Molecular cloning and expression of Bacillus anthracis Lethal Factor domain 1 gene in Escherichia coli

زمینه و هدف: سیاه‌زخم (آنتراکس) یک بیماری مشترک بین انسان و دام است. عامل ایجاد کننده بیماری باکتری باسیلوس آنتراسیس می‌باشد که آنتی‌ژن حفاظت‌کننده (PA) و ناحیه یک فاکتور کشنده (LFD1) ایمونوژن‌های قوی این باکتری بوده و همواره به عنوان کاندیدای واکسن علیه باسیلوس آنتراسیس در نظر گرفته شده‌اند. هدف این مطالعه تولید آنتی‌ژن ناحیه یک فاکتور کشنده(LFD1) در باکتری Escherichia coli می‌باشد. روش بررسی: در این مطالعه تجربی آزمایشگاهی ژن LFD1 از پلاسمید pXO1 شناسایی و با واکنش PCR تکثیر شد. با جایگاه‌های آنزیمی BamH I و Xho Iدر وکتور (pGEM-T easy) همسانه‌سازی شد و بعد از جداسازی به وکتور بیانی pET28a(+) زیرهمسانه‌سازی گردید. این وکتور به باکتری E. coli-BL21 (DE3) تراریخت (ترانسفورم) شد. بیان ژن LFD1 تحت القای ایزوپروپیل-β -ِD -I-گالاکتوپیرانوزید (IPTG) انجام و پروتئین مورد نظر بیان شد. یافته‌ها: ژن ناحیه یک فاکتور کشنده (LFD1) کلون شده در وکتور بیانی pET28a(+) به وسیله‌ی توالی یابی، PCR و هضم به وسیله آنزیم‌های با اثر محدود تأیید گردید. همچنین پروتئین نوترکیب تولید شده به وسیله سدیم دودسیل سولفات پلی آکریل آمید ژل (SDS-PAGE) و لکه‌گذاری وسترن تایید گردید. نتیجه‌گیری: با توجه به ایمونوژن بودن پروتئین LFD1، پروتئین نوترکیب تولید شده در این تحقیق را می‌توان به‌صورت مجزا یا ترکیبی با یاورها و یا انتقال دهنده‌ها در طراحی واکسن برای بیماری سیاه‌زخم استفاده نمود

On the impact of the galaxy window function on cosmological parameter estimation

One important source of systematics in galaxy redshift surveys comes from the estimation of the galaxy window function. Up until now, the impact of the uncertainty in estimating the galaxy window function on parameter inference has not been properly studied. In this paper, we show that the uncertainty and the bias in estimating the galaxy window function will be salient for ongoing and next-generation galaxy surveys using a simulation-based approach. With a specific case study of cross-correlating Emission-line galaxies from the DESI Legacy Imaging Surveys and the Planck CMB lensing map, we show that neural network-based regression approaches to modelling the window function are superior in comparison to linear regression-based models. We additionally show that the definition of the galaxy overdensity estimator can impact the overall signal-to-noise of observed power spectra. Finally, we show that the additive biases coming from the window functions can significantly bias the modes of the inferred parameters and also degrade their precision. Thus, a careful understanding of the window functions will be essential to conduct cosmological experiments.Comment: 13 pages, 12 figures, complementary paper to an upcoming paper on Cross-Correlation of ELGs and Planck CMB lensing, accepted for publication in MNRA

The Angiogenic Paracrine Potential of Mesenchymal Stem Cells

Tissue engineering and regenerative medicine are branches of biomedical sciences that facilitate the use of cells and biocompatible scaffolds in favor of tissue restoration. In this regard, restoration and maintenance of angiogenesis and blood supplementation could be an effective strategy for injured tissue removal, accelerating healing rate, and successful transplantation of cells and scaffolds into target sites. It has been elucidated that mesenchymal stem cells have the potency to promote angiogenesis via paracrine activity and trans-differentiation into the endothelial lineage. In this chapter, we highlighted the paracrine property of mesenchymal stem cells to modulate angiogenesis in the target tissues

Hydrogen Peroxide Preconditioning Promotes Protective Effects of Umbilical Cord Vein Mesenchymal Stem Cells in Experimental Pulmonary Fibrosis

Purpose Idiopathic pulmonary fibrosis (IPF) is a progressive lung disorder with few available treatments. Mesenchymal stem cell therapy (MSCT), an innovative approach, has high therapeutic potential when used to treat IPF. According to recent data, preconditioning of MSCs can improve their therapeutic effects. Our research focuses on investigating the anti-inflammatory and antifibrotic effects of H2O2-preconditioned MSCs (p-MSCs) on mice with bleomycin-induced pulmonary fibrosis (PF). Methods Eight-week-old male C57BL/6 mice were induced with PF by intratracheal (IT) instillation of bleomycin (4 U/kg). Human umbilical cord vein-derived MSCs (hUCV-MSCs) were isolated and exposed to a sub-lethal concentration (15 pM for 24 h) of H2O2 in vitro. One week following the injection of bleomycin, MSCs or p-MSCs were injected (IT) into the experimental PF. The survival rate and weight of mice were recorded, and 14 days after MSCs injection, all mice were sacrificed. Lung tissue was removed from these mice to examine the myeloperoxidase (MPO) activity, histopathological changes (hematoxylin-eosin and Masson\u27s trichrome) and expression of transforming growth factor beta 1 (TGF-β1) and alpha-smooth muscle actin (α-SMA) through immunohistochemistry (IHC) staining. Results Compared to the PF+MSC group, p-MSCs transplantation results in significantly decreased connective tissue () and collagen deposition. Additionally, it is determined that lung tissue in the PF+pMSC group has increased alveolar space () and diminished expression of TGF-β1 and α-SMA. Conclusion The results demonstrate that MSCT using p-MSCs decreases inflammatory and fibrotic factors in bleomycin-induced PF, while also able to increase the therapeutic potency of MSCT in IPF

The protective effect of Liza klunzingeri protein hydrolysate on carbon tetrachloride-induced oxidative stress and toxicity in male rats

Objective(s): Today, consumers are looking for food products providing health benefits in addition to meeting the basic nutritional needs of the body. This study aimed to evaluate the antioxidant and cytotoxic effects of Liza klunzingeri protein hydrolysate both in vivo and in vitro. Materials and Methods: Fish protein hydrolysate (FPH) was prepared using enzymatic hydrolysis with papain. In vitro antioxidant activity was assessed using five different antioxidant assays. The cytotoxic effect on 4T1 cell line was evaluated using the MTT assay. The distribution of the molecular weight of FPH was measured using HPLC. In the in vivo study, CCl4-exposed Wistar rats were orally treated with FPH (150, 300, and 600 mg/kg) or gallic acid (50 mg/kg) for 28 consecutive days. Results: Enzymatic hydrolysis gave hydrolysate rich in low molecular weight peptides (<1000 Da) with strong free radicals (ABTS, DPPH, and OH) scavenging activity and cytotoxicity. Treatment of CCl4-exposed rats with all doses of FPH significantly lowered serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). FPH at doses of 300 and 600 mg/kg significantly decreased lipid peroxidation and improved total antioxidant capacity in serum, liver, and kidney of the CCl4 exposed rats. All doses of L.klunzingeri protein hydrolysate reduced CCl4-induced nitric oxide production of the kidney. Liver histopathological damage caused by CCl4 also ameliorated with all doses of FPH. Conclusion: L. klunzingeri protein hydrolysate can be considered as a functional food to alleviate oxidative stress. Keywords Author Keywords:Antioxidant activity; Cytotoxicity; Liza klunzingeri; Oxidative stress; Protein hydrolysat

Angular clustering properties of the DESI QSO target selection using DR9 Legacy Imaging Surveys

The quasar target selection for the upcoming survey of the Dark Energy Spectroscopic Instrument (DESI) will be fixed for the next 5 yr. The aim of this work is to validate the quasar selection by studying the impact of imaging systematics as well as stellar and galactic contaminants, and to develop a procedure to mitigate them. Density fluctuations of quasar targets are found to be related to photometric properties such as seeing and depth of the Data Release 9 of the DESI Legacy Imaging Surveys. To model this complex relation, we explore machine learning algorithms (random forest and multilayer perceptron) as an alternative to the standard linear regression. Splitting the footprint of the Legacy Imaging Surveys into three regions according to photometric properties, we perform an independent analysis in each region, validating our method using extended Baryon Oscillation Spectroscopic Survey (eBOSS) EZ-mocks. The mitigation procedure is tested by comparing the angular correlation of the corrected target selection on each photometric region to the angular correlation function obtained using quasars from the Sloan Digital Sky Survey (SDSS) Data Release 16. With our procedure, we recover a similar level of correlation between DESI quasar targets and SDSS quasars in two-thirds of the total footprint and we show that the excess of correlation in the remaining area is due to a stellar contamination that should be removed with DESI spectroscopic data. We derive the Limber parameters in our three imaging regions and compare them to previous measurements from SDSS and the 2dF QSO Redshift Survey.This research is supported by the Director, Office of Science, Office of High Energy Physics of the U.S. Department of Energy under contract no. DE-AC02-05CH11231, and by the National Energy Research Scientific Computing Center, a DOE Office of Science User Facility under the same contract; additional support for DESI is provided by the U.S. National Science Foundation, Division of Astronomical Sciences under contract no. AST-0950945 to the NSF’s National Optical–Infrared Astronomy Research Laboratory; the Science and Technology Facilities Council of the United Kingdom; the Gordon and Betty Moore Foundation; the Heising-Simons Foundation; the French Alternative Energies and Atomic Energy Commission (CEA); the National Council of Science and Technology, Mexico; the Ministry of Economy of Spain, and by the DESI Member Institutions. ADM was supported by the U.S. Department of Energy, Office of Science, Office of High Energy Physics, under Award Number DE-SC0019022

Topical Licorice for Aphthous: A Systematic Review of Clinical Trials

Background: Recurrent aphthous stomatitis (RAS) is the most common ulcerative disease that affects oral mucosa. The coating agents, topical analgesics, and topical steroids are usually used as treatment methods. Glycyrrhiza glabra has been used for RAS treatment based on its anti-inflammatory, antioxidant, and immunomodulatory properties. In this study, a systemic review on the therapeutic effect of topical licorice on RAS management was performed. Methods: Science Direct, Scopus, Cochrane databases, PubMed Google Scholar, and ResearchGate were searched up to September 2021 to find all English randomized clinical trials studying the effect of G. glabra, or its compositions on RAS. Meta-analysis was not conducted because of data heterogeneity. Articles were reviewed qualitatively, and only those with a Jadad score ≥3 were included. Animal studies, in vitro, review papers, non-English papers, and case reports were excluded.Results: Six studies with 314 subjects were included after screening. The result showed licorice has significant effects on RAS pain reduction, ulcer size, and healing time. Its effectiveness is related to its dose-dependent anti-inflammatory and antioxidant effects through several mechanisms. It also has antibacterial effects against Streptococci mutans and Porphyromonas gingivalis as another mechanism of action in RAS treatment. In addition, licorice can elevate the epidermal growth factor (EGF) level compared to the control group, which has an essential role in oral mucosal tissue integrity. Conclusion: Licorice extract has been used in different dosage forms, including paste, patch, and mouthwash with concentrations of 1% or 5%. The healing time after licorice therapy is expected to be within 4-8 days. Licorice did not show any adverse effect in the intervention groups, indicating its effectiveness and safety in RAS treatment