, diagnostic methods, and treatment approaches. It emphasizes the complexity of managing TB meningitis and the importance of considering various factors, including immune responses, diagnostic accuracy, and tailored treatment strategies, particularly in the context of HIV co-infection.

Different Concentrations of Produced Recombinant TNF-α Protein Caused Different Cellular Viability in Breast Cancer Cell Lines

Background: Breast cancer is a prevalent malignancy among women globally. The implementation of early detection methods and improved treatment strategies have shown a considerable decrease in the financial burden associated with breast cancer mortality. One of the inflammatory cytokines is tumor necrosis factor (TNF), produced during chronic inflammation, apoptosis, tumor proliferation, and angiogenesis. The main objectives of this study were to produce recombinant TNF-α protein and assess its impact on the MCF-7 and MDA-MB-468 cancer cell lines. Methods: The present study employed the SHuffle® T7 express strain of E. coli to synthesize recombinant TNF-α protein, primarily due to its advantageous characteristics of cost-effectiveness and high productivity. The molecular weight of the protein was determined using SDS-PAGE and Coomassie blue staining techniques, and its presence was subsequently verified through Western blot analysis. Subsequently, the cells were treated with the produced recombinant protein, and the cellular viability was assessed using the MTT assay. Then, the Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) technique was employed to quantify the expression level of the Caspase-3 gene, known as an apoptotic factor, and the interleukin-1 (IL-1) gene, recognized as a pro-inflammatory factor, in both cell lines. Results: Lower quantities of recombinant TNF-α protein were shown to cause cellular cytotoxicity, according to the findings of the MTT test, while cellular proliferation was observed at increased levels of recombinant TNF-α protein. The results of the qRT-PCR test exhibited a lack of identifiable patterns in both cell lines. Conclusion: The findings indicate that cellular viability is significantly decreased at a minimal concentration of 5-10 ng/mL of TNF-α, whereas it is noticeably elevated at concentrations higher than 10 ng/mL. Furthermore, gene expression analysis revealed a significant reduction in IL-1 expression in both cell lines after administering a high dose of TNF-α.

Sp1 Regulates the M1 Polarization of Microglia Through the HuR/NF-κB Axis after Spinal Cord Injury

The M1 polarization of microglia, followed by the production of pro-inflammatory mediators, hinders functional recovery after spinal cord injury (SCI). Our previous study has illuminated that specificity protein 1 (Sp1) expression is increased following SCI, whereas the function and regulatory mechanism of Sp1 during M1 polarization of microglia following SCI remain unknown. RNA binding protein, HuR, has been shown to be up-regulated in the injured spinal cord through analysis of the GEO database. Further investigation using Chip-Atlas data suggests a binding between Sp1 and HuR. Emerging evidence indicates that HuR plays a pivotal role in neuroinflammation after SCI. In this research, Sp1 and HuR levels in mice with SCI and BV2 cells treated with lipopolysaccharide (LPS) was determined by using quantitative real-time polymerase chain reaction and Western blotting techniques. A series of in vitro assays were performed to investigate the function of Sp1 during M1 polarization of microglia. The association between Sp1 and its target gene HuR was confirmed through gene transfection and luciferase reporter assay. Enhanced expression of HuR was observed in both SCI mice and LPS-treated BV2 cells, while Sp1 knockdown restrained M1 polarization of microglia and its associated inflammation by inhibiting the NF-κB signaling pathway. Silencing Sp1 also suppressed microglia activation and its mediated inflammatory response, which could be reversed by overexpression of HuR. In conclusion, silencing Sp1 restrains M1 polarization of microglia through the HuR/NF-κB axis, leading to neuroprotection, and thus promotes functional restoration following SCI.

Inhibition of Alzheimer's disease by 4-octyl itaconate revealed by RNA-seq transcriptome analysis

AimsThis study aimed to examine the therapeutic effects and response mechanisms of 4-OI in Alzheimer's disease (AD). MethodsIn this study, network pharmacology was employed to analyze potential targets for AD drug therapy. Immunofluorescence and quantitative reverse transcription polymerase chain reaction (qRT-PCR) techniques were utilized to detect inflammatory phenotypes in a 4-OI-resistant mouse microglia cell line (BV2). We conducted four classical behavioral experiments, namely the open field test, new object recognition test, Y maze test, and Morris water maze, to assess the emotional state and cognitive level of APPswe/PS1dE9 (referred to as APP/PS1) mice after 4-OI treatment. Hematoxylin and eosin (HE) staining, along with immunofluorescence staining, were performed to detect amyloid (Aβ) deposition in mouse brain tissue. To explore the potential molecular mechanisms regulating the effects of 4-OI treatment, we performed RNA-SEQ and transcription factor prediction analyses. Additionally, mouse BV2 cells underwent Western blotting analysis to elucidate potential molecular mechanisms underlying the observed effects. ResultsWe discovered that 4-OI exerts an inhibitory effect on neuroinflammation by promoting autophagy. This effect is attributed to the activation of the AMPK/mTOR/ULK1 pathway, achieved through enhanced phosphorylation of AMPK and ULK1, coupled with a reduction in mTOR phosphorylation. Furthermore, 4-OI significantly enhances neuronal recovery in the hippocampus and diminishes Aβ plaque deposition in APP/PS1 mice, improved anxiety in mice, and ultimately led to improved cognitive function. ConclusionsOverall, the results of this study demonstrated that 4-OI improved cognitive deficits in AD mice, confirming the therapeutic effect of 4-OI on AD.

The Association Between Genetic Variants in ACE1and ACE2 Genes with Susceptibility to COVID-19 Infection.

Angiotensin-converting enzyme 2 (ACE2) receptors facilitate the entry of the causative virus severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) into target cells. Some ACE gene variants have been suggested to be involved in COVID-19 pathogenesis. So, the aim was to assess the association between ACE1 rs4646994 and ACE2 rs2285666 genes polymorphisms and the susceptibility and severity of COVID-19. This case-control study was conducted on 197 patients with COVID-19 and 197 healthy controls. ACE-1 insertion/deletion (I/D) (rs4646994) and ACE2 rs2285666 genes polymorphisms were determined by the amplification refractory mutation system- polymerase chain reaction (ARMS-PCR) technique. The DD genotype of ACE1 I/D polymorphism was associated with increased susceptibility to COVID-19 infection (p = 0.012), whereas the ID genotype of this polymorphism was associated with decreased susceptibility (p = 0.003) (significance level = 0.017). There was no significant association in allele and genotype distribution of ACE2 rs2285666 polymorphism between cases and controls. The ACE1 I/D polymorphism may be considered as a risk factor for COVID-19 susceptibility.

Usefulness of polymerase chain reaction tests in Chagas disease studies

The Polymerase Chain Reaction (PCR) test is a highly sensitive, specific, and rapid diagnostic tool for Chagas disease. Chagas disease is caused by the protozoan flagellate Trypanosoma cruzi and is endemic to the Americas. While conventional serological methods are still used in the diagnosis of Chagas disease, they are being gradually replaced by molecular methods like PCR. PCR can detect the parasite’s DNA in blood or tissue samples from humans and animals, including asymptomatic infections and animal reservoirs. In a study conducted on a colony of New World monkeys, PCR analysis was found to be superior to conventional screening tools for trypanosome infection, although false negatives can still occur. In clinical studies, PCR has been used to assess the effectiveness of Nifurtimox and Benznidazole in treating acute and chronic Chagas patients. However, the presence of low-grade and intermittent parasitemia in peripheral blood, even in the absence of treatment, renders PCR an unreliable test for evaluating successful treatment. Based on this limiting factor, among others, we do not believe that PCR is an appropriate gold standard test for Chagas in clinical and preclinical studies. Other diagnostic methods, such as serological and biomarker tests, should be used in conjunction with PCR techniques for more accurate diagnosis of Chagas.

Insulin Ameliorates Neurodegeneration in an Experimental Animal Model of Brain Aging by a Decrease of GFAP Expression and Apoptosis Inhibition

Background: Aging is one of the factors that will cause gradual changes in brain tissue and its destruction. One of the protective factors against aging is insulin. Objectives: Our study investigated insulin's neuroprotective effect by preventing oxidative stress-mediated neuronal damage. Methods: A total of 48 adult male NMRI (the Naval Medical Research Institute) mice were divided into 2 groups: Control and insulin. Insulin was administered intraperitoneally (IP) for 8 weeks to the control group and for 12 weeks to the insulin group. In this study, animals were treated with insulin, and insulin's role in the aging process was evaluated. Then, brain tissue was extracted for evaluation of oxidative stress by assessment of glutathione disulfide (GSH) and reactive oxygen species (ROS) levels, the expression of the glial fibrillary acidic protein (GFAP) as a biomarker for astrogliosis, stereological study, and the real-time polymerase chain reaction (PCR) technique evaluated the level of apoptosis biomarker. Results: The results of stereological parameters show that the volumes of the brain cortex and the number of neurons improved in the insulin group by aging as compared to the control group. The results of real-time PCR showed that the expression level of apoptotic markers decreased in the insulin group compared to the control group. In the insulin group, GSH levels were higher than in the control group, and the control group produced more ROS than the insulin group. As compared with insulin, the control group expressed more GFAP protein and increased the number of glial cells by aging. Conclusions: The results showed that insulin could have neuroprotective effects against aging changes and reduce apoptosis and cell death.

The impact of MYD88 and PIM1 in mature large B-cell non-Hodgkin lymphomas: Defining element of their evolution and prognosis.

Sequence studies of the entire exome and transcriptome of lymphoma tissues have identified MYD88 and PIM1 as involved in the development and oncogenic signaling. We aimed to determine the frequency of MYD88 and PIM1 mutations, as well as their expressions in conjunction with the clinicopathological parameters identified in mature large B-cell non-Hodgkin lymphomas. The ten-year retrospective study included 50 cases of mature large B-cell lymphoma, diagnosed at the Pathology Department of the Emergency County Hospital of Constanţa and Săcele County Hospital of Brasov. They were statistically analyzed by demographic, clinicopathological, and morphogenetic characteristics. We used a real-time polymerase chain reaction technique to identify PIM1 and MYD88 mutations as well as an immunohistochemical technique to evaluate the expressions of the 2 genes. Patients with lymphoma in the small bowel, spleen, brain, and testis had a low-performance status Eastern Cooperative Oncology Group (P = .001). The Eastern Cooperative Oncology Group performance status represented an independent risk factor predicting mortality (HR = 9.372, P < .001). An increased lactate dehydrogenase value was associated with a low survival (P = .002). The international prognostic index score represents a negative risk factor in terms of patient survival (HR = 4.654, P < .001). In cases of diffuse large B-cell lymphoma (DLBCL), immunopositivity of MYD88 is associated with non-germinal center B-cell origin (P < .001). The multivariate analysis observed the association between high lactate dehydrogenase value and the immunohistochemical expression of PIM1 or with the mutant status of the PIM1 gene representing negative prognostic factors (HR = 2.066, P = .042, respectively HR = 3.100, P = .004). In conclusion, our preliminary data suggest that the oncogenic mutations of PIM1 and MYD88 in our DLBCL cohort may improve the diagnosis and prognosis of DLBCL patients in an advanced stage.

Genetic identification of the sticktight flea Echidnophaga gallinacea (Westwood) infesting chickens in the Al-Baha region, Saudi Arabia

Stick-tight fleas, Echidnophaga gallinacea (Westwood), are major ectoparasites of domesticated chickens and can cause serious diseases and even death if left untreated. In the present study the flea samples were collected from three traditional chicken-raising farms in the Al-Baha region. The samples were examined under a stereomicroscope and identified using classification keys. After extracting DNA from the insects, the polymerase chain reaction technique was used to identify the hereditary gene, cytochrome oxidase, present in the insect biopics. The gene was purified, its nucleotide sequence was obtained, and the accession number (OR161051) has been assigned in GenBank. After determining its nucleotide sequence, it was compared with other insects in GenBank, where it was found to be identical (99.82%) to the E. gallinacea isolate from Thailand recorded in GenBank [OQ291364 and MW492259 (https://www.ncbi.nlm.nih.gov/nucleotide/MW492259.1?report=genbank&log$=nucltop&blast_rank=1&RID=VJR9ZAEE013)]. The match was 94.33% with Echidnophaga iberica from Spain (KF479239) and 93.97% with Echidnophaga oschanini from China (KU880666). The phylogenetic tree also showed similarities between the Saudi Arabian isolates and other isolates. This genetic study of sticktight fleas is the first of its kind in the Kingdom of Saudi Arabia, therefore will be valuable for assessing the prevalence and geographical distribution of this parasite.

Genotoxicity Assessment of Quinoin, a Ribosome Inactivating Protein from Quinoa Seeds, in the Teleost Danio rerio.

Ribosome inactivating proteins (RIPs) are N-glycosylases found in various plants that are able to specifically and irreversibly inhibit protein translation, thereby leading to cell death. Their cytotoxic properties have attracted attention in the medical field in the context of developing new anticancer therapies. Quinoin is a novel toxic enzyme obtained from quinoa seeds and classified as a type 1 RIP (Chenopodium quinoa Willd.). Recently, quinoin was found to be cytotoxic to normal fibroblasts and keratinocytes in vitro, as well as to several tumor cell lines. The aim of this study was to evaluate the in vitro and in vivo genotoxicity of quinoin in a zebrafish model. We evaluated its ability to induce DNA fragmentation, genomic instability, and reactive oxygen species (ROS) generation by means of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) reaction, randomly amplified polymorphic DNA (RAPD) Polymerase Chain Reaction (PCR) technique, and dichlorofluorescine (DCF) assay, respectively. Quinoin was found to cause genomic damage in zebrafish, as shown by DNA fragmentation, polymorphic variations leading to genomic instability, and oxidative stress. Interestingly, longer quinoin treatment caused less damage than shorter treatments. This study demonstrated ROS-mediated genotoxicity of quinoin toward the zebrafish genome. The reduced damage observed after longer quinoin treatment could indicate the activation of detoxification mechanisms, activation of repair mechanisms, or the loss of protein activity due to enzymatic digestion. In order to clarify the genotoxic actions of quinoin, further investigations of the response pathways to DNA damage are needed. Overall, the ability of quinoin to cause breaks and instability in DNA, together with its clear cytotoxicity, make it an interesting candidate for the development of new drugs for cancer treatment.

Effects of mouthwash on periodontal pathogens and glycemic control in patients with type 2 diabetes mellitus

Periodontitis is known to be associated with type 2 diabetes mellitus (T2DM), and gargling with mouthwash is known to reduce the incidence of periodontitis by inhibiting periodontal pathogens. However, the effects of mouthwash on oral and systemic conditions in patients with T2DM remain unknown. In this study, we investigated the effects of gargling with mouthwash on the number of red complex species, including Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, and HbA1c levels in patients with T2DM. Patients were instructed to gargle with water for 6 months, followed by gargling with mouthwash containing chlorhexidine gluconate for the subsequent 6 months. At each clinic visit, saliva was collected and bacterial DNA was extracted to detect red complex species using the polymerase chain reaction technique. The HbA1c level was determined using a blood sample. The number of red complex species significantly decreased in younger or male patients who gargled with mouthwash. Furthermore, HbA1c levels significantly decreased in younger patients or patients with higher HbA1c levels who gargled with mouthwash. These results suggest that gargling with mouthwash reduces the number of red complex species and improves the hyperglycemic status in patients with T2DM, especially younger patients.

Healthcare Provider N95 Respirator Contamination Worn Behind Face Shields With SARS-CoV-2 During Routine Clinical Care of Patients With COVID-19.

N95 respirator contamination with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during clinical care of patients with coronavirus disease 2019 is poorly understood. We performed a prospective observational study on healthcare provider's (HCP's) N95 respirators' and face shields' SARS-CoV-2 contamination during aerosol-generating procedures on SARS-CoV-2-positive patients housed in a COVID-19-specific unit. Medical masks worn on top of HCP's N95 respirators, and under face shields, during study aerosol-generating procedures were used as surrogates to detect contamination to avoid waste. Thirty-three HCPs were studied, and a total of 33 mask and 27 face shields were sampled. Masks were cut into 9 pieces and face shields were sampled twice, front and back, to determine locality of contamination; however, no positive samples were identified using standard polymerase chain reaction techniques with a CT value up to 40. All 9 mask piece samples were then pooled, as were face shield samples, using centrifugal concentration with polyethersulfone membranes. Once pooled and concentrated, overall, 9 (15%) samples were positive via real-time polymerase chain reaction: 5 from masks (15.2%) and 4 from face shields (14.8%).

Histomolecular profile of Helicobacter pylori strains circulating in Brazzaville (Congo).

Helicobacter pylori (Hp) infection is a real public health problem in the Congo. We aimed study the histomolecular profile of Hp strains circulating in Brazzaville, Congo, in order to contribute to the improvement of Hp-infected patients in the country. This was an analytical-transversal study carried out from January to November 2020 (i.e. a study period of 11 months) in the endoscopy centers of Brazzaville as well as the molecular biology and anatomopathology laboratories of Pointe-Noire and Oyo. It involved 100 symptomatic patients over the age of 18 referred for upper GI endoscopy. These patients underwent gastric biopsies for histopathological analysis according to the Sydney classification and molecular analysis using the real-time polymerase chain reaction (PCR) technique. The frequency of Hp infection was determined using real-time PCR. PCR was also used to identify the Hp strains and assess their tropism in the gastric mucosa. Digestive symptoms, endoscopic lesions, and histopathological lesions associated with HP infection were studied. The incidence of Hp infection was 91%, with a female predominance of 52.75% and an average age of 46.32 years. Endoscopy revealed normal mucosa (56.14%), ulcerated lesions (12.28%), and gastritis (22.81%) in infected patients. Histopathologically, the lesions were chronic atrophic gastritis (91%), with inflammatory activity (16.46%), intestinal metaplasia (16.46%), and adenocarcinoma (3.3%). Cag A strains were present in 85.71% of cases and had no preferential tropism in the gastric mucosa. Strains carrying the Cag A gene were present in severe and serious endoscopic and histopathological lesions. The prevalence of Hp infection is 91% in the Brazzaville population. Cag A strains circulate in high proportions and are implicated in the occurrence of severe lesions of the gastric mucosa.

Molecular study of blaVIM and blaIMP genes in Acinetobacter baumannii strains isolated from burn patients in Duhok City, Iraq.

Acinetobacter baumannii (A. baumannii) is an opportunistic pathogenic bacterium mainly associated with hospital acquired infections and in immunocompromised individuals who stay in hospitals for a long time. In recent years, it has become increasingly resistant to many different types of antibiotics. The production of the metallo-beta-lactamase (MBL) enzyme is one of the primary causes of this resistance. This study aimed to detect the presence of MBL genes that belong to the verona integrin metallo-β-lactamase (bla-VIM) and imipenemase (bla-IMP) groups in the isolates of Acinetobacter baumannii from burn patients. One hundred and seventeen (117) isolates of A. baumannii were obtained from patient specimens using traditional methods followed by using the VITEK 2 (BioMérieux, Les Pennes-Mirabeau, France) identification system. Metallo β-lactamases were detected in the imipenem-resistant strains by using imipenem disks on Muller-Hinton agar. The polymerase chain reaction (PCR) technique was utilized to examine 117 isolates for the detection of MBLs encoding genes such as bla-VIM, and bla-IMP. Imipenem resistance was detected in 78.6% of the patients. The PCR assays of the isolates identified bla-VIM-1, bla-VIM-2, bla-IMP-1 and bla-IMP-2 genes at the rates of 17%, 40.1%, 29.9% and 4.2%, respectively. The findings suggest that the majority of A. baumannii isolates harbour one or more of the detected genes, signifying that the production of MBLs plays a pivotal role in resistance mechanisms.

Pathogenic variant c.35delG of the GJB2 gene associated with nonsyndromic prelingual deafness

Introduction: the pathogenic variant c.35delG of the GJB2 gene is the most frequently observed in all populations, associated with nonsyndromic autosomal recessive prelingual prelingual sensorineural deafness, since 2001 is available in the National Network of Medical Genetics the study of this mutation. Objective: to describe the presence of the pathogenic variant c.35delG of the GJB2 gene associated with nonsyndromic prelingual deafness, with evidence of autosomal recessive inheritance. Methods: a descriptive cross-sectional study was carried out on 379 cases registered with isolated prelingual hearing loss between 2001 and 2023; for the identification of the c.35delG mutation, the polymerase chain reaction technique was used, with enzymatic digestion, and its genotype and frequency were described. Results: the pathogenic variant c.35delG of the GJB2 gene was found in 121 of those studied (31,91 %), 59 in homozygosis and 62 in heterozygosis. The allele frequency found among the positive cases was 0,743. Conclusion: the pathogenic variant c.35delG in individuals with nonsyndromic prelingual deafness of possible autosomal recessive inheritance is found in a high proportion