Role of MTHFR 677C>T and 1298A>C gene polymorphisms on renal toxicity caused by lead exposure in wastewater treatment plant workers.

Environmental and occupational lead (Pb) exposures continue to pose major public health problems. Wastewater treatment plant (WWTP) workers proved are exposing to high Pb concentrations in sludge departments. The aim of the work was to investigate the role of MTHFR C677T and MTHFR A1298C gene polymorphisms on alteration of oxidative stress and homocysteine levels in WWTP workers exposed to high Pb concentrations, and study its relations with renal functions. The study included 90 WWTP workers from Abu-Rawash WWTP. Homocysteine, creatinine, urea, malondialdehyde (MDA), and total antioxidant capacity (TAC) were measured. Polymorphisms of MTHFR C677CT and MTHFR A1298C genes were studied using PCR/RFLP. Urine Pb concentrations were also measured. About 32.2% of the workers were with detectable Pb levels. Pb, homocysteine, and MDA levels were significantly higher among workers carrying TT polymorphism compared to other MTHFR C677T gene polymorphisms, while TAC was significantly lower among them compared to other polymorphisms. The same results were found among workers carrying CC compared to other MTHFR A1298C gene polymorphisms. WWTP workers carrying MTHFR 677TT and MTHFR 1298CC are more susceptible to elevation of homocysteine and the urinary Pb compared to the workers with the other polymorphisms. Furthermore, those workers were found to have increase in urea and creatinine. Therefore, MTHFR C677T and MTHFR A1298C gene polymorphisms could be used for prediction of the susceptibility to the risk of kidney impairments among WWTP workers in the sludge departments caused by their exposure to high Pb in their workplace.

A Clinical Evaluation of Circulating MiR-106a and Raf-1 as Breast Cancer Diagnostic and Prognostic Markers.

OBJECTIVE: MicroRNAs (MiRNAs) regulate mammalian cell growth, differentiation, and apoptosis by altering the expression of other genes and serve multiple roles in tumorigenesis and progression. Proto-oncogene serine/threonine-protein kinase (RAF-1) functions as a part of the MAPK/ERK signal transduction pathway. The present study aim was to prospectively evaluate MicroRNA 106a (MiR-106a) and RAF-1 as a diagnostic and prognostic factor in early prediction of breast cancer (BC), recurrence and early detection of distant metastasis as well as to analyses the statistical correlation between MiR-106a and RAF-1 levels and clinical-pathological parameters including tumor size, lymph node, histological type and grading. METHODS: Sera and plasma of 30 normal women and 50 women with breast carcinoma were assayed for MiR-106a by RT-qPCR as well as levels of Hb, WBCs and platelets count and RAF-1 by solid phase enzyme-linked immunosorbent assay (ELISA). RESULTS: The patients' characteristics, they were classified according to grade into 8% grade I, 66% grade II, 22% grade III and 4% grade IV. The stages were classified according to the TNM system as stage II was the highest percentage 66%, while the lowest percentage was 10% for stage I and 24% for stage III. Also, Hb% and RAF-1 levels were significantly decreased in breast cancer patients as compared with healthy control. On the other hand, MiRNA-106a gene expression was non-significantly increased in positive lymph node metastasis patients (FC=3.66) when compared to patients with negative lymph node metastasis (FC=3.51). In addition, MiR-106a was significantly up-regulated in breast cancer patients with a fold of change 3.63 when compared to control samples. CONCLUSION: Expression of MiR-106a gene can be used as a diagnostic and prognostic noninvasive biomarker which can stimulates breast cancer cell invasion and proliferation through downregulation of Raf-1 levels.
.

Assessment of the Anti-Cancer Efficiency of Silver Moringa oleifera Leaves Nano-extract against Colon Cancer Induced Chemically in Rats.

BACKGROUND: Colorectal cancer (CRC) categorized as the most common type of gastrointestinal cancers affected both genders equally. Chemotherapeutic drugs became limited due to their deleterious side effects. Therefore, efficiency of M. oleifera leaves extract increased by incorporating silver nanoparticles (Ag-NPs) then studied against colon cancer induced by azoxymethane (AOM) in rats. METHODS: Different hematological and biochemical measurements in addition to specific tumor and inflammatory markers were quantified. Histopathological examination for Colonic tissues was performed. Native proteins and isoenzyme patterns were electrophoretically detected in addition to assaying expression of Tumor Protein P53 (TP53) and Adenomatous Polyposis Coli (APC) genes in colonic tissues. RESULTS: M. oleifera nano-extract restored levels of the hematological and biochemical measurements in addition to levels of tumor and inflammatory markers to normalcy in both of nano-extract simult- and post-treated groups. Also, it minimized severity of the histopathological alterations in the simult-treated group and prevented it completely in the post-treated group. The lowest similarity index (SI%) values were noticed with electrophoretic protein (SI=61.54%), lipid (SI=0.00%) and calcium (SI=75.00%) moieties of protein patterns, catalase (SI=85.71%), peroxidase (SI=85.71%), α-esterase (SI=50.00%) and ß-esterase (SI=50.00%) isoenzymes in addition to altering the relative quantities of total protein and isoenzyme bands in colon of cancer induced group. Moreover, levels of TP53 and APC gene expression increased significantly (P≤0.05) in colon cancer induced group. The nano-extract prevented the qualitative and quantitative alterations in the different electrophoretic patterns in addition to restoring levels of the gene expressions to normalcy in both of simult- and post-treated groups. CONCLUSION: M. oleifera nano-extract exhibited ameliorative effect against the biochemical, physiological and molecular alterations induced by AOM in nano-extract simult- and post-treated groups.
.

Clinical Potentials of miR-576-3p, miR-613, NDRG2 and YKL40 in Colorectal Cancer Patients.

INTRODUCTION: Colorectal cancer (CRC) is the most common type of gastrointestinal tract cancers. This investigation aim was to assess the expression of miR-576-3p and miR-613 in CRC patients in addition to NDRG2 and YKL40 serum levels determination to decide their diagnostic and prognostic significance. METHODS: Sixty early diagnosed CRC patients prior to any treatment in addition to twelve healthy subjects were enrolled in this study. Blood samples were taken from subjects and allowed for clotting and centrifugation, then the collected sera were stored at -80ºC till it were used for detection of our molecular biomarkers. The mature miRNAs expressions (miR-576-3p and miR-613) were detected in serum by qRT-PCR, while NDRG2 and YKL40 serum levels were determined by ELISA. In addition, the correlation of the measured parameters with the clinicopathological data of the patients was investigated. RESULTS: The study results showed that both miRNA-576-3p and miRNA-613 were down-regulated in CRC patients with fold change 0.33, 0.36; respectively. A significant positive correlation was observed between miR-576-3p and miR-613 (r = 0.75, p < 0.001). NDRG2 serum levels were decreased in patients compared to the control group but the decrease wasn't statistically significant. On the other hand, it was observed that YKL40 serum level was significantly increased in CRC patients compared to control (p-value < 0.001). Furthermore, YKL40 showed a very high diagnostic value (AUC = 0.97, specificity = 91.7%, sensitivity = 96%, p-value = 0.0001). CONCLUSION: The observations of this investigation concluded that, the expressions of miR-576-3p and miR-613 in addition to YKL40 serum levels determinations may help in the diagnosis of CRC.

Expression and clinical significance of SNAI1 and ZEB1 genes in acute myeloid leukemia patients.

Acute myeloid leukemia (AML) is the most common type of acute leukemia in adults, it represents nearly 32% of all new cases of leukemia. This study aimed to evaluate the SNAI1 and ZEB1 genes expression in AML patients and determine their diagnostic and prognostic significance. We determined the expression of SNAI1 and ZEB1 genes and serum E-cadherin levels in early diagnosed patients with AML. Sixty early diagnosed AML patients and 20 healthy subjects were enrolled in this study, SNAI1 and ZEB1 genes expression was determined by Real-time PCR while E-Cadherin serum levels were determined by ELISA. The results of this study demonstrated that, all AML patients positively expressed the SNAI1 gene with fold change 2.6. While, the ZEB1 expression was positive in 56.7% of the patients with fold change 1.8. SNAI1 and ZEB1 genes were highly expressed in M5 subtype (FC = 13.8 and 9.3, respectively). On the other hand, serum E-cadherin concentrations of the AML patients showed decrease when compared with those of the control but the decrease was not reach to the significance level. The findings of this study suggest inclusion of SNAI1 and ZEB1 genes expression in the cluster of potential genetic biomarkers to be studied in AML cases as diagnostic and prognostic markers.

Improving the catalytic, kinetic and thermodynamic properties of Bacillus subtilis KU710517 milk clotting enzyme via conjugation with polyethylene glycol.

Milk clotting enzyme (MCE) produced by Bacillus subtilis KU710517 was conjugated to several activated polysaccharides. Among all the conjugates, the enzyme conjugated with polyethylene glycol (PEG) exhibited the highest retained activity (551U/mg protein) with a recovered activity of 95.3%. The activation energy of PEG-conjugated enzyme was calculated as 24.56kJ·mol-1which was lower than that of the native one (29.27kJ·mol-1) however, the temperature quotient (Q10) was about 1.08 for the two forms of the enzyme. The calculated half-life times of PEG-conjugated enzyme at 55 and 60°C were 317.78 and 128.6min respectively, whereas at the same temperatures the native enzyme had lower half-life times (53 and 19.6min respectively). The data of thermodynamic analysis for substrate catalysis including the specificity constant (Vmax/Km), turnover number (kcat), catalytic efficiency (kcat/Km), enthalpy of activation (ΔH*), free energy of activation (ΔG*), free energy for transition state formation ΔG*E-T and free energy of substrate binding ΔG*E-S were determined for both native and PEG-conjugated enzyme. In addition, the thermodynamic parameters for irreversible inactivation (ΔH, ΔG, ΔS) were evaluated. The calculated results indicated that the catalytic properties after the PEG-conjugation were significantly improved.