Production of monoclonal antibody against recombinant bovine sex-determining region Y (SRY) and their preferential binding to Y chromosome-bearing sperm

Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms.     

The response of wildtype and mutant cultivars of soybean to salt stress - comparing vegetative and reproductive phases on the basis of leaf biochemical contents,RWC, and stomatal conductance

Soybean, as a major oilseed crop, is cultivated in many parts of the world including saline-affected regions. A four-replicated factorial experiment based on a completely randomized design with five salinity levels (control, 50, 100, 150, and 200 mM NaCl) and five wild-type and new mutant cultivars of soybean (Williams, Clark, M-4, M-7, and M-9) was done at two vegetative and reproductive phases to study leaf oxidants, antioxidants and osmolytes under salinity stress. The vegetative phase demonstrated higher K⁺:Na⁺ ratio, cell membrane stability, relative water content (RWC) and stomatal conductance to H₂O (gs) accompanied with lower oxidative stress, antioxidants and osmolytes contrasting to reproductive phase. At both phenological phases, saline stress stimulated antioxidative defense owing to the enhancement of oxidative stress. The results indicated that mutants possessed a better antioxidative system than their wild-types.     

Association Study of rs1333040 and rs1004638 Polymorphisms in the 9p21 Locus with Coronary Artery Disease in Southwest of Iran

Background:

Coronary artery disease (CAD) is a multifactorial and heterogenic disease. Recently, genome-wide association studies have reported that rs1333040 (C/T) and rs1004638 (A/T) single nucleotide polymorphisms (SNPs) in the 9p21 locus have very strong association with CAD. This study aimed to examine these associations in Southwest of Iran.

Methods:

Blood samples were collected from 200 CAD patients and 110 healthy individuals with no CAD. The association of two SNPs with CAD was evaluated by PCR and restriction fragment length polymorphism.

Results:

Chi-square test showed no association between rs1333040 SNP and CAD (X²: 4.66, df: 2, P=0.09). Also, there was no association between rs1004638 SNP and CAD (X²: 0.27, df: 2, P=0.88).

Conclusion:

No association was observed between rs1333040 and rs1004638 SNPs in the 9P21 region and CAD in Southwest of Iran. Key Words: Coronary artery disease, Single nucleotide polymorphisms, Genetic association study, Iran     

Effect of vermicompost on some morphological,physiological and biochemical traits of bean (Phaseolus vulgaris L.) under salinity stress

Vermicompost can play an effective role in plant growth and development and also in reducing harmful effects of various environmental stresses on plants due to its porous structure, high water storage capacity, having hormone-like substances and plant growth regulators and also high levels of macro and micro nutrients. The aim of this study was to investigate the effects of vermicompost and salinity interactions on some morphological and physiological features and concentration of mineral elements of bean (Phaseolus vulgaris L. cv. Light Red Kidney) cultivar. A factorial experimental with five different volumetric ratios of vermicompost and sand, including to: 0:100; 10:90; 25:75; 50:50 and 75:25 and four levels of salinity [20, 40, 60 and 80 mmol l^?1 sodium chloride (NaCl)], and control was conducted base on Randomized Complete Block Design with three replications. Bean seeds were sowed in plastic pots, the seedlings being sampled 42 days old (flowering stage).The results showed that vermicompost had significant effect on all studied traits under stress and non-stress (p ≤ 0.05).In this experiment, the vermicompost significantly increased the photosynthetic rate and concentrations of potassium (K⁺) and calcium (Ca²)⁺in leaf and root tissues. In salinity levels of 20, 40 and 60 mmol l^?1NaCl, all subjected ratios of vermicompost and in 80 mmol l^?1NaCl the ratios of 10% and 75% vermicompost, significantly ameliorated negative effects of salinity. In both stress and non-stress conditions, using 10% volume of vermicompost is recommended to improve the growth of bean plants.     

Comparing invasive and non-invasive of isolated Shigella flexneri by electron microscopy of cell culture, SDS-PAGE and Congo red method

BACKGROUND: The aim of this study was to compare invasive and non-invasive strains of Shigella flexneri isolated from Tehran by a 120 kDa protein band by SDS-PAGE, electron microscopy of cell culture and Congo red dye methods. METHODS: S. flexneri strains were isolated by standard bacterial methods from fecal specimens of children attending to the 3 children's hospitals. Phenotype analysis for screening virulent of strains of S. flexneri was done on a plate of tryptic soy agar contained 0.003% Congo red dye. Whole membrane protein preparations were used to examine the protein profiles of the inner and outer membrane of these Gram-negative bacteria. The protein mixture was electrophoresed through a polyacrylamide gel. The gel was stained with Coomassie brilliant blue R250 and destained with ethanol and acetic acid. HeLa cell culture was done by two-step preparations: one for light microscopy and the other for electron microscopy. RESULTS: Some of S. flexneri (46%) were Congo red positive colonies. S. flexneri with negative Congo red phenotype could not enter the HeLa cell culture. A 120 kDa protein band was found in 46% of these bacteria which could enter into HeLa cell culture. Pseudopod structures which facilitate bacterial cell-to-cell spread were readily identified by electron microscopy. DISCUSSION: Since the existence of 120-kDa protein band was corresponded to enter of S. flexneri into the HeLa cell culture and correlated with Congo red dye positive, for identification of invasive and non-invasive S. flexneri strains, the use of a 120-kDa protein band by SDS-PAGE or a simple, rapid and very cheap Congo red dye method is recommended. Because, there are some deaths due to Shigella sp. in our country, notification on the isolation of these bacteria in both children hospitals laboratories and private clinical laboratories is important.     

Improved immunogenicity of Newcastle disease virus inactivated vaccine following DNA vaccination using Newcastle disease virus hemagglutinin-neuraminidase and fusion protein genes

The present study describes the development of DNA vaccines using the hemagglutinin-neuraminidase (HN) and fusion (F) genes from AF2240 Newcastle disease virus strain, namely pIRES/HN, pIRES/F and pIRES-F/HN. Transient expression analysis of the constructs in Vero cells revealed the successful expression of gene inserts in vitro. Moreover, in vivo experiments showed that single vaccination with the constructed plasmid DNA (pDNA) followed by a boost with inactivated vaccine induced a significant difference in enzyme-linked immunosorbent assay antibody levels (p < 0.05) elicited by either pIRES/F, pIRES/F+ pIRES/HN or pIRES-F/HN at one week after the booster in specific pathogen free chickens when compared with the inactivated vaccine alone. Taken together, these results indicated that recombinant pDNA could be used to increase the efficacy of the inactivated vaccine immunization procedure.     

Effects of the nitrification inhibitor nitrapyrin and tillage practices on yield-scaled nitrous oxide emission from a maize field in Iran

Nitrification inhibitors can effectively decrease nitrification rates and nitrous oxide(N₂O)emission while increasing crop yield under certain conditions.However,there is no information available on the effects of nitrification inhibitors and tillage practices on N₂O emissions from maize cropping in Iran.To study how tillage practices and nitrapyrin(a nitrification inhibitor)affect N₂O emission,a split factorial experiment using a completely randomized block design with three replications was carried out in Northeast Iran,which has a cold semiarid climate.Two main plots were created with conventional tillage and minimum tillage levels,and two nitrogen(N)fertilizer(urea)management systems(with and without nitrapyrin application)were created as subplots.Tillage level did not have any significant effect on soil ammonium(NH₄⁺)and nitrate(NO₃^-)concentrations,cumulative amount and yield-scaled N₂O emission,and aboveground biomass of maize,whereas nitrapyrin application showed significant effect.Nitrapyrin application significantly reduced the cumulative amount of N₂O emission by 41%and 32%in conventional tillage and minimum tillage practices,respectively.A reduction in soil NO₃^-concentration by nitrapyrin was also observed.The average yield-scaled N₂O emission was 13.6 g N₂O-N kg^-1N uptake in both tillage systems without nitrapyrin application and was significantly reduced to 7.9 and 8.2 g N₂O-N kg^-1N uptake upon the application of nitrapyrin in minimum tillage and conventional tillage practices,respectively.Additionally,nitrapyrin application increased maize biomass yield by 4%and 13%in the minimum tillage and conventional tillage systems,respectively.Our results indicate that nitrapyrin has a potential role in reducing N₂O emission from agricultural systems where urea fertilizers are broadcasted,which is common in Iran due to the practice of traditional farming.