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     

Dietary ginger administration attenuates oxidative stress and immunosuppression caused by oxytetracycline in rainbow trout (Oncorhynchus mykiss)

In the present study, potential ameliorative effects of dietary ginger (GN) were investigated on antioxidant and immune responses of rainbow trout (Oncorhynchus mykiss) during oxytetracycline (OX) administration. As a 2 × 3 factorial design, the fish were orally treated with OX (a daily dose of 100 mg/kg) and GN (either 10 or 20 g/kg diet) for 10 days. Then, blood samples were taken from each treatment to monitor plasma lysozyme, complement (ACH50), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione‐S‐transferase (GST) activities, and reduced glutathione (GSH), malondialdehyde (MDA), total immunoglobulin (Ig) and globulin levels. OX treatment significantly decreased SOD (30%), GPx, (10%) and lysozyme (23%) activities, and GSH (19%) levels; however, it increased GST (16%) activity and MDA (28%) levels. Ten grams GN per kg levels significantly decreased SOD (35%), CAT (13%), GST (20%) and MDA (30%), but increased GSH (30%), lysozyme (48%) and globulin (16%). Twenty grams GN per kg diet significantly decreased SOD (26%) and MDA (17%), but increased lysozyme (31%) levels. Interaction effects of dietary GN and OX were observed on plasma MDA and GPx levels, as 10 g GN per kg diet prevented the OTC‐induced changes in these parameters. Moreover, 20 g GN per kg diet prevented the OX‐induced change in GPx activity and mitigated the MDA elevation by 20%. It is concluded that GN administration at 10 g/kg diet is beneficial in mitigating oxidative stress and immunosuppression of rainbow trout during OX administration.     

Comparison of genetic variation among accessions of <Emphasis Type="Italic">Aegilops tauschii</Emphasis> using AFLP and SSR markers

Genetic diversity of 54 accessions of Aegilops tauschii from five countries was assessed using sequence-tagged microsatellites (or simple sequence repeats, SSRs) and amplified fragment length polymorphisms (AFLPs). In the case of AFLP analysis, a total of 256 amplification products obtained, 234 of them were polymorphic across all the 54 accessions. A total of 224 fragments were obtained from the 24 SSR primers and 219 of fragments were polymorphic across all the genotypes screened. Based on both AFLP and SSR markers, the highest percentage of polymorphisms were obtained in Iranian and accessions of unknown origin. The highest polymorphic information content (PIC) value was observed for SSRs (0.82) while the highest marker index (MI) value was for AFLPs (8.5) reflecting the hyper-variability of the first and the distinctive nature of the second system. Principal co-ordinate analysis (PCO) revealed congruent patterns of genetic relationships for both data sets, but did not group accessions strictly according to their geographical origins. Poor correlation was found between AFLP and SSR marker loci. This low association may be due to low number of AFLP and SSR markers. These results show that molecular markers can help to organize the genetic variability and expose useful diversity for breeding purposes.     

Preference and performance of <Emphasis Type="Italic">Plutella xylostella</Emphasis> (L.) (Lepidoptera: Plutellidae) on canola cultivars

The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is an important insect pest of canola, Brassica napus L., in Ardabil, Iran. Host plant resistance is an essential component of integrated management of P. xylostella. We investigated the preferences and performance of P. xylostella on nine commercial cultivars of canola, namely Zarfam, RGS003, Adder, Okapi, Opera, Hyola401, Ebonite Option500 and Elite under greenhouse conditions at 23 ± 1°C, 50 ± 5% RH and 14L:10D. In free-choice situation, oviposition was lowest on Opera (23.5) and highest on Zarfam (44.7). In the life table study, fecundity of new generation female moths was lowest on Opera (95.4) and highest on Adder (145.7). Survival from egg to adult was significantly lower on Opera, Option500 and Hyola401 than on other tested cultivars. The intrinsic rate of natural increase (r _m) and population growth rate (λ) were lowest on Opera and highest on Zarfam. The generation time (T) was shortest on Zarfam (17.2 days) and longest on Hyola401 (19.9 days) whereas doubling time (DT) was longest on Opera (3.9 days) and shortest on Zarfam (3.0 days). Our results clearly suggest that Opera was the most resistant host among the tested cultivars and has the potential to be used in the integrated management of P. xylostella.     

Exogenously applied hexaconazole ameliorates salinity stress by inducing an antioxidant defense system in Brassica napus L. plants

The ability of hexaconazole (HEX) to ameliorate salinity stress was studied in canola plants (Brassica napus L.). Canola seedlings were subjected to sodium chloride (NaCl) treatment. A treatment with 200 mM NaCl reduced growth parameters, chlorophyll content and protein content as well as increased the proline (Pro) content in canola plants. In addition, NaCl stress increased the endogenous, nonenzymatic antioxidants and the activity of antioxidant enzymes, such as peroxidase (POX; EC 1.11.1.7), superoxide dismutase (SOD; EC 1.15.1.1) and catalase (CAT; EC 1.11.1.6). When these plants were treated with a combination of NaCl and 50 mg L⁻¹ HEX, the inhibitory effects of NaCl stress were decreased by increasing the root growth, shoot growth, dry weight (DW), chlorophyll content, protein content and antioxidant enzyme activity by ameliorating the salinity injury. These results suggested that HEX has an important role in the enhancement of plant antioxidant systems and resistance to salinity in canola plants.     

Maternal age influence on diapause induction in sexual and asexual Trichogramma brassicae

In this study, the effect of maternal age on prepupae diapause of progeny in sexual (Wolbachia free) and asexual (Wolbachia infected) Trichogramma brassicae was studied at two temperatures. Maternal generation (G₁) of both populations was reared at 10L:14D photoperiod, 14 and 20 °C, separately. Mated females from G₁ were supplied with eggs of Ephestia kuehniella, each 48 h from one day after emergence to death. The tendency to diapause in progeny (G₂) was determined by rearing at 10 °C. According to the results, reproductive modes, maternal temperature and age have significant effect on the mean number of eggs laid by G₁ and the diapause percentage of G₂. Offspring whose maternal generation was reared at 20 °C had higher diapause percentage than those reared at 14 °C. Diapause percentage in sexual population was higher than the asexual, which demonstrates a negative effect of Wolbachia on this process. Wolbachia infection modifies the effect of maternal age on the diapause percentage in the offspring at 20 °C but not at 14 °C. The result can be due to lower Wolbachia density at 14 °C and greater Wolbachia density at 20 °C.     

COMPARATIVE EVALUATION OF HYDROPIC DEGENERATION OF THE LIVER IN DAIRY CATTLE THROUGH BIOCHEMISTRY, ULTRASONOGRAPHY AND DIGITAL ANALYSIS

Ultrasonography of the liver of 181 Holstein-Friesian cows was performed and blood samples were collected for analysis. The hepatic ultrasonograms were evaluated and the echoes were analyzed digitally. After slaughter, liver specimens were taken and examined histopathologically. Of the 181 animals, 120 had a normal liver and 61 had hydropic degeneration of the liver, diagnosed through histopathologic examination. Diagnostic accuracy rates for hydropic degeneration were determined based on the following test positive conditions: a) for biochemical analysis—high levels of aspartate aminotransferase, alanine aminotransferase, total bilirubin and non-esterified fatty acids; b) for ultrasonography—presence of dark pattern and blurring of edges; and c) for digital analysis—low echo means at 1 cm and 3 cm from the hepatic surface. Digital analysis had the highest overall specificity, accuracy and positive predictive values for hydropic degeneration, followed by ultrasonography. The results suggest that ultrasonography and digital analysis of hepatic ultrasonograms can be used for diagnosis of hydropic degeneration of the liver in place of biochemical analysis.     

EVALUATION OF FATTY INFILTRATION OF THE LIVER IN DAIRY CATTLE THROUGH DIGITAL ANALYSIS OF HEPATIC ULTRASONOGRAMS

Ultrasonography of the liver of 49 Holstein-Friesian cows was performed, liver specimens were taken, examined microscopically and the fatty occupying rate (FOR) was calculated. Echoes from the hepatic B-mode ultrasonograms were quantified as histogram mean (Emean) and histogram mode (Emode) of echo amplitudes within various areas at a depth of 1–9 cm from the hepatic surface. Of the 49 animals, 26 had a normal liver and 23 had fatty infiltration of the liver, diagnosed through histopathological examination. Fatty occupying rate ranged from 1.7 to 64.5%, with 11 animals having 1–15% FOR (mild fatty infiltration), 6 having 15.1–30% FOR (moderate fatty infiltration) and 6 having > 30% FOR (severe fatty infiltration). At 1 cm, severe fatty infiltration had higher Emean and Emode than normal liver (p < 0.05). At 7 cm and 9 cm, moderate and severe fatty infiltration had lower Emeans and Emodes than normal liver (p < 0.001). The results suggest that digital analysis of hepatic ultrasonogram can be useful in the evaluation of the degree of fatty infiltration of the liver in dairy cattle.