NEW CELLULAR & MOLECULAR BIOTECHNOLOGY JOURNAL
Year:2012 | Volume:2 | Issue:8|Papers Count:14

Mycoplasmataceae family found in different hosts such as humans, animals, plants and insects. Mycoplasma are one of the smallest and simplest bacteria, which have a free life and most of them are normal flora of male and female, moreover they are pathogens species. Genital mycoplasma and ureaplasma infections can lead to infertility. Sticking ability of these microorganisms to epithelial cells of urogenital system, sperm and erythrocyte make them able to establish infection in the human urogenital tract. These infections are often asymptomatic and will have pathological effects on reproductive health. The influence of infection on the volume, PH, motility, morphology and sperm count in men and pass through the birth canal in women is cause of infertility factors. There are a variety of statistics from different regions of the world about the role of infections in infertility since 39 percent in developed to 85 percent in African countries. In additional, according to existing reports, the prevalence of Ureaplasma in infertile couples (42 to 95 percent) is more than fertile couples (23 to 25 percent). In spite of lack of diagnosis, prevention and treatment, these infections remain continuously and ultimate to a risky consequences such as pelvic inflammatory disease (PID) and infertility. Therefore, the identification of these bacteria in asymptomatic infertile men and women is important and microbial screening for infertile couples, especially in young age should be essential.

Aim and Background. Stem cells have self-renewal potential and differentiating ability into adult somatic cell types. In this study the mesenchymal stem cells of human umbilical cord Wharton’s jelly were isolated. After proliferation, they were differentiated into blood cells via treating them by rat lung tissue extract which is rich in G-CSF, M-CSF, GM-CSF, IL-6 cytokines essential for various steps of differentiation.Materials and Methods. Human umbilical cord Wharton’s jelly tissue was cut into 0.5 cm, and then cultured in T25 flask. After second passage, mesenchymal stem cells were treated with continuous lung tissue extract for differentiation. The cells cultured in 1: 3, 1: 1and 3: 1 experimental volume groups of DMEM: extract ratio.Results. Blood cells were observed at 1: 3, 3: 1 and 1: 1 DMEM: extract experimental groups on the seventh day. Mesenchymal stem cells were confirmed by flow cytometric analysis (CD90, CD105). Positive cells were considered as mesenchymal stem cell in human Wharton’s jelly. Microscopic analysis of cells after Giemsa staining verified the presence of blood cells.Conclusion. Human Wharton’s jelly mesenchymal stem cells were isolated by tissue culture methods. Also mouse cytokines could selectively induce human stem cells to adult human blood cells.

Aim and Background: Due to evolving in stress habitats, Halophyte plants are the best options to describe the fundamentals of molecular physiology of salt tolerance and breeding of crops. Therefore, because of genetic relationship of Aeluropus littoralis with rice, wheat and barley, the present study was performed to investigate leaves proteome pattern changes in response to different salt treatment using proteomics methods Materials and Methods. To examine the effect of salinity on the Aeluropus proteome pattern, salt treatment in 3 levels (100, 200 and 300 mM NaCl) was applied. The mature leaves located in the middle part of plant were harvested for protein extraction after 21 days of salt exposure. Proteins were extracted according to TRIZOL approach and were separated by 2-DE using a nonlinear pH 4-7 IPG strips (24 cm) and 12.5 % poly acryl amide gel.Results: Statistical analysis revealed that among 550 repeatable detected spots, at least 95 protein spots showed a significant change during salinity levels. In addition, the most proteins with increased and decreased expression levels were obtained in comparison between control and 200 mM NaCl levels and between 200 & 300 mM NaCl treatments, respectively. Based on cluster analysis the whole responsive proteome was classified into 10 classes.Conclusion: Evaluation of these co-expression classes revealed that A. littoralis salinity responsive proteome follows four distinct expression patterns. These results showed that study of individual protein expression changes alone will not be fruitful. But also the identification of co-expression proteins collection and study of proteins gregarious changes pattern in response to different levels of stress was more importance and create better understanding.

Aims and Background: Cardiovascular complications are the main cause for much mortality among diabetics. Many studies show that cardiovascular diseases are developed after intravascular thrombosis in type 2 diabetics (T2D). Our previous studies show a significant hypercoagulable state and a prolongation in coagulation pathways. The current study tends to study the reasons of prolonged coagulation cascade in T2D patients and the possible relations between calcium and other minerals in blood plasma of T2D patients.Materials and Methods: In the present study, sixty T2Ds without any acute and chronic disorders were selected randomly and compared with 30 healthy subjects. In relation to the results of both groups, the following factors were studied: FBS, TG, calcium (Ca), Na, K, phosphate ions (PO4-3) and specific activity of coagulation factors: I, II, V, VII, VIII, IX, X, and XI with PT and APTT. All these factors were examined using laboratory methods and analyzed through Pearson correlation.Results: The obtained results show that the factors of FBS, TG, II, IX, X and XI increase significantly in T2Ds in comparison with healthy subjects. The T2Ds show hypocalcaemia and hyperphosphatemia in comparison with healthy subjects. Also, the results show that APTT is prolonged in T2Ds and the Pearson correlation coefficient does not show a significant correlation between Ca, Na, K, PO4-3 and coagulation factors in T2Ds.Conclusion: Hypercoagulable state, hypocalcaemia and hyperphosphatemia in the T2Ds suggest that heterogeneity has existed even in early stages in T2Ds. Although, there is an increase in hypercoagulable specific activities, the hypocalcaemia and its regulatory effect on the coagulation process might be considered as the reason for APTT prolongation. On the other hand, the hypocalcaemia and its compensatory mechanism cause hypercoagulable in order to compensate for the ineffectiveness of the coagulation process. Therefore, a sudden rise in plasma calcium concentration may increase the risk of intravascular thrombosis and cardiovascular complication in diabetics.

Aim and Background: The importance of effective parameters on biosorption of Cu and Mn ions from Sarcheshme Copper Mine Acid Mine Drainage (AMD) was investigated using regression analysis.Materials and Methods: Bacillus Thuringiensis was isolated from Sarcheshme AMD water and sorption parameters of that AMD water were investigated.Results: Correlation between amount of biosorbent and amount of Cu removed (0.99) was greater than the correlation between amount of biosorbent and Mn removed (0.98) from wastewater. Furthermore, increasing pH increased Cu and Mn ions removal up to 19 and 9 times, respectively. It was also observed that increasing contact time raised the Cu and Mn uptake 4.83 and 1.5 times, respectively.Conclusion: Results showed that parameters like amount of biosorbent, pH, temperature and contact time have the most impact on amount of biosorption respectively and variation of effective parameters have more effects on Cu removal from wastewater.

Aim and Background: The toxic effects of cadmium on growth parameters and cell division in Meristem of different plants have been demonstrated. However, the molecular mechanisms of this process are not completely understood until now. The aim of this study was to investigate the effect of cadmium on the expression of CDK-A and cyclin B1 genes in root tip of wheat seedlings for the identification of molecular cadmium limiting mechanism.Materials and Methods: Treated wheat root tips with 0, 5, 10, 15, 20, 30, 60, 90 and 120 mg/l concentrations of cadmium used for RNA extraction and RT-PCR with primers specific for cyclin B1 and CDK-A genes.Results: Resuls showed that expression of cyclin B1 gene at cadmium concentrations ranging from 5-15mg/l was invariant. But in higher concentrations (more than 15 mg/l) limiting effect of cadmium was quite clear. Also expression of CDK-A gene at 60, 90 and 120 mg/l concentrations of cadmium affected by limiting effect of cadmium.Conclusion: The decrease in root and coleoptiles growth of wheat seedlings treated with different concentration of cadmium depends on the expression of cyclin B1 and CDK-A genes and other unknown factors.

Aim and Background: Bacteria of the species Bacillus pumilus produce a large number of biological compounds which are active against bacteria and fungi. This study was designed with the aim of isolation, screening and optimization of antimicrobial production of Bacillus pumilus against four pathogenic microorganisms.Materials and Methods: For the initial screening of antimicrobial activity against four pathogenic microorganisms, agar disk diffusion method and TSB medium and for the production of antimicrobial compounds, synthetic medium and agar well diffusion method is used. To obtain the maximum production of antimicrobial compounds, pH (6-9), incubation period (0-72 hours) and glucose concentration (1-5%) are well considered. Data were analyzed by SPSS and ANOVA tests.Results: Maximum production of antimicrobial compounds by standard and isolated strain of B. pumilus was observed after 48 hours of incubation at 37oC, pH 7 and 3% glucose against four microbial strains.Conclusion: B. pumilus species might be a potential candidate for the production of antimicrobial compounds in the future.

Aim and Background: The purpose of this research is to have increased level of stability and fatality rate of cisplatin by application of encapsulation process in nanoarchaeosome. Success of this research is proved by examining the results obtained from MTT tests on Breast cancer cells in MCF-7 cell line.Materials and Methods: After cultivation of MCF-7 cell line in culture room, fatality rate of encapsulated cisplatin was compared with normal cisplatin by performing the related MTT tests. Output numbers from Elisa Reader explained the death rate of cancer cells.Results: During the MTT tests performed on MCF-7 cell line, effect rate of encapsulated cisplatin was compared with normal cisplatin. Examining the figures resulted from Elisa Reader output numbers, shows that stability and fatality rate of encapsulated drug is extremely higher than normal drug.Conclusion: Nanoarchaeosome is used for encapsulating of cisplatin. This process causes increasing the stability of drug in addition to the rise of fatality rate. This was clearly shown by examining the Breast cancer cells in MCF-7 cell line.

Aim and Background: Metalloproteases have numerous industrial applications such as peptide and aspartame synthesis. However, their application is limited by instability as a result of autolysis.. In the present study, following purification, biochemical properties of a metalloprotease (elastase) from Pseudomonas aeruginosa have been investigated.Materials and Methods: Recombinant protein has been purified from E. coli by affinity chromatography. Temperature dependence of enzyme activity and optimum temperature has been measured using a plot of enzyme activity versus different temperatures. Optimum temperature of enzyme activity has been determined within a mix buffer solvent by activity measurements at different pHs, and the inactivation rate, t1/2 and thermodynamic parameters of inactivation have been determined using irreversible thermoinactivation.Results: Optimum temperature was 60oC in the presence of 0, 5 and 10 mM CaCl2, and optimum pH was 8.5. Inactivation rate was increased with elevation of temperature from 55 to 80oC. Half-life of thermion activation (t1/2) was 150, 112, 47, 32, 18, 8 and 4 min at 55, 60, 65, 70, 75 and 80oC, respectively. Ea,  DH#, DS# and DG# was 37.70, 37.02, 23.61 kcal/mol, respectively, and k inactivation was 4.49´103 s-1.Conclusion: Pseudomonas aeruginosa elastase can be considered between thermophilic and mesophilic enzymes. Optimum temperature did not alter in different calcium concentrations which imply that elastase stability is Ca-independent.

Aim and Background: CEL I, is isolated from celery and the first eukaryotic nuclease is known to have cleavage DNA with high specificity at sites of the two DNA strands in a heteroduplex DNA molecule. The aim of this study was cloning, expression and purification CEL I endonuclease and a simple method for the detection of mutations.Materials and Methods: CEL I mRNA was used for RT-PCR. CEL I cDNA was synthesized and cloned into T-vector pTZ57R, then CEL I subcloned into pET32a expression vector. Recombinant plasmid was transformed into BL2I bacterial cells. Expression of recombinant plasmid was analyzed by western blot technique. Protein was purified by affinity chromatography. In first study, Recombinant enzyme was active in mutation detection in Product PCR normal and mutant BRCA1 gene.Results: The recombinant (pET32a/CEL I) was successfully expressed in BL2I cells. Western blot analysis showed that the successfully expressed CEL I in the cells transfected. The first study showed that recombinant enzyme is known to nick mismatch sites of the two DNA strands in a heteroduplex DNA molecule.Conclusion: The expression of CEL I in BL2I cells was strong. In vitro, purification protein was successful. Recombinant nucleases CEL I known to nick mismatch sites in a heteroduplex DNA molecule.

Aim and Background: Peroxidases are one of the most important enzymes participating in many physiological processes in plants. Saffron, (Crocus sativus L), is a valuable plant which has been grown since ancient times to date. But there is a lack of sufficient biochemical and physiological information about the stages of the saffron growth, development and the important enzymes involved. Peroxidases in Crocus sativus L. can be a marker of distinct physiological ordevelopmental processes, as shown before by laboratory studies.Materials and Methods: In this study, the Crucus Sativus L. corms in two different months –July and November as dormant and waking stages of saffron growth, sampled and Corms were used for the enzyme extraction. It was partially purified with ion-exchange chromatography.Results: The results showed that amount of total protein in waking corms is about two times in comparison with the dormant corms. The high activity of the plant in waking state corresponds with more protein content.Conclusion: This difference might be partially due to the different patterns of the enzymeglycosylation and surface electrostatic charges.

Aims and Background: The development of prostate cancer is dependent on heredity, androgenic influences, and exposure to environmental agents. The carcinogenic compounds require biotransformation by metabolizing enzymes and carcinogenesis of the prostate involves the associations between genetic polymorphisms of different metabolizing enzymes and prostate cancer. CYP2D6 belongs to the cytochrome P450 super family of enzymes and plays an important role in the metabolism of clinically used drugs including antidepressants. The polymorphic cytochrome P450 (CYP) enzymes are of particular interest in prostate cancer susceptibility because of their ability to metabolize both endogenous and exogenous compounds, including dietary constituents.Materials and Methods: Association between CYP2D6 gene polymorphisms and prostate cancer was studied in a case-control study on North Indian population comprising 100 patients and an equal number of age-matched control men. For this purpose, the PCR products were exposed to restriction enzyme through RFLP method.Results: Relative to men homozygous for the wild-type allele in CYP2D6 gene, those heterozygous for the B allele had an odd ratio of 1.78 (95% CI, 0.76-4.17, P=0.18) for patients, and for homozygous individuals, it was 1.95 (0.55-6.93, P=0.30).Conclusion: No association was observed between B allele of CYP2D6 gene polymorphism and prostate cancer risk.