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

Flow Cytometric Analysis of Inflammatory Cells in Experimental Acute Pancreatitis

 Background: Accumulating evidence indicates that inflammatory cells migrate into the pancreas tissue and play an important role in the pathogenesis of acute pancreatitis (AP). The aim of this study was to establish a flow cytometric method to enumerate these infiltrating cells in the pancreas of an experimental AP.Materials and Methods: Twelve hours after inducing of AP, mice pancreatic tissues were cut into small fragments and single cells were prepared by mechanical dissociation. The isolated cells were stained with either anti-mouse CD45-PerCP or isotype antibody and analyzed by flow cytometry. Using side scatter (SSC)/CD45 gating we were able to identify inflammatory cells from non-inflammatory cells.Results: The mean percentage of leukocytes was 5.9±1.6 in the control group whereas, it was 26.7±8.1 in the AP. Moreover, we found that the percentage of lymphocytes, monocytes and granulocytes were 1.1±0.2, 0.9±.04 and 2.9±1.8 of total pancreatic cells, respectively, in the control mice. In contrast to lymphocytes, the percentage of monocytes and granulocytes were significantly increased in the AP group and it was 3±1.3 and 18.2±3.2 for monocytes and granulocytes, respectively.Conclusion: Quantitative flow cytometric analysis is feasible and provides a reliable and rapid assay to determine the number and percentage of inflammatory cells in experimental AP

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

Biosynthesis and recovery of rod-shaped tellurium nanoparticles and their bactericidal activities

In this study, a tellurium-transforming Bacillus sp. BZ was isolated from the Caspian Sea in northern Iran. The isolate was identified by various tests and 16S rDNA analysis, and then used to prepare elemental tellurium nanoparticles. The isolate was subsequently used for the intracellular biosynthesis of elemental tellurium nanoparticles. The biogenic nanoparticles were released by liquid nitrogen and purified by an n-octyl alcohol water extraction system. The shape, size, and composition of the extracted nanoparticles were characterized. The transmission electron micrograph showed rod-shaped nanoparticles with dimensions of about 20 nm � 180 nm. The energy dispersive X-ray and X-ray diffraction spectra respectively demonstrated that the extracted nanoparticles consisted of only tellurium and have a hexagonal crystal structure. This is the first study to demonstrate a biological method for synthesizing rod-shaped elemental tellurium by a Bacillus sp., its extraction and its antibacterial activity against different clinical isolates

Effects of Coenzyme Q10 Level on Clinical Parameters in Cystic Fibrosis Patients

Background: Cystic Fibrosis (CF) is a hereditary autosomal recessive genetic disorder that can affect many organs including the lungs and the digestive system. We aimed to assess the effects of coenzyme Q10 level on clinical parameters in cystic fibrosis patients. Materials and Methods This cross-sectional study was performed on 40 patients (13 females and 19 males) with cystic fibrosis who were admitted in Masih Daneshvari Hospital in Tehran- Iran, in 2017. A researcher-made questionnaire was distributed among them, and then the height, weight and arm circumference of the patients were measured and also their serum Q10 levels were evaluated. Then, the pulmonary function was evaluated using a 6-minute respiration test and a spirometry test. Finally, their total scores were calculated based on Shwachman score. Results: The age range of participants was from 6 to 27 years old. Anthropometric indices such as weight (p=0.02, r=-0.408), age (p=0.016, r=-0.422), height (p=0.002, r=-0.520), birth weight (p=0.0113, R=-0.286), and height at birth (p=0.037, r=-0.37) had a significant negative relationship with coenzyme Q10. There was relationship between anthropometric indices and spirometry tests, including the association of FEV1 (act) with weight (p=0.00), and arm circumference (p=0.00) which were determined. Conclusion: According to the results, there was no direct relationship between muscle mass and Q10. Besides, the mean age of Q10 was greater than that of in age group of 9-16 years old, which can be considered for nutrition or childhood reserves for coenzyme Q10 production, which is decreased due to increasing age and lack of intake or loss of body power for the production of coenzyme Q10

Assessing diverse evidence to improve conservation decision-making

Meeting the urgent need to protect and restore ecosystems requires effective decision‐making through wisely considering a range of evidence. However, weighing and assessing evidence to make complex decisions is challenging, particularly when evidence is of diverse types, subjects, and sources, and varies greatly in its quality and relevance. To tackle these challenges, we present the Balance Evidence Assessment Method (BEAM), an intuitive way to weigh and assess the evidence relating to the core assumptions underpinning the planning and implementation of conservation projects, strategies, and actions. Our method directly tackles the question of how to bring together diverse evidence whilst assessing its relevance, reliability, and strength of support for a given assumption, which can be mapped, for example to a Theory of Change. We consider how simple principles and safeguards in applying this method could help to respectfully, and equitably, include more local forms of knowledge when assessing assumptions, such as by ensuring diverse groups of individuals contribute and assess evidence. The method can be flexibly applied within existing decision‐making tools, platforms, and frameworks whenever assumptions (i.e., claims and hypotheses) are made. This method could greatly facilitate and improve the weighing of diverse evidence to make decisions in a range of situations, from local projects to global policy platforms

Incidence and risk factors for infantile colic in Iranian infants

AIM: To assess the incidence of infantile colic and its association with variable predictors in infants born in a community maternity hospital, Tehran, Iran. METHODS: In this prospective cohort study, mothers who gave birth to live newborns between February 21 and March 20, 2003 at the hospital were invited to join to the study. For every infant-mother dyad data were collected on infant gender, type of delivery, gestational age at birth, birth weight, birth order, and mother’s reproductive history. Then mothers were given a diary to document the duration of crying/fussiness behaviors of their infants for the next 12 wk. We scheduled home visits at the time the infants were 3 mo of age to collect the completed diaries and obtain additional information on infants’ nutritional sources and identify if medications were used for colic relief. Cases of colic were identified by applying Wessel criteria to recorded data. Chi-square and Mann-whitney U tests were used to compare proportions for non-parametric and parametric variables, respectively. RESULTS: From 413 infants, follow-up was completed for 321 infants. In total, 65 infants (20.24%) satisfied the Wessel criteria for infantile colic. No statistical significance was found between colicky and non-colicky infants according to gender, gestational age at birth, birth weight, type of delivery, and, infant’s feeding pattern. However, firstborn infants had higher rate for developing colic (P = 0.03). CONCLUSION: Colic incidence was 20% in this population of Iranian infants. Except for birth order status, no other variable was significantly associated with infantile colic