Journal of Pathobiology Reaearch
Volume:18 Issue: 2, 2015

To evaluate the effect of mouse fallopian tube cell co-culture on the proliferation of human endometrial stromal cells. The endometrial cells were collected from the uterus under hysteroscopy. Then the cells were isolated withcollagenase type 3 and passed within the filters with 150 and 40 micrometer sizes respectivelyand then the isolated cells were cultured in DMEM/F12 medium. Percentage of CD90 positive cells were evaluated using flowcytometry at the end of fourth sub culture.. The endometrial cells then were co-cultured with mitomycin C-treated cells of mouse fallopian tube as experimental group and were cultured without feeder layer as control group. The proliferation rate of cells in both co-cultured and control groups were compared using MTT assay The endometrial cells were adhered to the floor of plate after 24 hours and they have spindle shape after 72 hours similar to the fibroblast. The rate of CD90 positive cells at the fourth passage was % 94.26 ± 0.08. The proliferation rate of co-cultured and control groups were 1.1 ± 0.015 and 1.17 ± 0.17 respectively and there was no significant difference between these groups. Co-culture of mouse fallopian tube cells with endometrial stem cells had no effect on their proliferation rate.

Because of the host immunological system response against the minor blood groups of donor, transfusion in some cases, especially for patients require repeated transfusions (such as people with thalassemia), can be considered as a significant problem. A proposed way is coating the surface antigens on red blood cells (RBCs) by covalent binding of methoxy polyethylene glycol (mPEG). The aim of this study was to determine the storage time of PEGylated cells before injection and its effective time at in vivo condition. Methods and materials: mPEG activated by succinimidyl valerate (SVA) was used for PEGlaytion the cells. The stability of the created coating at in vitro condition was investigated using 3

counting the number of free cells, flow cytometry and qualitative investigation. Using electron microscopic investigations, the appropriate concentration of mPEG for rabbit RBC PEGylation was determined. The effective time of PEGylated rabbit RBCs was determined using flow cytometric analysis, after injection. Also, the serum biochemical properties were investigated 24 hours after injection.

The appropriate concentration of 15 mg/mL for rabbit RBC PEGlytion was determined. Forty eight hours after injection, %83 of the cells were alive in the host circulatory system and could keep their polymeric coating.

The time of 18 days was obtained as appropriate time for storage the PEGylated RBCs at in vitro condition. Also, the effective time of 14 days was determined for PEGylated RBCs, by tracking the cells at in vivo condition. Investigating the serum biochemical properties of rabbits, 24 hours after injection, showed that RBC coating, significantly inhibited the stimulation of the host immunological system and destruction the cells.

Eukaryotic proteins generally have signal peptides, which are not only crucial for their secretion efficiencies but are important for their expression levels. The coagulation factor IX (FIX) is a glycoprotein that plays a fundamental role in a blood coagulation pathway. Reduced levels or dysfunctional FIX are associated with hemophilia B. To improve the hFIX secretion efficiency in a heterologous expression system, in this study we investigated the function of the human prothrombin signal peptide. With this aim, we used SignalP, and PrediSi programs for in silico evaluation of the signal peptides efficiency before being examined experimentally. Using molecular techniques, we amplified and joined the coding region of the human prothrombin signal peptide to the cDNA of mature hFIX. The chimeric fragment was examined for transient expression in mammalian cell line (HEK293T) in comparison with the native hFIX, under a CMVp regulation. Using the neural network-based prediction programs, we evaluated the scores for cleavage position and secretion efficiency of the human prothrombin and hFIX signal peptides. The expression efficiencies of hFIX expressed by the recombinant cells was analyzed by RT-PCR, ELISA. In silico analysis predicted the human prothrombin signal peptide with the high score more efficient than the native hFIX signal peptide. These data was confirmed by the results obtained from the expression analysis in RNA and protein level of the rhFIX by RT-PCR and ELISA, respectively. The present study showed that, the signal peptide derived from the human prothrombin has the potential for efficient secretion of hFIX, which was evidenced by the results taken from a transient expression system and it was in consistent with in silico analysis. This replacement can be evaluated in stable state.

Cutaneous leishmaniasis is an endemic disease in some parts of Iran.Using Pentavalent antimony compounds as first line treatment have been reported are associated with limitations and several adverse complications. So, attempt to find a new and effective compound has been under consideration.In the present study,the effect of Achillea biebersteinii Afanas that is a native plant in Iran against Leishmania major promastigote and amastigote growth in vitro condition was investigated. This experimental study was carried out in Tarbiat Modares University at 1392.The essential oil ofAchillea biebersteinii Afan plan textracted by steam distillation method and was analysis by gas chromatography mass spectrograph. Then effect of different concentrations of 10%, 15%, 25% and 50% of the oil on growth of promastigotes stage of Leishmania and infected macrophages‌‌ contain amastigote were evaluated in vitro conditions. Effectiveness of theoil on promastigotes and amastigote was assessed by direct counting and MTT assay. In all of tests, each well of plate containing culture media and parasites without drug was considered as control group.The data was statistically nalyzed with ANOVAtest. The MTT results indicated significant differences among the number of parasites in control and case groups treated with10%, 15%, 25% and 50% of the oil within 24, 48 and 72 h after culture.The concentration of 50% of the oil killed 66% of the macrophages contain amastigotes after 72 h. Achillea biebersteinii Afanoilis found to be effective in killing Leishmania major promastigotes and infected macrophages contain amastigotes.So, we propose to study the extract in vivo for treatment of cutaneous leishmaniasis lesions.

Drug delivery systems related to different cancer therapy is now expanding. Chitosan (CS) is currently receiving enormous interest for medical and pharmaceutical applications due to its biocompatibility in animal tissues. In this study two nanogels were prepared from Chitosan. Some of the critical factors such as controlling the release, adsorption and specially targeting drug delivery are considered while preparing the nanogels. Phosphorylated chitosan (PCS) and Myristilated chitosan (MCS)nanogeles were prepared by reacting Chitosan with tripolyphosphate (TPP) and Myristate as a cross linking agents respectively and then were loaded with Doxorubicin (DOX). The Nanogels were characterized by different techniques such as Scaning Electron Microscopy, Dynamic Light Scattering and Fourier-transform infrared. The cytotoxicity of free DOX, MCS nanogels and DOX loaded MCS were evaluated by MTT assay. The results of DOX loading and releasing of the nanogels showed high loading capacity and drug loading efficiency of about 97%. Results indicated slow release of about 16-28% of DOX from PCS within 5 days and 18-40% from MCS within 15 days. The DOX and MCS-DOX showed the same toxic effect on the prostate cancer cells (LNCaP). Both PCS and MCS nanogels were qualified on the base of size, loading and releasing capacity.

Prostate cancer after lung cancer is the second cause of cancer-associated death in men. In recent years, targeted therapy of cancer has attracted researchers attention. The targeted therapy lead to decrease in side effects of drugs. Studies have indicated that targeting peptides for cancer cells represent valuable tools for diagnostic and therapeutic. Recently, phage display peptide libraries have been used for identifying targeting peptides to a variety of cancer cells. In the current study, we aim to isolate peptides targeting to PC3 cells (human prostate adenocarcinoma cells). Four rounds of subtractive panning on control cells including 5637 (bladder), Huh-7 (liver) and SW480 (colon) and AGS (stomach) and human fibroblast normal and four rounds of positive panning on PC3 (target cell) were performed. Polyclonal phage ELISA was exploited to evaluate the process of enrichment during biopanning. Subsequently, phage clones were randomly picked out from titer plates, amplified by using plaque-PCR and their genomic DNA was sequenced. Bioinformatic analysis was conducted for further characterization of isolated peptides. Several rounds of panning resulted in the enrichment of some peptides. The results of polyclonal phage ELISA are indicated that the biopanning process was successfully done. Also, in silico analysis has shown the presence of several consensus amino acid motifs in peptides. The peptides identified through biopanning can be considered as a potential specific binders to PC3 cells. Peptides with specificity binding to target cells can be used for targeted gene and drug delivery to malignant tumor cells. Further analysis of these peptides are required to show their capacity for targeted delivery of various genes and drugs into prostate cancer cells.

Diabetes mellitus is a common metabolic disorder that is one of the important factors in the etiology of injuries Oxidant know the purpose The aim of the present study, was the effect of swimming training with Arbutin on total oxidant status (TOS) and total antioxidant status(TAS) the kidney tissue of Alloxan-induced diabetic rats. For this purpose, 42 male Wistar rats with an average weight of 195 to 220 g were randomly divided into 6 groups(7 rats per group) of control, diabetic, Arbutin, diabetic-Arbutin, diabetic -swimming training and diabetic - combinatorial. Swimming training protocol consisted of 5 day/week for 6 weeks and 5-36 min/day. Diabetes was induced whit alloxan (90 mg/k g, intraperitoneally, ip) in rats and Arbutin (50 mg/kg, subcutaneously) was administered for 5 days/week. The rats were killed 48 h after the last treatments and kidney TOS and TAS levels were evaluated. A one-way analysis of variance was used to data analysis. Six weeks of supplementation with Arbutin, swimming training and combination of swimming training and Arbutin were associated with an significant decrease in TOS (P

A pure nucleotide pool is an essential precursor of correct DNA replication and prevention of mutagenesis and abnormality in a living cell. Inosine triphosphatase (ITPase) is a critical enzyme to remove any deaminated rough purine nucleotides such as inosine from nucleotide pool. It has been shown that the rate of substitution mutation in genomic DNA increase by abnormal function and expression of ITPA gene. The object of this study was comparisons the ITPA gene expression between human adenocarcinoma tumor with the normal margins tissues. the expression of ITPA gene was examined in 24 pairs of gastric adenocarcinoma tumor and their normal adjacent tissues using quantitative Real-Time PCR technique. the analysis gene expression showed reduction in tumor tissue in comparison with their adjacent normal tissues. The reduction in ITPA gene expression is especially significant in higher grade samples. with regard to the biological function of ITPA gene in omitting the rough deaminated purines from the nucleotides pool, it seems that lower expression of this gene can be considered as a risk factor for development and progression of gastric cancer. Also abnormality in this gene increase the frequency of mutation in genomic DNA and make a suitable background for higher genomic instability.