The Role of Calcium in Differentiation of Human Adipose-Derived Stem Cells to Adipocytes

Differentiation process of mesenchymal stem cells (MSCs) into adipocyte is involved in obesity. Multiple factors such as Ca²⁺ play important roles in different stages of this process. Because of the complicated roles of Ca²⁺ in adipogenesis, the aim of present investigation was to study the influx and efflux of Ca²⁺ into and out of the cells during adipogenesis. Adipose-derived MSCs were used to differentiate into adipocytes. MSCs were exposed to 2.5 mM Ca²⁺ or 1.8 mM Ca²⁺ plus calcium ionophore, A23187, for 3 days. Lipid staining, triglycerides (TG) content, and glyceraldehyde phosphate dehydrogenase (GAPDH) activity were evaluated to confirm the efficiency of the differentiation. Gene expression of GLUT4, PPARγ2, RAR-α, and calreticulin, as well as the protein levels of GLUT4 and PPARγ2 were determined. Ca²⁺ and in particular Ca²⁺ plus A23187 significantly lowered the efficiency of differentiation accompanied by decrease in intracellular TG deposits, GAPDH activity and alleviation of gene, and protein levels of GLUT4 and PPARγ2. While calreticulin and RAR-α were remarkably upregulated in A23187 group. This study showed the inhibitory effects of calcium in adipogenesis. Additionally, it indicated the greater inhibitory effect of calreticulin and RAR-α in controlling adipogenesis by higher levels of calcium.     

Stimulated and Unstimulated Saliva Levels of Calcium and Magnesium in Giardiasis

Giardia lamblia causes malabsorption. The aim of this study was to evaluate serum and saliva calcium and magnesium levels in patients with giardiasis. Thirty patients with giardiasis as a case and 30 person without giardiasis as a control group were enrolled. The stimulated and unstimulated whole saliva and serum calcium and magnesium levels were assayed by Arsenazo reaction and xylidyl blue complex methods, respectively. Mean calcium and magnesium level was low in serum and stimulated saliva of case group than that of controls. However, they were higher in the unstimulated saliva of the case group. It is suggested that patients suffering from giardiasis have low calcium and magnesium levels, and they lose the most of calcium and magnesium by saliva during unstimulated condition.     

Orbital Dirofilariasis in Iran: A Case Report

Dirofilariasis is a common parasitic disease in both domestic and wild animals around the world, with canines as the principal reservoir host and mosquitoes as the vector. Human ophthalmic dirofilariasis is an uncommon condition, but there have been reports from many parts of the world, including Africa, Australia, the Americas, Europe, and Asia. Ophthalmic involvement with Dirofilaria may present itself as periorbital, subconjunctival, orbital, or intraocular infections. In this report, we present a case of orbital dirofilariasis with lateral rectus muscle involvement. To our knowledge, this is the first orbital dirofilariasis case reported in Iran. Although debulking of the tumor usually leads to resolution, our patient showed an episode of recurrence after biopsy. Complete recovery was achieved after a spontaneous discharge, without the need for systemic medication.     

Effects of the exogenous application of auxin and cytokinin on carbohydrate accumulation in grains of rice under salt stress

Phytohormones, such as auxin and cytokinin, are known to be involved in the regulation of plant responses to salinity stress and counteract the adverse effect of stress conditions. This work investigated the effects of the exogenous spraying of indole-3-acetic acid (IAA) and kinetin (KIN) during the reproductive phase on grain yield by examining the 1000-grain weight and filled-grain percentage as well as the changes in starch, total soluble sugars, sucrose, glucose and fructose concentrations in the grains of two rice cultivars under salt stress. The results indicated that the applied IAA and KIN led to an increased grain yield, 1000-grain weight and filled-grain percentage for both rice cultivars under salt stress. The storage starch content in the grain of the salt-sensitive cultivar was more than that in the salt-tolerant cultivar under IAA application compared with KIN, whereas a decrease in the total soluble sugar content was observed with both IAA and KIN treatments, in comparison to the non-hormone treatment. Interestingly, this study showed that IAA led to a much higher increase in the sucrose content in grain, as compared to the KIN. Furthermore, this experiment suggests that glucose and fructose may play important roles during salt stress because there were clearly higher concentrations of these sugars in the grain of the stressed cultivars under IAA and KIN application: it appears that their accumulation was the earliest response detected during the grain-filling period in rice. Finally, this work indicated that an increase in the rice grain yield, 1000-grain weight and filled-grain percentage are associated with an increase in the contents of starch, sucrose, glucose and fructose in grain caused by the application of IAA and KIN.     

Intraperitoneal Aminoguanidine Improves Sciatic Nerve Ischemia–Reperfusion Injury in Male Sprague-Dawley Rats

The present work was designed to investigate the potential protective effects of post-ischemic treatment with aminoguanidine (AG) on sciatic nerve ischemia/reperfusion (I/R) injury in rat. Seventy-two rats were divided into 12 groups (n = 6). We used ischemia model in these groups by occluding the right common iliac and femoral arteries for 3 h with a silk suture 6-0 using slipknot technique. Treatment groups (2, 4, 6, 8, 10, and 12) received 150 mg/kg AG intraperitoneally 24 h after induction of ischemia. After certain time intervals of reperfusion (2, 4, 7, 14, and 28 days), the function of the hind limb was assessed using behavioral scores based on gait, racing reflex, toe spread, pinch sensitivity, paw position, and grasp. After euthanasia, sciatic nerves were removed at the end of reperfusion times and sections were cut at 5 μm, then were stained for light microscopy studies and graded for ischemic fiber degeneration (IFD), edema, and apoptosis. Maximal behavioral deficit occurred at 7 days of reperfusion. The comparison of behavioral score pertaining to the control and AG groups revealed significant differences and showed also a better time course in recovery (P < 0.05). Other than 3 and 4 groups, the amount of edema in AG treatment groups showed significant differences compared with control groups (P < 0.05). IFD was also significantly decreased in the AG treatment groups than controls. Most importantly, I/R-induced apoptosis were improved significantly on the 4th, 7^th, and 14th days of reperfusion in AG-treated groups compared to controls. In conclusion, our findings suggest that post-ischemic administration of AG exhibits protective effect against sciatic nerve I/R injury.     

Aminoguanidine Changes Hippocampal Expression of Apoptosis-Related Genes,Improves Passive Avoidance Learning and Memory in Streptozotocin-Induced Diabetic Rats

Cognitive dysfunction occurs in patients with diabetes mellitus. The objective of this study was to examine whether bilateral intrahippocampal CA1 (intra-CA1) injection of aminoguanidine (AG) can either affect the Bcl-2 family gene expression or reduce the diabetic imposing abnormalities of passive avoidance learning (PAL) and memory. Rats were divided into five groups: control (C), control treated with normal saline (CS), control treated with AG (S-AG), diabetics (D), and diabetics treated with AG (D-AG). Diabetes mellitus was induced by a single intraperitoneal injection of streptozotocin (STZ) (50 mg/kg). AG (30 μg/rat) or vehicle was administered intra-CA1 bilaterally at the onset of hyperglycemia. PAL was assessed 7 weeks later. Animals were killed, and hippocampus was dissected following the behavioral test. The expressions of Bax, Bcl-2, and Bcl-xl mRNAs were measured using semiquantitative RT-PCR technique. The result of passive avoidance task showed that AG significantly improved the cognitive performance in diabetic rats. Moreover, AG treatment decreased the levels of Bcl-2 and Bcl-x_L expressions in diabetic group. The ratio of Bax/Bcl-2 and Bax/Bcl-x_L decreased significantly in AG-treated diabetic animals. In conclusion, initial treatment with AG by intra-CA1 micro-injection improves the impaired passive avoidance task in STZ-induced diabetic rats which may be related to the decreased Bax/Bcl-2 and Bax/Bcl-x_L ratios.     

Immunogenicity Evaluation of a Rationally Designed Polytope Construct Encoding HLA-A*0201 Restricted Epitopes Derived from Leishmania major Related Proteins in HLA-A2/DR1 Transgenic Mice: Steps toward Polytope Vaccine

Background

There are several reports demonstrating the role of CD8 T cells against Leishmania species. Therefore peptide vaccine might represent an effective approach to control the infection. We developed a rational polytope-DNA construct encoding immunogenic HLA-A2 restricted peptides and validated the processing and presentation of encoded epitopes in a preclinical mouse model humanized for the MHC-class-I and II.

Methods and Findings

HLA-A*0201 restricted epitopes from LPG-3, LmSTI-1, CPB and CPC along with H-2Kd restricted peptides, were lined-up together as a polytope string in a DNA construct. Polytope string was rationally designed by harnessing advantages of ubiquitin, spacers and HLA-DR restricted Th1 epitope. Endotoxin free pcDNA plasmid expressing the polytope was inoculated into humanized HLA-DRB1*0101/HLA-A*0201 transgenic mice intramuscularly 4 days after Cardiotoxin priming followed by 2 boosters at one week interval. Mice were sacrificed 10 days after the last booster, and splenocytes were subjected to ex-vivo and in-vitro evaluation of specific IFN-γ production and in-vitro cytotoxicity against individual peptides by ELISpot and standard chromium-51(⁵¹Cr) release assay respectively. 4 H-2Kd and 5 HLA-A*0201 restricted peptides were able to induce specific CD8 T cell responses in BALB/C and HLA-A2/DR1 mice respectively. IFN-γ and cytolytic activity together discriminated LPG-3-P1 as dominant, LmSTI-1-P3 and LmSTI-1-P6 as subdominant with both cytolytic activity and IFN-γ production, LmSTI-1-P4 and LPG-3-P5 as subdominant with only IFN-γ production potential.

Conclusions

Here we described a new DNA-polytope construct for Leishmania vaccination encompassing immunogenic HLA-A2 restricted peptides. Immunogenicity evaluation in HLA-transgenic model confirmed CD8 T cell induction with expected affinities and avidities showing almost efficient processing and presentation of the peptides in relevant preclinical model. Further evaluation will determine the efficacy of this polytope construct protecting against infectious challenge of Leishmania. Fortunately HLA transgenic mice are promising preclinical models helping to speed up immunogenicity analysis in a human related mouse model.     

Fine-scale population genetic structure of Endangered Caspian Sea trout,Salmo caspius: implications for conservation

Hydrobiologia - Many populations of Caspian Sea trout (Salmo caspius)—a nationally endangered species in Iran—have been extirpated or depleted due to anthropogenic impacts. The Lar...