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     

Reconsidering betaine as a natural anti-heat stress agent in poultry industry: a review

Betaine is found ubiquitously in plants, animals, microorganisms, and rich dietary sources including seafood, spinach, and wheat bran. The chief physiological role of betaine is to function as a methyl donor and an osmolyte. Betaine also acts as an osmolyte, to maintain the avian’s cellular water and ion balance to improve the avian’s capacity against heat stress via preventing dehydration and osmotic inactivation. It helps in maintaining the protective osmolytic activity, especially in heat-stressed birds. Betaine may promote various intestinal microbes against osmotic variations and thus improve microbial fermentation activity. Previous studies showed that dietary supplementation of betaine in poultry diets could positively affect nutrients’ digestibility, reduce abdominal fat weight, and increase breast meat yield. In addition, betaine has been reported to protect internal organs and boost their performance. Its inclusion in poultry diet is sparing essential amino acids like choline and methionine. In addition, it may play an important role in lean meat production by positively affecting the lipid metabolism with increased fatty acids catabolism and thus reducing carcass fat deposition. The aim of this review article was to broaden the knowledge regarding betaine and its importance in the poultry industry to cope with the heat stress problem. Moreover, it should be added to the diet as a natural anti-stressor through different routes (water/feed) to overcome the heat stress problem. However, further studies need to be conducted at the genetic and molecular basis to elucidate the mechanism behind the betaine as a natural anti-heat agent to decrease the heat stress problem in the poultry industry.     

Effects of steckling weight and planting density on sugar beet (Beta vulgaris L.) monogerm seed yield and qualitative traits

In order to determine the effects of steckling weight and planting density on sugar beet monogerm (Cv. 9597) seed quantitative and qualitative characteristics, this experiment was carried out in Ardabil Agricultural Research Station-Iran (38 degrees 30'N and 48 degrees 30'E) during 2 years (2002-2003). In the study, steckling weight (main-plot) in three levels including 100, 100-200 and 200-300 g and planting density (sub-plot) in three levels including 40, 50 and 60 cm were used in a split-plot experiment based on the RCBD (Randomized Complete Blocks Design) with 5 replications. The measured characteristics were morphological and qualitative (velocity and uniformity of germination) characters, seed size distribution and monogerm seed percentage. According to the results of the experiment, the effect of planting density on the auxiliary branches No. and seed yield was significant at the probability levels of 1 and 5%, respectively. Seed yield increased by 43% as planting density was increased from 65 x 40 to 65 x 60 cm. The increase in steckling weight led to the increase in standard seed portion (with 3.5-4.5 mm in diameter), germinable standard seed yield (11%) and decrease in germination velocity and germination uniformity; and increase in planting density was accompanied by increase in > 4.5 mm seed percentage, velocity and uniformity of germination and decrease in germinable standard seed yield. Eventually, the highest seed yield (1208 kg ha(-1)) was obtained by using steckling weight of 200-300 g and planting density of 65 x 60 cm.     

Diminished gene expression of cardiac heat shock proteins in pulmonary hypertensive chickens

1. The relative expression of heat shock protein (HSP) genes (HSP60, HSP70 and HSP90) was performed using quantitative real-time PCR on tissue from the heart ventricles to investigate the effect of 3,5,3′-l-triiodothyronine (T₃)-induced pulmonary hypertension in broiler chickens.

2. The ratio of the right ventricle to total ventricle (index of pulmonary hypertension) was increased in the treated groups at 12 and 42 d of age compared to controls but was significant only at 42 d.

3. The HSP genes were expressed in the right and left ventricles of control but T₃-treated broilers at 12 and 42 d of age. The relative amounts of HSP60 and HSP90 gene expression in the right ventricle of treated groups were significantly increased at 12 d and decreased at 42 d of age compared to controls.

4. Variations of HSP60 and HSP90 mRNAs in the left ventricle were not significant. The relative amount of HSP70 mRNA expression in the right and left ventricles of treated groups was significantly decreased at 42 d of age compared to controls. HSP70 mRNA expression did not differ between the right and left ventricles at 12 d of age.

5. It is concluded that gene expression of HSPs (i.e. HSP60 and HSP90) was upregulated in the heart of chickens developing pulmonary hypertension syndrome, probably to delay the pathological process of disease. The right ventricle from hearts of pulmonary hypertensive chickens showed considerable reductions of HSP60, HSP70 and HSP90, which is evidence of a loss of compensatory responsiveness in dilated heart.     

Identifying invasive fish species threats to RAMSAR wetland sites in the Caspian Sea region—A case study of the Anzali Wetland Complex (Iran)

Risk screening tools play a crucial role in identifying potential high-risk non-native (NN) fish species. In this study, potentially invasive NN fish species in the Anzali Wetland Complex (AWC), which is located on the south coast of the Caspian Sea (Iran), were identified using the Aquatic Species Invasiveness Screening Kit (AS-ISK). Twenty-nine freshwater fish species were screened of which 13 exist in the AWC and 16 in close proximity to it (“horizon” species). Receiver operating characteristic curve analysis showed that AS-ISK could discriminate reliably between non-invasive and invasive fish species for the AWC. Mean threshold scores were 3.25 for the Basic Risk Assessment (BRA) and 11.75 for the BRA + CCA (BRA + Climate Change Assessment), and these, respectively, classified 89.7% and 86.2% of the species as high risk. The CCA resulted in an increase in the BRA scores for 86.2% of the species, suggesting the need to account in future NN species management for a likely increased invasiveness of those species under future climate conditions. These results suggest that AS-ISK could prove an effective tool for identifying potentially invasive NN freshwater fishes in other wetlands of the Caspian Sea basin.     

Graphite Furnace Atomic Absorption Spectrometry After Dispersive Liquid–Liquid Microextraction for the Determination of Selenium in the Anodic Slime

In the present study, a method based on dispersive liquid–liquid microextraction (DLLME) followed by graphite furnace atomic absorption spectrometry (GFAAS) determination was proposed for the determination of selenium by using ammonium pyrrolidine dithiocarbamate (APDC) as the chelating reagent. The main factors influencing the DLLME were investigated systematically. Under the optimal conditions, the limit of detection for Se(IV) was 0.02 ng mL^?1. The relative standard deviation was 4.1% (C_Se(IV) = 0.2 ng mL^?1, n = 8) with an enhancement factor of 135.8-fold from only 5 mL of the water sample. The proposed method was successfully applied to the determination of Se(IV) in anodic slime and electrolyte samples. In order to validate the proposed method, a Certified Reference Material (trace elements in water, 1643e, NIST) was analyzed, and the determined value obtained was in good agreement with the certified value.     

The Effect of Human Platelet-Rich Plasma on Adipose-Derived Stem Cell Proliferation and Osteogenic Differentiation

Background: The cultured mesenchymal stem cells (MSC) have been used in many clinical trials; however, there are still some concerns about the cultural conditions. One concern is related to the use of FBS as a widely used xenogeneic supplement in the culture system. Human platelet-rich plasma (hPRP) is a candidate replacement for FBS. In this study, the effect of hPRP on MSC proliferation and osteogenic differentiation has been evaluated. Methods: Human adipose-derived stem cells (hADSC) were expanded. Cells from the third passage were characterized by flow cytometric analysis and used for in vitro experiments. Resazurin and alizarin red stains were used for cell proliferation and osteogenic differentiation assays, respectively. Results: Treatment with hPRP resulted in a statistically significant increase in cell proliferation compare to the negative control group (P<0.001). Cell proliferation in the 15% hPRP group was also significantly higher than that in the 10% hPRP group (P<0.05). Additionally, it caused less osteogenic differentiation of the hADSC compared to the FBS (P<0.001), but in comparison to negative control, it caused acceptable mineralization (P<0.001). Conclusion: These findings indicate that hPRP not only improves the proliferation but also it can be a suitable substitution in osteogenic differentiation for clinical purposes. However, the clinical application value of hPRP still needs more investigation. Key Words: Platelet-Rich Plasma, Adipose tissue, Stem Cells, Cell differentiation, Cell proliferation