Diverse Expression Patterns of EBV Oncogenes (LMP2A, EBV-Encoded microRNA, and EBV-encoded dUTPase) in EBV Associated Gastric Carcinoma and their Association with Viral Loads.

The chromogenic in situ hybridization (CISH) test is the gold standard for detecting Epstein-Barr virus (EBV)-associated gastric carcinoma (GC). Real-time (RT) PCR method is also a sensitive test that can detect the viral load in samples. As such, three EBV oncogenes were investigated in this study. RNA extraction and cDNA synthesis were performed on GC tissues of nine patients, who were previously confirmed to have EBVGC subtype. In addition, 44 patients that had positive RT-PCR but negative CISH results were also included as the control group. TaqMan RT-PCR analysis was performed to determine the expression of EBV-encoded microRNAs, and the expression of EBV-encoded dUTPase, as well as LMP2A, was analyzed by SYBR Green RT-PCR. EBV-encoded microRNAs and LMP2A were identified in 2 out of 9 (22%) EBVGC subtypes. In addition, EBV-encoded dUTPase was detected in 4 out of 9 (44.5%) EBVGC subtypes. EBV-encoded dUTPase was also expressed in a sample of the control group. The expression of LMP2A, EBV-encoded microRNAs, and EBV-encoded dUTPase viral oncogenes in patients with high EBV viral loads indicates that these expressions correlate with viral loads. Our findings indicate that the EBV-encoded dUTPase gene may have a role in EBVGC patients' non-response to treatment and might be considered a Biomarker-targeted therapy.

Surveillance of Highly Pathogenic Avian Influenza Viruses (H5Nx Subtypes) in Wild Birds in Iran, 2014-2019.

After the emergence of the highly pathogenic avian influenza viruses (HPAIV) subtypes H5N6 in 2013 and H5N8 in 2014, a surveillance study using molecular epidemiology approaches was carried out during 2014 - 2019 in Iran to discover any potential introduction or outbreak of HPAIV in wild bird populations. All sick and dead wild birds found in nature, or in cases of an outbreak, a collection of representative samples was tested using the specific molecular methods for HPAIV H5 subtypes. Additionally, wild bird species in wetlands, several zoos, zoological gardens, or rehabilitation centers were tested for HPAIV. During the active surveillance plan, several individual and outbreak cases of HPAIV and orthoavulaviruses were identified. In general, more than 900 fecal materials, cloacal and oropharyngeal swabs, and/or tissue samples were collected from apparently healthy live birds representing several different species and families. In addition, tissue and swab samples were collected and investigated from any reported wild birds' mortality cases in different parts of Iran in the framework of this study. No positive bird was found among apparently healthy live birds; however, the highly pathogenic influenza viruses of H5N1, H5N2, H5N6, and H5N8 were found in individual dead birds or mass die-off cases.

First detection of mobilized colistin resistance mcr-1 gene in Escherichia coli isolated from livestock and sewage in Iran.

Currently, few studies have investigated the mechanisms of resistance to colistin in Iran. The aim of this study was to investigate mcr-harbouring Escherichia coli dissemination in livestock and sewage in Iran. A total of 115 samples from cows (n = 38), chickens (n = 47) and urban sewage samples (n = 30) were collected. The presence of genes including mcr1-6 and ampC ß-lactamase (bla MOX, bla CIT, bla DHA, bla ACC, bla EBC, bla FOX) for colistin-resistant isolates was investigated by multiplex PCR method. Genetic association of colistin-resistant strains was also evaluated by ERIC PCR. Sixty-five isolates were identified as E. coli. Meaningless were resistant to colistin. The highest (26.1%) and lowest (3.07%) resistance were shown to ampicillin and meropenem respectively. Among the three colistin-resistant isolates, 2 (66%) were multidrug resistant, with one of them being mcr-1 positive and the other one positive for DHA ampC ß-lactamase gene. No mcr2-6 genes were found. Minimum inhibitory concentration of mcr-producing isolate was 4 mg/L by microbroth dilution. This study reports, first the detection of mcr-1 in E. coli from farm animals in Iran, a finding that is indicative of a global distribution of this plasmidic element and threatning the use of colistin as a last resort antibiotic. No clonal relationship was observed between the colistin-resistant E. coli isolates by ERIC-PCR. Monitoring the presence of these strains in animal sources help as to controlling the spread of resistance genes from animal to human is vital.

Identification of microorganisms in irreversible pulpitis and primary endodontic infections with respect to clinical and radiographic findings.

OBJECTIVES: The aim of this study was to evaluate the composition of microbiota of irreversible pulpitis and primary endodontic infections with respect to clinical and radiographic findings by performing cultures and 16s rDNA sequencing in Iranian patients. MATERIAL AND METHODS: In this prospective cross-sectional study, samples were collected from 41 root canals for 4 main groups of patients. Bacterial identification was performed by the polymerase chain reaction (PCR) and 16s rDNA sequencing of aerobic and anaerobic cultivable colonies taken from patients' culture plates. Additionally, the presence of 13 bacterial species and 3 nonbacterial species was also explored using PCR and species-specific primers. RESULTS: Sixteen microbial species, 1 fungus (Candida albicans), and 1 virus (Herpes simplex virus) were discovered and isolated. Species with the highest prevalence were Dialister invisus (68.3%), Porphyromonas gingivalis (58.8%), Streptococcus salivarius (58.5%), and Treponema denticola (56.1%). Lysinibacillus fusiformis (19.1%) was detected in the root canals for the first time. Candida albicans was seen in 11 cases (26.8%). Herpes simplex virus (HSV) was seen in 4 patients (9.8%). CONCLUSIONS: Our results suggest that Gram-negative anaerobic oral bacteria are the majority of the microbes in primary endodontic infections. Various combinations of bacterial species were related to different clinical and radiographic conditions. Lysinibacillus fusiformis was detected for the first time in primary endodontic infections. CLINICAL RELEVANCE: The results of this investigation might help clinicians choose to identify suspected endodontic pathogens in the etiology of each form of pulpal and periradicular diseases to determine the best therapeutic measures.

Stem Cell-Based Therapies and Tissue Engineering of Trachea as Promising Therapeutic Methods in Mustard Gas Exposed Patients.

Tissue engineering and cell-based therapies are promising therapeutic approaches in structural and functional defects of the trachea. Researchers have focused on these approaches to overcome the complications related to such diseases. Patients exposed to mustard gas suffer from massive damage to the respiratory system. Current treatment plans are only palliative and include anti-inflammatory drugs, broncholytics, long-acting ß2-agonists, and inhaled corticosteroids. As mustard gas exposure leads to chronic airway inflammation, it seems that tracheobronchomalacia, because of chronic inflammation and weakness of the supporting cartilage, is an important factor in the development of chronic and refractory respiratory symptoms. The previous studies show that regenerative medicine approaches have promising potential to improve the life quality of patients suffering from tracheal defects. It seems that the engineered tracheal graft may improve the respiratory function and decrease symptoms in patients who suffer from asthma-like attacks due to mustard gas exposure. There are several successful case reports on the transplantation of stem cell-based bioartificial grafts in structural airway diseases. Therefore, we hope that the reconstruction of tracheobronchial structure can lead to a decrease in respiratory difficulties in mustard gas-exposed patients who suffer from tracheomalacia. In the present review, we summarize the main aspects of tracheal tissue engineering and cell-based therapies and the possibilities of the application of these approaches in mustard gas-exposed patients.

Paediatric burn injuries requiring hospitalization in Fars, Southern Iran.

The aim of this study was to determine the demographic characteristics of burnt children in need of hospitalization, causes of burns and associated complications in Fars province, Iran. This is a retrospective cross-sectional study. Files of all children under 15 years of age who were hospitalized in the only burn referral centre of Fars province were evaluated. Data regarding age, gender, location (urban, rural), burn surface area (BSA), cause of burn, length of hospital stay and complications were extracted from patients' files. Data were analyzed using SPSS, version 22 and the Chi-square test. A p-value of less than 0.05 was considered as statistically significant. A total of 122 patients (54.9% males) were studied. Children from rural areas were hospitalized 1.4 times more often than urban children. Overall, 31.2% of admissions occurred in winter. Burning with hot liquids (scalding) was the most common cause of burns (56.6%, n = 69). Mean BSA was 12.29 ± 21.18% and mean length of hospital stay was 7.59 ± 12.78 days. Burn complications were seen in 19 cases (15.6%). One child died due to inhalational thermal injury. This study showed that burns mostly occur in boys, in the winter and in rural areas of Fars province. Furthermore, scald burns are the most common type of burn injury. Since a significant number of children suffer from permanent complications following burn injuries, special planning is needed to prevent this type of injury.

Le but de cet étude rétrospective était de déterminer les caractéristiques démographiques, les causes et les complications des brûlures pédiatriques ayant nécessité une hospitalisation à Fars (Iran). Les dossiers des enfants (<15 ans) hospitalisés dans le CTB de Fars ont été évalués. Les données concernant l'âge, le sexe, l'habitat (rural ou urbain), la surface brûlée (SB), la cause, la durée de séjour et les complications ont été recueillies et analysées par X² au moyen de SPSS 22, avec un seuil de significativité à p≤ 0,05. Cent vingt deux dossiers (54,9% de garçons) ont été étudiés. Les rapports enfants ruraux/urbains était de 1,4/1. Il y avait un pic d'admissions en hiver (31,4%). Un ébouillantement était le mécanisme le plus fréquent (69 fois soit 56,6%). La surface brûlée moyenne était de 12,29 (+/- 21,18%), et la durée moyenne d'hospitalisation était de 7,59 +/- 12,78. Un enfant est mort, dans un contexte d'inhalation de fumées. Dix neuf séquelles (15,6%) ont été observées. Les brûlures (qui sont plus souvent des ébouillantements) touchent donc plutôt des garçons, ruraux, pendant l'hiver. Dans la mesure où un nombre significatif d'enfants brûlés gardent des séquelles, des actions de prévention spécifiques sont nécessaires.

Naloxone added to bupivacaine or bupivacaine-fentanyl prolongs motor and sensory block during supraclavicular brachial plexus blockade: a randomized clinical trial.

BACKGROUND: In this study, the effect of naloxone on duration of supraclavicular brachial plexus block was evaluated. It was hypothesized that naloxone can increase the duration of neural blockade. METHODS: Sixty-eight patients scheduled for surgery under supraclavicular brachial plexus block were randomly assigned to receive 30 ml bupivacaine (Group C); 30 ml bupivacaine with 100 µg of fentanyl (Group F); 30 ml bupivacaine with 100 ng naloxone (Group N); or 30 ml bupivacaine with 100 µg of fentanyl and 100 ng naloxone (Group N + F). Sensory and motor blockade were recorded at 5, 15, and 30 min following the block, and every 10 min following the end of surgery. Duration of sensory and motor block was considered to be the time interval between the complete block and the first postoperative pain and complete recovery of motor functions. RESULTS: Sensory and motor onset times were the same in all groups. The duration of sensory and motor block in Group C (11.3 ± 1.7 h and 4.56 ± 1.0 h) and Group F (12.8 ± 3.3 h and 5.1 ± 2.0 h) were less than in the other groups (18.1 ± 2.2 h and 6.18 ± 1.0 h in Group N, and 15.8 ± 2.9 h and 6.53 ± 1.1 h in Group N + F, P < 0.0001). CONCLUSION: Addition of naloxone to bupivacaine in supraclavicular brachial plexus block prolonged the duration of the neural blockade.

Design of a multiplex PCR method for detection of toxigenic-pathogenic in Vibrio cholerae.

OBJECTIVE: To study virulence and regulatory genes (hlyA, ctxB, tcpI) in clinical strains of Vibrio cholerae (V. cholerae), simultaneously. METHODS: Three important genes, tcpI, hlyA and ctxB were used for detection of toxigenic and pathogenic V. cholera by chain reaction assay method. RESULTS: According to the results of the PCR, the incidence of hlyA, tcpI, and ctxB genes in clinical isolates was obtained as 94.7% (72 sample), 90.8% (69 sample), and 92.1% (70 sample), respectively. Five strains possessed all genes except ctxB, six strains possessed all genes except tcpI, four strains possessed all genes except hlyA, one strain possessed only hlyA and 60 strains contained a combination of three genes, Including hlyA, ctxB and tcpI. CONCLUSIONS: Result show that this method could be reliable to detect toxigenic-pathogenic strains of V. cholerae in Iran.

Chemical cleavage of mismatches in heteroduplexes of the rpoB gene for detection of mutations associated with resistance of Mycobacterium tuberculosis to rifampin.

In order to detect mutations in the core region of the RNA polymerase B (rpoB) subunit gene of Mycobacterium tuberculosis that are known to be associated with resistance to rifampin, we applied rapid chemical cleavage of mismatches (CCM) to heteroduplexes formed between the DNA of M. tuberculosis H37Rv and strains resistant to rifampin. DNA fragments amplified from normal and mutant rpoB genes by polymerase chain reaction were mixed, denatured and re-annealed to create heteroduplexes containing mispaired bases reactive to modification by hydroxylamine (cytosine mismatches) or osmium tetroxide (thymine mismatches) and cleavage of DNA by piperidine at the position of modified base. The cleaved products and the heteroduplexes were separated by polyacrylamide-urea gel electrophoresis and detected by autoradiography. The position of mutations was confirmed by DNA sequencing of the amplified DNA fragments. The results suggest further applicability of the CCM method as a means to screen M. tuberculosis isolates for mutations associated with drug resistance.