Hydrocortisone Reduces Toll-Like Receptor 4 Expression on Peripheral CD14+ Monocytes in Patients Undergoing Percutanoues Coronary Intervention

Bacground: Evidence from several lines of investigations suggests that Toll-like receptor 4 (TLR4) is involved in atherosclerosis as a bridge between innate and acquired immunity. Percutaneous coronary intervention (PCI) can trigger inflammation through activation of human TLR4 (hTLR4) on monocytes. Hydrocortisone as an anti-inflammatory and immuno-suppressant agent has multiple mechanisms of action. In this study, we aimed at assessing the effects of hydrocortisone on monocyte expression and activity of hTLR4 in patients underwent PCI. Methods: Blood samples were taken from a total of 71 patients with chronic stable angina who were scheduled for a PCI, before the intervention. Thirty patients received 100 mg hydrocortisone prior to the procedure. Control group was composed of 41 patients underwent PCI without receiving hydrocortisone. Blood collection was repeated 2 and 4 h after PCI. The expression of hTLR4 on the surface of CD14⁺ monocytes and the serum levels of TNF-α and IL-1β were measured using flowcytometry and Sandwich ELISA. Results: Compared with controls, hydrocortisone significantly reduced monocyte expression of hTLR4 in test group (P<0.01). In addition, it had a significant effect on reduction of serum concentrations of TNF-α and IL-1β in test group in a time-dependent manner (P<0.01). Conclusion: In this study, hydrocortisone was able to reduce the hTLR4/CD14 positive monocytes and its related pro-inflammatory cytokines, thus it can decrease inflammatory responses following PCI. Key Words: Toll-like receptor 4, Cytokines, Hydrocortisone     

Vitamin D Deficiency Impairs Spatial Learning in Adult Rats

Background: Through its membrane and intracellular receptors, vitamin D regulates many vital functions in the body including its well known actions on musculoskeletal system. Growing body of evidences demonstrate that vitamin D undergoes some of behavioral aspects of neurocognition. The present study was designed to evaluate the effect of food regimens without vitamin D or with a supplement of 1,25(OH)₂D3 on spatial performance of adult rats. Methods: The animals were trained in the Morris water maze to find a hidden platform. The time spent and the distance traveled to find the platform, speed of navigation and the percentage of unsuccessful trials were considered for assessment of the task learning. Results: Our findings indicated that the vitamin D-deprived rats had a significant lower performance compared to both the controls and the animals receiving 1,25(OH)₂D3 supplementation. Concerning the unsuccessful trials, lack of vitamin D resulted in the highest failures in the maze navigation. The regimen with additional 1,25(OH)₂D3 did not considerably influence learning of the maze task. Conclusion: We concluded that while vitamin D deficiency deteriorates the spatial task learning, the 1,25(OH)₂D3 supplementation did not effectively underlie the maze performance. Key Words: Vitamin D, Maze learning, Dietary supplements     

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     

Aberrant Promoter Methylation Profile of Niemann-Pick Type C1 Gene in Cardiovascular Disease

Background: The protein of Niemann-pick type C1 (NPC1) gene promotes the egress of cholesterol from late endosomes and lysosomes to other cellular compartments and contributes to a process known as reverse cholesterol transport. This study aimed to examine whether promoter methylation of NPC1 is associated with risk of cardiovascular disease (CVD). Methods: Fifty CVD patients and 50 healthy subjects as the control group were recruited in this study. Promoter methylation of NPC1 gene was defined using a nested-methylation specific polymerase chain reaction method. Statistical analyses were done using the chi-square, t-test or ANOVA tests. Results: Our study showed that the frequency of semi-methylated promoter (methylated/unmethylated status) was significantly higher in CVD patients than that in controls (OR = 6.521, 95% CI = 2.211-19.215, P = 0.008). However, a completely methylated promoter (methylated/methylated status) was not detected in any subjects in either of the two groups tested. Additionally, the analysis of clinical data according to the methylation status of NPC1 gene demonstrated that serum levels of total cholesterol, total triglycerides, high low-density lipoprotein cholesterol (LDL-C) and low high-density lipoprotein cholesterol (HDL-C) are influenced by NPC1 methylation, so that subjects with a completely unmethylated promoter (Unmethylated/unmethylated status) held lower levels of total triglycerides, total cholesterol, LDL-C and higher levels of HDL-C. Conclusion: Our findings propose that the NPC1 promoter methylation is a probable mechanism that can result in reduced/impaired NPC1 expression/activity and may thus contribute to progression of CVD. Key Words: Niemann-pick type C1 (NPC1), Promoter methylation, cardiovascular disease     

Radiographic measurement of vertebral heart size in healthy stray cats

The aims of this study were to determine vertebral heart size (VHS) in stray cats and to compare different radiographic views. This study was performed on 50 adult stray cats. All cats were short-haired and non-obese and were considered to be healthy based on physical examination and electrocardiography. Left and right lateral, dorsoventral and ventrodorsal radiographs were taken. The long and short axes of the heart were measured in millimetres. The thoracic vertebral length spanned by each dimension was measured caudally from the fourth thoracic vertebra. Mean+/-SD and the correlation coefficient between the measurements were calculated with standard statistical software. The sum of the long and short axes of the heart expressed as VHS was 7.3+/-0.49 vertebrae in right lateral, 7.3+/-0.55 vertebrae in left lateral, 7.5+/-0.68 vertebrae in dorsoventral and 7.5+/-0.53 vertebrae in ventrodorsal. The differences between right and left lateral as well as dorsoventral and ventrodorsal views were not significant (P>0.05). Absolute measurements and vertebral heart scale values were slightly smaller than those reported in the literature for mixed population of cats. It is, therefore, important to take the breed in to account.     

Quercetin attenuates H2O2‐induced toxicity of rooster semen during liquid storage at 4°C

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H₂O₂ (40 μM) and combination groups which incubated with constant dose of H₂O₂ (40 μM) plus various levels of quercetin (20, 40 and 80 μM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 μmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 μmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 μmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co‐administration of quercetin (especially at 40 and 80 μM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide‐mediated damage during cold liquid storage of rooster semen.     

Karyotype study on 15 populations of Eremostachys laciniata Bunge in Iran

A karyotype study was performed on 15 populations of Eremostachys laciniata Bunge from Iran in order to quantify the extent of cytological variation for use in breeding programmes. Ten mitotic root tip cells at the metaphase stage were prepared from each population by the squash method. The chromosomes of suitable mitotic cells were counted and various parameters, including long and short arm lengths, chromosome lengths, total length of the haploid chromosome complement, arm ratios, and the centromeric index, were measured. All 15 populations were diploid with 2n = 2x = 22 and were differentiated by their karyotypic parameters. All 15 E. laciniata populations occupied Class 1A of Stebbins’ karyotype classification, indicating the presence of a primitive symmetrical karyotype. The size of the mitotic chromosomes was medium, and mean chromosome lengths ranged from 3.84 to 4.77 μm. The shortest chromosome lengths were observed in the Ajabshir population and the longest in the Heydarabad population. The Heydarabad population also had the longest total chromosome length and the Ajabshir population had the shortest total chromosome length. The centromeric indices of the chromosome complements varied from 39.3% to 43.7%. Metacentric chromosomes were the most common in all populations, whereas submetacentric chromosomes were rare. The A₁ index varied from 0.215 (Areshtanab) to 0.337 (Til). The A₂ index ranged from 0.116 (Til) to 0.143 (Kaleybar), and the karyotype asymmetry index varied between 0.051 and 0.097. The results of a principal component analysis (PCA) showed that the main elements of the first three main principal components accounted for 97% of the total variance. Cluster analysis by Ward’s method indicated that all 15 populations could be classified into three clusters. Finally, the results of the karyotypic parameters showed that the Til, Heydarabad, and Marand populations had higher levels of karyotypic heterogeneity and may be useful when developing breeding programmes.     

Analysis of the Phenylpropanoid Enzyme Activities and Products in the Soft- and Hard-Seeded Pomegranate Genotypes During Fruit Development

To find out more about the biochemical aspects of seed-softness trait in pomegranate (Punica granatum L.), some of the phenylpropanoid pathway enzymes as well as their products were evaluated in the various soft- and hard-seeded genotypes during fruit development. Results showed that peroxidase (POD) activity decreased in all studied genotypes during fruit growth, while increasing at the last stages only in the hard-seeded genotypes. Although laccase (LAC) activity did not show high differences during fruit ripening and in different genotypes, but its general trend was increasing. Phenylalanine ammonia-lyase (PAL) activity decreased during the fruit development and approximately diminished for the hard-seeded genotypes. All genotypes had lignin components in their seeds and amount of lignin increased during the growth season. Total phenolic decreased drastically and antioxidant capacity increased gradually showing an opposite trend and probable competitive effect for the same substrate during fruit development. Considerable differences that were observed in the POD activity at the last sampling stage, and higher activity that was observed in POD than LAC, may indicate that POD is more important for polymerization of lignin in the pomegranate seed and this enzyme might have some role in the development of seed softness trait in pomegranate. Also, this is the first report about lignin detection in the seeds of soft-seeded pomegranate genotypes, and represents a different mechanism (such as monolignol conformation in the final polymer) from lignin content for development of the soft-seeded pomegranate genotype.     

Effects of chronic high stocking density on liver proteome of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

The main aim of the present study was to assess the effects of chronic high stocking density on liver proteome of rainbow trout. Rainbow trout juveniles (42.6 ± 2.3 g average body weight) were randomly distributed into six tanks at two stocking densities (low stocking density (LD) = 20 kg m^?3 and high stocking density (HD) = 80 kg m^?3). Both treatments were performed in triplicate tanks for a period of 60 days. High stocking density caused a reduction in the growth performance compared with LD fish. Lysozyme activity increased with stocking density, while serum complement activity presented the opposite pattern. Serum cortisol and total protein levels did not show significant differences (P > 0.05) between experimental groups. The fish reared at high stocking density showed significantly lower osmolality and globulin values but higher albumin level. The HD group had significantly higher activities of catalase, glutathione peroxidase and superoxide dismutase, and malondialdehyde content in the liver when compared to the LD group. Comparative proteomics was used to determine the proteomic responses in livers of rainbow trout reared at high stocking density for 60 days. Out of nine protein spots showing altered abundance (>1.5-folds, P < 0.05), eight spots were successfully identified. Two proteins including apolipoprotein A-I-2 precursor and mitochondrial stress-70 protein were found to increase in HD group. The spots found to decrease in the HD group were identified as follows: 2-peptidylprolyl isomerase A, two isoforms of glyceraldehydes-3-phosphate dehydrogenase, an unnamed protein product similar to fructose-bisphosphate aldolase, 78 kDa glucose-regulated protein, and serum albumin 1 protein.