Effect of pancreatic extract on insulin secreting cell differentiation from mouse bone marrow mesenchymal stem cells

زمینه و هدف: با دیابت نوع یک در نتیجه ی تخریب خود ایمنی سلول های بتای جزایر پانکراس ایجاد می شود. مطالعات اخیر نشان می‌دهد بسیاری از انواع سلول های بنیادی می توانند به عنوان منابع احتمالی برای به دست آوردن سلول های قابل پیوند تولید کننده انسولین (IPCs) در نظر گرفته شوند. در این مطالعه تمایز سلول های بنیادی مزانشیمی مغز استخوان به سلول های مولد انسولین با استفاده از عصاره پانکراس موش مورد بررسی قرار گرفت. روش بررسی: در این مطالعه تجربی آزمایشگاهی از سلول های بنیادی مزانشیمی مغز استخوان موش کوچک آزمایشگاهی برای تمایز به سلول های انسولین ساز استفاده شد. سلول های تمایز یافته با استفاده از رنگ اختصاصی دیتیزون و آنتی بادی های ضد انسولین- پروانسولین و ضد رسپتور بتای انسولین مورد ارزیابی قرار گرفتند. همچنین بیان ژن اختصاصی سلول های پانکراسی یعنی pdx-I در این سلول ها با روش RT-PCR بررسی شد. یافته ها: سلول های تمایز یافته مشتق از روش مستقیم مورفولوژی مشابه با سلول های بتای پانکراس نشان دادند. سلول های دیتیزون مثبت به صورت دستجات قرمز ارغوانی دیده شدند. نتایج بررسی RT-PCR بیان ژن اختصاصی سلول های بتا (pdx-I) را در سلول های تمایز مستقیم نشان داد. ایمنوفلورسنس وجود نشانگرهای اختصاصی سلول های بتا را دراین سلول ها به اثبات رساند. نتیجه گیری: نتایج این مطالعه نشان داد که سلول های بنیادی مزانشیمی مغز استخوان می توانند در حضور عصاره پانکراس به سلول های مولد انسولین تمایز یابند؛ لذا استفاده از این نتایج تولید سلول های بتا از سلول های بنیادی در شرایط آزمایشگاهی را تسهیل می کند

Efficient and simple production of insulin-producing cells from embryonal carcinoma stem cells using mouse neonate pancreas extract, as a natural inducer

An attractive approach to replace the destroyed insulin-producing cells (IPCs) is the generation of functional β cells from stem cells. Embryonal carcinoma (EC) stem cells are pluripotent cells which can differentiate into all cell types. The present study was carried out to establish a simple nonselective inductive culture system for generation of IPCs from P19 EC cells by 1–2 weeks old mouse pancreas extract (MPE). Since, mouse pancreatic islets undergo further remodeling and maturation for 2–3 weeks after birth, we hypothesized that the mouse neonatal MPE contains essential factors to induce in vitro differentiation of pancreatic lineages. Pluripotency of P19 cells were first confirmed by expression analysis of stem cell markers, Oct3/4, Sox-2 and Nanog. In order to induce differentiation, the cells were cultured in a medium supplemented by different concentrations of MPE (50, 100, 200 and 300 µg/ml). The results showed that P19 cells could differentiate into IPCs and form dithizone-positive cell clusters. The generated P19-derived IPCs were immunoreactive to proinsulin, insulin and insulin receptor beta. The expression of pancreatic β cell genes including, PDX-1, INS1 and INS2 were also confirmed. The peak response at the 100 µg/ml MPE used for investigation of EP300 and CREB1 gene expression. When stimulated with glucose, these cells synthesized and secreted insulin. Network analysis of the key transcription factors (PDX-1, EP300, CREB1) during the generation of IPCs resulted in introduction of novel regulatory candidates such as MIR17, and VEZF1 transcription factors, as well as MORN1, DKFZp761P0212, and WAC proteins. Altogether, we demonstrated the possibility of generating IPCs from undifferentiated EC cells, with the characteristics of pancreatic β cells. The derivation of pancreatic cells from EC cells which are ES cell siblings would provide a valuable experimental tool in study of pancreatic development and function as well as rapid production of IPCs for transplantation.Marzieh Ebrahimie, Fariba Esmaeili, Somayeh Cheraghi, Fariba Houshmand, Leila Shabani, Esmaeil Ebrahimi

Isolation and identification of bacterial and fungal microflora from Litopenaeus vannamei in Choibdeh, Abadan

Bacterial and fungal microflora of Litopenaeus vannamei cultured in Choibdeh, Abadan was studied. For this purpose, PLs before and after stocking and those shrimps persisting on food tray from June to August, 2006 were taken randomly. Live samples transferred to microbiology laboratory of South Aquaculture Research Center, Ahwaz. Special culture media (e.g. Tryptic Soy Agar + 1.5-2% Nacl & Sabouraud Dextrose Agar + 1.5-2% Nacl) were used for bacterial and fungal culture. We isolated 10 bacterial species of which Vibrio alginolyticus (36.92%) had high abundance among bacterial species. We also isolated and identified three fungal species including Aspergillus niger (66.66%) A. fumigatus (16.66%) and Fusarium sp. (16.66%). A. niger was predominant among fungal species. All bacterial and fungal species that were identified were opportunistic

Evaluation of the immunity factors (THC, TPP, PO, SOD, POD) of shrimp fed with the algae Gracilaria corticata compared to shrimp fed without algae and exposed to white spot virus

White spot disease (WSD) is one of the shrimp deadliest viral diseases that causes heavy losses on all shrimp of Penaeid family. Most invertebrates are lacking adaptive immune system and its defense is the innate immune system that is as cellular and humoral, but a like immune system against white spot virus in shrimp was been detected. In this research, control and prevention of white spot disease in shrimp using algae Gracilaria corticata, investigated. About 300 vannamei shrimp were divided to 4 groups and fed by normal pellet and algae extract in 14 days. At the end of the fourteenth day half of the shrimp were challenged with acute white spot virus. After the fourteenth day in the days 0, 3, 9, 18 and 25 sampling were done from the hemolymph of survived shrimps and survival and immune factors were evaluated. Based on results, in the challenge test, shrimps that fed with algae extract has a significant survival rate than shrimp fed with commercial diet. Increased the Immune Factors from day one to day 25 observed during the test. Greatest amount of Immune Factors THC, TPP, SOD, POD and PO in T1 group were observed in day 25 of tests. This situation was also true for group2 (T2), but its rate significantly was less than group 1(T1)

The synergism between DHODH inhibitors and dipyridamole leads to metabolic lethality in acute myeloid leukemia

Dihydroorotate Dehydrogenase (DHODH) is a key enzyme of the de novo pyrimidine biosynthesis, whose inhibition can induce differentiation and apoptosis in acute myeloid leukemia (AML). DHODH inhibitors had shown promising in vitro and in vivo activity on solid tumors, but their effectiveness was not confirmed in clinical trials, probably because cancer cells exploited the pyrimidine salvage pathway to survive. Here, we investigated the antileukemic activity of MEDS433, the DHODH inhibitor developed by our group, against AML. Learning from previous failures, we mimicked human conditions (performing experiments in the presence of physiological uridine plasma levels) and looked for synergic combinations to boost apoptosis, including classical antileukemic drugs and dipyridamole, a blocker of the pyrimidine salvage pathway. MEDS433 induced apoptosis in multiple AML cell lines, not only as a consequence of differentiation, but also directly. Its combination with antileukemic agents further increased the apoptotic rate, but when experiments were performed in the presence of physiological uridine concentrations, results were less impressive. Conversely, the combination of MEDS433 with dipyridamole induced metabolic lethality and differentiation in all AML cell lines; this extraordinary synergism was confirmed on AML primary cells with different genetic backgrounds and was unaffected by physiological uridine concentrations, predicting in human activity

18F-NaF and 18F-FDG as molecular probes in the evaluation of atherosclerosis

The early detection of atherosclerotic disease is vital to the effective prevention and management of life-threatening cardiovascular events such as myocardial infarctions and cerebrovascular accidents. Given the potential for positron emission tomography (PET) to visualize atherosclerosis earlier in the disease process than anatomic imaging modalities such as computed tomography (CT), this application of PET imaging has been the focus of intense scientific inquiry. Although 18F-FDG has historically been the most widely studied PET radiotracer in this domain, there is a growing body of evidence that 18F-NaF holds significant diagnostic and prognostic value as well. In this article, we review the existing literature on the application of 18F-FDG and 18F-NaF as PET probes in atherosclerosis and present the findings of original animal and human studies that have examined how well 18F-NaF uptake correlates with vascular calcification and cardiovascular risk

An iPS-derived in vitro model of human atrial conduction

Atrial fibrillation (AF) is the most common arrhythmia in the United States, affecting approximately 1 in 10 adults, and its prevalence is expected to rise as the population ages. Treatment options for AF are limited; moreover, the development of new treatments is hindered by limited (1) knowledge regarding human atrial electrophysiological endpoints (e.g., conduction velocity [CV]) and (2) accurate experimental models. Here, we measured the CV and refractory period, and subsequently calculated the conduction wavelength, in vivo (four subjects with AF and four controls), and ex vivo (atrial slices from human hearts). Then, we created an in vitro model of human atrial conduction using induced pluripotent stem (iPS) cells. This model consisted of iPS-derived human atrial cardiomyocytes plated onto a micropatterned linear 1D spiral design of Matrigel. The CV (34-41 cm/s) of the in vitro model was nearly five times faster than 2D controls (7-9 cm/s) and similar to in vivo (40-64 cm/s) and ex vivo (28-51 cm/s) measurements. Our iPS-derived in vitro model recapitulates key features of in vivo atrial conduction and may be a useful methodology to enhance our understanding of AF and model patient-specific disease

Analyzing three-player quantum games in an EPR type setup

We use the formalism of Clifford Geometric Algebra (GA) to develop an analysis of quantum versions of three-player non-cooperative games. The quantum games we explore are played in an Einstein-Podolsky-Rosen (EPR) type setting. In this setting, the players' strategy sets remain identical to the ones in the mixed-strategy version of the classical game that is obtained as a proper subset of the corresponding quantum game. Using GA we investigate the outcome of a realization of the game by players sharing GHZ state, W state, and a mixture of GHZ and W states. As a specific example, we study the game of three-player Prisoners' Dilemma.Comment: 21 pages, 3 figure

A study on immunological factors in white spot vaccinated shrimp, Litopenaeus vannamei in comparison to none vaccinated groups

The aim of this study was to evaluate the efficacy of white spot virus vaccine produced by gamma irradiation in the face of Litopenaeus vannamei in comparison with Gracilaria corticata and Saccharomyces cerevisiae Seven hundred and twenty healthy shrimp SPF L. vannamei subadult with average weight of 10±1.02 g were collected and divided into 8 groups. The first group (T1) was fed with commercial pellet as control. The second group (T2) was fed with S. cerevisiae added to shrimp feed (1 g/Kg), the third group (T3) G. corticata so that algae Gracilaria were dried and added to shrimp feed at the rate of 1500 mg per kg and finally, the fourth group (T4) was vaccination group which the shrimp were exposed to the vaccine and injected intramuscularly gamma irradiant WSSV (1µl/gbw) for 10 days. The shrimps of all groups were then injected with WSSV and maintained for 25 days. Results indicated that the survival rates for groups T4, T3 T2 and T1 were 57.05±3.52%, 22.5±0.5%, 15±1.05% and 00.0±0%, respectively. Ultimately, at the end of the study the shrimp group T4 showed higher hematological data: THC, TPP, SOD, POD and PO. The study concluded that gamma irradiant WSSV is effective immunostimulants in shrimp L. vannamei and the immunity has better performances than those of the G. corticata and S. cerevisiae

Ancient Migratory Events in the Middle East: New Clues from the Y-Chromosome Variation of Modern Iranians

Knowledge of high resolution Y-chromosome haplogroup diversification within Iran provides important geographic context regarding the spread and compartmentalization of male lineages in the Middle East and southwestern Asia. At present, the Iranian population is characterized by an extraordinary mix of different ethnic groups speaking a variety of Indo-Iranian, Semitic and Turkic languages. Despite these features, only few studies have investigated the multiethnic components of the Iranian gene pool. In this survey 938 Iranian male DNAs belonging to 15 ethnic groups from 14 Iranian provinces were analyzed for 84 Y-chromosome biallelic markers and 10 STRs. The results show an autochthonous but non-homogeneous ancient background mainly composed by J2a sub-clades with different external contributions. The phylogeography of the main haplogroups allowed identifying post-glacial and Neolithic expansions toward western Eurasia but also recent movements towards the Iranian region from western Eurasia (R1b-L23), Central Asia (Q-M25), Asia Minor (J2a-M92) and southern Mesopotamia (J1-Page08). In spite of the presence of important geographic barriers (Zagros and Alborz mountain ranges, and the Dasht-e Kavir and Dash-e Lut deserts) which may have limited gene flow, AMOVA analysis revealed that language, in addition to geography, has played an important role in shaping the nowadays Iranian gene pool. Overall, this study provides a portrait of the Y-chromosomal variation in Iran, useful for depicting a more comprehensive history of the peoples of this area as well as for reconstructing ancient migration routes. In addition, our results evidence the important role of the Iranian plateau as source and recipient of gene flow between culturally and genetically distinct population