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     

Effects of foliar spray of two kinds of zinc oxide on the growth and ion concentration of sunflower cultivars under salt stress

This study investigated the effects of foliar application of normal and nano-sized zinc oxide on the response of sunflower cultivars to salinity. Treatments included five cultivars (‘Alstar’, ‘Olsion’, ‘Yourflor’, ‘Hysun36’ and ‘Hysun33’), two salinity levels [0 and 100 mM sodium chloride (NaCl)], and three levels of fertilizer application. Fertilizer treatments were the foliar application of normal and nano-sized zinc oxide (ZnO). Foliar application of ZnO in either forms increased leaf area, shoot dry weight, net carbon dioxide (CO₂) assimilation rate (A), sub-stomatal CO₂ concentration (Ci), chlorophyll content, Fv/Fm, and Zn content and decreased Na content in leaves. The extent of increase in chlorophyll content, Fv/Fm and shoot weight was greater as nano-sized ZnO was applied to the normal form. The results show that the nano-sized particles of ZnO compared to normal form has greater effect on biomass production of sunflower plants.     

Mechanism of ammonia transport by Amt/MEP/Rh: structure of AmtB at 1.35 A

The first structure of an ammonia channel from the Amt/MEP/Rh protein superfamily, determined to 1.35 angstrom resolution, shows it to be a channel that spans the membrane 11 times. Two structurally similar halves span the membrane with opposite polarity. Structures with and without ammonia or methyl ammonia show a vestibule that recruits NH4+/NH3, a binding site for NH4+, and a 20 angstrom-long hydrophobic channel that lowers the NH4+ pKa to below 6 and conducts NH3. Favorable interactions for NH3 are seen within the channel and use conserved histidines. Reconstitution of AmtB into vesicles shows that AmtB conducts uncharged NH3.     

Aldosterone-to-renin and cortisol-to-adrenocorticotropic hormone ratios in healthy dogs and dogs with primary hypoadrenocorticism

In dogs with primary hypoadrenocorticism, hypocortisolism and hypoaldosteronism usually are present, but these deficiencies also may occur in isolated forms. The diagnosis is commonly made by measuring plasma cortisol concentration before and after stimulation with ACTH, thereby ignoring aldosterone. In search of an alternative approach that would include assessment of glucocorticoid and mineralocorticoid production, 2 pairs of endocrine variables were measured: (1) plasma concentration of cortisol and ACTH, and (2) plasma aldosterone concentration and plasma renin activity. In addition, the cortisol-to-ACTH ratio (CAR) and the aldosterone-to-renin ratio (ARR) were calculated. Reference intervals were established in a population of 60 healthy dogs. In these dogs, CAR ranged from 1.1 to 26.1 and ARR ranged from 0.1 to 1.5. The variables were compared with those of 22 dogs with spontaneous primary hypoadrenocorticism. Plasma concentration of cortisol and ACTH in both groups of dogs overlapped, whereas CAR did not. Similarly, plasma aldosterone concentration and plasma renin activity overlapped, whereas ARR did not. These observations indicate that measurement of these endogenous variables (in one blood sample) allows the specific diagnoses of primary hypocortisolism and primary hypoaldosteronism.     

Estimation of genetic parameters for body weight and egg production traits in Mazandaran native chicken

Native chicken breeding station of Mazandaran was established in 1988 with two main objectives: genetic improvement through selection programs and dissemination of indigenous Mazandarani birds. (Co)variance components and genetic parameters for economically important traits were estimated using (bi) univariate animal models with ASREML procedure in Mazandarani native chicken. The data were from 18 generations of selection (1988?C2009). Heritability estimates for body weight at different ages [at hatch (bw1), 8 (bw8), 12 (bw12) weeks of ages and sex maturation (wsm)] ranged from 0.24?±?0.00 to 0.47?±?0.01. Heritability for reproductive traits including age at sex maturation (asm); egg number (en); weight of first egg (ew1); average egg weight at 28 (ew28), 30 (ew30), and 32 (ew32) weeks of age; their averages (av); average egg weight for the first 12?weeks of production (ew12); egg mass (em); and egg intensity (eint) varied from 0.16?±?0.01 to 0.43?±?0.01. Generally, the magnitudes of heritability for the investigated traits were moderate. However, egg production traits showed smaller heritability compared with growth traits. Genetic correlations among egg weight at different ages were mostly higher than 0.8. On the one hand, body weight at different ages showed positive and relatively moderate genetic correlations with egg weight traits (ew1, ew28, ew30, ew32, ew12, and av) and varied from 0.30?±?0.03 to 0.59?±?0.02. On the other hand, low negative genetic correlations were obtained between body weight traits (bw1, bw8, bw12, and wsm) and egg number (en). Also, there is low negative genetic correlation (?24?±?0.04 to ?29?±?0.05) between egg number and egg weight. Therefore, during simultaneous selection process for both growth and egg production traits, probable reduction in egg production due to low reduction in egg number may be compensated by increases in egg weight.     

Study on Fruitset and Pollen-Compatibility Status in Sweet Cherry (Prunus avuim L.) Cultivars

Prunus avium L. is one of the most important temperate zone fruits in the world. Most of the cherry cultivars always have difficulties of fertilization and fruit set due to self-incompatibility, so they need suitable and compatible pollinizers for commercial fruit production. In this experiment, pollination compatibility of cherry cultivars of ‘Napoleon’, ‘Burlat’, ‘Zhan’, and ‘Lambert’ was studied as both pollen recipients and donors. To determine the compatibility or incompatibility of pollinizers, percent of fruit set was calculated. This research was carried out as a factorial experiment in a randomized complete block design. The final average fruit set in studied cultivars was different under self or cross-pollination. The results showed that ‘Napoleon’, ‘Zhan’ and ‘Lambert’ cultivars are the suitable pollinizers for ‘Burlat’. Also, ‘Burlat’ is a cultivar which produced many fruits under self-pollination. Therefore, ‘Burlat’ can be used as monoculture for orchard establishment. Results showed that cultivar ‘Napoleon’ was cross-incompatible with ‘Lambert’. ‘Zhan’, ‘Napoleon’ and ‘Lambert’ cultivars are founded self-incompatible and require cross-pollinations to obtain fruits.

     

Rapid screening of toxigenic vibrio cholerae O1 strains from south Iran by PCR-ELISA

BACKGROUND: The ability to sensitively detect Vibrio cholera with PCR-ELISA method represents a considerable advancement over alternative more time-consuming methods for detection of this pathogen. The aim of this research is to evaluate the suitability of a PCR-enzyme-linked immunosorbent assay for sensitive and rapid detection of V. cholera O1. METHODS: The 398-bp sequence of a gene that codes for the cholera toxin B subunit was amplified by PCR. The digoxigenin-labeled amplified products were coated on microplates and detected by ELISA. The PCR product was also hybridized with biotin labelled probe and detected by ELISA using streptavidin. RESULTS AND CONCLUSION: The specificity of the PCR was determined using 10 bacterial strains and 50 samples from south Iran. The detection limit was 0.5 pg of the genomic DNA and five bacterial cells. Adaptation of PCR into PCR-ELISA assay format facilitates specific and sensitive detection and diagnosis of human cholera disease. We conclude that this PCR-ELISA is a diagnostic method that specifically detects toxin genes in V. cholera O1 strains. It is more rapid and less cumbersome than other diagnostic methods for detection of toxicity in these strains.     

Localization of herpes simplex virus type 1 DNA in latently infected BALB/c mice neurons using in situ polymerase chain reaction

Background: Herpes simplex virus type-1 (HSV-1) establishes a lifelong latent infection in neurons following primary infection. The existence of latent HSV-1 DNA in the trigeminal ganglia of infected BALB/c mice was examined using a direct in situ PCR technique, based on Digoxigenin-11-dUTP detection system with anti-digoxigenin-peroxidase and 3,3''-diaminobenzidine (DAB) substrate. Methods: Eight-week-old male BALB/c mice were inoculated via the eye by 10⁴ plaque forming unit of wild type Iranian isolates of HSV-1. After establishment of latency, trigeminal ganglia were removed and examined using in situ PCR to detect HSV-1 genome. Finally, the results of in situ PCR were verified by a two-round PCR method, using amplification cocktail of in situ reaction, as a template for a conventional gel base PCR. Results and Conclusion: The results suggest that a direct in situ PCR method using a peroxidase and DAB detection system is a useful means for detection of latent HSV-1 DNA in the latently infected ganglia. Key Words: Herpes simplex virus-1, Latency, In situ PCR, two-round PCR, Trigeminal ganglia