EXPRESSION LEVELS OF FOXO-1, P27KIP1, MIR-27, MIR-186 AND AKT1/AKT-P PROTEINS IN WOMEN WITH ENDOMETRIAL CANCER AND HYPERPLASIA: IMPLICATIONS FOR THE HUMAN REPRODUCTIVE SYSTEM.

Objectives: Despite extensive research on endometrial cancer (EC) and endometrial hyperplasia, there is still a gap in understanding the molecular mechanisms underlying their development and progression. The aim of this study was to investigate the expression levels of FOXO-1, P27Kip1, miR-27, and miR-186, and Akt1, Akt-P proteins in patients with EC and endometrial hyperplasia compared to control subjects. Subjects and methods: Samples of the endometrial tumor (n=30), normal (control) (n=30) and endometrial hyperplastic (n=30) tissue were obtained from patients referring to Arash and Imam Khomani hospitals, Tehran, Iran. Expression levels of genes and microRNAs were evaluated by qRT- PCR. Western blot analysis was applied for protein evaluation. The data were analyzed using t-test, Mann -Whitney U, Pearson correlation coefficient analysis, ANCOVA and ANOVA. Results: There was significant decrease in FOXO-1 in EC tissue compared to control tissue (p<0.05). Significant increase was observed in expression of miR-27 in patients with EC (p<0.001) and hyperplasia (p<0.01), whereas miR-186 expression level increased significantly only in patients with EC (p<0.05). P27Kip1 expression level did not significantly change in patients with EC and hyperplasia. There was a significant association between expression levels of miR-27 with FOXO-1 and P27Kip1 in patients with EC. Western blot analysis revealed higher endometrial AKT1-P protein levels in patients with EC and hyperplasia than control subjects (p<0.05). Conclusions: Our findings suggest that FOXO-1, miR-27, miR-186, and Akt1-P/Akt1 protein have the potential to serve as tissue biomarkers for early diagnosis, prognosis, and progression of EC in the human reproductive system.

Helicobacter pylori infection and iron deficiency in patients with coronary artery disease.

The aim of this study was to investigate whether impact of the seropositivity to Helicobacter pylori (H pylori) infection on ferritin and iron levels is an independent risk factor for atherosclerosis in patients with cardiovascular disease. The anti H pylori IgG, IgA levels, serum ferritin and iron concentration of 86 patients with cardiovascular disease and 64 participants free of cardiovascular disease as control subjects were determined by ELISA assay. The results of present study showed that seropositivity to H pylori IgG and IgA levels of coronary artery disease (CAD) patients was higher than controls and CAD patients with negative anti H pylori IgG and IgA significantly. A significant negative correlation was found between seropositivity to H pylori IgG and IgA, ferritin and iron levels of CAD patients with seronegativity and seronegativity to H pylori IgG and IgA in comparison with controls. The achieved results from present study suggest that the involvement of H pylori infection in atherosclerosis process is based on the chronic inflammation which might facilitate the CAD-related pathologies. Moreover, impact of the presence of H pylori infection on reduction of the ferritin and iron levels of CAD patients as a risk factor independent of other classic factors including lipid profiles and inflammatory factors was remarkable.

Association of serum lipid indices and statin consumption with periodontal status.

OBJECTIVES: Periodontal and cardiovascular diseases share some common underlying mechanisms. Hyperlipidemia is a major risk factor for cardiovascular diseases. This study sought to assess the association of hyperlipidemia and statin consumption with periodontal status. MATERIALS AND METHODS: This cross-sectional study was conducted on 150 participants including 50 individuals with normal lipid profile (group C), 50 hyperlipidemic patients without drug therapy (group N), and 50 hyperlipidemic patients on drug therapy for a minimum of 3 months (group S). Periodontal parameters including plaque index (PI), clinical attachment level (CAL), bleeding on probing (BOP), and pocket depth (PD) were measured for all teeth except for the third molars. Serum levels of total cholesterol (TC), HDL, LDL, and triglycerides (TGs) were measured. RESULTS: The mean values of CAL and PD were significantly higher in the two hyperlipidemic groups compared with the C group (P < 0.005). Also, CAL and PD had significant associations with serum levels of TGs, LDL, and TC (P < 0.0001); PI in the group S was significantly lower than that in the other groups (P < 0.005). CONCLUSIONS: Hyperlipidemic patients showed higher values of periodontal parameters compared with the statin-treated and control groups. Lower PI in the group S may indicate the anti-inflammatory effect of statin.

The effect of Morus alba leaves extract and powder on resistin levels and liver transaminase enzymes activities in diabetes.

The current study was designed to investigate the changes of the resistin, aspartate aminotransferase (AST), alanine aminotransferase (ALT) levels of diabetic rats after treatment with Morus alba leaves flavonoid extract (MLE) and Morus alba leaves powder (MLP). Thirty male wistar rats in five groups including control and diabetic groups were included. Diabetic groups consisted of diabetic control, sham and treated group with MLE and MLP. Type 2 diabetes was induced in rats by administration of streptozotocin (STZ) and - nicotinamide. The serum concentrations of resistin and insulin in the study groups were identified by ELISA. ALT and AST activities were assayed by spectrophotometer. For the first time, it was shown that the uptake of MLE and MLP by diabetic rats could significantly decrease the serum fasting blood sugar (FBS), resistin levels and enzymes activity of ALT and AST and increases the concentration of serum insulin significantly (P<0.05). in comparison with the sham group and diabetic control. The results showed that there was no significant difference between the anti-diabetic and inflammatory properties of MLE and MLP. In this study, the possible protective effect of MLE and MLP administration was evaluated against destructive effect of STZ on liver and pancreas function in diabetic rats. The results showed that these effects may play an important role in the regulating of adipokines secretion such as resistin and insulin secretion which are involved in the control of diabetes and obesity. MLE and MLP treatment could be useful agents in combination with other therapies in diabetes improvement.

Human colon cancer HT-29 cell death responses to doxorubicin and Morus Alba leaves flavonoid extract.

The mechanistic basis for the biological properties of Morus alba flavonoid extract (MFE) and chemotherapy drug of doxorubicin on human colon cancer HT-29 cell line death are unknown. The effect of doxorubicin and flavonoid extract on colon cancer HT-29 cell line death and identification of APC gene expression and PARP concentration of HT-29 cell line were investigated. The results showed that flavonoid extract and doxorubicin induce a dose dependent cell death in HT-29 cell line. MFE and doxorubicin exert a cytotoxic effect on human colon cancer HT-29 cell line by probably promoting or induction of apoptosis.

Prevalence of beta lactamase producing species of pseudomonas and acinetobacter in pediatric burn patients.

Burn wound infection is a major cause of morbidity and mortality in burn victims. Pseudomonas and Acinetobacter species are among the most common organisms complicating burn wounds. Presence of extended spectrum ß-lactamase (ESBL) and metallo-ß-lactamase (MBL) genes plays an important role in spreading ß-lactam resistant strains of these organisms and is a serious condition in the treatment of the affected patients. As a result, we aimed to determine the prevalence of SHV, TEM, PER and VIM ß-lactamases in Pseudomonas and Acinetobacter species isolates from burn wound swabs of children with burn injury. In this descriptive observational study, 107 Pseudomonas and Acinetobacter isolates collected from burn patients were subjected to PCR assay. Using PCR method and DNA sequencing, the existence of SHV-, TEM-, PER- and VIM-type ß-lactamase encoding genes were determined. Out of the 107 Pseudomonas and Acinetobacter isolates, 66 (77.6%) were ESBL positive, 26.2% were positive for SHV gene, 37.4% were positive for TEM gene, 14% were positive for PER gene and 15.9% of them harbored VIM gene. More than half of the Pseudomonas and Acinetobacter strains in our pediatric burn unit harbor ß-lactamase encoding genes that make them resistant to a wide range of ß-lactam antibiotics. Consequently, it is suggested to choose an appropriate antibiotic regimen based on the antibiogram pattern of the strains.

Les infections cutanées sont une cause majeure de morbidité et de mortalité chez les brûlés. Pseudomonas et Acinetobacter sont parmi les micro-organismes les plus communs chez les brûlés. La présence des gènes codant les ß-lactamases à spectre étendu (BLSE) et métallo-ß-lactamases (MBL) joue un rôle important dans la dissémination des souches résistantes et obère le traitement des patients infectés. Nous avons étudié la prévalence des gènes encodant pour des enzymes des groupes SHV, TEM, PER et VIM dans des isolats de Pseudomonas et Acinetobacter chez les brûlés pédiatriques, grâce à des techniques de PCR. Dans cette étude observationnelle descriptive, 107 isolats de Pseudomonas et Acinetobacter, obtenus chez des patients brûlés ont été étudiés. Plus des 3/4 des souches de Pseudomonas et Acinetobacter expriment une BLSE (26.2% SHV; 37.4% TEM; 14% PER; 15.9% VIM), ce qui les rend résistants à de nombreuses ß-lactamines. Il est donc suggéré de choisir un traitement antibiotique approprié, basé sur l'antibiogramme des souches infectantes.

Fluoride concentration in potable groundwater in rural areas of Khaf city, Razavi Khorasan Province, northeastern Iran.

Long-term exposure to high concentrations of fluoride is associated with several adverse effects on human including dental and skeletal fluorosis. We studied all the groundwater wells located in rural areas of Khaf city, Razavi Province, northeastern Iran between 2009 and 2010. Fluoride concentration of water samples was measured by SPADNS method. We found that in rural areas the fluoride concentration ranged from 0.11 to 3.59 ppm-the level was less than the permissible limit in 31% of studied samples, higher than the permissible limit in 4% of the samples, and within the optimum limit of 1 to 1.5 ppm in 65% of water samples.

Angiotensin II induces NF-κB, JNK and p38 MAPK activation in monocytic cells and increases matrix metalloproteinase-9 expression in a PKC- andRho kinase-dependent manner

Angiotensin II (ANG II), the main effector of the renin-angiotensin system, is implicated in endothelial permeability, recruitment and activation of the immune cells, and also vascular remodeling through induction of inflammatory genes. Matrix metalloproteinases (MMPs) are considered to be important inflammatory factors. Elucidation of ANG II signaling pathways and of possible cross-talks between their components is essential for the development of efficient inhibitory medications. The current study investigates the inflammatory signaling pathways activated by ANG II in cultures of human monocytic U-937 cells, and the effects of specific pharmacological inhibitors of signaling intermediates on MMP-9 gene (MMP-9) expression and activity. MMP-9 expression was determined by real-time PCR and supernatants were analyzed for MMP-9 activity by ELISA and zymography methods. A multi-target ELISA kit was employed to evaluate IκB, NF-κB, JNK, p38, and STAT3 activation following treatments. Stimulation with ANG II (100 nM) significantly increased MMP-9 expression and activity, and also activated NF-κB, JNK, and p38 by 3.8-, 2.8- and 2.2-fold, respectively (P < 0.01). ANG II-induced MMP-9 expression was significantly reduced by 75 and 67 percent, respectively, by co-incubation of the cells with a selective inhibitor of protein kinase C (GF109203X, 5 µM) or of rho kinase (Y-27632, 15 µM), but not with inhibitors of phosphoinositide 3-kinase (wortmannin, 200 nM), tyrosine kinases (genistein, 100 µM) or of reactive oxygen species (α-tocopherol, 100 µM). Thus, protein kinase C and Rho kinase are important components of the inflammatory signaling pathways activated by ANG II to increase MMP-9 expression in monocytic cells. Both signaling molecules may constitute potential targets for effective management of inflammation.

Angiotensin II induces NF-κB, JNK and p38 MAPK activation in monocytic cells and increases matrix metalloproteinase-9 expression in a PKC- andRho kinase-dependent manner.

Angiotensin II (ANG II), the main effector of the renin-angiotensin system, is implicated in endothelial permeability, recruitment and activation of the immune cells, and also vascular remodeling through induction of inflammatory genes. Matrix metalloproteinases (MMPs) are considered to be important inflammatory factors. Elucidation of ANG II signaling pathways and of possible cross-talks between their components is essential for the development of efficient inhibitory medications. The current study investigates the inflammatory signaling pathways activated by ANG II in cultures of human monocytic U-937 cells, and the effects of specific pharmacological inhibitors of signaling intermediates on MMP-9 gene (MMP-9) expression and activity. MMP-9 expression was determined by real-time PCR and supernatants were analyzed for MMP-9 activity by ELISA and zymography methods. A multi-target ELISA kit was employed to evaluate IκB, NF-κB, JNK, p38, and STAT3 activation following treatments. Stimulation with ANG II (100 nM) significantly increased MMP-9 expression and activity, and also activated NF-κB, JNK, and p38 by 3.8-, 2.8- and 2.2-fold, respectively (P < 0.01). ANG II-induced MMP-9 expression was significantly reduced by 75 and 67%, respectively, by co-incubation of the cells with a selective inhibitor of protein kinase C (GF109203X, 5 µM) or of rho kinase (Y-27632, 15 µM), but not with inhibitors of phosphoinositide 3-kinase (wortmannin, 200 nM), tyrosine kinases (genistein, 100 µM) or of reactive oxygen species (α-tocopherol, 100 µM). Thus, protein kinase C and Rho kinase are important components of the inflammatory signaling pathways activated by ANG II to increase MMP-9 expression in monocytic cells. Both signaling molecules may constitute potential targets for effective management of inflammation.

Safety profile of morphine following surgery in neonates.

OBJECTIVE: To determine the effect of morphine on duration of mechanical ventilation, apnoea and hypotension among full-term neonates who underwent thoracic or abdominal surgery in a level III neonatal intensive care unit. METHOD: Medical records of 82 infants were reviewed retrospectively and data including patient demographics, clinical diagnosis, type of surgery, postoperative opioid administration, duration of mechanical ventilation, hypotension, apnoea and pain scores (premature infant pain profile (PIPP) score) were collected. RESULT: Sixty-two neonates (76%) received morphine following surgery as a continuous intravenous infusion during the postoperative period. Linear regression analysis showed that morphine dosage and duration were significantly associated with the duration of mechanical ventilation. An increase in morphine infusion rate by 10 microg kg(-1) h(-1) was associated with an increase in the duration of mechanical ventilation by 24 h (P<0.0001) and an increase in morphine duration of 1 hour was associated with a longer duration of mechanical ventilation by 38 min (P<0.0001). Logistic regression analysis showed no association between morphine infusion rate or duration and hypotension. Apnoea was not associated with morphine dosage or duration of infusion in neonates receiving morphine following extubation. Score on the PIPP correlated significantly with morphine infusion rate across time (r=0.47, P<0.01). CONCLUSION: Postoperative morphine dose and duration may prolong the duration of mechanical ventilation but there are no significant dose-dependent effects on other parameters including apnoea or hypotension following extubation in term neonates. More research is needed to determine the safety profile of morphine for management of pain in non-ventilated neonates.

Interaction of a low mobility group protein, LMG160, with deoxyribonucleic acid.

A fraction of low mobility group (LMG) nonhistone protein designated LMG(160) was isolated from rat liver chromatin by preparative gel electrophoresis and its interaction with DNA was studied using thermal denaturation and DNA-cellulose affinity chromatography techniques. The results showed that LMG(160) with an isoelecteric point of 5-5.5 was bound to DNA and decreased its melting temperature. Increasing ionic strengths decreased this effect. DNA-cellulose affinity chromatography showed the affinity of LMG(160) to double stranded DNA was higher than that to single stranded DNA, since it required 0.6 M NaCl for elution. The results suggest that LMG(160) protein preferentially binds to double stranded DNA destabilizes it and the binding is electrostatic.

Modeling complex disease with demographic and environmental covariates and a candidate gene marker.

We randomly chose replicates 28 and 29 of the simulated data sets of Genetic Analysis Workshop 12 to model the dependence of affection status on covariates, quantitative traits, and genes using all living pedigree members. First we explored the relationship of affection status to demographic and environmental factors using logistic regression and the Cox proportional hazards models. In the second stage of our analyses the generalized transmission disequilibrium test (GTDT) was applied to nuclear families with at least two affected siblings to select single markers and high-risk alleles, which were tested in the population association analyses including all pedigree members. Multiple logistic regression models were fitted to investigate the joint contributions of genetic and nongenetic factors and a block-recursive modeling approach was adopted to study inherent hierarchical dependence structure in the data. We found that allele 2 on marker 35 of chromosome 6 is associated with higher risk compared with the other 3 alleles of this marker. In addition to this significant genetic effect, age at exam and four of the five quantitative traits (QT1, QT2, QT4, and QT5) had a significant association with the disease. Our results were obtained without knowledge of the true disease generating models.

Evaluation of the contribution of environmental factors.

For the simulated data of GAW11, the roles of two environmental factors, E1 and E2, were investigated. Logistic regression analyses measuring the association between outcome (either mild or severe disease versus no disease) and E1 and E2 exposure indicated that E1 was a risk factor for disease (either mild or severe) but that E2 was not associated with outcome. Linkage analyses were performed for strata defined by E1 and E2 exposure. A specific disease locus was identified in these stratified analyses where this locus would not have been identified with an unstratified linkage analysis. Finally, stratified generalized transmission disequilibrium test analyses yielded several false positive results.

A comparison of some allele-sharing based linkage analysis methods for detecting complex trait loci.

Using randomly selected sib pairs from a subset of the GAW11 simulated data in Problem 2, we compared the results of some linkage analysis methods based on allele sharing. One method was the Haseman-Elston test for a binary disease outcome (unaffected vs. mild or severe). The other methods, which analyzed the trinary ordered outcome unaffected/mild/severe were the Haseman-Elston test, an extended Haseman-Elston incorporating sib-pair sums, variance components analysis, and regression analysis. Our analysis was done without knowledge of the generating model.