Isolation and characterization of a novel denitrifying bacterium with high nitrate removal: Pseudomonas stutzeri

The aim of this study was to isolate and characterize a high efficiency denitrifier bacterium for reducing nitrate in wastewater. Six denitrifier bacteria with nitrate removal activities were isolated from a petrochemical industry effluent with high salinity and high nitrogen concentrations without treatment. The isolated bacteria were tested for nitrate reomoval activity. One of the bacterium displayed the highest reduction of nitrate. The strain was preliminarily identified using biochemical tests and further identified based on similarity of PCR-16S rRNA using universal primers. Biochemical and molecular experiments showed that the best bacterium with high nitrate removal potential was Pseudomonas stutzeri, a member of the alpha subclass of the class Proteobacteria. The extent of nitrate removal efficiency was 99% at 200 mg/L NO[3] and the nitrite content of the effluent was in the prescribed limit. The experiments showed the ability of Pseudomonas stutzeri to rapidly remove nitrate under anoxic conditions. The strain showed to be potentially good candidate for biodenitrification of high nitrate solutions

[Effect on alpha hemoglobin chain expression in K562 cell line]

MicroRNAs [miRNA] are small noncoding RNA molecules that transcribed by RNA polymerase II. After biogenesis, these molecules act by incorporation into the RNA-induced silencing complex [RISC]. MiRNAs are involved in multiple physiological and pathological processes such as proliferation, differentiation, apoptosis and cancer. Recently several studies reported down regulation of mir-150 during erythropoesis. Since hemoglobin expression is valuable indicator of erythroid differentiation we evaluated the mir-150 downregulation effect on alpha chain expression by Quantitative RT-PCR. K562cells were grown in RPMI1640 in standard condition. K562 cells were transfected by microRNA 150 Inhibitor using transfection kit .Mir-150 downregulation was confirmed by miRNA Real time PCR, followed by Q-RT-PCR to investigate the alpha chain expression changes. By relative QRT-PCR the alpha chain expression was increased 10 folds in comparison to untransfected and scramble cells. Furthermore, the differences were statistically significant [P<0.05] Elevation of alpha chain expression in our study showed that mir-150 downregulation has a crucial role in erythroid differentiation and can introduce as a novel marker in alpha thalassemia. Further researches to find out the detail mechanism and miRNAs genes target could improve our knowledge about miRNAs potential in management of diseases and their applications in gene therapy and regenerative medicine

[Differentiation of somatic unrestricted cord blood stem cells into chondrocyte cells]

To isolate and purify unrestricted somatic stem cells from human umbilical cord blood and evaluation of their differentiation into chondrocyte in vitro. In this study cells from human umbilical cord blood were isolated and plated in flask. Colonies were performed after one week. To determine the kind of cells, 100000 cells were analysed with flowcytometry. Twenty thousands [200000] of cells were plated in 6 wells that coated with poly-L-lysine II and incubated in chondrogenic medium to analyze the differentiation of these cells into cartilage. After 24 hs the first pletted cells were formed that continued to be existing to 21 days. The culture of cells were exchanged into chondrogenic culture every two days. At the end of differentiation period and 3 weeks the cells were analyzed by Alcian blue, immunohistichemistry and RT-PCR In addition these cells were passaged 50 times and their karyotyping analyzed. In early days of primary cultures, the number of spindle cells were increased and almost purified in second passage. The differentiation by RT-PCR analysis showed high production of collagen II, aggrican, BMP-6 and collagen that all are the specific genes of chondrocye cells, and.histochemistry assay showed that the methachromatic matrix was accumulated between the cells and expression of collagenll was confirmed. Karyotyping analysis showed high passages for these cells that was expected. Cultured USSC Differentiated into a chondroblast cell linage potential source for cell transplantation for rheumatoid arthirits as soon as cord blood is better source for mesenchymal Stem cells against bone marrow

Detection of N-RAS gene mutations in codons 12, 13 and 61 in patients with acute myeloid leukemia

Acute myeloid leukemia [AML] comprises a heterogenic group of malignant disorders involving cell maturation arrest at an undifferentiated stage in bone marrow. Activation of N-RAS proto-oncogene due to point mutations plays a major role in AML malignancy. Since there was no report on the frequency of N-RAS gene mutations in Iranian AML patients, therefore, we decided to determine its frequency and compare the results with age, sex and FAB subtypes. In this descriptive study, 60 de novo AML patients from Tehran Shariati hospital, hematology-oncology and bone marrow transplantation center were screened for the mutations of N-RAS gene at codons 12, 13 and 61. DNA was extracted from peripheral blood samples before the start of chemotherapy. The above mentioned codons were amplified by PCR and analyzed by restriction endnuclease enzymes. We were able to detect mutations in 12 out of 60 [20%] patients. Most of the mutations were detected in men with an age over 40 years old. The frequency of mutations for codons 12, 13 and 61 were 15%, 11.6% and 5% respectively. Most of the mutations [33.3%] were found to happen in AML-M4 FAB subtype. We could not detect any mutation in AML-MO, M6 and M7. We detected mutations in 20% of our AML patients. In general, the frequency of the mutations we found was in agreement with the results of other studies. However, a study with more patients and a wider range of age using a combination of PCR-RFLP and direct gene sequencing is highly recommended

[Comparison of serum leptin in major beta -talasemia patients and normal subjects]

Leptin, is a adipocyte-derived hormone. Exogenous leptin allows the recovery of the reproductive function. In humans, leptin correlates positively with body mass index [BMI]. The aim of the study was to investigate the association of leptin with toxic effects of iron overload. In a cross sectional study in 2006, we compared the serum leptin level of thalasemic patients with normal group. Blood samples were collected from 219 patients with Cooley's anemia, [119 males, 100 females] and 137 normal subjects [86 males, 51 females]. Leptin was measured by a commercial ELISA kit. Data were analyzed by SPSS software. Mean serum leptin level was 5.33 +/- 5.02 ng/ml in thalassaemic males. It was significantly lower than controls [9.43 +/- 7.8 ng/ml] [P<0.001]. Thalassaemic females had lower leptin levels [12.12 +/- 11.4 ng/ml] than normal females subjects [14.6 +/- 13.1 ng/ml] [P<0.001]. Furthermore, the physiologically positive BMI/leptin relationship disappeared in thalassaemic patients. It seems that the adipocytes of thalassaemic patients are unable to maintain adequate leptin production. These results suggest that adipose tissue dysfunction can be considered as one of the endocrinepathies affecting thalassaemic patients