INPP5A phosphatase is a synthetic lethal target in GNAQ and GNA11-mutant melanomas.

Activating mutations in GNAQ/GNA11 occur in over 90% of uveal melanomas (UMs), the most lethal melanoma subtype; however, targeting these oncogenes has proven challenging and inhibiting their downstream effectors show limited clinical efficacy. Here, we performed genome-scale CRISPR screens along with computational analyses of cancer dependency and gene expression datasets to identify the inositol-metabolizing phosphatase INPP5A as a selective dependency in GNAQ/11-mutant UM cells in vitro and in vivo. Mutant cells intrinsically produce high levels of the second messenger inositol 1,4,5 trisphosphate (IP3) that accumulate upon suppression of INPP5A, resulting in hyperactivation of IP3-receptor signaling, increased cytosolic calcium and p53-dependent apoptosis. Finally, we show that GNAQ/11-mutant UM cells and patients' tumors exhibit elevated levels of IP4, a biomarker of enhanced IP3 production; these high levels are abolished by GNAQ/11 inhibition and correlate with sensitivity to INPP5A depletion. Our findings uncover INPP5A as a synthetic lethal vulnerability and a potential therapeutic target for GNAQ/11-mutant-driven cancers.

Investigating the effect of eye drops based on iodine nanoparticles in the treatment of corneal ulcers in rabbit eyes.

BACKGROUND: Various organisms, such as bacteria, viruses, and fungi, can cause corneal ulcers. One of the leading causes of vision loss and disability worldwide is corneal ulceration. Practical, accessible, and affordable treatment for this disease seems essential. MATERIALS AND METHODS: Fifteen New Zealand rabbits infected with Staphylococcus aureus (ATCC 25923) corneal ulcers were randomly divided into three groups of five for the present study. (I, II, and III). Group I was used as the control group (without treatment). The second group received an iodine solution (1.25%) without a nanoparticle structure (betadine). The third group received an iodine solution with a nanoparticle structure used as eye drops. Drops in the corneal ulcer group were used five times daily for 14 days. Microbial counts and disease severity scores were measured on the first, second, fifth, and fourteenth days and compared between groups separately for each disease. RESULTS: The results showed that the changes in microbial load were significant in the group that received betadine and nanoparticles. The microbial load was further reduced when using iodine nanoparticles than betadine. The betadine and nano-iodine groups significantly reduced the severity of the disease in rabbits with corneal ulcers (p < 0.05). The average changes in disease severity score were 4.8 ± 1.3, -2.6 ± 0.89, and -2.22 ± 1.22 in the untreated, nano iodine, and betadine groups, respectively. However, a significant increase in disease severity was observed in the untreated group (p = 0.001). It shows a significant difference (p < 0.001) between the nano iodine, betadine, and untreated groups. However, the difference in disease severity changes between nano iodine and non-nano iodine groups was insignificant. CONCLUSION: Nanoparticle iodine is more effective than non-nanoparticle iodine in reducing bacterial load. In reducing the severity of the disease, both types of iodine were superior to no treatment. But there was no apparent difference between the two groups treated with iodine.

In vitro and in vivo antibiofilm activity of the synthetic antimicrobial peptide WLBU2 against multiple drug resistant Pseudomonas aeruginosa strains.

BACKGROUND: The global crisis of antibiotic resistance increases the demand for the novel promising alternative drugs such as antimicrobial peptides (AMPs). Here, the antibiofilm activity of the WLBU2 peptide against Pseudomonas aeruginosa (P. aeruginosa) isolates was investigated in this study. METHODS: Two clinical MDR and carbapenem resistant P. aeruginosa (CRPA) isolates, and standard P. aeruginosa ATCC 27,853 were investigated. The MIC and MBC of WLBU2 were determined. The MBIC was determined to evaluate inhibitory activity of WLBU2 on biofilm formation and MBEC to dispersal activity on preformed biofilm. The relative expression levels of biofilm-associated genes including rhlI, rhlR, lasI and lasR were analyzed using RT-qPCR. In vivo evaluation of inhibitory effect of WLBU2 on biofilm formation was performed in the murine models of P. aeruginosa biofilm-associated subcutaneous catheter infection. RESULTS: MIC and MBC of WLBU2 for both MDR and ATCC 27,853 P. aeruginosa strains were 8 and 16 µg/mL, respectively, while both the MIC and MBC against the CR strain were 4 µg/mL. MBIC was estimated to be 64 µg/ml for all strains. MBEC against MDR and ATCC 27,853- P. aeruginosa strains was 128 µg/ml and against CRPA was 64 µg/ml. The bacterial adhesion to a static abiotic solid surface (the surface in the polypropylene microtiter wells) was significantly inhibited at 1/4× MIC in all P. aeruginosa strains and at 1/8× MIC in CRPA strain (P < 0.05). Following treatment with WLBU2 at 1/8× MIC, significant inhibition in biofilm formation was observed in all isolates (P < 0.05). Results of the colorimetric assay showed that WLBU2 at 4× MIC was able to disperse 69.7% and 81.3% of pre-formed biofilms on abiotic surface produced by MDR and standard (ATCC 27,853) P. aeruginosa, respectively (P < 0.03), while a 92.2% reduction in the CRPA biofilm was observed after treatment with 4× MIC WLBU2 (P < 0.03). The expression levels of all genes in isolates treated with 1/2 MIC of WLBU2 were down-regulated by more than four-fold compared to the untreated isolates (P < 0.05). WLBU2 significantly inhibited biofilm formation in murine catheter-associated CRPA infection model at 1/4×MIC, 1/2×MIC, and 1×MIC by 33%, 52%, and 67%, respectively. CONCLUSION: Considering relatively strong inhibitory and eradication potency of WLBU2 on the P. aeruginosa biofilms in in vitro and in vivo conditions, the peptide can be considered as a promising candidate for designing an antibiofilm drug.

Tumor-targeted induction of intrinsic apoptosis in colon cancer cells by Lactobacillus plantarum and Lactobacillus rhamnosus strains.

BACKGROUND: Colorectal cancer is one of the widespread and lethal types of malignancies. Recently, antineoplastic attributes of probiotics have attracted lots of attention. Here, we investigated anti-proliferative potential of the non-pathogenic strains Lactobacillus plantarum ATCC 14,917 and Lactobacillus rhamnosus ATCC 7469 on human colorectal adenocarcinoma-originated Caco-2 cells. METHODS AND RESULTS: Caco-2 and HUVEC control cells were treated with ethyl acetate extracts of the two Lactobacillus strains to assess cell viability by MTT assay. Annexin/PI staining flow cytometry, and caspase-3, -8 and - 9 activity assays were performed to determine the type of cell death induced in extract-treated cells. Expression levels of apoptosis-related genes were evaluated by RT-PCR. Extracts from both L. plantarum and L. rhamnosus specifically targeted the Caco-2 cells and not HUVEC controls, and significantly affected the viability of the colon cancer cell line in a time- and dose-dependent manner. This effect was shown to occur through activation of the intrinsic apoptosis pathway, as indicated by the increased caspase-3 and - 9 activities. While there are limited and conflicting data about the mechanisms underlying the specific antineoplastic attributes of Lactobacillus strains, we clarified the overall induced mechanism. The Lactobacillus extracts specifically down-regulated the expression of the anti-apoptotic bcl-2 and bcl-xl, and simultaneously up-regulated the pro-apoptotic bak, bad, and bax genes in treated Caco-2 cells. CONCLUSIONS: Ethyl acetate extracts of L. plantarum and L. rhamnosus strains could be considered as targeted anti-cancer treatments specifically inducing the intrinsic apoptosis pathway in colorectal tumor cells.

Occurrence of diarrheagenic Escherichia coli pathotypes from raw milk and unpasteurized buttermilk by culture and multiplex polymerase chain reaction in southwest Iran.

BACKGROUND: In developing countries including Iran, there are limited data on diarrheagenic Escherichia coli (DEC) contamination in milk and unpasteurized buttermilks. This study aimed to determine the occurrence of DEC pathotypes by culture and multiplex polymerase chain reaction (M-PCR) in some dairy products from southwest Iran. METHODS AND RESULTS: In this cross-sectional study (September to October 2021), 197 samples (87 unpasteurized buttermilk and 110 raw cow milk) were collected from dairy stores of Ahvaz, southwest Iran. The presumptive E. coli isolates were primarily identified using biochemical tests and then confirmed by PCR of uidA gene. The occurrence of 5 DEC pathotypes: enterotoxigenic E. coli (ETEC), enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), and enteroinvasive E. coli (EIEC) were investigated using M-PCR. Overall, 76 (76/197, 38.6%) presumptive E. coli isolates were identified by biochemical tests. Using uidA gene, only 50 isolates (50/76, 65.8%) were confirmed as E. coli. DEC pathotypes were detected in 27 of 50 (54.0%) E. coli isolates (74.1%, 20/27 from raw cow milk and 25.9%, 7/27 from unpasteurized buttermilk). The frequency of DEC pathotypes was as follows: 1 (3.7%) EAEC, 2 (7.4%) EHEC, 4 (14.8%) EPEC, 6 (22.2%) ETEC, and 14 (51.9%) EIEC. However, 23 (46.0%) E. coli isolates had only the uidA gene and were not considered DEC pathotypes. CONCLUSION: Possible health risks for Iranian consumers can be attributed to the presence of DEC pathotypes in dairy products. Hence, serious control and prevention efforts are needed to stop the spread of these pathogens.

Comparison of Antifungal Activity of Jaftex Mouthwash and Nystatin Suspension against the Growth of Candida albicans.

Objective: Oral candidiasis is an opportunistic fungal infection in the oral cavity caused by an overgrowth of Candida species, especially Candida albicans. Various herbal agents have been designed to target Candida albicans. The aim of this study was to evaluate the antifungal activity of Jaftex mouthwash and nystatin suspension on the growth of Candida albicans. Materials and methods:In the present in vitro study, a standard strain of Candida albicans was prepared in the form of lyophilized ampoules. Jaftex mouthwash was prepared with an active ingredient (10 g per 100 cc) of aqueous extract of oak fruit hull (Jaft), Zataria multiflora and Satureja bachtiarica. Nystatin oral suspension (100,000 IU/mL) was also prepared. Both mouthwashes were serially diluted using the two-fold serial dilution method (Jaftex: eight-fold dilutions; nystatin suspension: nine-fold dilutions). A volume of 10 ìL of each dilution of Jaftex mouthwash and nystatin suspension was placed on the discs that were linearly inoculated on culture medium and stored in an incubator for 24 hours at 37 °C. The minimum inhibitory concentration (MIC) of the two antifungal agents was determined using the modified E-test. Data were analyzed using SPSS Version 26.0. A p-value less than 0.05 was considered statistically significant. Results:The mean MIC values of Jaftex mouthwash and nystatin suspension were 0.0625 (mg/mL) and 0.0015 (mg/mL), respectively. There was a significant difference between the antifungal effect of Jaftex mouthwash and nystatin suspension on the growth of Candida albicans. Nystatin showed the lowest MIC and greater antifungal activity compared with Jaftex mouthwash. Conclusion:Nystatin increasingly suppressed the growth of Candida albicans. Jaftex mouthwash inhibited the growth of Candida albicans. Since nystatin may show allergic reactions, Jaftex mouthwash can be used as an alternative to nystatin. Due to the synergistic effect of nystatin with thymol, Jaftex mouthwash can be prescribed with nystatin.

Beta-lactamase determinants and molecular typing of carbapenem-resistant classic and hypervirulent Klebsiella pneumoniae clinical isolates from southwest of Iran.

This study investigated the molecular epidemiology of carbapenem-resistant classic Klebsiella pneumoniae (CR-cKp) and carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKp) isolates in southwestern Iran. From 2019 to 2021, 136 (88.9%) cKp and 17 (11.1%) hvKp isolates were identified using biochemical tests and polymerase chain reaction (PCR). Antibiotic resistance, beta-lactamases, and clonal relatedness of carbapenem-resistant isolates were investigated using disk diffusion, PCR, and enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR), respectively. The different markers of hvKp isolates were as follows: string test (35.3%, n = 6/17), magA (11.8%, n = 2/17), rmpA (11.8%, n = 2/17), rmpA2 (52.9%, n = 9/17), iucA (52.9%, n = 9/17), and peg344 (35.3%, n = 6/17). Also, 55.1% (n = 75/136) of cKp and 47.1% (n = 8/17) of hvKp isolates were CR-cKp and CR-hvKp, respectively. All CR-hvKp (100.0%, n = 8) isolates were MDR. Colistin, tetracycline, and tigecycline were the most effective antibiotics. The occurrence of beta-lactamase genes in 75 CR-cKp and 8 CR-hvKp isolates was as follows: bla NDM (41.3, 25.0%), bla IMP (4.0, 0.0%), bla VIM (8.0, 0.0%), bla GES (14.7, 25.0%), bla OXA-48-like (20.0, 0.0%), bla CTX-M (26.7, 12.5%), bla SHV (24.0, 12.5%), bla TEM (10.7, 0.0%), bla FOX (6.7, 0.0%), bla DHA (6.7, 0.0%), bla CMY (5.3, 0.0%), bla LAT (12.0, 0.0%), and bla ACT (8.0, 0.0%). ERIC-PCR showed a high diversity among isolates. In this study, the occurrence of MDR CR-hvKp isolates harboring bla NDM and bla GES was detected for the first time in southwestern Iran. To prevent the spread of CR-hvKp and reduce selection pressure, long-term surveillance and more effective treatment strategies should be implemented.

MOLECULAR ASSESSMENT OF FECAL LACTOBACILLI POPULATIONS IN CHILDREN WITH FUNCTIONAL CONSTIPATION.

BACKGROUND: Investigation of the gut-specific bacterial strains including lactobacilli is essential for understanding the bacterial etiology of constipation. OBJECTIVE: This study aimed to compare the prevalence and quantity of intestinal lactobacilli in constipated children and healthy controls. METHODS: Forty children fulfilling Rome IV criteria for functional constipation and 40 healthy controls were recruited. Fecal samples were analyzed using species-specific polymerase chain reaction followed by random amplified polymorphic DNA-PCR and quantitative real-time PCR. RESULTS: Totally, seven different species of lactobacilli were detected. Out of 80 volunteers, 65 (81.3%) were culture and species-specific PCR positive from which 25 (38.46%) constipated children and 40 (61.54%) healthy subjects. The most prevalent species were L. paracasei 21 (32.3%) followed by L. plantarum 18 (27.7%) among both healthy and patient groups. Analysis of the RAPD dendrograms displayed that strains isolated from constipated and non-constipated children have similarity coefficients of more than 90%. The qPCR assays demonstrated constipated children had a lower amount of total lactobacilli population (per gram of feces) than healthy controls. CONCLUSION: Our findings showed that the mere existence of various species of Lactobacillus in the gut does not enough to prevent some gastrointestinal disorders such as functional constipation, and their quantity plays a more important role.

Avaliação molecular de populações fecais de lactobacilos em crianças com prisão de ventre funcional / Molecular assessment of fecal lactobacilli populations in children with functional constipation

ABSTRACT Background: Investigation of the gut-specific bacterial strains including lactobacilli is essential for understanding the bacterial etiology of constipation. Objective: This study aimed to compare the prevalence and quantity of intestinal lactobacilli in constipated children and healthy controls. Methods: Forty children fulfilling Rome IV criteria for functional constipation and 40 healthy controls were recruited. Fecal samples were analyzed using species-specific polymerase chain reaction followed by random amplified polymorphic DNA-PCR and quantitative real-time PCR. Results: Totally, seven different species of lactobacilli were detected. Out of 80 volunteers, 65 (81.3%) were culture and species-specific PCR positive from which 25 (38.46%) constipated children and 40 (61.54%) healthy subjects. The most prevalent species were L. paracasei 21 (32.3%) followed by L. plantarum 18 (27.7%) among both healthy and patient groups. Analysis of the RAPD dendrograms displayed that strains isolated from constipated and non-constipated children have similarity coefficients of more than 90%. The qPCR assays demonstrated constipated children had a lower amount of total lactobacilli population (per gram of feces) than healthy controls. Conclusion: Our findings showed that the mere existence of various species of Lactobacillus in the gut does not enough to prevent some gastrointestinal disorders such as functional constipation, and their quantity plays a more important role.

RESUMO Contexto: A investigação das cepas bacterianas específicas do intestino, incluindo lactobacilos, é essencial para a compreensão da etiologia bacteriana da prisão de ventre. Objetivo: Este estudo teve como objetivo comparar a prevalência e a quantidade de lactobacilos intestinais em crianças constipadas e controles saudáveis. Métodos: Foram recrutadas quarenta crianças que preenchem os critérios de Roma IV para prisão de ventre funcional e 40 controles saudáveis. As amostras fecais foram analisadas utilizando-se uma reação da cadeia de polimerase específica da espécie, seguida por DNA polimórfico amplificado aleatório e PCR quantitativo em tempo real. Resultados: Foram detectadas sete espécies diferentes de lactobacilos. Dos 80 voluntários, 65 (81,3%) eram cultura em PCR específico de espécies, dos quais 25 (38,46%) crianças constipadas e 40 (61,54%) indivíduos saudáveis. As espécies mais prevalentes foram L. paracasei 21 (32,3%) seguidas por L. plantarum 18 (27,7%) entre grupos saudáveis e de pacientes. A análise dos dendrogramas do RAPD mostrou que cepas isoladas de crianças constipadas e não constipadas têm coeficientes de similaridade superiores a 90%. Os ensaios qPCR demonstraram que as crianças constipadas apresentavam uma quantidade menor de população total de lactobacilos (por grama de fezes) do que os controles saudáveis. Conclusão: Nossos achados mostraram que a mera existência de várias espécies de Lactobacillus no intestino não é suficiente para prevenir alguns distúrbios gastrointestinais, como a prisão de ventre funcional, e sua quantidade desempenha um papel mais importante.

Evaluation of the effect of Jaftex herbal mouthwash on the growth of Candida albicans and Candida tropicalis.

BACKGROUND: Increased resistance of Candida albicans to standard antifungal agents has caused special attention to medicinal plants. The aim of this study was to evaluate the effect of Jaftex mouthwash on the growth of C. albicans and Candida tropicalis. METHODS AND MATERIAL: In this in vitro study, standard strains of C. albicans and C. tropicalis were used. Jaftex mouthwash was prepared with the active ingredient (10g/100cc) of aqueous extract of oak fruit hull (Jaft), Zataria multiflora, and Satureja bachtiarica. The mouthwash was diluted in half, 8 different concentrations were obtained. 10 µl volume of each dilution was poured on discs mounted linearly on the culture medium inoculated with the target fungus. After 24 h, due to the slow growth rate of these fungi, the Petri dishes were incubated at 37°C and the mean minimum inhibitory concentration (MIC) was determined for each fungus. The modified E. test method was used to measure the MIC of Jaftex mouthwash for the two fungi. The experiment was repeated three times for each fungus and the mean value was measured. RESULTS: The mean value of MIC for C. albicans and C. tropicalis was 0.0625 (mg/mL) and 0.0833 (mg/mL), respectively. Candida albicans appeared to be more sensitive to Jaftex, but no statistically significant difference was observed. CONCLUSION: Jaftex mouthwash inhibits the growth of C. albicans and C. tropicalis. The use of this mouthwash is recommended for treatment of oral candidiasis.

Occurrence of the Legionella species in the respiratory samples of patients with pneumonia symptoms from Ahvaz, Iran; first detection of Legionella cherrii.

BACKGROUND: This study aimed to investigate the occurrence of Legionella species in the respiratory samples of patients with pneumonia symptoms from Ahvaz, Iran by culture and the real-time PCR of 23S-5S rRNA gene spacer region. METHODS AND RESULTS: A total of 123 clinical respiratory samples including 63 pleural aspirates, 57 bronchoalveolar lavage (BAL), and 3 sputum were collected from 65 males and 58 females with pneumonia symptoms. All samples were cultured on the Modified Wadowsky-Yee (MWY) agar. The Legionella species was identified by routine bacteriological tests. The presence of the 16S-23S rRNA spacer region gene was investigated by real-time PCR. The Legionella species were differentiated by sequencing of 16S-23S rRNA gene. A total of 2 (1.6%) BAL specimens were positive for Legionella species by culture method. No Legionella spp. were identified in pleural aspirates and sputum samples by the culture method. Using real-time PCR, 9 (7.3%) samples including 6 BAL, 1 sputum, and 2 pleural aspirates were positive for legionella species. These species were detected in 3 (5.2%) females and 6 males (9.2%). The results of sequencing showed that eight species were L. pneumophila while one was L. cherrii. Also, the 2 isolates that were identified by culture method, were confirmed as L. pneumophila by sequencing. CONCLUSIONS: The results showed that using the real-time PCR has a more efficacy for detecting of Legionella species in respiratory samples. Also, L. pneumophila was the most prevalent species circulating in the southwest region of Iran. So, periodic monitoring programs is recommended to prevent epidemics due to this bacterium.

3D structure prediction, dynamic investigation and rational construction of an epitope-masked thermostable bovine hyaluronidase.

Hyaluronidase (HAase) from bovine testes (BTH) has long been used in broad pharmaceutical areas, while it is associated with drawbacks in aspects of solubility, immunogenicity and pharmacokinetics. These issues can be addressed by gaining structural insights and designing rational modifications to the enzyme structure, as proposed in this study. A 3D structural model was built for HAase and underwent 40 ns of molecular dynamic simulation to examine its thermostability under normal, melting, and extreme conditions. The enzyme activity of BTH was measured against temperature and pH by kinetic assays. The interaction of bovine HAase with HA and chondroitin was defined by molecular docking. Furthermore, immunogenic properties of the enzyme were explored by immunoinformatics. Thermal effects on bovine HAase structural model and the HAase interactions with its substrates were described. We identified some B- and T-cell epitopes and showed that the protein could be recognized by human immune receptor molecules. Epitope masking by adding polyethylene glycol (PEG) to amine groups of residues presenting on the surface of the protein structure was adopted as a surface modification to enhance pharmacological properties of BTH. Assays showed that PEGylated BTH had higher thermostability and similar activity compared to the native enzyme.

In vitro evaluation of the antibacterial effects of Cinnamomum zeylanicum essential oil against clinical multidrug-resistant Shigella isolates.

As there are little data about the antimicrobial effects of the cinnamon essential oils (EO) against multidrug-resistant (MDR) Shigella species, this study aimed to evaluate the antibacterial activities of Cinnamomum zeylanicum EO against the clinical MDR Shigella isolates. Totally 50 MDR Shigella isolates including 17 (34%) S. flexneri, 20 (40%) S. sonnei, and 13 (26%) S. boydii were collected. The isolates were identified by standard phenotypic and molecular methods. The MDR phenotypes were determined as resistant to three antibiotic classes using disc diffusion. The C. zeylanicum EO was analyzed by gas chromatography/mass spectrometry (GC/MS). The minimum inhibitory concentration (MIC) of cinnamon EO was evaluated by microtiter broth dilution. The most Shigella isolates 38% (n = 19) were resistant to six antibiotics. The ampicillin-amikacin-cefotaxime-erythromycin-ciprofloxacin-cotrimoxazole resistotype was the most prevalent pattern detected in five S. sonnei, four S. boydii, and three S. flexneri isolates. The result of GC/MS revealed the cinnamaldehyde (84.8%) as the main ingredient of C. zeylanycum EO. The most susceptible strain to the C. zeylanycum EO was S. boydii (MIC range = 0.15-0.62 µl/ml) followed by S. flexneri (MIC range = 0.07-1.25 µl/ml), and S. sonnei (MIC range = 0.15-1.25 µl/ml). The observed ranges of MIC and MBC values of cinnamon EO against Shigella spp. were 0.07-1.25 µl/ml and 0.31-1.25 µl/ml, respectively. The antibacterial effects of cinnamon EO in this study may increase the hope of finding suitable plant compounds to treat infections caused by MDR Shigella isolates.

A randomized clinical trial evaluating the immunomodulatory effect of convalescent plasma on COVID-19-related cytokine storm.

Evaluating the effect of convalescent plasma (CP) on some cytokine storm indices in severe COVID-19 patients. Totally, 62 patients were randomly assigned into two groups for this clinical trial. Patients in the intervention group received one unit (500 mL) plasma on the admission day plus standard drugs while the controls merely received standard treatments. Eventually, primary and secondary outcomes were evaluated. In the CP group, compared with controls, the mean levels of lymphocytes and IL-10 significantly increased while the levels of IL-6, TNF-α, and IFN-γ decreased (p < 0.05). The length of in-hospital stay, and mortality rate did not significantly reduce in the CP group compared with controls (p > 0.05) while WHO severity scores remarkably improved (p = 0.01), despite the higher frequency of underlying diseases among the CP group (66.7%) vs. controls (33.3%). Although CP has a remarkable immunomodulatory and antiviral potential to improve the cytokine storm and disease severity in COVID-19 patients, it did not considerably affect the mortality rate.

Quantification of Intestinal Lactobacillus Species in Children with Functional Constipation by Quantitative Real-Time PCR.

BACKGROUND: Functional constipation is a common and annoying gastrointestinal disorder among children worldwide in which the intestinal microbiota composition plays a fundamental role. This study aimed to compare the quantity of main intestinal Lactobacillus species in constipated children and healthy controls. MATERIALS AND METHODS: Fecal samples were collected from 40 children fulfilling Rome IV criteria for functional constipation and 40 healthy volunteers. Quantitative real-time polymerase chain reaction (qPCR) method with species-specific primers was used to investigate seven main Lactobacillus species in fecal samples. RESULTS: Lactobacillus strains of the patient group were different from the healthy controls, main differences being noticeable decrease in the population quantity of Lactobacillus reuteri (mean 102.61 CFU/gram feces) and lower abundance of Lactobacillus fermentum (p<0.0001). CONCLUSION: Lactobacillus species might be quantitatively altered in childhood functional constipation. Our findings on the different species of Lactobacillus population showed significantly decreased quantity in the patient group compared with the healthy subjects.

Isolation and identification of potential probiotic Lactobacillus species from feces of infants in southwest Iran.

OBJECTIVES: To evaluate the potential probiotic properties of Lactobacillus strains isolated from feces of infants and also to determine their antimicrobial activity against some enteropathogenic bacteria. METHODS: The Fecal samples were prepared from 120 infants aged less than 24 months. In total, 105 Lactobacillus strains were identified by phenotypic tests. Thirty isolates were randomly selected to study their potential probiotic properties. These isolates were examined for resistance to acid (pH: 2.5, 2 h) and bile (oxgall 0.3%, 8 h), adhesion to HT-29 cells, antibiotic susceptibility, and antimicrobial activities. RESULTS: On basis of 16S rRNA sequencing, 30 isolates identified as Lactobacillus fermentum (n = 11; 36.7%), Lactobacillus plantarum (n = 9; 30%), Lactobacillus rhamnosus (n = 6; 20%), and Lactobacillus paracasei (n = 4; 13.3%). All tested strains survived at acid and bile conditions. Six Lactobacillus strains revealed high adherence to HT-29 cells. Three strains including the L. fermentum (N2, N7), and the L. plantarum (N20) showed good probiotic potential and inhibited the growth of Yersinia enterocolitica ATCC 23715, Shigella flexneri ATCC 12022, Salmonella enterica ATCC 9270, and enteropathogenic Escherichia coli (EPEC) ATCC 43887. The antibiotic resistance test showed that all the isolates were susceptible to tetracycline, and chloramphenicol. CONCLUSIONS: Lactobacillus strains like L. fermentum (N2, N7), and the L. plantarum (N20), could be potential probiotic, but further in vitro and in vivo studies on these probiotic strains are still required.

Coexistence of genes encoding aminoglycoside modifying enzymes among clinical Acinetobacter baumannii isolates in Ahvaz, Southwest Iran.

Aminoglycosides are widely recommended for treatment of Acinetobacter baumannii infections in combination with ß-lactams or quinolones. This cross-sectional study was aimed to investigate the coexistence of aminoglycoside modifying enzyme (AME) genes among A. baumannii isolates from clinical samples in Ahvaz, Iran. A total of 85 clinical A. baumannii isolates typed by ERIC-PCR were investigated for the presence of AME genes, including ant(3″)-Ia, aac(6')-Ib, aac(3')-Ia, ant(2″)-Ia, and aph(3')-VIa by PCR. The resistance rates to aminoglycoside agents were evaluated by disk diffusion. In this study, 84 out of 85 A. baumannii isolates were resistant to at least one of the aminoglycosides and harbored at least one AME gene. The most common gene encoding AMEs was aph (3')VIa, followed by aac(3')-Ia, ant(3″)-Ia, ant (2″)-Ia, and aac(6')-Ib. The aminoglycoside-resistant genotypes were completely matched to resistant phenotypes to each one of the aminoglycoside agents. There was a clear association between AME gene types and the phenotype of resistance to aminoglycosides with their ERIC-PCR types. Our findings highlight the coexistence of AME genes and clonal dissemination of multiresistant A. baumannii in hospital setting.

Association Between Biofilm Formation, Structure, and the Expression Levels of Genes Related to biofilm formation and Biofilm-Specific Resistance of Acinetobacter baumannii Strains Isolated from Burn Infection in Ahvaz, Iran.

BACKGROUND: The ability of biofilm formation is an effective way for Acinetobacter baumannii survival from stressed conditions. This present study was aimed to evaluate the association between biofilm formation, structure, the expression levels of genes related to biofilm formation and biofilm-specific resistance of A. baumannii strains isolated from burn infections in Ahvaz, Iran. METHODS: In this study, we assessed the antibiotic susceptibilities, ERIC-PCR typing, capacity of biofilm formation and biofilm structure of 64 A. baumannii isolates collected from burn infections. The distribution and the expression levels of genes involved in the biofilm formation including bap, ompA, abaI, pgaA and csuE were assessed by PCR and real-time PCR, respectively. RESULTS: We classified A. baumannii isolates in 14 clonal types of ERIC-PCR. Most A. baumannii isolates were resistant to all antibiotics tested except to tigecycline and colistin and had the biofilm formation capability but with different capacities. There was a significant inverse relationship between resistance to antibiotic agents and biofilm formation. The biofilm matrix of 50 strains consisted of polysaccharides together with DNA or proteins. The genes involved in the biofilm formation were detected in both biofilm-forming and non-biofilm forming; however, the expression levels of these genes were higher in biofilm producers compared with non-producers. CONCLUSION: The biofilm cells exhibited dramatically decreased susceptibility to antibiotic agents; hence, they have great significance for public health. Therefore, the determination of antibiotic susceptibilities in biofilm and planktonic mode, molecular typing, and capacity of biofilm formation in clinical setting is essential.

Concurrent detection of cagA, vacA, sodB and hsp60 virulence genes and their relationship with clinical outcomes of disease in Helicobacter pylori isolated strains of southwest of Iran.

BACKGROUND AND OBJECTIVES: Helicobacter pylori is a Gram-negative spiral-shaped bacterium that contaminates more than half of the world's inhabitants, and infection with this bacterium is associated with some gastric disorders. Also, 5% to 10% of H. pylori genes are specific to this bacterium and many bacterial virulence factors fall into this group. The cagA, vacA, sodB and hsp60 are among important virulence factors of H. pylori. MATERIALS AND METHODS: A gastric biopsy specimen was taken from 341 gastric patients and cultivated on a Colombia agar plate, containing various antibiotics, such as vancomycin, amphotericin B, and trimethoprim & polymyxin B, and incubated for 3 to 10 days under microaerophilic conditions at 37°C. PCR was used to detect the ureC, cagA, vacA, sodB and hsp60 genes. RESULTS: In this study, 131 isolates were identified as H. pylori. The prevalence of cagA, vacA, sodB and hsp60 were 74%, 100%, 92.4% and 96.2%, respectively. The correlation between the clinical forms of the disease and the virulence genes were analyzed by statistical tests and no significant correlation was found. CONCLUSION: The obtained results are similar to some studies conducted in different parts of the world and is different in other cases. This discrepancy is due to the difference in the type of gastric disorders, sample size and methodology.

Genomic Evolutionary Patterns of Leiomyosarcoma and Liposarcoma.

PURPOSE: Leiomyosarcoma and liposarcoma are common subtypes of soft tissue sarcoma (STS). Patients with metastatic leiomyosarcoma or dedifferentiated liposarcoma (DDLPS) typically have worse outcomes compared with localized leiomyosarcoma or well-differentiated liposarcoma (WDLPS). A better understanding of genetic changes between primary/metastatic leiomyosarcoma and between WDLPS/DDLPS may provide insight into their genetic evolution. EXPERIMENTAL DESIGN: We interrogated whole-exome sequencing (WES) from "trios" of normal tissue, primary tumor, and metastatic tumor from individual patients with leiomyosarcoma (n = 9), and trios of normal tissue, well-differentiated tumor, and dedifferentiated tumor from individual patients with liposarcoma (n = 19). Specifically, we performed mutational, copy number, and tumor evolution analyses on these cohorts and compared patterns among leiomyosarcoma and liposarcoma trios. RESULTS: Leiomyosarcoma cases harbored shared drivers through a typical parent/child relationship where the metastatic tumor was derived from the primary tumor. In contrast, while all liposarcoma cases shared the characteristic focal chromosome 12 amplicon, most paired liposarcoma cases did not share additional mutations, suggesting a divergent evolutionary pattern from a common precursor. No highly recurrent genomic alterations from WES were identified that could be implicated as driving the progression of disease in either sarcoma subtype. CONCLUSIONS: From a genomic perspective, leiomyosarcoma metastases contain genetic alterations that are also found in primary tumors. WDLPS and DDLPS, however, appear to divergently evolve from a common precursor harboring 12q amplification, rather than as a transformation to a higher-grade tumor. Further efforts to identify specific drivers of these distinct evolutionary patterns may inform future translational and clinical research in STS.