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Background and Objective: Several strains of the Echinococcus granulosus have been described based on morphological characters, intermediate host specificity and/or genetic analysis of mitochondrial and nuclear DNA. The aim of this study was to characterize different E.granulosus isolates by using sequences of mitochondrial atp6 gene. Materials and Methods: In this study, Sixty infected liver and lungs of cattle, sheep and goats were collected from the abattoir of Varamin city-Iran during 2008. Protoscoleces were removed from each fertile cyst and DNA extracted. New and specific primers were designed for two existing genotypes (G1 and G6) of E. granulosus known to occur in Iran and applied in PCR reactions. Results: The new primers selectively amplified the G1 and G6 genotypes of E. granulosus with specific bands of 708 and 705 bp respectively. The G1 genotype was identified in all fertile cyst samples. Conclusion: This study showed that the new primer pairs which specifically amplify portions of the mitochondrial atp6 gene of the G1 and G6 strains of Echinococcus granulosus are proper molecular marker for investigating genetic variation in a number of isolates of E. granulosus from a range of hosts (sheep, goats, cattle) in Iran. The result of sequenced samples showed that our sequences were the same as those reported previously for these strains.

Human-generated waste, including infectious healthcare waste, poses significant risks to public health and the environment. The COVID-19 pandemic has increased the global production of infectious waste, emphasizing the need for safe and sustainable waste management practices. While autoclaves are commonly used for on-site disposal, alternative methods like ozone gas and UV-C radiation offer environmentally friendly options that effectively eliminate pathogens without leaving toxic residues. Inadequate waste management can contribute to disease transmission, while open burning releases harmful pollutants. This study investigated the effectiveness of different disinfection agents - ozone gas and UV-C radiation - on infectious solid waste contaminated with bacteria. The bacterial indicators examined were Staphylococcus aureus , Bacillus subtilis , and Pseudomonas aeruginosa . The experimental methods included operating each ozone and UV-C radiation individually and simultaneously using ozone gas and UV-C radiation. The study also investigated exposure times and various concentrations of ozone gas. The findings demonstrated that the simultaneous application of ozone gas and UV-C radiation was the most effective method for decontaminating infectious solid waste and targeting the selected bacteria. The concentration of ozone gas ranged from 165 to 5000 ppm, depending on generation time and treatment chamber volume, while exposure times varied from 1 to 180 minutes. In applying UV-C rays, complete elimination of S. aureus was observed after 60 minutes up to 6-log, while the reduction of B. subtilis and P. aeruginosa were 2-log and 3-log, respectively. Ozone gas had the ability to inactivate all strains, but when ozone gas and UV-C rays were used simultaneously, this process was accelerated and improved. The total reduction in the bacterial load was 8-log. Considering the increase in population and the subsequent increase in waste generation, adopting an environmentally friendly waste management method can be very advantageous. Implications : This study highlights the effectiveness of simultaneously applying ozone gas and UV-C radiation for decontaminating infectious solid waste, offering an environmentally friendly alternative to traditional thermal treatments like autoclave and incineration. By optimizing ozone concentrations and exposure times, this method reduces disease transmission risks and minimizes environmental impact. These findings are crucial, especially during outbreaks such as the COVID-19 pandemic, providing scalable, sustainable waste management solutions for healthcare facilities. Implementing these techniques can protect public health and the environment, setting a new standard for safe infectious waste disposal worldwide, mitigating hazardous pollutants, and reduce the exposure risk of bio-hazardous residues.

Cardiovascular disease (CVD) is one of the main causes of death in the world. The increased level of blood cholesterol is significantly correlated to CVD incidents. Statins are a group of drugs that decrease the synthesis of cholesterol in the liver by inhibiting the final enzyme of the pathway named HMG-CoA reductase. Several investigations showed that different patients give different responses to the administration of statin drugs according to their genetic background. In this research study, using Genome-Wide Association Studies (GWAS) data analysis methods, such as the SimpleM statistical approach and genomic connection matrix, we tried to discover the novel candidate SNPs that were involved in response to statin drugs. The investigation was carried out using 3,221 cardiovascular patients' data about genotypes and phenotypes of two important parameters including total cholesterol, and LDL level, in response to statin administration. Functional annotation of nearest genes to candidate SNPs was also carried out by using comprehensive databases and tools such as BioMart-Ensembl, UCSC, NCBI, and WebGestalt software. Our results represented eight novel SNPs (rs10820084, rs4803750, rs10989887, rs1966503, rs17502794, rs10785232, rs484071, rs4785621) significantly associated with statin response in different individual cardiovascular patients for the first time. In addition, the groups of genes that are close to the SNPs were also represented and evaluated in detail. Our results illustrated that some of the genes such as BAAT, BCL3, and CMTM6 have a direct functional impact on cholesterol level or LDL biosynthesis which confirmed the effects of neighbor SNPs on the response to statin drugs. Today, finding the loci, genes, and molecular mechanisms involved in the response to drugs is of great importance in pharmacogenomics and personalized medicine., (© 2024. The Author(s).)

The association between oral dysbiosis and celiac disease (CD) remains poorly understood, as does the impact of CD-associated dysbiosis on disease development or exacerbation. This study aims to investigate alterations in salivary microbial composition among children with CD. In this cross-sectional study, saliva samples from 12 children with active CD (A-CD group), 14 children with CD on a gluten-free diet (GFD), and 10 healthy control (HC) children were analyzed using DNA sequencing targeting the 16S ribosomal RNA. Both patients in A-CD and GFD groups showed a significant increase (p = 0.0001) in the Bacteroidetes phylum, while the Actinobacteria phylum showed a significant decrease (p = 0.0001). Notably, the Rothia genus and R.aeria also demonstrated a significant decrease (p = 0.0001) within the both CD groups as compare to HC. Additionally, the control group displayed a significant increase (p = 0.006) in R.mucilaginosa species compared to both CD patient groups. Distinct bacterial strains were abundant in the saliva of patients with active CD, indicating a unique composition of the salivary microbiome in individuals with CD. These findings suggest that our approach to assessing salivary microbiota changes may contribute to developing noninvasive methods for diagnosing and treating CD., (© 2024. The Author(s).)

Drug resistance has become a matter of great concern, with many bacteria now resist multiple antibiotics. This study depicts the occurrence of antibiotic-resistant bacteria (ARB) and resistance patterns in five full-scale hospital wastewater treatment plants (WWTPs). Samples of raw influent wastewater, as well as pre- and post-disinfected effluents, were monitored for targeted ARB and resistance genes in September 2022 and February 2023. Shifts in resistance profiles of Escherichia coli , Pseudomonas aeruginosa , and Acinetobacter baumannii antimicrobial-resistant indicators in the treated effluent compared to that in the raw wastewater were also worked out. Ceftazidime (6.78 × 10 5 CFU/mL) and cefotaxime (6.14 × 10 5 CFU/mL) resistant species showed the highest concentrations followed by ciprofloxacin (6.29 × 10 4 CFU/mL), and gentamicin (4.88 × 10 4 CFU/mL), in raw influent respectively. WWTP-D employing a combination of biological treatment and coagulation/clarification for wastewater decontamination showed promising results for reducing ARB emissions from wastewater. Relationships between treated effluent quality parameters and ARB loadings showed that high BOD 5 and nitrate levels were possibly contributing to the persistence and/or selection of ARBs in WWTPs. Furthermore, antimicrobial susceptibility tests of targeted species revealed dynamic shifts in resistance profiles through treatment processes, highlighting the potential for ARB and ARGs in hospital wastewater to persist or amplify during treatment., Competing Interests: The authors declare there is no conflict., (© 2024 The Authors This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC BY 4.0), which permits copying, adaptation and redistribution, provided the original work is properly cited (http://creativecommons.org/licenses/by/4.0/).)

Antibody drug conjugates (ADCs) with twelve FDA approved drugs, known as a novel category of anti-neoplastic treatment created to merge the monoclonal antibody specificity with cytotoxicity effect of chemotherapy. However, despite many undeniable advantages, ADCs face certain problems, including insufficient internalization after binding, complex structures and large size of full antibodies especially in targeting of solid tumors. Camelid single domain antibody fragments (Nanobody®) offer solutions to this challenge by providing nanoscale size, high solubility and excellent stability, recombinant expression in bacteria, in vivo enhanced tissue penetration, and conjugation advantages. Here, an anti-human CD22 Nanobody was expressed in E.coli cells and conjugated to Mertansine (DM1) as a cytotoxic payload. The anti-CD22 Nanobody was expressed and purified by Ni-NTA resin. DM1 conjugated anti-CD22 Nanobody was generated by conjugation of SMCC-DM1 to Nanobody lysine groups. The conjugates were characterized using SDS-PAGE and Capillary electrophoresis (CE-SDS), RP-HPLC, and MALDI-TOF mass spectrometry. Additionally, flow cytometry analysis and a competition ELISA were carried out for binding evaluation. Finally, cytotoxicity of conjugates on Raji and Jurkat cell lines was assessed. The drug-to-antibody ratio (DAR) of conjugates was calculated 2.04 using UV spectrometry. SDS-PAGE, CE-SDS, HPLC, and mass spectrometry confirmed conjugation of DM1 to the Nanobody. The obtained results showed the anti-CD22 Nanobody cytotoxicity was enhanced almost 80% by conjugation with DM1. The binding of conjugates was similar to the non-conjugated anti-CD22 Nanobody in flow cytometry experiments. Concludingly, this study successfully suggest that the DM1 conjugated anti-CD22 Nanobody can be used as a novel tumor specific drug delivery system., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In type 1 diabetes, the immune system destroys pancreatic beta cells in an autoimmune condition. To overcome this disease, a specific monoclonal antibody that binds to pancreatic beta cells could be used for targeted immunotherapy. Protein tyrosine phosphatase receptor N (PTPRN) is one of the important surface antigen candidates. Due to its high sequence homology among mammals, so far, no single-chain monoclonal antibody has been produced against this receptor. In this study, we developed a novel single-chain variable fragment (scFv) against the PTPRN extracellular domain. To this aim, ostrich species was used as a host is far phylogenetically birds from mammals to construct a phage display library for the first time. An ostrich-derived scfv phage display library was prepared and biopanning steps were done to enrich and screen for isolating the best anti-PTPRN binders. An scFv with appropriate affinity and specificity to the PTPRN extracellular domain was selected and characterized by ELISA, western blotting, and flow cytometry. The anti-PTPRN scFv developed in this study could be introduced as an effective tool that can pave the way for the creation of antibody-based targeting systems in cooperation with the detection and therapy of type I diabetes., (© 2024. The Author(s).)

Objective: Type 1 diabetes (T1Ds) is an autoimmune disease in which the immune system invades and destroys insulin-producing cells. Nevertheless, at the time of diagnosis, about 30-40% of pancreatic beta cells are healthy and capable of producing insulin. Bi-specific antibodies, chimeric antigen receptor regulatory T cells (CAR-Treg cells), and labeled antibodies could be a new emerging option for the treatment or diagnosis of type I diabetic patients. The aim of the study is to choose appropriate cell surface antigens in the pancreas tissue for generating an antibody for type I diabetic patients., Materials and Methods: In this bioinformatics study, we extracted pancreas-specific proteins from two large databases; the Human Protein Atlas (HPA) and Genotype-Tissue Expression (GTEx) Portal. Pancreatic-enriched genes were chosen and narrowed down by Protter software for the investigation of accessible extracellular domains. The immunohistochemistry (IHC) data of the protein atlas database were used to evaluate the protein expression of selected antigens. We explored the function of candidate antigens by using the GeneCards database to evaluate the potential dysfunction or activation/hyperactivation of antigens after antibody binding., Results: The results showed 429 genes are highly expressed in the pancreas tissue. Also, eighteen genes encoded plasma membrane proteins that have high expression in the microarray (GEO) dataset. Our results introduced four structural proteins, including NPHS1, KIRREL2, GP2, and CUZD1, among all seventeen candidate proteins., Conclusion: The presented antigens can potentially be used to produce specific pancreatic antibodies that guide CARTreg, bi-specific, or labeling molecules to the pancreas for treatment, detection, or other molecular targeted therapy scopes for type I diabetes.

Background: Helicobacter pylori outer membrane vesicles (OMVs) are nano-sized structures, which have been recently suggested to play a crucial role in H. pylori pathogenesis. There are growing evidence indicating the relationship of H. pylori infection with extra-gastroduodenal diseases, especially liver-related disorders. This study was aimed to investigate the effects of H. pylori-derived OMVs on autophagy in hepatic stellate cells (HSCs)., Material and Methods: A selection of five clinical strains of H. pylori with different virulence genotypes were included. The OMVs were isolated by ultracentrifugation and characterized by scanning electron microscopy (SEM) and dynamic light scattering (DLS). The protein concentration of OMVs was measured by BCA assay. MTT assay was used to determine the viability of LX-2 cells (human HSCs) treated with OMVs. The expression level of MTOR, AKT, PI3K, BECN1, ATG16 and LC3B genes was assessed in OMVs-treated LX-2 cells using quantitative real-time PCR. Moreover, immunocytochemistry was performed to evaluate the protein expression of MTOR and LC3B autophagy markers., Results: H. pylori strains produced round shape nano-vesicles ranging from 50 to 500 nm. Treatment of HSCs with H. pylori-derived OMVs at concentration of 10 μg/mL for 24 h significantly elevated the expression of autophagy inhibitory markers (PI3K, AKT, and MTOR) and suppressed the mRNA expression level of autophagy core proteins (BECN1, ATG16 and LC3B). Immunocytochemistry also presented a substantial reduction in the concentration of LC3B autophagy core protein, and a marked elevation in the amount of MTOR autophagy inhibitory protein., Conclusion: This study revealed that H. pylori-derived OMVs could potentially suppress autophagy flux in HSCs as a novel mechanism for H. pylori-mediated liver autophagy impairment and liver disease development. Further studies are required to elucidate the exact role of OMV-carried contents in liver autophagy, and liver-associated disorders., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Coeliac disease (CD) is associated with alterations in gut microbiota composition. This study evaluated the effects of probiotics on gut microbiota composition and clinical symptoms of treated CD patients. In this double-blind, placebo-controlled trial study, 31 CD patients that were randomly classified as probiotics (n = 15) and placebo (n = 16) groups received 10 9  colony-forming units/capsule for 12 weeks. Fecal samples were collected before and after probiotics, or placebo administration and the changes in intestinal microbiota were assessed by quantitative real-time PCR. Probiotic administration improved the patients' clinical symptoms when compared to the placebo group. Fatigue score was significantly reduced by the intake of probiotic supplements (P = 0.02). Except for Staphylococcus spp., the relative abundances of Bacteroidetes , Lactobacillus spp., Bifidobacterium spp., Clostridium cluster I, Enterobacteriaceae , and Firmicutes were higher in probiotics group. Accordingly, a 12-week multi-strain probiotic treatment regimen may modify the composition of intestinal microbiota and improve GI symptoms in CD patients., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (© 2023 The Authors.)

Chimeric antigen receptor (CAR) T cells and natural killer (NK) cells are genetically engineered immune cells that can detect target antigens on the surface of target cells and eliminate them following adoptive transfer. Recent progress in CAR-based therapies has led to outstanding clinical success in certain patients with leukemias and lymphomas and offered therapeutic benefits to those resistant to conventional therapies. The universal approach to stable CAR transgene delivery into the T/NK cells is the use of viral particles. Such approaches mediate semi-random transgene insertions spanning the entire genome with a high preference for integration into sites surrounding highly-expressed genes and active loci. Regardless of the variable CAR expression level based on the integration site of the CAR transgene, foreign integrated DNA fragments may affect the neighboring endogenous genes and chromatin structure and potentially change a transduced T/NK cell behavior and function or even favor cellular transformation. In contrast, site-specific integration of CAR constructs using recent genome-editing technologies could overcome the limitations and disadvantages of universal random gene integration. Herein, we explain random and site-specific integration of CAR transgenes in CAR-T/NK cell therapies. Also, we tend to summarize the methods for site-specific integration as well as the clinical outcomes of certain gene disruptions or enhancements due to CAR transgene integration. Also, the advantages and limitations of using site-specific integration methods are discussed in this review. Ultimately, we will introduce the genomic safe harbor (GSH) standards and suggest some appropriate safety prospects for CAR integration in CAR-T/NK cell therapies., (© 2023. The Author(s).)

Introduction: Helicobacter pylori is a prevalent pathogenic bacterium that resides in the human stomach. Outer membrane vesicles (OMVs) are known as nanosized cargos released by H. pylori , which have been proposed to have a key role in disease progression, pathogenesis, and modulation of the immune system. There are multiple evidences for the role of H. pylori in extragastroduodenal illnesses especially liver-related disorders. However, the precise mechanism of H. pylori extragastroduodenal pathogenesis still remains unclear. In the current study, we aimed to determine the impact of H. pylori -isolated OMVs on hepatic stellate cell (HSC) activation and expression of liver fibrosis markers., Materials and Methods: Five H. pylori clinical strains with different genotype profiles were used. Helicobacter pylori OMVs were isolated using ultracentrifugation and were analyzed by scanning electron microscopy (SEM) and dynamic light scattering (DLS). Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis was applied to determine protein components of H. pylori -derived OMVs. Cell viability of LX-2 human hepatic stellate cell line exposed to OMVs was measured by MTT assay. LX-2 cells were treated with OMVs for 24 h. The gene expression of α -SMA, E-cadherin, vimentin, snail, and β -catenin was analyzed using quantitative real-time PCR. The protein expression of α -SMA, as a well-studied profibrotic marker, was evaluated with immunocytochemistry., Results: Our results showed that H. pylori strains released round shape nanovesicles ranging from 50 to 500 nm. Totally, 112 various proteins were identified in OMVs by proteomic analysis. The isolated OMVs were negative for both CagA and VacA virulence factors. Treatment of HSCs with H. pylori -derived OMVs significantly increased the expression of fibrosis markers., Conclusions: In conclusion, the present study demonstrated that H. pylori -derived OMVs could promote HSC activation and induce the expression of hepatic fibrosis markers. Further research is required to elucidate the definite role of H. pylori -derived OMVs in liver fibrosis and liver-associated disorders., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2023 Shahin Bolori et al.)

Background and Aims: Individuals with celiac disease (CD), non-celiac wheat sensitivity (NCWS), and irritable bowel syndrome (IBS), show overlapping clinical symptoms and experience gut dysbiosis. A limited number of studies so far compared the gut microbiota among these intestinal conditions. This study aimed to investigate the similarities in the gut microbiota among patients with CD, NCWS, and IBS in comparison to healthy controls (HC)., Materials and Methods: In this prospective study, in total 72 adult subjects, including CD (n = 15), NCWS (n = 12), IBS (n = 30), and HC (n = 15) were recruited. Fecal samples were collected from each individual. A quantitative real-time PCR (qPCR) test using 16S ribosomal RNA was conducted on stool samples to assess the relative abundance of Firmicutes, Bacteroidetes, Bifidobacterium spp., and Lactobacillus spp., Results: In all groups, Firmicutes and Lactobacillus spp. had the highest and lowest relative abundance respectively. The phylum Firmicutes had a higher relative abundance in CD patients than other groups. On the other hand, the phylum Bacteroidetes had the highest relative abundance among healthy subjects but the lowest in patients with NCWS. The relative abundance of Bifidobacterium spp. was lower in subjects with CD (P = 0.035) and IBS (P = 0.001) compared to the HCs. Also, the alteration of Firmicutes to Bacteroidetes ratio (F/B ratio) was statistically significant in NCWS and CD patients compared to the HCs (P = 0.05)., Conclusion: The principal coordinate analysis (PCoA), as a powerful multivariate analysis, suggested that the investigated gut microbial profile of patients with IBS and NCWS share more similarities to the HCs. In contrast, patients with CD had the most dissimilarity compared to the other groups in the context of the studied gut microbiota., (© 2023. The Author(s).)

Infertility is one of the major health problems of patients suffering from bacterial infections. Given the high percentage of infertility, the aim of this study was to investigate the prevalence of Chlamydia trachomatis , Mycoplasma genitalium, Neisseria gonorrhoeae and Ureaplasma urealyticum in fertile and infertile women. In the prospective study, 65 infertile patients and 54 pregnant women referred to Mahdieh Hospital in Tehran were included. After transferring of vaginal swabs to the laboratory, DNA extraction and Polymerase Chain Reaction (PCR) were performed using specific primers. Of the 65 vaginal swab specimens, the prevalence of U. urealyticum, M. genitalium , C. trachomatis and N. gonorrhoeae were as 15 (23.1%), 11 (16.9%), 9 (13.8%) and 4 (6.2%), respectively; However, these rate in fertile group was as 6 (11.1%), 3 (5.5%), 5 (9.2%) and 1 (1.8%), respectively. Bacterial infections were higher in infertile group; therefore, these bacterial agents may be associated with female infertility. Timely control and treatment of infections caused by these organisms, together with other factors, can be important in prevention and treatment of the women's infertility and thereby community health.Impact Statement What is already known on this subject? Infertility is one of the most common reproductive health issues in Iran. Female reproductive system is a suitable environment for the growth of many pathogens, which may disrupt any stage of foetal formation, implantation or growth. Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium and Ureaplasma urealyticum are the most important microorganisms that have been considered in the infertility. What do the results of this study add? The prevalence of C. trachomatis, M. genitalium, N. gonorrhoeae, M. genitalium and U. urealyticum were higher in infertile women, but there was no statistically significant compared to pregnant women. These results suggest that timely control and treatment of infections caused by these organisms, along with other factors, can be used to prevent and treat women infertility and community health. What are the implications of these findings for clinical practice and/or further research? Based on the results, designing and implementing national control programs to prevent subsequent complications is thought to be necessary. Comprehensive analyses of the overall prevalence of these bacteria, particularly in developing countries (including Iran), may help to carry out such a strategy.

Chimeric antigen receptor (CAR) therapy is a method directing T lymphocytes against antigens on the surface of tumors, increasing target cell elimination. Genetic engineering enhances the capability of immune cells to detect new antigens expressed on cell surfaces. CAR T cell therapy is a significant breakthrough for treating human malignancies; however, different side effects (e.g., cytokine release syndrome) restrict its application. Improving design and using various combined receptors enhance the performance of these cells. This review discusses limitations and risk factors associated with CAR T cell therapy. We also review some alternative approaches for developing the next generation of CAR T cells., (© 2022 New York Academy of Sciences.)

Compared to the growing body of literature on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection and quantification in sewage, there are limited studies reporting on correlations between the viral loads in sewage and the prevalence of infected patients. The present work is a part of the regular monitoring effort for SARS-CoV-2 in wastewater influents from seven wastewater treatment plants (WWTPs) in Tehran, Iran, starting from late September 2020 until early April 2021. These facilities cover ~64% of the metropolis serving >5000,000 M individuals. The study set out to track the trends in the prevalence of COVID-19 in the community using wastewater based epidemiology (WBE) and to investigate whether these measurements correlate with officially reported infections in the population. Composite sewage samples collected over 16 h were enriched by polyethylene glycol precipitation and the corresponding threshold cycle (Ct) profiles for CDC 'N' and 'ORF1ab' assays were derived through real time RT-qPCR. Monte Carlo simulation model was employed to provide estimates of the disease prevalence in the study area. RNA from SARS-CoV-2 was detectable in 100% ('N' assay) and 81% ('ORF1ab' assay) of totally 91 sewage samples, with viral loads ranging from 40 to 45,000 gene copies/L. The outbreak of COVID-19 positively correlated (R 2  = 0.80) with the measured viral load in sewage samples. Furthermore, sewage SARS-CoV-2 RNA loads preceded infections in the population by 1 to 2 days, which were in line with public adherence with and support for government instructions to contain the pandemic. Given the transient presence of human host-restricted infections such as SARS-CoV-2, these results provide evidence for assessment of the effectiveness of coordinated efforts that specifically address public health responses based on wastewater-based disease surveillance against not only COVID-19 but also for future infectious outbreaks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The pathogenesis of celiac disease (CD) is significantly influenced by gut microbiota. Daily nutritional profile influences the diversity of gut microbiota. This study was aimed to compare the abundance of gut microbiota in CD patients compared to normal control (NC), and to investigate the impact of nutritional factors on their fecal microbiota diversity. In this study, a selected panel of intestinal bacteria was assessed in 31 confirmed CD patients adhering to gluten-free diet (GFD) for more than 6 months and in 20 NC subjects. Stool samples were collected from each participant, DNA was extracted, and absolute quantitative real-time PCR (qPCR) was carried out. The gut microbiota including Bacteroidetes, Bifidobacterium, Clostridium, Staphylococcus, Enterobacteiaceae, Firmicutes, and Lactobacillus were assessed. The quantities of fruits, vegetables, meat, liquids, sugar and gluten-free candy/bread consumption were evaluated using a questionnaire. The proportion of Bifidobacterium, Firmicutes, and Lactobacillus in CD cases was significantly lower than NC (P < 0.005). Significant correlation coefficients between Bifidobacterium and Lactobacillus (P < 0.001), and also Firmicutes and Lactobacillus (P < 0.001) were recorded. Moreover, a significant association between medium amount of meat and bean consumptions and low abundance of Lactobacillus and Firmicutes (P = 0.024 and P = 0.027, respectively), and also high amount of bean consumptions and low abundance of Lactobacillus (P = 0.027) in CD were observed. The results showed that meat and bean consumptions could reduce the beneficial bacteria including Firmicutes and Lactobacillus in CD patients. Therefore, changes in the gut microbiota abundance may contribute to dietary changes and unimproved CD symptoms., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Introduction: Pyrazinamide (PZA) susceptibility testing plays a critical role in determining the appropriate treatment regimens for multidrug-resistant tuberculosis. We conducted a systematic review and meta-analysis to evaluate the diagnostic accuracy of sequencing PZA susceptibility tests against culture-based susceptibility testing methods as the reference standard. Methods: We searched the MEDLINE/PubMed, Embase, and Web of Science databases for the relevant records. The QUADAS-2 tool was used to assess the quality of the studies. Diagnostic accuracy measures ( i.e. , sensitivity and specificity) were pooled with a random-effects model. All statistical analyses were performed with Meta-DiSc (version 1.4, Cochrane Colloquium, Barcelona, Spain), STATA (version 14, Stata Corporation, College Station, TX), and RevMan (version 5.3, The Nordic Cochrane Centre, the Cochrane Collaboration, Copenhagen, Denmark) software. Results: A total of 72 articles, published between 2000 and 2019, comprising data for 8,701 isolates of Mycobacterium tuberculosis were included in the final analysis. The pooled sensitivity and specificity of the PZA sequencing test against all reference tests (the combination of BACTEC mycobacteria growth indicator tube 960 (MGIT 960), BACTEC 460, and proportion method) were 87% (95% CI: 85-88) and 94.7% (95% CI: 94-95). The positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio, and the area under the curve estimates were found to be 12.0 (95% CI: 9.0-16.0), 0.17 (95% CI: 0.13-0.21), 106 (95% CI: 71-158), and 96%, respectively. Deek's test result indicated a low likelihood for publication bias ( p  = 0.01). Conclusions: Our analysis indicated that PZA sequencing may be used in combination with conventional tests due to the advantage of the time to result and in scenarios where culture tests are not feasible. Further work to improve molecular tests would benefit from the availability of standardized reference standards and improvements to the methodology.

Wastewater-based epidemiology (WBE) approaches to detect SARS-CoV-2 in municipal wastewater can provide unique information on the incidence or prevalence of COVID-19 in community. However, there are several technical challenges coupled with sewage sampling for SARS-CoV-2, including intermittent shedding of viruses, sampling time, volume, and frequency. Sampling schemes thus may need to be tailored to reach out highly sensitive, accurate, and reliable results. Herein, we compared the accuracy and threshold cycle (Ct) profiles of SARS-CoV-2 in Moore swabs, composite (16-h), and grab samples taken from sewage manholes (n = 17) at the Middle Eastern city of Tehran, Iran, on two occasions (November 2020 and May 2021). Samples were concentrated by polyethylene glycol precipitation and the corresponding Ct values for CDC 'N' and 'ORF1ab' assays were derived by means of real time RT-qPCR. Overall, the Moore swabs performed equal to samples composited over 16 h for qualitative monitoring, and 34/34 (100%) were positive for SARS-CoV-2. The 'N' assay showed the highest detection frequency as compared to 'ORF1ab'. The mean Moore swab Ct profiles were more consistent with 16 h composite sampling as compared with corresponding grab samples, providing hints as to the best sampling protocol to adopt when planning a sewage monitoring campaign particularly under WBE. Furthermore, our analyses on local differences showed somewhat higher virus copy numbers in the southern areas. The experimental design of this study revealed that the Moore swab and composite samples are more sensitive than grab samples, suggesting that the collection of grab samples may be inappropriate for characterizing total number of viral RNA copies in sewage samples. Given the transiently presence of human host-restricted infections such as SARS-CoV-2 and the simplicity and affordability of Moore swab, the method is well suited for disease surveillance in resource poor regions struggling with limited capacity for clinical testing., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Objective: MicroRNAs (miRNAs) are short non-coding RNAs that play a role in post-transcriptional regulation of gene expression. Hsa-miR-11181 was originally introduced as a regulator of genes involved in some brain tumours. Due to the high expression of Hsa-miR-11181 in limited glioblastoma brain tumours, in this study we intend to assess the expressions of Hsa-miR-11181 and Has-miR11181-3p in brain tumour tissues and attribute new target genes to these miRNAs., Materials and Methods: In this experimental study, total RNA from brain tissue samples was extracted for real-time quantitative polymerase chain reaction (RT-qPCR) analysis after cDNA synthesis. In order to confirm a direct interaction of Hsa-miR-11181 with two target genes, the 3' UTR of AKT2 and transforming growth factor-beta receptor 1 ( TGFBR1 ) were cloned separately for assessment by the dual luciferase assay., Results: RT-qPCR analysis indicated that both Hsa-miR-11181-5p and Has-miR11181-3p specifically up-regulated in higher grades of glioma tumours versus other brain tumour types. Consistently, lower expression levels of AKT2 and TGFBR1 were detected in higher grade gliomas compared to other types of brain tumours, which was inverse to the level of expression detected for the heparin-binding EGF-like growth factor ( HBEGF ) gene. The results of the dual luciferase assay supported a direct interaction of Hsa-miR-11181 with the 3' UTR sequences of the AKT2 and TGFBR1 genes., Conclusion: Overall, our data suggest that miR-1118 is a potential molecular biomarker for discrimination of glioma brain tumours from other brain tumour types., Competing Interests: There is no conflict of interest in this study., (Copyright© by Royan Institute. All rights reserved.)

The etiological agent for novel coronavirus (COVID-19, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), not only affects the human respiratory system, but also the gastrointestinal tract resulting in gastrointestinal manifestations. The high rate of asymptomatic infected individuals has challenged the estimation of infection spread based on patients' surveillance, and thus alternative approaches such as wastewater-based epidemiology (WBE) have been proposed. Accordingly, the number of publications on this topic has increased substantially. The present systematic review thus aimed at providing state-of-the-knowledge on the occurrence and existing methods for sampling procedures, detection/quantification of SARS-CoV-2 in sewage samples, as well as anticipating challenges and providing future research direction to improve the current scientific knowledge. Articles were collected from three scientific databases. Only studies reporting measurements of virus in stool, urine, and wastewater samples were included. Results showed that improving the scientific community's understanding in these avenues is essential if we are to develop appropriate policy and management tools to address this pandemic pointing particularly towards WBE as a new paradigm in public health. It was also evident that standardized protocols are needed to ensure reproducibility and comparability of outcomes. Areas that require the most improvements are sampling procedures, concentration/enrichment, detection, and quantification of virus in wastewater, as well as positive controls. Results also showed that selecting the most accurate population estimation method for WBE studies is still a challenge. While the number of people infected in an area could be approximately estimated based on quantities of virus found in wastewater, these estimates should be cross-checked by other sources of information to draw a more comprehensive conclusion. Finally, wastewater surveillance can be useful as an early warning tool, a management tool, and/or a way for investigating vaccination efficacy and spread of new variants.

Background and Objective: Recently, dietary restriction of fermentable carbohydrates (a low-FODMAP diet) in combination with a gluten-free diet (GFD) has been proposed to reduce the symptoms in irritable bowel syndrome (IBS) patients. Different studies reported that IBS has been associated with dysbiosis in the gut microbiota. Additionally, a few studies have reported inflammation in the gastrointestinal (GI) system of adults with IBS. In this study, we aimed to investigate the effects of low FODMAP-gluten free diet (LF-GFD) on clinical symptoms, intestinal microbiota diversity, and fecal calprotectin (FC) level in Iranian patients with IBS., Design: In this clinical trial study, 42 patients with IBS (Rome IV criteria) underwent LF-GFD intervention for 6 weeks. Symptoms were assessed using the IBS symptom severity scoring (IBS-SSS), and fecal samples were collected at baseline and after intervention and analyzed by quantitative 16 S rRNA PCR assay. The diversity of gut microbiota compared before and after 6 weeks of dietary intervention. FC was also analyzed by the ELISA method., Results: Thirty patients (mean age 37.8 ± 10.7 years) completed the 6-week diet. The IBS-SSS was significantly (P = 0.001) reduced after LF-GFD intervention compared to the baseline. Significant microbial differences before and after intervention were noticed in fecal samples. A significant increase was found in Bacteroidetes, and the Firmicutes to Bacteroidetes (F/B) ratio was significantly (P = 0.001) decreased after the dietary intervention. The value of FC was significantly decreased after 6 weeks of dietary intervention (P = 0.001)., Conclusions: Our study suggests that patients with IBS under an LF-GFD had a significant improvement in IBS symptoms severity, with reduced FC level following normalization of their gut microbiota composition. Further rigorous trials are needed to establish a long-term efficacy and safety of this dietary intervention for personalized nutrition in IBS. Clinical Trial Registry Number: IRCT20100524004010N26., (© 2021. The Author(s).)

Introduction: Carbapenem-resistant Enterobacteriaceae (CRE) represents an urgent threat worldwide. We aimed to investigate the frequency of carbapenem-resistant Klebsiella pneumoniae and Escherichia coli in Iran. Materials and Methods: PubMed/Medline, Embase, Scopus, Web of Sciences, and Iranian databases were searched to find potentially relevant articles. Statistical analyses were performed using STATA version 14. Results: Forty-nine studies fulfilled the inclusion criteria. The pooled rates of resistance to carbapenem in K. pneumoniae and E. coli were 24.0% (95% confidence interval [CI] 18.0-31.0) and 5.0% (95% CI 2.0-8.0), respectively. bla OXA-48 gene was the most common cause of carbapenem resistance in K. pneumoniae and E. coli. Conclusions: CRE is prevalent in Iran, which confers the importance of strength prevention and control measures.

Cardiac fibrosis describes the inappropriate proliferation of cardiac fibroblasts (CFs), leading to accumulation of extracellular matrix (ECM) proteins in the cardiac muscle, which is found in many pathophysiological heart conditions. A range of molecular components and cellular pathways, have been implicated in its pathogenesis. In this review, we focus on the TGF-β and WNT signaling pathways, and their mutual interaction, which have emerged as important factors involved in cardiac pathophysiology. The molecular and cellular processes involved in the initiation and progression of cardiac fibrosis are summarized. We focus on TGF-β and WNT signaling in cardiac fibrosis, ECM production, and myofibroblast transformation. Non-coding RNAs (ncRNAs) are one of the main players in the regulation of multiple pathways and cellular processes. MicroRNAs, long non-coding RNAs, and circular long non-coding RNAs can all interact with the TGF-β/WNT signaling axis to affect cardiac fibrosis. A better understanding of these processes may lead to new approaches for diagnosis and treatment of many cardiac conditions. Video Abstract.

Introduction: Carbapenem-resistant Acinetobacter baumannii (CRAB) is recognized to be among the most difficult antimicrobial-resistant gram-negative bacilli to control and treat. An understanding of the epidemiology of CRAB and the mechanisms of resistance to carbapenems is necessary to develop strategies to curtail their spread. Methods: Electronic databases were searched from January 1995 to December 2017 for all studies, which: (1) provide data on the frequency and antibiotic resistance profile of the isolated A. baumannii and (2) describe the mechanisms of carbapenem resistance in detail. Results: Sixty-eight studies were found referring to mechanisms of carbapenem resistance in clinical isolates of A. baumannii , and 56 studies were found referring to the frequency of CRAB. The pooled frequency of carbapenem resistance was 85.1% (95% confidence interval [CI]: 82.2-88.1) in 8,067 clinical isolates of A. baumannii . Resistances due to bla OXA23 (55.3%), bla OXA24 (41.4%), and bla OXA58 (5.2%) genes were the most prevalent reported mechanisms of resistance to carbapenem, respectively. Conclusions: Our data warn that CRAB will rise if the current situation remains uncontrolled. Better control infection strategies and antibiotic managements, particularly in the health care systems, are needed to limit the spread of this pathogen.

Background: Neglected parasitic diseases (NTDs) like cutaneous leishmaniasis (CL) have caused high mortality and morbidity rate in developing countries. This disease is considered as one of the six major tropical diseases, and has a great importance in HIV infected individuals as an opportunistic infection in those areas that both infections are endemic. This study evaluated the therapeutic effects of the Urtica dioica L (U. dioica) aqueous extract as an anti-leishmanial herbal drug in-vitro and in-vivo, and in addition to that, evaluated two vital immune system cytokines including gamma interferon (IFN-γ) and interleukin-4 (IL-4) plus nitric oxide (NO) and arginase activity against Leishmania major (L. major) infected mice., Methodology/principal Findings: In-vitro anti-leishmanial activity of U. dioica aqueous extract was determined using MTT method and also Parasite Rescue Transformation Assay. Also, the footpad lesion size and parasite load in BALB/c mice infected with L. major were quantified for in-vivo assessment. Furthermore, for evaluating the immune responses, the levels of IFN-γ, IL-4, NO and arginase were measured in the BALB/c mice. These results indicated that U. dioica extract significantly reduced the L. major promastigotes viability. According to the in-vitro cytotoxicity assay of the extract on Leishmania parasites (CC50) and infected macrophages (EC50), the extract had no toxicity to the macrophages, however it efficiently killed the L. major amastigotes. In addition, the lesion size, parasite load, IL-4, and ARG were decreased in the treated infected mice, however IFN-γ and NO were significantly increased., Conclusions/significance: This study established satisfactory results in Leishmania parasite clearing both in-vivo and in-vitro. Therefore, U. dioica extract can be considered as an effective and harmless herbal compound for killing the parasite without toxicity to the host macrophages. Furthermore, it also can treat the CL by switching the mouse immune response towards a cell-mediated response (Th1); hence, it may be identified as a perfect therapeutic herbal drug for CL treatment., Competing Interests: The authors have declared that no competing interests exist.

Extended-Spectrum Beta-lactamase producing Enterobacteriales (ESBL-PE) in fecal carriage have become a global health concern. Detection of putative virulent ESBL-producing E.coli (ESBL-EC) isolates among asymptomatic carriers is a threatening issue in public health. The aim of this study was to investigate the intestinal carriage of ESBL-EC, phylo-groups and clonal relatedness among putative virulent groups of ESBL-EC isolated from fecal carriages. A total of 120 rectal swabs; 50.8% (61/120) from inpatients of intensive care unit (ICU) and 49.2% (59/120) from outpatients were collected. The ESBL-EC screening was performed by using MacConkey agar supplemented with cefotaxime. PCR assays were applied for determination of phylo-groups, detection of ESBL and carbapenemase genes. Conjugation experiment, plasmid replicon typing and Multilocus Sequence Typing (MLST) were performed for putative virulent phylo-groups. Totally, of 120 studied individuals, 60.0% (72/120) were carrier for ESBL-EC. The rate of bla CTX-M-15 , bla TEM , bla SHV was 90.2% (65/72), 50.0% (36/72) and 5.5% (4/72), respectively. The frequency of phylo-groups A, B1, B2, C, D, and F were 20.8% (15/72), 6.9% (5/72), 20.8% (15/72), 2.7% (2/72), 13.8 (10/72) and 12.5% (9/72), respectively. In conjugation experiments, of 6 tested isolates, 5 had conjugative plasmids. The most prevalent plasmid types belonged to IncF incompatibility groups. The MLST analysis showed that the main sequence types among ESBL-EC isolates were ST769 and ST472. The current study provides novel information about the presence of the ESBL-EC isolates, particularly putative virulent phylo-groups among fecal carriages in Iran. Our data revealed that there was almost high ST heterogeneity among putative ESBL-EC isolates. In order to implementation of effective infection control program, detection of fecal carriage in appropriate time typically at the beginning of admission to the hospital is recommended., (Copyright © 2019 Elsevier B.V. All rights reserved.)