Prevalence of non-alcoholic fatty liver disease in Kurdistan province, Iran, 2013-2014

Background: Generally, 15% to 40% of the world populations are suffering from non-alcoholicfatty liver disease [NAFLD]. The aim of this study was to conduct an epidemiological evaluation of NAFLD and its related factors in the west of Iran

Materials and Methods: This cross-sectional study was conducted for 12 months, from July 2013 to July 2014, in Sanandaj city, Kurdistan province. In this study, multistage cluster sampling method was used. The general characteristics of the subjects including their age, sex, body mass index, history of diabetes, hypertension, and heart disease were recorded. All the subjects underwent abdominal ultrasonography. Besides, patients with fatty liver underwent blood tests [lipid profile,liver function test, fasting blood sugar,hepatitis B virus antigen, and hepatitis C virus antibody]. Statistical analysis was performed using descriptive statistics and logistic regression test

Results: A total of 410 patients were included in the study; of whom 145 [35%] had NAFLD. The prevalence of fatty liver in men [43%] was twice more than that in women [22%]. The severity of fatty liver disease increased with increasing blood sugar [OR = 3.214, 95% CI: 1.357, 7.612], triglycerides, and total cholesterol [OR = 2.897, 95% CI: 1.245, 6.736]

Conclusion: Findings of this study show that, the prevalence of NAFLD in the west of Iranis similar to the highest rates reported from other countries and the prevalence was much higher than other Asian countries. It is recommended to implement fast and effective interventions to control fatty liver disease

Soluble CD93 as a Novel Biomarker in Asthma Exacerbation

Asthma research is shifting from studying symptoms and lung functions to the narrow-focus cellular profiles protein analysis, biomarkers, and genetic markers. The transmembrane glycoprotein CD93 is involved in endothelial cell migration, angiogenesis, leukocytes extravasation, apoptosis, innate immunity and inflammation. Relationships between the serum level of soluble CD93 (sCD93) and acute myocardial infarction/premature MI/inflammatory arthritis/skin sclerosis have recently been reported. We hypothesized that sCD93 would be elevated during the acute phase of asthma. We measured the serum level of sCD93 in 57 patients with asthma exacerbation and 57 age-and gender-matched healthy controls. Additionally, sCD93 was reassessed at the time of discharge from the hospital. Clinical characteristics and peak expiratory flow (PEF) of the patients were assessed. The primary outcome was the comparison of serum level of sCD93 between asthmatics and healthy subjects. The sCD93 values ranged from 128 to 789 ng/mL in asthmatics (345.83±115.81) and from 31 to 289 ng/mL in control subjects (169.46±62.43). The difference between the 2 groups was statistically significant (P<0.001). The association between sCD93 and asthma remained significant after adjusting for age, sex, and BMI. The differences between asthmatics and controls remained significant on the last day of hospital stay. The association between sCD93 and PEF was not significant. In conclusion, the serum level of soluble CD93 is increased in patients with asthma exacerbation. It also showed that serum levels of sCD93 decreased with treatment of asthma attack. The clinical usefulness of determination of sCD93 as a biomarker of asthma requires further studies.

Autophagy-modulated human bone marrow-derived mesenchymal stem cells accelerate liver restoration in mouse models of acute liver failure

Background: Mesenchymal stem cells [MSCs] have been recently received increasing attention for cell-based therapy, especially in regenerative medicine. However, the low survival rate of these cells restricts their therapeutic applications. It is hypothesized that autophagy might play an important role in cellular homeostasis and survival. This study aims to investigate the regenerative potentials of autophagy-modulated MSCs for the treatment of acute liver failure [ALF] in mice

Methods: ALF was induced in mice by intraperitoneal injection of 1.5 ml/kg carbon tetrachloride. Mice were intravenously infused with MSCs, which were suppressed in their autophagy pathway. Blood and liver samples were collected at different intervals [24, 48 and 72 h] after the transplantation of MSCs. Both the liver enzymes and tissue necrosis levels were evaluated using biochemical and histopathological assessments. The survival rate of the transplanted mice was also recorded during one week

Results: Biochemical and pathological results indicated that 1.5 ml/kg carbon tetrachloride induces ALF in mice. A significant reduction of liver enzymes and necrosis score were observed in autophagy-modulated MSC-transplanted mice compared to sham [with no cell therapy] after 24 h. After 72 h, liver enzymes reached their normal levels in mice transplanted with autophagy-suppressed MSCs. Interestingly, normal histology without necrosis was also observed

Conclusion: Autophagy suppression in MSCs ameliorates their liver regeneration potentials due to paracrine effects and might be suggested as a new strategy for the improvement of cell therapy in ALF

Advances in hematopoietic stem cell mobilization and peripheral blood stem cell transplantation

Hematopoietic stem/progenitor cells [HSPCs] which give rise to different blood cell types are present within the bone marrow microenvironment, especially in flat bones such as skull, vertebrae, pelvis and chest. Interacting factors such as stromal derived factor-1/CXCR4, very late antigen-4/vascular cell adhesion molecule-1, Lymphocyte function-associated antigen-1/ intercellular adhesion molecule-1 retain the cells in the microenvironment. Any factor affecting these links may lead to migration and mobilization of HSPCs into peripheral blood. Several factors are involved in hematopoietic stem cells [HSC] mobilization such as granulocyte-colony stimulating factor, sphingosine-1-phosphate, hepatocyte growth factor, complement system, plasminogen system and matrix metalloproteinases. In bone marrow transplantation, HSC is transferred to the recipient from bone marrow of the donor, which can be performed in two ways. In the first method, Jamshidi needle is used for aspiration of bone marrow to extract hematopoietic cells usually from the hip. The second method uses mobilizer factors such as granulocyte-colony stimulating factor and granulocyte-macrophage colony-stimulating factor to mobilize the HSC into peripheral blood. Mobilized hematopoietic stem cells are suitable for the bone marrow transplantation in leukemias such as chronic myeloid leukemia, acute myeloid leukemia, chronic lymphocyte leukemia, Hairy cell leukemia, etc

Down-regulation of the autophagy gene, ATG7, protects bone marrow-derived mesenchymal stem cells from stressful conditions

BACKGROUND: Mesenchymal stem cells (MSCs) are valuable for cell-based therapy. However, their application is limited owing to their low survival rate when exposed to stressful conditions. Autophagy, the process by which cells recycle the cytoplasm and dispose of defective organelles, is activated by stress stimuli to adapt, tolerate adverse conditions, or trigger the apoptotic machinery. This study aimed to determine whether regulation of autophagy would affect the survival of MSCs under stress conditions. METHODS: Autophagy was induced in bone marrow-derived MSCs (BM-MSCs) by rapamycin, and was inhibited via shRNA-mediated knockdown of the autophagy specific gene, ATG7. ATG7 expression in BM-MSCs was evaluated by reverse transcription polymerase chain reaction (RT-PCR), western blot, and quantitative PCR (qPCR). Cells were then exposed to harsh microenvironments, and a water-soluble tetrazolium salt (WST)-1 assay was performed to determine the cytotoxic effects of the stressful conditions on cells. RESULTS: Of 4 specific ATG7-inhibitor clones analyzed, only shRNA clone 3 decreased ATG7 expression. Under normal conditions, the induction of autophagy slightly increased the viability of MSCs while autophagy inhibition decreased their viability. However, under stressful conditions such as hypoxia, serum deprivation, and oxidative stress, the induction of autophagy resulted in cell death, while its inhibition potentiated MSCs to withstand the stress conditions. The viability of autophagy-suppressed MSCs was significantly higher than that of relevant controls (P<0.05, P<0.01 and P<0.001). CONCLUSION: Autophagy modulation in MSCs can be proposed as a new strategy to improve their survival rate in stressful microenvironments.

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

Role of BCR-ABL P190 in Diagnosis and Prognosis of ALL patients

Acute lymphoblastic leukemia[ALL] is due to early stage arrest of lymphoblast development. The translocation of Philadelphia [Ph] chromosome occurs as a result of the BCR-ABL fusion gene, which constitutively produced activated tyrosine kinase. This gene fusion is an important indicator for prognosis in ALL and is associated with poor overall survival and remission duration. BCR-ABL could interfere in establishment of ALL. Therefore, in this study, we will try to investigate most pathological aspects involved in BCR-ABL fusion. Strategies for genetic alterations in B-ALL pathogenesis are discussed. Then, the main cytogenetic changes and genetic subtypes for ALL are highlighted. Moreover, intermediate reactions between cancer stem cells [CSC] related to ALL, its niche and microenvironment is discussed. The main objective in this review is to understand the principle prognosis in ALL to introduce new approaches and treatment alternatives

immunosuppressive activity of amniotic membrane mesenchymal stem cells on T lymphocytes

Mesenchymal Stem Cells [MSCs] are isolated from different sources like placenta. The placenta and its membranes like Amniotic Membrane [AM] are readily available and easy to work with. There is only limited knowledge on the immunomodulatory properties of human Amniotic Membrane-derived Mesenchymal Stem Cells [hAM-MSCs]. The aim of this study was to survey the suppressive activity of hAM-MSCs on T lymphocytes in vitro. Human AMs were obtained after caesarean section births from healthy women. After enzymatic digestion, cells were cultured and hAM-MSCs were obtained. In addition, human T lymphocytes were isolated and co-cultured with hAM-MSCs for 72 hr in the presence or absence of phytohemagglutinin [PHA]. Subsequently, proliferation of T cells was analyzed using BrdU and subsequently flow cytometry technique. Besides, the production of IL-4 and IFN-gamma was examined by ELISA method. Additionally, the expression of activation markers [CD38, HLA-DR] was studied on T lymphocytes by flow cytometry technique. It was revealed that hAM-MSCs could significantly suppress the proliferation of T lymphocytes [p

Anti-tumor activity of ferulago angulata boiss. extract in gastric cancer cell line via induction of apoptosis

Ferulago angulata Boiss. known in Iran as Chavir, has some bioactive compounds having antioxidant activity. Because of its antioxidant activities, it sounded Chavir extract can be a good candidate for finding chemopreventive agents having inductive apoptosis properties on cancer cells. In this study, the cytotoxic effects and proapoptotic activities of Chavir's leaf and flower extracts were investigated on human adenocarcinoma gastric cell line [AGS]. The ferric reducing antioxidant power [FRAP] assay was used to determine antioxidant activity of the extract. Cytotoxic effects of the extract were performed by trypan blue and neutral red assays. For apoptosis detection, we used Annexin V staining, flow cytometry and DNA fragmentation assays. The FRAP assay results showed that antioxidant activity of leaf extract was higher than flower extract. Cytotoxicity and apoptosis-inducing activity of flower and leaf extracts changed coordinately, indicating the cytotoxicity of chavir extracts is due probably to induce apoptosis. Our results revealed that the cytotoxic effects of F. angulate Boiss. extracts on AGS cell line is close to some other plant extracts such as Rhus verniciflua Stokes [RVS] and Scutellaria litwinowii. This is the first study on cytotoxic and apoptosis-inducing effects of chavir leaf and flower extracts against AGS cell line. The Further investigation can be identification of the agent[s] by which these effects is observed

Production of cloned mice by nuclear transfer of cumulus cells

Over the past several years, mammals have been successfully cloned by either the splitting of an early stage embryo or nuclear transfer of adult somatic cells [NT] into oocytes. Although it has been 15 years since the generation of the first cloned mammals from somatic cells by NT, the success rate for producing live offspring by this technique is low regardless of the cell type and animal species used. However, these techniques have the potential to be important tools for future research in basic biology. In the present study, we described our experiences in producing successfully cloned mouse using NT method and piezo-actuated micromanipulator. B6D2F1 mice, 8-12 weeks old, were superovulated with injections of 5 IU of pregnant mare serum gonadotropin and 5 IU of human chorionic gonadotropin administered 48 hr apart. Enucleation and donor nuclei cumulus cell injection were performed with a piezo-actuated micromanipulator after which activation and trichostatin A treatment were used for reconstructed oocytes. Two-cell stage cloned embryos that developed in the mWM medium were transferred into the oviducts of pseudopregnant NMRI mice. Of 367 oocytes collected, 131 [69%] developed into 2-cell stage embryos. Of these, 5 [1%] live pups were successfully delivered. We used NMRI foster mother to raise the pups by lactation. One adult cloned mouse was mated, after which she delivered and raised normal offspring. For mouse cloning, the present study also successfully tested the capability of somatic cell nuclear transfer SCNT using a piezo unit

Vitrectomy for posterior segment intraocular foreign bodies, visual and anatomical outcomes

To evaluate the visual and anatomic results and determine the prognostic factors after pars plana vitrectomy and posterior segment intraocular foreign body [IOFB] removal. This retrospective study reviews the patients' charts of 48 consecutive patients with posterior segment IOFB who underwent pars plana vitrectomy and IOFB removal over a 4-year period, recently. Association between visual outcome and various preoperative, operative, and postoperative variables was statistically analyzed. Data were analyzed with the paired t-test and the chi square test. Statistical significance was indicated by P < 0.05. The mean interval between the time of injury and IOFB removal was 24 +/- 43.1 days and 27 [53%] eyes underwent IOFB removal within 7 days of the injury. Nine [19.1%] patients achieved a visual acuity of 20/40 or better. An improvement of visual acuity of at least three lines occurred in 21 [44.6%] eyes and the vision remained unchanged in 15 [31.9%] eyes. Postoperative retinal detachment occurred in five [10.6%] eyes. Visual improvement was more likely to occur in eyes with lower levels of presenting visual acuity [P = 0.2]. Visual improvement was not associated with an entry site and IOFB location, lens injury, time to surgery, and pre- and post-operative retinal detachment. At the end of follow up, anatomical success was achieved in 97.9% of eyes. High anatomical success could be achieved after the removal of posterior segment IOFBs by vitrectomy, despite a delay in surgery. Poor visual outcome may be mainly due to the initial ocular injury

Natural oils enhance IL-10 and IFN-alpha production by human PBMCs cultured with malassezia furfur

Malassezia furfur is alipophilic yeast that causes skin disease. To evaluate the level of IL-10, IFN-alpha and IL-12P70 in co-incubation of M. furfur grown on different forms of natural oils with PBMCs of healthy individuals. PBMCs were obtained from blood samples of normal volunteers. M. furfur was cultured in different culture media containing almond oil, fish oil, walnut oil, full-fat milk, and a fat-free medium; and the yeasts grown were harvested and used for co-incubation with PBMCs in vitro. The IFN-alpha, IL-10, and IL-12P70 levels were measured at different time intervals using ELISA methods. Generally, IFN-alpha and IL-10 levels in the coincubation of yeasts with walnut oil group [WOG] and fish oil group [FOG] were higher than those in the almond oil group [AOG] and full-fat milk group [FFMG]. Although the IL-12P70 was higher in groups such as AOG, FOG, and WOG; the increase was not statistically significant. The results demonstrated that the type of fat used by M. furfur in the culture media can influence the immune response and increases IFN-alpha and IL-10 levels in an early time point of the culture system

New synthesis of aminorhodanin and condensed derivatives

Aminorhodanins are heterocyclic compounds of dithiocarbazoyl which include sulphur and nitrogen in their structures. Different research studies have been performed on rhodanin and it-s condensed derivatives and several reports have been issued indicating their antimicrobial properties. In the present work, first a new method has been developed for production of aminorhodanin, which include one more nitrogen atom in third position compare to rhodanins molecule, then condensed reactions with aldehydes have been investigated. The probable biological properties of produced compounds will be investigated in another research study

Pasteurization of IgM-enriched immunoglobulin

Human plasma proteins are important for therapy or prophylaxis of human diseases. Due to the preparation of human plasma proteins from human plasma pools and risk of contamination with human viruses, different viral reduction treatments such as: pasteurization, solvent/detergent, dry heat treatment, steam treatment, beta-propiolactone/UV and nanofiltration have been implemented. As pasteurization can be performed for liquid protein, this method [a 10-hour heat treatment of the aqueous solutions at 60°C] was introduced into the manufacturing procedure of IgM-enriched immunoglobulin, to improve its safety further. The efficiency of this method for inactivation of viruses was evaluated by the use of Foot-and-Mouth Disease Virus [a non-enveloped virus] and Infectious Bovine Rhinotracheitis [IBR] Virus [a lipid-enveloped virus]. Pasteurization inactivated Foot-and-Mouth Disease Virus by 7 log10 and for IBR Virus by 5log10. These findings show a significant added measure of virus safety associated with pasteurization of IgM-enriched immunoglobulin preparation

Preparation of enriched immunoglobulin M and immunoglobin A from human plasma

As IgM and IgA-enriched preparations are needed to complete the immunotherapeutic spectrum, a simple procedure is described for the preparation of IgM and IgA-enriched immunoglobulins. Fraction III which was prepared by cold ethanol fractionation was treated by octanoic acid followed by ethanol precipitation and ion-exchange chromatography using Sephadex DEAE A-50 and 0.1 M tris-0.35M NaC1 buffer, pH 8.1, resulting in recovery of 85% IgM, 84% IgA and 33% IgG. The comparison of our results with immunoglobulins' percentage in plasma indicates that IgM and IgA-enrichment was obtained by three times

Solvent-detergent treatment of IgM-enriched immunoglobulin

Viral safety of human plasma products plays a key role in their safe uses. Solvent- detergent [SD] virus-inactivation method has gained widespread popularity in the manufacture of biological products. This treatment which inactivates lipid-enveloped viruses effectively consists of incubation of a plasma protein solution in the presence of a non-volatile organic solvent and a detergent. In this study, IgM-enriched immunoglobulin was incubated at 24 °C for 6 h under slow stirring in the presence of tri[n-butyl] phosphate [0.3% w/w] as solvent and tween 80 [1% w/w] as detergent. After completion of the inactivation process and removal of the solvent-detergent, the ability of SD-treatment to remove Infectious Bovine Rhinotracheitis [IBR] virus [a lipid-enveloped virus] and Foot-and-Mouth Disease virus [a non-enveloped virus] were evaluated by "virus spiking studies" using a scaled down process. Reduction factor of 4 log was obtained for the SD-treatment of IgM-enriched immunoglobulin spiked with IBR virus. No virus inactivation was observed in the SD-treated IgM-enriched immunoglobulin, spiked with Foot-and-Mouth Disease virus. It was concluded that treatment of IgM-enriched immunoglobulin with TNBP-TWEEN 80 may be considered as an efficient lipid-enveloped virus inactivation step in the manufacture of this product