Enhancing maturity in 3D kidney micro-tissues through clonogenic cell combinations and endothelial integration.

As an advance laboratory model, three-dimensional (3D) organoid culture has recently been recruited to study development, physiology and abnormality of kidney tissue. Micro-tissues derived from primary renal cells are composed of 3D epithelial structures representing the main characteristics of original tissue. In this research, we presented a simple method to isolate mouse renal clonogenic mesenchymal (MLCs) and epithelial-like cells (ELCs). Then we have done a full characterization of MLCs using flow cytometry for surface markers which showed that more than 93% of cells expressed these markers (Cd44, Cd73 and Cd105). Epithelial and stem/progenitor cell markers characterization also performed for ELC cells and upregulating of these markers observed while mesenchymal markers expression levels were not significantly increased in ELCs. Each of these cells were cultured either alone (ME) or in combination with human umbilical vein endothelial cells (HUVECs) (MEH; with an approximate ratio of 10:5:2) to generate more mature kidney structures. Analysis of 3D MEH renal micro-tissues (MEHRMs) indicated a significant increase in renal-specific gene expression including Aqp1 (proximal tubule), Cdh1 (distal tubule), Umod (loop of Henle), Wt1, Podxl and Nphs1 (podocyte markers), compared to those groups without endothelial cells, suggesting greater maturity of the former tissue. Furthermore, ex ovo transplantation showed greater maturation in the constructed 3D kidney.

Alteration of gut bacteria composition among individuals with asymptomatic Blastocystis infection: A case-control study.

The gut microbiota consists a diverse and complex ecosystem that is involved in beneficial functions as well as potentially harmful conditions for human. Blastocystis sp. is a common parasite of the digestive tract of animals and humans; however, limited data is available concerning the association of asymptomatic Blastocystis infection and gut bacteria composition. Hence, in this cross-sectional study, the gut bacteria composition of twenty asymptomatic Blastocystis sp. positive and twenty Blastocystis sp. negative individuals was assessed with real time PCR. The case and control groups were matched for age and sex. Both groups were negative for other gastrointestinal infections and did not have any gastrointestinal symptoms. The subtype of ten Blastocystis sp. isolates was assessed based on sequencing. Sequencing of ten Blastocystis sp. isolates revealed the ST1, ST2, and ST3 subtypes in 40%, 30%, and 30% of the isolates. The relative expression of each bacteria in the case than control group revealed that the expression level of Bifidobacterium group (P < 0.033), Peptostreptococcus productus (P < 0.014), Lactobacillus/Enterococcus group (P < 0.001), and Escherichia coli (P < 0.001) were significantly upregulate in the Blastocystis sp. carriers than the control group, while the relative amounts of Bacteroides fragilis (P < 0.001) and Enterococcus sp. (P < 0.001) were significantly downregulated in the case than the control group. Taken together, the results of this study have shown that asymptomatic Blastocystis infection could alter the composition of gut bacteria in healthy individuals.

Genetic characterization of Toxoplasma gondii isolates from human spontaneous aborted fetuses in Jahrom, southern Iran.

Toxoplasma gondii (T. gondii) is an intracellular protozoan that infects the fetus through the placenta and leads to severe complications in the fetus. One of the complications of congenital toxoplasmosis is spontaneous abortion. The prevalence of toxoplasmosis infection was investigated among spontaneously aborted fetuses (SAFs), and the genotypes of parasite isolates were determined in the present study. Placentas from 330 samples of SAFs were collected in Jahrom (Fars province) from February to September 2018. DNA was extracted from each placental tissue. The T. gondii infection was detected using nested polymerase chain reaction (Nested-PCR) assay based on a 529 bp repeat element (RE) gene. Afterward, Toxoplasma was genotyped using PCR-restriction fragment length polymorphism (PCR-RFLP) based on the GRA6 gene. The frequency of T. gondii infection was found to be 14.5% (48 out of 330 samples). Genotyping of nine T. gondii isolates revealed that all belonged to genotype II. Statistically, the prevalence of T. gondii infection was significantly correlated with the education levels of the mothers and the age of the fetus (P < 0.05). The lowest prevalence of Toxoplasma infection belonged to mothers with university education and the highest frequency of infection was observed among the fetuses in the age group of 8-9 weeks. The findings of the present study suggest a significant role for toxoplasmosis in SAFs in Jahrom city.

A dietary pattern rich in fruits and dairy products is inversely associated to gestational diabetes: a case-control study in Iran.

BACKGROUND: Gestational diabetes mellitus (GDM) causes many problems for mother and her neonate. A healthy diet plays an important role in preventing GDM. This study aimed to investigate the relationship between major dietary patterns and the GDM. METHODS: 386 healthy and 306 GDM pregnant women (total 693) completed this case-control study. Basic information and anthropometric indices were recorded, and a food frequency questionnaire was completed. For extracting major dietary patterns, the principal component analysis was performed. Multivariable logistic regression models were used to examine whether specific dietary patterns are associated to the GDM. RESULTS: Four dietary patterns were identified: "fruits and dairy products", "red meat and plant-based foods", "snacks and high-fat foods" and "carbohydrate-rich foods". Among these major extracted dietary patterns, "fruits and dairy products" showed an inverse association to the GDM (odds ratio adjusted for confounders: 0.50, confidence interval: 0.284-0.882, p-trend = 0.019, for highest vs. lowest quartile). CONCLUSIONS: It seems using a healthy dietary pattern such as "fruits and dairy products" may decrease GDM risk.

Case-control study on the association between the GATA2 gene and premature myocardial infarction in the Iranian population.

In recent decades, due to the high prevalence of coronary artery disease (CAD) and myocardial infarction (MI), numerous studies have attempted to elucidate genetic contributing factors in these complex disorders. A very interesting gene in this regard is GATA-binding protein 2 (GATA2), an important regulator of various gene expressions in vascular endothelial cells. Accordingly, the association of different GATA2 polymorphisms with CAD and MI has already been evaluated. Rs2713604 is a genetic marker whose association with CAD has not been reproduced in previous studies. Considering the importance of replicating the initial association, the present case-control study aimed to examine the association of this intronic variant with premature MI in a sample of the Iranian population. In this study, 193 participants from Jahrom Hospital (Jahrom, Iran) were consecutively recruited during a 1.5-year period, and, following blood sampling, genomic DNA was extracted. We then proceeded to genotype rs2713604 using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and statistically analyzed the data. After adjustment for hyperlipidemia, hypertension, and type 2 diabetes mellitus, the results of the multivariate regression analysis showed no significant association between rs2713604 and premature MI. Interestingly, the risk allele (A-allele) of rs2713604 displayed a slightly higher frequency among controls compared to cases.

Study of the mechanisms of crocetin-induced differentiation and apoptosis in human acute promyelocytic leukemia cells.

Crocetin, the major carotenoid in saffron, exhibits potent anticancer effects. However, the antileukemic effects of crocetin are still unclear, especially in primary acute promyelocytic leukemia (APL) cells. In the current study, the potential antipromyelocytic leukemia activity of crocetin and the underlying molecular mechanisms were investigated. Crocetin (100 µM), like standard anti-APL drugs, all-trans retinoic acid (ATRA, 10 µM) and As2 O 3 (arsenic trioxide, 50 µM), significantly inhibited proliferation and induced apoptosis in primary APL cells, as well as NB4 and HL60 cells. The effect was associated with the decreased expressions of prosurvival genes Akt and BCL2, the multidrug resistance (MDR) proteins, ABCB1 and ABCC1 and the inhibition of tyrosyl-DNA phosphodiesterase 1 (TDP1), while the expressions of proapoptotic genes CASP3, CASP9, and BAX/BCL2 ratio were significantly increased. In contrast, crocetin at relatively low concentration (10 µM), like ATRA (1 µM) and As 2 O 3 (0.5 µM), induced differentiation of leukemic cells toward granulocytic pattern, and increased the number of differentiated cells expressing CD11b and CD14, while the number of the immature cells expressing CD34 or CD33 was decreased. Furthermore, crocetin suppressed the expression of clinical marker promyelocytic leukemia/retinoic acid receptor-α ( PML/RARα) in NB4 and primary APL cells, and reduced the expression of histone deacetylase 1 ( HDAC1) in all leukemic cells. The results suggested that crocetin can be considered as a candidate for future preclinical and clinical trials of complementary APL treatment.

Kaempferol increases apoptosis in human acute promyelocytic leukemia cells and inhibits multidrug resistance genes.

Acute promyelocytic leukemia (APL) is one of the most life-threatening hematological malignancies. Defects in the cell growth and apoptotic pathways are responsible for both disease pathogenesis and treatment resistance. Therefore, pro-apoptotic agents are potential candidates for APL treatment. Kaempferol is a flavonoid with antioxidant and anti-tumor properties. This study was designed to investigate the cytotoxic, pro-apoptotic, and differentiation-inducing effects of kaempferol on HL-60 and NB4 leukemia cells. Resazurin assay was used to determine cell viability following treatment with kaempferol (12.5-100 µM) and all-trans retinoic acid (ATRA; 10 µM; used as a positive control). Apoptosis and differentiation were also detected using propidium iodide and NBT staining techniques, respectively. Furthermore, the expression levels of genes involved in apoptosis (PI3 K, AKT, BCL2, BAX, p53, p21, PTEN, CASP3, CASP8, and CASP9), differentiation (PML-RAR and HDAC1), and multi-drug resistance (ABCB1 and ABCC1) were determined using quantitative real-time PCR. The protein expressions of Bax/Bcl2 and casp3 were confirmed using Western blot. The results showed that kaempferol decreased cell viability and increased subG1 population in the tested leukemic cells. This effect was associated with decreased expression of Akt, BCL2, ABCB1, and ABCC1 genes, while the expression of CASP3 and BAX/BCL-2 ratio were significantly increased at both gene and protein levels. Kaempferol promoted apoptosis and inhibited multidrug resistance in a concentration-dependent manner, without any differential effect on leukemic cells. In conclusion, this study suggested that kaempferol may be utilized as an appropriate alternative for ATRA in APL patients.

Epigallocatechin-3-gallate enhances differentiation of acute promyelocytic leukemia cells via inhibition of PML-RARα and HDAC1.

The use of all-trans retinoic acid (ATRA) has dramatically improved the treatment and survival rate of patients with acute promyelocytic leukemia (APL). However, toxicity and resistance to this drug are major problems in the treatment of APL with ATRA. Earlier studies have suggested that the green tea polyphenol epigallocatechin gallate (EGCG) induces cell death in hematopoietic neoplasms without adversely affecting normal cells. In the present study, the potential therapeutic effect of EGCG in APL and the underlying molecular mechanisms were investigated. EGCG (100 µM) significantly inhibited proliferation and induced apoptosis in HL-60 and NB4 cells. This effect was associated with decreased expressions of multidrug resistance proteins ABCB1, and ABCC1, whereas the expressions of pro-apoptotic genes CASP3, CASP8, p21, and Bax/Bcl-2 ratio were significantly increased. EGCG, at 25 µM concentration, induced differentiation of leukemic cells towards granulocytic pattern in a similar manner to that observed for ATRA (1 µM). Furthermore, EGCG suppressed the expression of clinical marker PML/RARα in NB4 cells and reduced the expression of HDAC1 in leukemic cells. In conclusion, the results suggested that EGCG can be considered as a potential treatment for APL.

Association of OX40L gene polymorphism with multiple sclerosis in Iranians.

Introduction: The exact etiology of multiple sclerosis is unknown but recent studies indicated a link between tumor necrosis factor superfamily member 4 and the disease. Polymorphisms located in the regulatory region of the gene may affect its phenotype. Hence, we aimed to investigate the association of promoter polymorphisms of the gene with multiple sclerosis and also to estimate the frequency of haplotypes in the patients and healthy subjects. Methods: Two hundred age- and sex-matched subjects including 100 patients and 100 healthy subjects were investigated in the study. Genotype and allele distributions of rs3850641, rs1234313, and rs10912580 polymorphisms in the promoter region of the gene were investigated by polymerase chain reaction-restriction fragment length polymorphism. In addition, haplotype frequencies estimation and linkage disequilibrium analysis were performed by SNPStats web tool. Results: The distribution of AA, AG and GG genotypes of rs3850641 was significantly different between the patient and healthy groups (P = 0.009). In addition, frequencies of A and G alleles of rs3850641 were different between the groups (P < 0.001). Also the distribution of rs3850641 genotypes was different between the women of the both groups (P = 0.007). Our analysis revealed that rs3850641 AG (Odds ratio = 0.393, 95 % confidence interval = 0.170-0.907, P = 0.029) and GG (Odds ratio = 0.373, 95 % confidence interval = 0.168-0.830, P = 0.016) genotypes were associated with decreased risk of the disease. However, rs1234313 genotype and allele distributions were not different between the groups. The distribution of rs10912580polymorphism. AA, AG, and GG genotypes was significantly different between the groups (P = 0.007). rs10912580 AG genotype was associated with low risk of the disease (Odds ratio = 0.252, 95 % confidence interval = 0.102-0.623, P = 0.003). The distribution of haplotypes was statistically different between the patient and healthy groups (P < 0.001). A-G-A was the most frequent haplotype among the patients and the estimated frequency was higher than that of the control group (0.5527 versus 0.3739). Conclusion: The distribution of rs3850641 and rs10912580 genotypes was different between the patients and healthy subjects. Moreover, rs3850641 AG and GG genotypes and also rs10912580 AG genotype were associated with low risk of the disease in Iranians. Further studies with large groups are recommended to show whether genotype variation in the patients could alter the response to treatment or not.

Association between OX40L polymorphism and type 2 diabetes mellitus in Iranians.

INTRODUCTION: Diabetes mellitus (DM) is one of the leading causes of morbidity and mortality worldwide. It is a multifactorial disease that genetic and environmental factors contribute to its development. The aim of the study was to investigate the association of OX40L promoter gene polymorphisms with type 2 diabetes mellitus (T2DM) in Iranians. MATERIALS AND METHODS: Three hundred and sixty-eight subjects including 184 healthy subjects and 184 T2DM patients were enrolled in our study. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied to detect genotype and allele frequencies of rs3850641, rs1234313 and rs10912580. In addition, SNPStats web tool was applied to estimate haplotype frequency and linkage disequilibrium (LD). RESULTS: The distribution of tested polymorphisms was statistically different between the T2DM patients and healthy subjects (P < 0.01). rs1234313 AG (OR = 0.375, 95% CI = 0.193-0.727, P = 0.004) and rs10912580 AG (OR = 0.351, 95% CI = 0.162-0.758, P = 0.008) genotypes were associated with the decreased risk of T2DM in Iranians. Moreover, our prediction revealed that AAG (OR = 0.46, 95% CI= (0.28-0.76), P = 0.0028) and GAG (OR = 0.24, 95% CI= (0.13-0.45), P < 0.0001) haplotypes were related to the reduced risk of the disease. However, the tested polymorphisms had no effect on biochemical parameters and body mass index (BMI) in the patient group (P > 0.05). CONCLUSION: Our findings revealed that OX40L promoter gene polymorphisms are associated with T2DM. Moreover, genotype and allelic variations were related to the decreased risk of T2DM in Iranians. Further studies are recommended to show whether these polymorphic variations could affect OX40/OX40L interaction or OX40L phenotype.

Detection of Leishmania RNA Virus 2 (LRV2) among Clinical Isolates of Leishmania Major in Four Endemic Regions of Iran.

PURPOSE: Leishmania RNA viruses (LRV) are double-stranded RNA viruses (dsRNA viruses) that play a role in the pathogenesis of Leishmania parasites. Cutaneous leishmaniasis (CL) is endemic in various parts of Iran. Our aimed was to investigate presence of LRV among the Leishmania major isolates in four endemic regions of Iran. METHODS: In a cross-sectional study, we assessed the presence of LRV1 and LRV2 in 181 clinical isolates of L. major from four endemic cities in Iran using reverse transcription polymerase chain reaction (RT-PCR). After RNA extraction and cDNA synthesis, RT-PCR tests were conducted with LRV1 and LRV2 specific primers. Human beta-actin and kmp genes served as internal and external controls, respectively, and the Allele ID software was used to optimize melting curves. RESULTS: LRV2 was detected in 27.6% (50 out of 181) of L. major isolates, while no LRV1 was found. We did not observe a statistically significant difference in the presence of LRV2 based on age group, number, or location of lesions. CONCLUSION: This study confirms the presence of LRV2 in clinical isolates of L. major from endemic regions of Iran. Further researches with larger sample sizes is recommended to explore the association between LRV and clinical symptoms as well as treatment response.

Promotion of cardiac microtissue assembly within G-CSF-enriched collagen I-cardiogel hybrid hydrogel.

Tissue engineering as an interdisciplinary field of biomedical sciences has raised many hopes in the treatment of cardiovascular diseases as well as development of in vitro three-dimensional (3D) cardiac models. This study aimed to engineer a cardiac microtissue using a natural hybrid hydrogel enriched by granulocyte colony-stimulating factor (G-CSF), a bone marrow-derived growth factor. Cardiac ECM hydrogel (Cardiogel: CG) was mixed with collagen type I (ColI) to form the hybrid hydrogel, which was tested for mechanical and biological properties. Three cell types (cardiac progenitor cells, endothelial cells and cardiac fibroblasts) were co-cultured in the G-CSF-enriched hybrid hydrogel to form a 3D microtissue. ColI markedly improved the mechanical properties of CG in the hybrid form with a ratio of 1:1. The hybrid hydrogel demonstrated acceptable biocompatibility and improved retention of encapsulated human foreskin fibroblasts. Co-culture of three cell types in G-CSF enriched hybrid hydrogel, resulted in a faster 3D structure shaping and a well-cellularized microtissue with higher angiogenesis compared to growth factor-free hybrid hydrogel (control). Immunostaining confirmed the presence of CD31+ tube-like structures as well as vimentin+ cardiac fibroblasts and cTNT+ human pluripotent stem cells-derived cardiomyocytes. Bioinformatics analysis of signaling pathways related to the G-CSF receptor in cardiovascular lineage cells, identified target molecules. The in silico-identified STAT3, as one of the major molecules involved in G-CSF signaling of cardiac tissue, was upregulated in G-CSF compared to control. The G-CSF-enriched hybrid hydrogel could be a promising candidate for cardiac tissue engineering, as it facilitates tissue formation and angiogenesis.

Association of gastric inhibitory polypeptide receptor (GIPR) gene polymorphism with type 2 diabetes mellitus in iranian patients.

INTRODUCTION: Gastric inhibitory polypeptide receptor (GIPR) encodes a G-protein coupled receptor for gastric inhibitory polypeptide (GIP), which was demonstrated to stimulate insulin secretion. Relation of GIPR gene variation to impaired insulin response has been suggested in previous studies. However, little information is available regarding GIPR polymorphisms and type 2 diabetes mellitus (T2DM). Hence, the aim of the study was to investigate single nucleotide polymorphisms (SNPs) in the promoter and coding regions of GIPR in Iranian T2DM patients. MATERIALS AND METHODS: Two hundred subjects including 100 healthy and 100 T2DM patients were recruited in the study. Genotypes and allele frequency of rs34125392, rs4380143 and rs1800437 in the promoter, 5' UTR and coding region of GIPR were investigated by RFLP-PCR and Nested-PCR. RESULTS: Our finding indicated that rs34125392 genotype distribution was statistically different between T2DM and healthy groups (P = 0.043). In addition, distribution of T/- + -/- versus TT was significantly different between the both groups (P = 0.021). Moreover, rs34125392 T/- genotype increased the risk of T2DM (OR = 2.68, 95%CI = 1.203-5.653, P = 0.015). However, allele frequency and genotype distributions of rs4380143 and rs1800437 were not statistically different between the groups (P > 0.05). Multivariate analysis showed that the tested polymorphisms had no effect on biochemical variables. CONCLUSION: We concluded that GIPR gene polymorphism is associated with T2DM. In addition; rs34125392 heterozygote genotype may increase the risk of T2DM. More studies with large sample size in other populations are recommended to show the ethnical relation of these polymorphisms to T2DM.

Lack of association between CYP27B1 gene polymorphism and type 2 diabetes mellitus in Iranian patients.

Purpose: Type 2 diabetes mellitus (T2DM) is a global health problem with multiple etiological factors. Previous studies indicated that 1- alpha, 25-dihydroxyvitamin D3, a molecule that is produced by CYP27B1, could protect insulin-secreted cells from destruction by immune cells. The aim of the study was to investigate the CYP27B1 promoter gene polymorphism in T2DM. Methods: Two hundred subjects including 100 T2DM and 100 healthy individuals were recruited in the study. ARMS-PCR technique was used to identify rs10877012 genotypes in the 5' region of CYP27B1. Results: The frequency of CC, CA, and AA genotype was 61/50, 31/39, and 8/11, respectively in T2DM patients compared to healthy subjects. There was no significant difference between both groups in regarding to genotype and allele distribution (P > 0.05). CA (RR = 1.535, 95% CI = 0.841- 2.802) and AA (1.677, 95% CI = 0.627-4.490) genotypes had no association with increased risk of T2DM. In addition, CA + AA versus CC showed no increased risk for T2DM (RR = 0.639, 95% CI = 0.365-1.121). Conclusion: We found no association between rs10877012 polymorphism and T2DM. There was no increased risk of this polymorphism in T2DM. Further studies with large groups are suggested in other populations to better understand the relation of CYP27B1 gene variation, especially its ethnicity-dependent relation with T2DM.

Genotyping of Toxoplasma gondii Strains from Goats in Jahrom District, Southern Iran.

PURPOSE: Toxoplasma gondii is transmitted congenitally or acquired by consumption of food and water contaminated with cysts or oocysts. This study aimed at genotyping T. gondii strains from slaughtered goats in Jahrom. METHODS: A total of 561 specimens (heart, diaphragm, and tongue) from 187 slaughtered goats were collected from Jahrom slaughterhouse. After DNA extraction, the T. gondii strains were genotyped by the nested PCR-RFLP based on GRA6 and 3', and 5' ends of the SAG2 gene. RESULTS: T. gondii infection was present in 18.2% of cases. Among the examined organs, the diaphragm was more disposed to the infection (10.2%). Furthermore, infection rates of the heart and tongue were 8.6% and 3.7%, respectively. Concurrent infection in the heart and diaphragm, tongue and diaphragm, and heart and tongue were 3.2%, 0.5%, and 0.5%, respectively. In genotyping experiments, genotype I was the most frequent genotype of T. gondii (58.8%), followed by type II (23.5%), type III (11.8%), and a combination of type I and II (5.9%). CONCLUSIONS: The results of this study showed the presence of different genotypes of T. gondii in goats including three major and mixed genotypes. These results can be useful in toxoplasmosis control and prevention.

The effect of vitamin D on GATA3 gene expression in peripheral blood mononuclear cells in allergic asthma.

INTRODUCTION: Asthma is becoming a major health problem in many countries. Immune responses in allergic asthma, as the most prevalent asthmatic phenotype, are mediated mostly by a subtype of T lymphocytes referred to as the effector lineage of Type 2 Th cells (Th2). The development of Th2 cells is mainly governed by a zinc finger transcription factor, i.e., GATA-binding protein 3 (GATA3). Allergic asthma is a complex disease, and vitamin D deficiency has been named as a non-genetic risk factor for its development. Vitamin D, a steroid hormone belonging to the family of nuclear receptors, has shown significant immunosuppressive effects in previous studies. In this study, given its immunomodulatory properties, we aimed to investigate the effects of different concentrations of vitamin D on GATA3 gene expression in peripheral blood mononuclear cells (PBMCs), including Th2 cells, and compare GATA3 expression levels between PBMCs taken from allergic asthmatic patients and healthy controls. RESULTS: The total sample size was 40 and the quantitative real-time PCR (qPCR) procedure was applied to assess the mRNA expression levels of GATA3 in different groups. Collectively, our results demonstrated that the expression of GATA3 in PBMCs taken from patients with allergic asthma is lower than in that from healthy controls. In addition, in the control group, cells co-cultured with vitamin D had a significantly increased GATA3 expression. However, in the patient group, such an increase was only observed in cells treated with 10⁻7M-vitamin D. By contrast, incubation with vitamin D at the concentration of 10-6 M slightly decreased the expression of GATA3 among patients. CONCLUSION: In summary, it is likely that vitamin D should regulate GATA3 gene expression in the PBMCs in a dose-dependent manner. The impacts of this steroid hormone can also differ between the status of health and allergic asthma in either extent or direction.

PCR-based detection of Toxoplasma gondii from cattle in southern Iran.

OBJECTIVE: Toxoplasma gondii is a protozoan parasite that is widely prevalent in most warm-blooded vertebrates. Humans mainly become infected by eating raw or undercooked meat. This study was designed to investigate the infection of cattle with T. gondii in Jahrom, southern Iran. METHODS: Tissue samples consisting of heart, diaphragm, and tongue were collected from 125 slaughtered cattle. DNA samples were extracted from the homogenized tissues. T. gondii was detected and genotyped using nested-polymerase chain reaction (Nested-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on GRA6 and SAG2 (3', 5' terminal regions) genes, respectively. RESULTS: The prevalence of T. gondii DNA was 56% in cattle. The most infected tissue was the diaphragm (54.4%) followed by the heart (48.8%) and tongue (43.2%). Type II was the most prevalent genotype (70%) among T. gondii isolates. CONCLUSION: In this study, the high prevalence of T. gondii infection in cattle meat indicates the important role of cattle in the transmission of infection to humans. Therefore, incorporating the correct method of consuming meat in health education programs is crucial to prevent human infection.

Association of arginine vasopressin (AVP) promoter polymorphisms with preeclampsia.

OBJECTIVES: Preeclampsia (PE) is a disease of pregnancy characterized by early onset of maternal hypertension and proteinuria. New findings indicate that arginine vasopressin (AVP) may be a contributing factor to ignite PE. The aim of this study was to identify if there is any correlation between arginine vasopressin promoter polymorphisms and PE. STUDY DESIGN: Venous blood samples of 100 PE and 100 normal pregnant women were obtained for DNA extraction to identify the polymorphisms of AVP promoter by RFLP and nested-PCR techniques. MAIN OUTCOME: rs3729965 polymorphism of PE women was detected to have significant correlation with body mass index (BMI) (P = 0.028). RESULTS: Statistical analysis of three polymorphisms namely rs3729965, rs61138008 and rs3761249 of preeclamptic women (PEW) and none preeclamptic pregnant women (NPEW) revealed that rs3729965 genotypic distribution was significantly different between both groups (P = 0.04). Further analysis revealed that rs3729965 CT genotype of PEW had significant correlation to their BMI (P = 0.028). CONCLUSION: Polymorphic variants located on the promoter region of AVP are associated with PE. Thus we hypothesize that allelic variation may have a role in increasing the risk of developing PE.

Immunomodulatory Effects of Human Adipose Tissue-derived Mesenchymal Stem Cells on T Cell Subsets in Patients with Rheumatoid Arthritis.

Adipose-derived mesenchymal stem cells (Ad-MSCs) have been reported to suppress the effector T cell responses and have beneficial effects on various immune disorders, like rheumatoid arthritis (RA). This study was designed to investigate the effects of co-cultured Ad-MSCs on peripheral blood mononuclear cells (PBMCs) of RA patients and healthy individuals, through assessing transcription factors of T cell subsets. PBMCs from RA patients and healthy donors were co-cultured with Ad-MSCs with or without Phytohaemagglutinin (PHA). The quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression of T-box 21 (T-bet), GATA-binding protein-3 (GATA3), retinoid-related orphan receptor Î³t (ROR-γt) and forkhead box P3 (Foxp3). Based on the results, Ad-MSCs greatly upregulated Th2 and Treg cell transcription factors, i.e., GATA3 and Foxp3 (p<0.05), and downregulated Th1 and Th17 transcription factors, i.e., T-bet and RORγt (p<0.05). These results demonstrate that Ad-MSCs can result in an immunosuppressive environment through inhibition of pro-inflammatory T cells and induction of T cells with a regulatory phenotype. Therefore, they might have important clinical implications for inflammatory and autoimmune diseases such as RA.

Kaempferol increases apoptosis in human cervical cancer HeLa cells via PI3K/AKT and telomerase pathways.

Cervical cancer is one of the most frequent cancers in women worldwide. Defects in the apoptotic pathways are responsible for both the disease pathogenesis and its therapy resistance. It is thus a good candidate for treatment by pro-apoptotic agents. Kaempferol as a flavonoid has antioxidant and anti-tumor properties. Kaempferol has been shown to induce apoptosis and cell death in cancer cells. However, due to the problems in the treatment of cervical cancer, this study is designed to investigate the molecular mechanism by which kaempferol suppresses the growth of cervical cancer HeLa cell as compared with HFF cells (normal cells). Cells treated with kaempferol (12-100µM) and 5-FU (1-10µM), as the positive control, up to 72h. Cell viability was determined by MTT assay and real time PCR was used to investigate apoptosis and telomerase genes expression. The results showed that kaempferol decreased cell viability as concentration- and time-dependently. IC50 values were 10.48µM for HeLa and 707.00µM for HFF cells, as compared with 1.40µM and 16.38µM for 5-FU after 72h treatment, respectively. Also, kaempferol induced cellular apoptosis and aging through down-regulating the PI3K/AKT and hTERT pathways. This study suggests that kaempferol may be a useful adjuvant therapeutic agent in the treatment of cervical cancer.