The engineering, drug release, and in vitro evaluations of the PLLA/HPC/Calendula Officinalis electrospun nanofibers optimized by Response Surface Methodology.

A system based on poly(l-lactic acid) (PLLA) and hydroxypropyl cellulose (HPC) was considered in this study to achieve electrospun mats with outstanding properties and applicability in biomedical engineering. A novel binary solvent system of chloroform/N,N-dimethylformamide (CF/DMF:70/30) was utilized to minimize the probable phase separation between the polymeric components. Moreover, Response Surface Methodology (RSM) was employed to model/optimize the process. Finally, to scrutinize the ability of the complex in terms of drug delivery, Calendula Officinalis (Marigold) extract was added to the solution of the optimal sample (Opt.PH), and then the set was electrospun (PHM). As a result, the presence of Marigold led to higher values of fiber diameter (262 ± 34 nm), pore size (483 ± 102 nm), and surface porosity (81.0 ± 7.3 %). As this drug could also prohibit the micro-scale phase separation, the PHM touched superior tensile strength and Young modulus of 11.3 ± 1.1 and 91.2 ± 4.2 MPa, respectively. Additionally, the cumulative release data demonstrated non-Fickian diffusion with the Korsmeyer-Peppas exponent and diffusion coefficient of n = 0.69 and D = 2.073 × 10-14 cm2/s, respectively. At the end stage, both the Opt.PH and PHM mats manifested satisfactory results regarding the hydrophilicity and cell viability/proliferation assessments, reflecting their high potential to be used in regenerative medicine applications.

Runaway electrons and their interaction with tungsten wall: a comprehensive study of effects.

Runaway electrons are a notable phenomenon occurring during the operation of a tokamak. Proper material selection for the tokamak's first wall structure and plasma facing components, particularly in large sizes tokamaks like ITER and DEMO, is crucial due to the energy deposition of runaway electrons on plasma facing components during collision events, resulting in severe heat transfer and material damage in the form of melting, corrosion, and fracture. These runaway electrons also contribute to the production of photoneutrons through (γ, n) nuclear reactions, lead to material activation and require remote handling. In this study, using a Monte Carlo code and simulating the collision of runaway electrons with a tungsten target exposed to their radiation, the electron transport is investigated, and the energy deposition spectrum resulting from these collisions on the target is analyzed. The influence of incident angle and magnetic field on the energy deposition spectrum and the energy deposition per particle in the target is examined. With an increase in the incident angle of incoming electrons, the amount of energy deposited in the target rises and the energy deposition spectrum broadens. Moreover, applying a magnetic field, results the most significant increase in energy deposition for electrons with energies below 1 MeV in the tangential radiation case. The energy deposition spectrum resulting from each collision event in these interactions is determined. For electrons with energies below 5 MeV, multiple scattering and ionization processes are the primary contributors to energy deposition in the target. However, as the incident electron energy increases, the significance of multiple scattering and ionization diminishes, and the bremsstrahlung process becomes the most effective reaction in energy deposition. The energy deposition profile of electrons in the tungsten target indicates that higher incident electron energies lead to a shift of the maximum energy deposition location towards the inner layers of the target, and the energy deposition peak broadens. Analyzing the electrons transport inside the tungsten target reveals that a substantial portion of electrons with energies of 50-100 MeV passes through the wall and may exit from the back surface, potentially causing damage to equipment behind the tungsten wall. Additionally, secondary products of the reaction, such as photons, secondary electrons, and neutrons and their energy profiles are thoroughly studied. These secondary products can penetrate the target and activate materials in the equipment behind the plasma-facing components. For primary electrons below 1 MeV hitting tungsten, reflection process is significant. Analysis of primary and secondary runaway electrons in the tokamak's tungsten wall shows that electrons with energies of 0.1, 0.2, and 0.5 MeV predominantly interact within a first 0.1 mm layer, without passing through it. The secondary electrons can escape the tungsten target and impact other components, which making them an important consideration in runaway electron collisions with the tokamak wall. Produced photons, as one of the secondary products, also linearly increase with the rising energy of primary electrons. Also, the photoneutrons are produced only when runaway electrons with energies of 10 MeV and above collide with the target. These secondary products can penetrate the target and activate materials in the equipment behind the plasma-facing components.

Biomarkers for Assessment of Human Immunodeficiency Virus and its Co-Infection with Hepatitis B and Hepatitis C Viruses: A Comprehensive Review.

Recently, prevalence of hepatitis B virus (HBV), and hepatitis C virus (HCV) co-infection with Human immunodeficiency virus (HIV), has dramatically increased worldwide due to their shared routes of transmission. Compared to the sporadic infection with HIV, HBV, and HCV, concurrent infection with these agents increases the complications of these viruses. Furthermore, co-infection may also alter the therapeutic strategies against HIV. Accordingly, choosing appropriate biomarkers to detect these co-infections is one of the main concerns in the field of diagnostic pathology. Up to now, several markers have been introduced for the simultaneous diagnosis of HIV, HBV, and HCV. In this regard, serum adenosine deaminase activity (ADA), FibroTests, AST-to-Platelet Ratio Index (APRI), Fibrosis-4, Hyaluronic acid, and micro ribonucleic acids (MiR) have been investigated as potential biomarkers for diagnosis of HIV-HCV/HBV co-infections. This review summarizes diagnostic values of the current and emerging biomarkers in HIV patients concurrently infected with HBV and HCV.

Pathogenic CANVAS-causing but not nonpathogenic RFC1 DNA/RNA repeat motifs form quadruplex or triplex structures.

Biallelic expansions of various tandem repeat sequence motifs are possible in RFC1 (replication factor C subunit 1), encoding the DNA replication/repair protein RFC1, yet only certain repeat motifs cause cerebellar ataxia, neuropathy, and vestibular areflexia syndrome (CANVAS). CANVAS presents enigmatic puzzles: The pathogenic path for CANVAS neither is known nor is it understood why some, but not all expanded, motifs are pathogenic. The most common pathogenic repeat is (AAGGG)n•(CCCTT)n, whereas (AAAAG)n•(CTTTT)n is the most common nonpathogenic motif. While both intronic motifs can be expanded and transcribed, only r(AAGGG)n is retained in the mutant RFC1 transcript. We show that only the pathogenic forms unusual nucleic acid structures. Specifically, DNA and RNA of the pathogenic d(AAGGG)4 and r(AAGGG)4 form G-quadruplexes in potassium solution. Nonpathogenic repeats did not form G-quadruplexes. Triple-stranded structures are formed by the pathogenic motifs but not by the nonpathogenic motifs. G- and C-richness of the pathogenic strands favor formation of G•G•G•G-tetrads and protonated C+-G Hoogsteen base pairings, involved in quadruplex and triplex structures, respectively, stabilized by increased hydrogen bonds and pi-stacking interactions relative to A-T Hoogsteen pairs that could form by the nonpathogenic motif. The ligand, TMPyP4, binds the pathogenic quadruplexes. Formation of quadruplexes and triplexes by pathogenic repeats supports toxic-DNA and toxic-RNA modes of pathogenesis at the RFC1 gene and the RFC1 transcript. Our findings with short repeats provide insights into the disease specificity of pathogenic repeat motif sequences and reveal nucleic acid structural features that may be pathogenically involved and targeted therapeutically.

Impact of teaching, learning, and assessment of medical law on cognitive, affective and psychomotor skills of medical students: a systematic review.

BACKGROUND: It is necessary to improve medical students' legal cognitive, affective, and psychomotor skills to prevent further legal issues in the medical profession. Choosing the proper teaching and assessment methods is crucial in this matter. This study aimed to investigate the impact of teaching, learning, and assessment of medical law on the cognitive, affective, and psychomotor skills of medical students. METHODS: A systematic review was conducted in PubMed, Embass, and Web of Science databases, and Google Scholar search engine using MECIR and PRISMA, AMEE Guide 94 for 1980 to 2022.12.30. Nineteen articles met the inclusion criteria. MERSQI checklist was used to assess the quality of the articles, and URSEC (theoretical underpinning of the development, resources required, setting, educational methods employed, and content) used to assess the risk of educational bias. RESULTS: Internship courses called Medical Education Humanities and Society (MESH), clinical scenario design, seminars and small group discussions, web-based interactive training, legal training courses, PBL, and mind maps have been used to improve the medico-legal knowledge of medical students. MESH clerkship, simulation of a legal event, medico-legal advocacy program based on interdisciplinary education, group discussion, integration, and court-based learning used to improve student attitudes. Multidisciplinary training, small group discussions after the seminar, mock trial competition, and interdisciplinary education are used to teach psychomotor skills. All studies, except one on knowledge, reported positive effects of legal education on students' knowledge, attitudes, and legal performance. Written assessments were used for cognitive and affective domains, while performance was assessed by OSCE, simulated court, and evaluation of patient referrals. CONCLUSION: There are few studies to examine the cognitive, affective, and legal psychomotor skills of medical students. The texts have not yet fully explored the high level of affective and psychomotor domains, which is evidence of a gap in this sector. Recognizing that medico-legal problems are prevented through proper education and assessment, it is recommended that this area be considered a research priority and that effective educational policies are adopted.

Foliar-applied silicon and selenium nanoparticles modulated salinity stress through modifying yield, biochemical attribute, and fatty acid profile of Physalis alkekengi L.

Soil salinity is a major environmental problem owing to its negative impact on agricultural productivity and sustainability. Nanoparticles (NPs) have recently been highlighted for their ability to alleviate salinity stress. The current study aimed to alleviate salt stress by using silicon (Si) and selenium (Se) NPs on the growth and physiological attributes of Physalis alkekengi L. Plants were irrigated with saline water at 50, 100, and 200 mM NaCl, and Si NPs (200 mg L-1) and Se NPs (50 mg L-1) were sprayed on leaves three times in a pot experiment in 2022. Leaf chlorophyll (Chl) content, antioxidant capacity, and fatty acid (FA) profile of fruits were measured to find the effects of NPs and salinity in the plants. Salinity at 50 mM did not significantly differ from the control, but at 100-200 mM, salt stress had a substantial impact on the majority of traits. Compared with non-saline conditions, 200 mM NaCl led to decreases in shoot weight (40%), fruit weight (30%), Chl a (30%), Chl b (39%), anthocyanin (31%), ascorbic acid (16%), total phenolic content (TPC, 11%) but increases in total soluble solids (TSS, 79%), titration acidity (TA, 17%), and TSS/TA (52%) in plants without spraying the NPs. However, Si and Se NPs modulated salinity stress by increasing shoot and fruit weight, Chl content, anthocyanin, and TPC, and with decreasing TSS and TSS/TA. Salinity elevated polyunsaturated fatty acids (PUFAs) and lowered monounsaturated fatty acids (MUFAs). According to multivariate analysis, 50 mM and control were found to be in the same cluster, whereas 100 and 200 mM were shown to be in different clusters. Foliar application of Si and Se NPs at 200 and 50 mg L-1, respectively, can be recommended for mitigating salt stress at 100-200 mM NaCl in P. alkekengi L. Plants. Farmers can use the findings to increase the ability of Si and Se NPs to protect plants against salt.

Post-Traumatic Stress Disorder among Emergency Medical Technicians and its Relationship with Occupational Stress and Depression: Post-Corona Screening, Zanjan, 2022.

Objective: Emergency medical technicians (EMTs) are at risk of developing post-traumatic stress disorder (PTSD) as a result of seeing painful events involving suspected COVID-19 patients and being concerned about potentially infecting themselves and their families. Therefore, screening for these disorders is essential in the post-Corona era. This study aimed to investigate the prevalence of PTSD among EMTs and its relationship with occupational stress and depression when dealing with patients with suspected COVID-19. Methods: This cross-sectional study was conducted on EMTs at Zanjan University of Medical Sciences using a convenience sampling method. Data were collected using a demographic information questionnaire, PTSD checklist, occupational stress questionnaire, and the Goldberg depression scale. The data were analyzed using SPSS software. Statistical tests such as Pearson correlation and logistic regression analysis were used to evaluate the data. Result: 205 EMTs participated in this cross-sectional study. The mean and standard deviation of PTSD was 37.13±12.93 (17-85), and according to the cut-off (45), the prevalence of PTSD was 30.7%. There was a direct and significant association between the total PTSD and depression scores (r=0.435, p=0.001). Some occupational stress domains, such as demand (r=0.306, p=0.001), colleague support (r=0.149, p=0.033), and communication (r=0.293, p=0.001) had a significant association with PTSD. The domains of sadness in depression (OR=1.074, p=0.027) and demands in occupational stress (OR=1.872, p=0.029) were the most important predictors of PTSD. Among demographic variables, employment status was the most important protective factor for PTSD (OR=0.378, p=0.038). Conclusion: PTSD affected one-third of EMTs, and it had a significant relationship with various dimensions of depression and occupational stress. Due to the chronic nature of these diseases, policymakers are advised to prioritize psychological screening of EMTs as part of the post-Corona policy.

Effects of cigarette smoke condensate on reproduction in mice in vivo.

Smoking has dangerous and sometimes irreversible effects on various body tissues, including the reproductive system. We conducted this research to determine the in vivo effects of cigarette smoke condensate (CSC) on reproduction in mice. In this experimental in vivo study, 32 male and female NMRI mice were divided into four groups. The mice were injected with CSC (CSC-1R3F) for 28 days. The mice were mated 1 day after the last injection and observed daily for 1 week for the presence of a vaginal plug to track mating. We evaluated mating success rate, and sperm and oocyte quality, pregnancy outcome, childbearing status, and in vitro fertilization (IVF). The results showed a decrease in successful mating in female mice that received the CSC injections. CSC significantly influenced the number of offspring born to males. When the CSC was injected into male mice, there was a significant increase in the number of offspring compared with the group in which only the females received CSC injections. According to the results, there was a negative effect of CSC on morphological parameters in male and female mice. Also, successful IVF after exposure to CSC was significantly decreased in the female mice treated group. The results indicated that CSC significantly affected the number of offspring and fecundity success in females.

Effect of histone deacetylase 8 gene deletion on breast cancer cellular mechanism in vitro and in vivo study.

BACKGROUND: Triple-negative breast cancer (TNBC) is the most aggressive type of cancer without any approved targeted therapy. Epigenetic processes have a pivotal role in cancer cell progression and while histone deacetylase 8 (HDAC8) has been proven as a potential oncogene in breast cancer, its underlying molecular mechanism is not known. Therefore, the present study, aimed to evaluate the underlying mechanism of the HDAC8 carcinogenesis in breast cancer progression. METHODS: The potential role of HDAC8 in cancer cell processes such as apoptosis, invasion, migration, angiogenesis, and cancer stem cells (CSCs) markers were evaluated by using flow cytometry Annexin V-FITC/propidium iodide (PI), reverse transcription-polymerase chain reaction (RT-qPCR), Matrigel-coated transwell plates and wound healing assay on both cell lines. The impact of HDAC8 on tumor development was also studied using a breast cancer xenograft model. RESULTS: HDAC8 expression was significantly downregulated in the cell lines, post-transfection with KO-vector. Downregulation of HDAC8 dramatically decreased cell migration, angiogenesis, and invasion while inducing apoptosis in MDAMB-468 and MDA-MB-231 cell lines. HDAC8 knocked out TNBC cell lines had lower levels of cancer stemness markers, such as prominin-1 (CD133), CD44, BMI1, and Aldehyde dehydrogenase 1 (ALDH1). Additionally, the knockout of HDAC8 inhibited tumor growth in a breast cancer xenograft model. CONCLUSION: The findings show that knocking out HDAC8 retains several anticancer actions in BC cells, such as inducing apoptosis, reducing migration, invasion, angiogenesis and removing CSCs markers.

Effect of cigarette smoke condensate on mouse embryo development and expression of pluripotency and apoptotic genes in vitro.

The aim of the present study was to investigate the effect of cigarette smoke condensate (CSC) on in vitro development of mouse embryos. In total 3000 NMRI mice 2PN embryos were divided into six groups (n = 500). The test group was exposed to 20, 40, 80, 160 or 320 µg/ml of CSC. In the control group, CSC was not added to the culture medium during the development of 2PN embryos. The effects of 20 and 80 µg/ml of CSC on genes involved in pluripotency and apoptosis, and also, the aryl hydrocarbon receptor gene was assessed in the blastocysts. Our results showed that CSC had an adverse effect on the viability of mouse embryos at the concentrations of 80, 160 and 320 µg/ml compared with the control group (P < 0.05). In contrast, it had positive effects on the viability of mouse embryos at the concentrations of 20 and 40 µg/ml compared with the control group (P < 0.05). The 20 and 80 µg/ml concentrations of CSC increased the expression of pluripotency, apoptotic, and aryl hydrocarbon receptor genes in the blastocyst embryo stage compared with the control group (P < 0.05). It can be concluded that concentrations higher than 40 µg/ml of CSC have an adverse effect on mouse embryo development in the preimplantation stages. Also, 20 and 80 µg/ml concentrations of CSC have a significant effect on the expression of pluripotency, apoptotic, and the aryl hydrocarbon receptor genes in the blastocyst embryo stage compared with the control group.

Introducing a simple and cost-effective RT-PCR protocol for detection of DPYD*2A polymorphism: the first study in Kurdish population.

PURPOSE: Fluoropyrimidines, the major chemotherapeutic agents in various malignancies treatment, are metabolized by dihydropyrimidine dehydrogenase (DPD). DPD deficiency can lead to severe and sometimes fatal toxicity. In the present study, we developed a simple protocol to detect the DPYD*2A variant. Common side effects in patients treated with these drugs were also evaluated in a Kurdish population. METHOD: We established a reverse-transcriptase polymerase chain reaction (RT-PCR) technique for detection of DPYD*2A. Sanger sequencing was used to confirm the results. 121 Kurdish patients receiving fluoropyrimidine derivatives were enrolled, and clinical information regarding the dosage and toxicity was analyzed. RESULTS: Our RT-PCR method was able to detect one patient with heterozygous state for DPYD*2A (0.8%). The most observed adverse drug reactions were tingling, nausea, and hair loss. The frequency of patients with the toxicity of grade 3 or worse was 6.6%. CONCLUSION: This was the first study that detect DPYD*2A polymorphism in the Kurdish population. Our method was successfully able to detect the DPYD*2A variant and, due to its simplicity and cost-effectiveness, it may be considered as an alternative to the current methods, especially in developing countries. Our detected polymorphism rate at 0.8% is comparable with other studies. Despite the low rate of DPYD*2A polymorphism, pharmacogenetics assessment before beginning the treatment process is highly recommended due to its association with a high risk of severe toxicity.

miR-589-5p Inhibits Cell Proliferation by Targeting Histone Deacetylase 3 in Triple Negative Breast Cancer.

BACKGROUND: Histone deacetylase 3 (HDAC3) is a potential oncogene that is significantly up-regulated in patients with breast cancer. MicroRNAs (miRs) are a group of small non-coding and regulatory RNAs which have recently been proposed as promising molecules for breast cancer target therapy. In the current study, we investigated the impact of miR-589-5p/ HDAC3 axis on cancer cell development in triple negative breast cancer (TNBC) cells. METHODS: In-silico analysis determined that miR-589-5p potentially targets HDAC3. We evaluated the HDAC3 and mir-589-5p expression levels in clinical samples and breast cancer cell lines including MDA-MB-231, MDA-MB-468, MCF-7 and MCF-10A. HDAC3 was knocked out to investigate its role on cancer cell progression. Anti-cancerous role of the miR-589-5p was assessed using an expression vector. We evaluated possible alteration in the cell cycle progression, cell viability and cell proliferation, after transient transfection. RESULTS: HDAC3 was over-expressed in TNBC clinical samples and breast cancer cell lines compared to non-cancerous controls while miR-589-5p was down regulated in cancer cells. Suppression of HDAC3 decreased the cell viability, cell proliferation and colony formation. Similar results were observed after over-expression of the miR-589-5p. Dual-Luciferase reporter assay confirmed the direct targeting of HDAC3 by miR-589-5p. CONCLUSION: Our results showed that miR-589-5p mediates its anti-proliferative effects on breast cancer cells via targeting HDAC3. These findings suggest that the miR-589-5p/ HDAC3 axis could be considered as a possible therapeutic strategy in TNBC.

Relation between the circular and linear form of the Elongator Acetyltransferase Complex Subunit 3 in the progression of triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is the most aggressive type of breast cancer (BC) that hardly responds to common treatment. Recent studies show that circ-ELP3 (Elongator Acetyltransferase Complex Subunit 3 or hsa-circ-0001785) is involved in the pathogenesis of several malignancies. The present study aimed to evaluate the possible role of this circRNA in the progression of TNBC cells and the possible relation between the circular and linear forms of the ELP3. We evaluated the circ-ELP3 and its host gene expression level in clinical samples and breast cancer cell lines. Using an expression vector, hsa-circ-0001785 was upregulated to investigate its role on cancer cell progression. After a transient transfection, we evaluated possible alterations in the cell cycle progression, cell viability, and cell proliferation. Quantitative real-time polymerase chain reaction analyses verified that circ-ELP3 and its host gene were significantly upregulated in TNBC tissues and breast cancer cells. Overexpression of circ-ELP3 markedly increases the cell viability and proliferation and also the formation of colonies in transfected cells compared to the controls. Briefly, our results showed that Circ-ELP3 and its host gene were significantly upregulated in TNBC. Circ-ELP3 is involved in TNBC progression and may exert its effects by indirectly regulating of ELP3 expression.

Multi- and Transgenerational Effects of Developmental Exposure to Environmental Levels of PFAS and PFAS Mixture in Zebrafish (Danio rerio).

Per- and polyfluoroalkyl substances (PFASs) are ubiquitous in the environment and are tied to myriad health effects. Despite the phasing out of the manufacturing of two types of PFASs (perfluorosulfonic acid (PFOS) and perfluorooctanoic acid (PFOA)), chemical composition renders them effectively indestructible by ambient environmental processes, where they thus remain in water. Exposure via water can affect both human and aquatic wildlife. PFASs easily cross the placenta, exposing the fetus at critical windows of development. Little is known about the effects of low-level exposure during this period; even less is known about the potential for multi- and transgenerational effects. We examined the effects of ultra-low, very low, and low-level PFAS exposure (7, 70, and 700 ng/L PFOA; 24, 240, 2400 ng/L PFOS; and stepwise mixtures) from 0-5 days post-fertilization (dpf) on larval zebrafish (Danio rerio) mortality, morphology, behavior and gene expression and fecundity in adult F0 and F1 fish. As expected, environmentally relevant PFAS levels did not affect survival. Morphological abnormalities were not observed until the F1 and F2 generations. Behavior was affected differentially by each chemical and generation. Gene expression was increasingly perturbed in each generation but consistently showed lipid pathway disruption across all generations. Dysregulation of behavior and gene expression is heritable, even in larvae with no direct or indirect exposure. This is the first report of the transgenerational effects of PFOA, PFOS, and their mixture in terms of zebrafish behavior and untargeted gene expression.

The role of histone deacetylase 3 in breast cancer.

It has been recently revealed that Histone Deacetylase (HDAC) 3, a unique member of the HDACs family, can trigger and progress cancers by alternation in genes expression and proteins activity. Epigenetic modifications by HDACs have been studied well in various cancer cells. Recent studies have focused on the HDAC enzymes as a possible target in cancer therapy. There are significant documents on upregulation of HDAC3 in breast cancer (BC) cells which suggest an oncogenic role for this enzyme. Interestingly, some studies showed that HDAC3 inhibition could be considered as a promising target in breast cancer therapy, and thus far, several inhibitors from different nature have been introduced. In this review, we discussed the function and highlight the existing inhibitors of HDAC3 in BC pathogenesis and therapy.

Possible correlation between high circulatory levels of trimethylamine-N-oxide and 2177G>C polymorphisms of hepatic flavin containing monooxygenase 3 in Kurdish Population with non-alcoholic fatty liver disease.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is a multifactorial disorder with complicated pathophysiology. Trimethylamine-N-oxide (TMAO) has been thought to be correlated with the pathogenesis of NAFLD. The single nucleotide polymorphisms (SNPs) of hepatic flavin-containing monooxygenase 3 (FMO3) regulate the concentration of TMAO. This case-control study investigated the plasma levels of TMAO as well as its possible correlation with the frequency of specific genotype of FMO3 (-2650C>G, -2543T>A, -2177G>C, -2589C>T, -2106G>A polymorphisms) in Kurdish patients with NAFLD. METHODS AND RESULTS: In 85 confirmed NAFLD patients and 30 healthy individuals, triglycerides (TG), total cholesterol (Chol), low-density lipoprotein (LDL), high-density lipoprotein (HDL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities were measured. TMAO was also measured using the LC-MS/MS method. High-resolution melting analysis was applied to determine FMO3 genotypes. Plasma TMAO levels were significantly higher in patients (p = 0.030). A CC genotype with a frequency of 12.9% for SNP -2177G>C was found in Kurdish NAFLD patients. The distribution of the GC genotype was also significantly different (p = 0.017). CONCLUSIONS: The current results provide documentation for high circulatory levels of TMAO and its possible correlation with the presence of the specific genotype -2177G>C FMO3 in Kurdish NAFLD patients.

Fabrication and Characterization of a Three-Dimensional Fibrin Gel Model to Evaluate Anti-Proliferative Effects of Astragalus hamosus Plant Extract on Breast Cancer Cells.

BACKGROUND: Breast Cancer (BC) is a malignancy with high mortality among women. Recently, scaffold-based three-dimensional (3D) models have been developed for anti-cancer drug research. The present study aimed to investigate the anti-proliferative effects of Astragalus hamosus (A. hamosus) in 3D fibrin gel against MCF-7 cell line. We have also evaluated anti-proliferative effect of A. hamosus differences between 3D and 2D cultures. METHODS: The fibrin gel formulation was first optimized by testing the structural and mechanical properties. Then the cytotoxic effect of A. hamosus extract was assessed on MCF-7 cells by MTT assay. Cell apoptosis was evaluated using TUNEL method and flow cytometry. Cell cycle and proliferation were analyzed by flow cytometry. Apoptosis-related gene expression such as Bcl-2, caspase-3, -8 and -9 were quantified by real time-PCR. RESULTS: TUNEL staining showed a significant damage accompanied with cell apoptosis. Flow cytometry analysis revealed that apoptosis increased after treatment with A. hamosus extract in 3D culture model compared to 2D culture. The A. hamosus extract arrested cell cycle in the S and G2/M phases in 3D model while in the 2D culture G0/G1 phase was affected. Treatment with A. hamosus extract led to upregulation of the caspase-3, -8 and -9 genes and downregulation of the Ki-67 in the 3D-culture compared with the 2D culture. CONCLUSION: These results indicated that A. hamosus extract could be used as a therapeutic candidate for BC due to its anti-proliferative effects. Furthermore, 3D fibrin gel could be better than 2D-cultured cells in simulating important tumor characteristics in vivo, namely, anti-proliferative and anti-apoptotic features.

ARDS, Diffuse Alveolar Hemorrhage and Pericardial Effusion due to Anabolic-Androgenic Steroids Consumption: Legal and Ethical Policy in Medical Education.

Anabolic-androgenic steroids (AAS) are one of the ingredients of herbal and dietary supplements that are popular among sports trainers. AAS abuse predisposes everyone to several complications. Reviews of the literature on AAS users have shown mainly skin, renal, and hepatic complications. In this case report, we presented a case with simultaneous complications, including diffuse alveolar hemorrhage (DAH), acute respiratory distress syndrome (ARDS), pericardial effusion, gastrointestinal bleeding (GIB), and acute kidney injury (AKI). Given the potential for lethal complications and the consequences of ethical, civil, and criminal law, it seems that specific policies will be considered for the use of bodybuilding drugs. It is also suggested that this approach be added as a new part of the medical curriculum. Also, ARDS and DAH are unreported side effects in other studies, which is suggested to be considered by specialists.

Oral Administration of Probiotic Enterococcus durans to Ameliorate Experimental Autoimmune Encephalomyelitis in Mice.

Introduction: Probiotics, including lactobacilli, have immunomodulatory activities with promising effects on inflammatory diseases. In this study, we evaluate the effect of Enterococcus durans (Edu) and three various strains of lactobacilli (Lacto-mix), including L. rhamnosus, L. casei, and L. plantarum, to prevent Experimental Autoimmune Encephalomyelitis (EAE) features. Methods: C57BL/6 female mice were inoculated with Myelin Oigodendrocyte Glycoprotein (MOG35-55) in CFA (complete Freund's adjuvant) to induce EAE. Five groups (n=6 in each group) of animals received saline or probiotics by oral gavage with 200 µL of lactobacilli (1.5×108 CFU/mL) for 2 weeks before the immunization and during the test for one month. Results: Histopathological studies showed an increase in infiltration of inflammatory cells and destruction of the myelin membrane in the EAE group but a decrease in inflammatory cells in the probiotic-treated animals. Pro-inflammatory cytokines (Interleukin [IL]-17 and Interferon [IFN]-γ) concentration in the supernatant of the brain and spinal cord tissues showed a significant increase in the EAE compared with the normal saline group (P<0.01). While in the spinal cord tissue, there was a decrease in IL-17 in those animals treated with the Lactomix and Edu + Lacto-mix (P<0.01) and a significant decrease in IFN-γ in those animals that received Edu (P<0.05). Western blot analysis of matrix metalloproteinase-9 and myelin basic protein showed a decrease and increase in treatment and EAE groups, respectively, compared to the normal control group. Conclusion: Our data suggest that probiotic Enterococcus durans and Lacto-mix prevents EAE, but further studies are needed to clarify the exact mechanisms and their application in preclinical and clinical trials. Highlights: Dysfunction of the blood-brain barrier, migration of inflammatory cells into the Central Nervous System (CNS), and an increase in the pro-inflammatory factors, are the hallmarks in the pathogenesis of Multiple Sclerosis (MS) and Experimental Autoimmune Encephalomyelitis (EAE).The optimal effects of probiotic strains may involve the simultaneous use of more than one strain.Probiotic Enterococcus durans and Lacto-mix have a preventive effect against EAE. Plain Language Summary: Multiple Sclerosis (MS) is a myelin-degenerating autoimmune disease in the central nervous system. Experimental Autoimmune Encephalomyelitis (EAE), due to its similar clinical and pathologic features to MS, is widely used in many model studies of this disease. The microbiome refers to a genomic set of germs (bacteria, arches, fungi, and viruses), a commensal flora that lives in the intestine and niche of humans and other mammals. The microbiome affects the host's physiological system, especially the balance between health and disease. Additionally, the importance of the microbiome is evident in regulating the intestine-brain axis, or the coordination of the digestive and the central nervous system. In this regard, probiotics, including lactobacilli, have antioxidant and anti-inflammatory properties in vitro and in vivo. Probiotic strains have a wide range of health-improvement effects, and a combination of strains with specific properties provides a broader range of antimicrobial spectrum and stronger anti-inflammatory effects. Considering the critical role of probiotics in the immune system, this study aimed to investigate the possible role of Enterococcus durans alone or in combination with Lactobacillus mixture (L. rhamnosus, L. casei, and L. plantarum) on the EAE animal model of MS.

Toll-Like Receptor 4: A Macrophage Cell Surface Receptor Is Activated By Trimethylamine-N-Oxide.

OBJECTIVE: Trimethylamine-N-Oxide (TMAO) is considered as a risk factor for atherosclerosis which further leads to inflammation during atherosclerosis. The exact mechanism(s) by which TMAO induces the inflammatory reactions remains to be determined. TMAO can cause the endoplasmic reticulum (ER) stress that triggers activation of Toll-Like Receptors (TLRs). In macrophages, this process stimulates the production of proinflammatory cytokines. This study designed to evaluate the expression level of TLR4 in TMAO-treated macrophages. MATERIALS AND METHODS: In this experimental study, different concentrations of TMAO (37.5, 75, 150, and 300 µM) were exposed to murine macrophage (J774A.1 cell line) for 8, 18, 24, and 48 hours. The cells were also treated with 2.5 mM of 4-phenyl butyric acid as well as 2µg/ml of tunicamycin respectively as negative and positive controls for inducing ER-stress. We measured the viability of treated cells by the MTT test. Besides, the expression levels of TLR4 gene and protein were evaluated using western blotting and reverse transcription- quantitative polymerase chain reaction (RT-qPCR) analysis. One-Way ANOVA was used for statistical analysis. RESULTS: No cell death was observed in treated cells. The cells treated with 150 and 300 µM doses of TMAO for 24 hours showed a significant elevation in the protein and/or mRNA levels of TLR4 when compared to normal control or tunicamycin-treated cells. CONCLUSION: Our results may in part elucidate the mechanism by which TMAO induces the macrophage inflammatory reactions in response to the induction of ER stress, similar to what happens during atherosclerosis. It also provides documentation to support the direct contribution of TLR4 in TMAO-induced inflammation.