Effect of Food Deprivation on Formalin-Induced Nociceptive Behaviors and Beta-Endorphin and Sex Hormones Concentration in Rats

Background: The present study examined the possible role of endogenous opioidergic system in effect of food deprivation on formalin-induced nociceptive behaviors in male and female rats. Also, we investigated the effect of food deprivation on the plasma level of beta-endorphin and sex hormones. Methods: Food was withdrawn 48 h prior to performing the formalin test, but water continued to be available ad libitum. The formalin was injected into hind plantar paw. Results: There is significant difference between male and female control rats during phase 2B. Following 48-h food deprivation, both male and female rats exhibited enhanced nociceptive behavior in response to formalin. Food deprivation for 12 and 24 h increased and for 48 h decreased beta-endorphin level in male and female rats. Food deprivation for 24 h decreased testosterone level in male, while it had no significant effect on female rats and food deprivation for 48 h decreased testosterone level in both sexes. Food deprivation for 24 h increased estradiol level in female and that for 48 h had no significant effect on male and female rats. Conclusions: The present study demonstrates the existence of food deprivation for 48 h causes enhancement of nociception in the formalin test in male and female rats that has correlation with decrease in plasma beta-endorphin and testosterone levels. Key Words: Rat, Food Deprivation, beta-endorphin, Naloxone     

Quality Control of Widely Used Therapeutic Recombinant Proteins by a Novel Real-Time PCR Approach

Background:

Existence of bacterial host-cell DNA contamination in biopharmaceutical products is a potential risk factor for patients receiving these drugs. Hence, the quantity of contamination must be controlled under the regulatory standards. Although different methods such as hybridization assays have been employed to determine DNA impurities, these methods are labor intensive and rather expensive. In this study, a rapid real-time PCR test was served as a method of choice to quantify the E. coli host- cell DNA contamination in widely used recombinant streptokinase (rSK), and alpha interferon (IFN-α) preparations.

Methods:

A specific primer pair was designed to amplify a sequence inside the E. coli 16S rRNA gene. Serial dilutions of DNA extracted from E. coli host cells, along with DNA extracted from Active Pharmaceutical Ingredients of rSK, and IFN-α samples were subjected to an optimized real-time PCR assay based on SYBR Green chemistry.

Results:

The test enabled us to detect a small quantity of genomic DNA contamination as low as 0.0002 pg in recombinant protein-based drugs. For the first time, this study showed that DNA contamination in rSK and IFN-α preparation manufactured in Pasteur Institute of Iran is much lower than the safety limit suggested by the US FDA.

Conclusion:

Real-time PCR is a reliable test for rapid detection of host-cell DNA contamination, which is a major impurity of therapeutic recombinant proteins to keep manufacturers’ minds on refining drugs, and provides consumers with safer biopharmaceuticals. Key Words: DNA contamination, Real-time PCR, Streptokinase, Interferon-alpha (IFN-α)     

Identification of Sequence Variation in the Apolipoprotein A2 Gene and Their Relationship with Serum High-Density Lipoprotein Cholesterol Levels

Background:

Apolipoprotein A2 (APOA2) is the second major apolipoprotein of the high-density lipoprotein cholesterol (HDL-C). The study aim was to identify APOA2 gene variation in individuals within two extreme tails of HDL-C levels and its relationship with HDL-C level.

Methods:

This cross-sectional survey was conducted on participants from Tehran Glucose and Lipid Study (TLGS) at Research Institute for Endocrine Sciences, Tehran, Iran from April 2012 to February 2013. In total, 79 individuals with extreme low HDL-C levels (≤5^th percentile for age and gender) and 63 individuals with extreme high HDL-C levels (≥95^th percentile for age and gender) were selected. Variants were identified using DNA amplification and direct sequencing.

Results:

Screen of all exons and the core promoter region of APOA2 gene identified nine single nucleotide substitutions and one microsatellite; five of which were known and four were new variants. Of these nine variants, two were common tag single nucleotide polymorphisms (SNPs) and seven were rare SNPs. Both exonic substitutions were missense mutations and caused an amino acid change. There was a significant association between the new missense mutation (variant Chr.1:16119226, Ala98Pro) and HDL-C level.

Conclusion:

None of two common tag SNPs of rs6413453 and rs5082 contributes to the HDL-C trait in Iranian population, but a new missense mutation in APOA2 in our population has a significant association with HDL-C.     

Overweight and obesity among Iranian female adolescents in Rasht: more overweight in the lower social group

OBJECTIVE: This study aimed to investigate the frequency of overweight and obesity among Iranian adolescent girls aged 14-17 years and its possible association with maternal education in Rasht city in 2005. DESIGN: A cross-sectional survey on 1054 schoolgirls. SETTING: High schools in Rasht city, Iran. METHODS: A multistage sampling method was used and 1054 randomly selected schoolgirls were studied. Data on age and mother's years of schooling were collected by questionnaire and body weight and height were measured. Overweight and obesity were defined using age- and sex-specific body mass index (BMI) cut-off points proposed by the International Obesity Task Force (IOTF), and compared with 85th and 95th BMI percentile curves of the IOTF reference population, respectively. Data analyses included two groups based on mother's years of schooling: less educated ( < 12 years) and more educated ( > or = 12 years). RESULTS: The overall prevalence of overweight and obesity in this population was 21.9 and 5.3%, respectively. Results of logistic regression analysis showed that the risk of overweight was higher for lower age group (odds ratio = 2.0, 95% confidence interval 0.18-3.09). Overweight was more common in girls with less educated mothers than in girls with more educated mothers (26.0 vs. 19.8%, P = 0.03). CONCLUSION: These data indicate that overweight is highly prevalent among adolescent girls, especially in lower social groups in Rasht, and the rate is exceeding those reported in other parts of the country. Preventive strategies need to be adopted to combat the epidemic of overweight and obesity in this population.     

Molecular survey of <Emphasis Type="Italic">Toxoplasma</Emphasis> infection in sheep and goat from Fars province,Southern Iran

Toxoplasmosis is a widespread zoonotic disease that causes significant morbidity and mortality in human fetus and in immunocompromised patients. Moreover, it becomes a major cause of abortion in sheep and goats. Since consumption of meat of infected lamb and goat is considered as the main sources of human infection in Iran, this study was undertaken to determine the prevalence of Toxoplasma infection in edible tissues of sheep and goats in Shiraz in 2008. Samples of brain, tongue, liver, and muscles of neck, intercostals, and femoral were taken from 56 sheep and 22 goats and tested by PCR. The total prevalence of Toxoplasma infection among animals was found to be 33.3%. Five out of 22 goats (22.7%) and 21 out of 56 sheep (37.5%) were infected by Toxoplasma. Differences between the prevalence rate of infection among females (nine out of 14 = 46%) and males animals (12 out of 45 = 29.5%) was significant (P = 0.013). Furthermore, a positive correlation was found between the age of animals and the rate of infection; animals greater than 2 years old showed a higher rate of infection (47%) in comparison with those less than 2 years old (25%, P = 0.04). The highest infected tissue was tongue (21.8%) followed by brain (19.2%) and femoral and intercostal muscles (17.9%). This study demonstrated a high level of Toxoplasma infection in slaughtered animals in Shiraz and these should be considered as the main sources of infection for human population in the region.     

Effects of plant residues and calcite amendments on soil sodicity

The effects of wheat, potato, sunflower, and rape residues and calcite were evaluated in soil that received sodic water. These materials were added to a sandy‐loam soil at a rate of 5%, after which the treated soils were incubated for 1 month at field‐capacity moisture and a temperature of 25°C–30°C. Column leaching experiments using treated soils were then conducted under saturated conditions using water with three sodium‐adsorption ratios (SAR) (0, 10, 40) with a constant ionic strength (50 mmol L^–1). The results indicated that the application of plant residues to soils caused an increase in cation‐exchange capacity and exchangeable cations. Leaching experiments indicated that the addition of plant amendments led to increased Na⁺ leaching and decrease in exchangeable‐sodium percentage (ESP). The ESP of the control soil, after leaching with solutions with an SAR of 10 and 40, increased significantly, but the level of sodification in soils treated with plant residue was lower. Such decreases of soil ESP were greatly affected by the type of plant residues, with the order of: potato‐treated soil > sunflower‐treated soil > rape‐treated soil > wheat‐treated soil > calcite‐treated soil > control soil.     

Genetic analysis of Rutilus rutilus caspicus (Jakowlew 1870) populations in Iran by microsatellite markers

The Caspian roach (Rutilus rutilus caspicus) is regarded as one of the most important commercial species in the Caspian Sea. Despite the commercial and conservation importance of the species, information on genetic relationships and diversities at the molecular level in Rutilus rutilus caspicus is scarce. Six microsatellite loci were analysed to study the genetic variation in two major populations of Caspian roach in the Iranian waters of the Caspian Sea: the Anzali Wetland and Gorgan Bay. The F_st value between populations was 0.07, suggesting that the genetic differentiation was significant (P<0.01). Both the studied populations deviated from Hardy–Weinberg equilibrium proportions at a number of loci, mostly due to the deficiency of heterozygosities. The genetic distance between populations was 0.29, which indicates that the genetic difference among the studied populations is pronounced. The differences between both populations were not statistically significant (P>0.05), neither for average number of alleles per locus nor for observed heterozygosities. The data generated in this study provide useful information on the genetic variation and differentiation in populations of Caspian roach.     

The impact of land afforestation on carbon stocks surrounding Tehran,Iran

The city of Tehran,like many polluted metropolises of the world,has higher emissions of greenhouse gases than other cities in Iran,due to heavy consumption of fossil fuel and landuse changes.To estimate carbon sequestration in two 40 year-old stands of planted Cupressus arizonica and Fraxinus rotundifolia in degraded lands surrounding Tehran,sampling of above-and below-ground biomass,soil(at two depths of 0-15 and 15-30 cm),and leaf litter was done by systematic random sampling.The total carbon stocks of C.arizonica and F.rotundifolia stands were respectively 328.20 and 150.69 Mg·ha-1.The aboveground biomass with 233.16(71%)Mg·ha-1 in C.arizonica and88.16(58.50%)Mg·ha-1 in F.rotundifolia contributed the most shares to carbon sequestration.The diameter at breast height,total height,basal area,total volume,and biomass of C.arizonica were significantly(p0.01)higher than those of F.rotundifolia.Also the depth of 0-30 cm of soil contributed between 18.29%and 32.15%of total ecosystem carbon,respectively.The economic value of carbon sequestration in the two stands in 2011 was calculated at 3.5 and 2.5 million dollars,respectively.Our results indicate that afforestation of the degraded land surrounding Tehran would sequester more carbon than would continuously degraded land,the current status quo.These stands can absorb atmospheric CO2 at different rates,thus tree species selection and stand development should be considered in planning future afforestation projects.     

Serum concentration of sialic acids in naturally occurring ovine babesiosis

This study is designated to assess the effect of the severity of Babesia ovis infection on sialic acid concentration in blood sera in naturally infected sheep. Infected animals (diseased group) comprised 38 Iranian fat-tailed sheep, about 1–3 years old, naturally infected with B. ovis, divided into four subgroups with respect to parasitemia rates (low 0.1–0.3 %, moderate 0.4–0.9 %, high 1–2.5 %, and very high >2.5 %). The parasitological diagnosis was confirmed using PCR analysis. As a control group, ten clinically healthy sheep reared under the same management and environmental conditions were also sampled. Hematological parameters and the concentrations of total sialic acid (TSA), lipid-bound sialic acid (LBSA), and protein-bound sialic acid (PBSA) were measured in both groups. Compared to controls, sialic acid concentrations showed significant increase (p?Parasitemia rate was positively correlated with sialic acid concentrations. This study demonstrated that B. ovis infection induced marked and persistent elevations of serum sialic acid concentrations. It seems that increase of serum sialic acid concentrations during parasitemia alter receptor-ligand interactions, which are known to play important role in immune response. Furthermore, sialic acid would indirectly inhibit the action of leukocytes and consequently promote the evasion of the immune response and persistence of the parasite in the host. This factor could influence the parasite-host cell adhesion, but further detailed biochemical investigations are needed to precisely explain the exact role of sialic acid in invasion process of the parasite to the host cells.