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.     

Does streptobacillary arthritis reduce house mouse population density,activity and damage to sorghum crops?

An experiment to examine the effect of streptobacillary arthritis on mouse populations was conducted in maturing sorghum crops in mouse-proofed pens. There were three treatment pens and three control pens in which mice with and without clinical signs of streptobacillary arthritis were introduced, respectively. A constant mouse density (533 mice ha⁻¹) was introduced to the pens by releasing 12 mice in each pen. The sex ratio of introduced mice was also constant by releasing in each pen nine males and three females. Mean track index was lower in treatment pens than in control pens, indicating that streptobacillary arthritis reduced the activity of mice. However, this was not translated into a reduction in other population responses. Mean mouse density at harvest, percentage of female pregnant at harvest, yield loss caused by mice and a feeding index (oil card index) did not differ significantly between treatment and control groups. A plausible explanation for these findings was that mice with the clinical signs might be less active but were able to consume as much grains as mice without the clinical signs because food was in abundant supply.     

Assessment of morphological and functional changes in the mouse testis and epididymal sperms following busulfan treatment

BACKGROUND: Busulfan, a cytotoxic drug, which is currently used as a chemotherapeutic agent, has many side effects on different body organs. In this research, the effects of busulfan on sperm parameters and microstructure of mouse testis were investigated. METHODS: Busulfan was injected intrapretoneally at 10, 20, 30, 40 and 50 mg/kg and testes were removed after 4, 6 and 8 weeks, weighed and processed for light microscopic examination. Transverse and cross section diameters of testes, seminiferous tubules diameters, percentage of different types of tubules, epithelium thickness, spermatogenic cell numbers and capsule thickness as well as the sperm parameters in epididymis were measured. RESULTS: There was a significant decline in sperm numbers and marked changes in testes structures. Almost 8 weeks after the injection of drug, some of the changes are reversed. Accordingly, the changes in percent of normal tubules without sperm, abnormal tubules and capsule thickness were increased until 6 weeks of drug administration, the changes declined thereafter. CONCLUSION: In general, busulfan caused a decrease in all analyzed parameters (except capsule thickness, normal tubules without sperm and abnormal tubules), probably due to the arrest of spermatogenesis. Our results also revealed that some of the changes are reversible and dose dependent.     

Determining suitability of thermal development models to estimate temperature parameters for embryonic development of <Emphasis Type="Italic">Sitona lepidus</Emphasis> Gyll. (Coleoptera: Curculionidae)

The clover root weevil, Sitona lepidus Gyll. is a pest of clovers (Trifolium spp.), particularly white clover (T. repens). Larva feeding severely impairs the capacity of white clover to fix atmospheric nitrogen, by attacking the nitrogen-fixing root nodules. Development of the egg stage was studied at six different constant temperatures ranging from 9 to 30°C to improve the basis for phenological forecasts. Development occurred over the entire range of temperatures, although hatching percentage at 30°C was only 24.4% compared to 90.1% at 28.5°C. Developmental time decreased with increase in temperature from 9 to 28.5°C but increased markedly at 30°C. At 28.5°C, the embryo development period was the shortest (10.35 days); whereas at 9°C, this parameter was the longest (55.08 days). Using linear regression, the lower threshold temperature was calculated 4.38°C and the thermal constant was 236.45 degree-days. The relationship between constant temperature and developmental rate was evaluated using nine models. The suitability of the models was evaluated based on eight indicators (R ², RSS, R _adj², AIC, BIC, AICC, W _j , and Z _i ). Of the nonlinear models, the Logan-6, Sharpe and DeMichele, and Lactin models were the most accurate at calculating t _opt of 27, 27.50, and 28°C, respectively. The results suggest that of the three models (Lactin, Briere, and Hilbert and Logan), that estimated critical temperatures, (t _min, t _opt, and t _max), Lactin gave the most suitable fit of data. This response of S. lepidus to temperature can be used for developing phonological models to predict the timing of egg hatch which are important for management programs targeting the weevil.     

Plasma renin activity and plasma concentrations of aldosterone, cortisol, adrenocorticotropic hormone, and alpha-melanocyte-stimulating hormone in healthy cats

A pathogenetic role of the renin-angiotensin-aldosterone system has been implicated in cats in both systemic arterial hypertension and hypokalemic myopathy. Yet, measurement of plasma aldosterone concentrations (PACs) and plasma renin activity (PRA) has not unequivocally pointed to hyperaldosteronism as a cause of these conditions. To obtain appropriate reference ranges, this study included a large number (130) of healthy house cats of different breeds without a history of recent illness and plasma concentrations of urea and creatinine below the upper limit of the respective reference ranges. In addition, the pituitary-adrenocortical axis was studied by measuring plasma concentrations of adrenocorticotropic hormone (ACTH), alpha-melanocyte-stimulating hormone (alpha-MSH), and cortisol. Reference ranges for PACs (110-540 pmol/L; 40-195 pg/mL), PRA (60-630 fmol/L/s; 0.3-3 ng/mL/h), and the aldosterone to renin ratio (ARR) (0.3-3.8) were very similar to those established in the same laboratory for humans in a supine position. No breed differences were found. The ARRs in neutered cats were significantly higher than in intact cats, primarily because of low PRA in neutered cats. The ARRs of cats > or = 5 years of age were significantly higher than those of cats < 5 years of age. The plasma concentrations of ACTH, alpha-MSH, and cortisol did not correlate significantly with PAC. Thus, although blood sampling was performed in cats in nonstandardized positions and was associated with a wide variation of stress responses, the references ranges of PAC, PRA, and ARR were similar to the relatively narrow limits established for humans under standardized conditions. The effects of neutering and aging on PRA and ARR warrant further investigation.     

The Immunohistochemistry and Toluidine Blue Roles for Helicobacter pylori Detection in Patients with Gastritis

Background: Helicobacter pylori, which is associated with many upper gastrointestinal diseases, is found in half of the population of the world. Several special stains and immunohistochemistry stain for H. pylori are available. The need for and usefulness of immunohistochemical (IHC) technique has been debated for years. Toluidine blue is a simple stain for microbiological studies and is easily available in laboratories. Therefore, this study was conducted to compare hematoxylin and eosin (H&E), Giemsa and toluidine blue staining with immunehistochemistry for detection of H. pylori in patients with gastritis and also to correlate the results of these staining methods with pathological grading. Methods: We reviewed 54 consecutive gastric biopsy specimens stained by H&E and Giemsa as well as by toluidine blue and immunohistochemistry stains for H. pylori. Results: H. pylori was positively identified by IHC in 43 (79.63%) patients, while positive samples were found in 18 (33.33%), 24 (44.44%) and 33 (61.11%) patients using H&E, Giemsa and toluidine blue staining methods. Our results showed that classical histological staining methods are not sensitive enough to identify low numbers or coccoid forms of organism, while toluidine blue and immunohistochemistry play an important role in detection of H. pylori infection. Conclusion: Toluidine blue has been proved to be much more reliable than H&E and Giemsa in detection of H. pylori. In addition, in post treatment biopsies and in biopsies with unexplained chronic active gastritis without histological evidence of H. pylori should have immunohistochemistry done to detect possible low density or coccoid form of organisms.Key Words: Helicobacter pylori, Immunohistochemistry, Gastritis     

Kinetics of substrate oxidation and hydrogen peroxide production by Mycoplasma mycoides subsp. mycoides Large Colony (LC) type and Mycoplasma mycoides subsp. capri

Mycoplasma mycoides subsp. mycoides Large Colony (LC) type is a pathogen of goats causing contagious agalactia and respiratory disease, found on all continents where small ruminants are kept. It shares close genetic characteristics with M. mycoides subsp. capri. Substrate oxidation by 22 strains of M. mycoides subsp. mycoides LC from nine countries was compared with that of eight strains of M. mycoides subsp. capri from five countries. There was considerable similarity in the substrates used, but substrate saturation coefficients (K_s) varied for different substrates. Substrate utilization patterns and K_s values did not (1) significantly differentiate the LC strains from each other, (2) show any correlation with geographical origin, or (3) distinguish the LC strains from the capri strains. These results support previous studies justifying the reclassification of these subspecies as a single species.     

Producing Gelatin Nanoparticles as Delivery System for Bovine Serum Albumin

Background: Progress in the field of biology and biochemistry has led to the discovery of numerous bioactive peptides and proteins in the last few decades. Delivery of therapeutic proteins/peptides has received a considerable amount of attention in recent years. Methods: In this study, a two-step desolvation method was used to produce biodegradable hydrophilic gelatin nanoparticles (GNP) as a delivery system of protein model (BSA). The size and shape of the nanoparticles were examined by dynamic light scattering and scanning electron microscopy. Results: Particles with a mean diameter of 200-300 nm were produced and the percentage of entrapment efficiency was found to be 87.4. The optimum amount of theoretical BSA loading was obtained, the release of BSA was monitored in vitro, and the mechanism of release was studied. The BSA release profile showed a biphasic modulation characterized by an initial, relatively rapid release period, followed by a slower release phase. Conclusion: Results show that the two-step desolvation is an appropriate method for preparing GNP as a delivery vehicle for BSA. Key Words: Gelatin, Nanoparticles, Bovine serum albumin     

Screening drought-tolerant genotypes in bread wheat (Triticum aestivum L.) using different multivariate methods

Wheat (Triticum aestivum L.) is one of the most widely cultivated crops in rainfed areas of Iran, where drought is the main limiting factor on yield. The object of this study was the identification of drought-tolerant genotypes in bread wheat. Forty bread wheat genotypes were tested in separate experiments under drought stress and normal conditions in two years (2009–2010 and 2010–2011). Nine drought-tolerance/susceptibility indices including stress susceptibility index (SSI), mean productivity (MP), tolerance (TOL), stress tolerance index (STI), geometric mean productivity (GMP), yield index (YI), yield stability index (YSI), linear regression coefficient (β) and drought response index (DRI) were determined. Simultanously applied factor analysis used two factors instead of nine indices in this study. Mahdavi was recognized as the most drought-tolerant genotype in both years based on factor analysis. In this study an equation was developed for estimating the Stress Tolerance Score (STS). The results of the equation were identical to those of factor analysis in both years. The equation was much easier to use than factor analysis and is suggested as a screening tool for the identification of drought-tolerant genotypes. In this study, Mahdavi was the most drought-tolerant genotype also corresponding to this equation.     

Analysis of Aroma and Phenolic Components of Selected Achillea Species

Dried flowers and leaves of four different Achillea species grown wild in several provinces of Iran, including one species collected from three different locations of Isfahan province were analyzed for the headspace volatile components, total phenolics (TP) and tartaric esters (TE). Capillary gas chromatography/mass spectrometry (GC/MS) combined with a purge and trap method was used for quantification of aroma components. Over 70 compounds were determined in the samples. Flower samples from all species contained 2-methyl butanal, α-pinene, α-thujene, camphene, hexanal, β-pinene and 1,8-cineole; however, the major constituents of the aerial parts were determined as α-pinene, camphene, DL-limonene and 1,8-cineole. The largest number of aroma components were found in Achillea tenuifolia Lam. and Achillea millefolium L. In all species, except A. millefolium, the leaves contained more TP and TE than flowers. However, A. wilhelmsii from Semirom in Isfahan province showed the highest values for TP and TE.