Inter-Simple Sequence Repeat (ISSR) molecular marker for the isolation of five races of honey bee Apis mellifera from Khuzestan, Kurdistan, Markazi, Fars and Isfahan provinces was studied. DNA was extracted from the worker bees. The quality and quantity of extracted DNA were measured. Values obtained from the bands on agarose gel (1.5%) were scored and analyzed. Results showed that the bands are in the range between 250 and 1000 bp. Maximum number of bands were recorded for primer 1. Cluster analysis of races placed them in two main groups. In the first group were Kurdistan and Khuzestan. The second group includes Markazi, Fars and Isfahan, but this group has shown no relationship between the populations. It seems that ISSR molecular marker could separate different races of honey bee.

Previous investigations demonstrated that crude peroxidase (POD) obtained from onion solid wastes has the ability to catalyse the formation of the aurone aureusidin (ARS), using 2´,4´,6´,3,4-pentahydroxy chalcone (PHC) as substrate, although this reaction under physiological conditions is mediated by a polyphenol oxidase-like enzyme, called aureusidin synthase (AS). In this study, a crude onion POD preparation was used to study the effect of some critical factors affecting the reaction, including reaction time, pH and temperature. The optimal set of conditions was identified by deploying central composite factorial design and response surface methodology. The results obtained showed that the optimum values for pH and temperature were 5 and 20°C, respectively, while time was found to exert a statistically non-significant effect. These values were the same or very close to optimal conditions found for structurally different onion POD substrates. The outcome was discussed with regard to the applicability of the onion POD as a versatile tool of biocatalysis.

In temperate forests, formation of canopy gaps by windthrow is a characteristic natural disturbance event. Little work has been done on the effects of canopy gaps on soil properties and fauna, especially earthworms as ecosystem engineers. We conducted a study to examine the reaction of earthworms (density/biomass) and different soil properties (i.e., soil moisture, pH, organic matter, total N, and available Ca) to different canopy gap areas in 25-ha areas of Liresar district beech forest located in a temperate forest of Mazandaran province in the north of Iran. Soil samples were taken at 0-15, 15-30 and 30-45 cm depths from gap center, gap edge and closed canopy using core soil sampler with 81 cm2 cross section. The earthworms were collected simultaneously with the soil sampling by hand sorting method. Our study supports that the canopy gap will create a mosaic of environmental conditions. Earthworm's density and biomass tended to be higher in small canopy gaps compared with the other canopy gap areas. Earthworm's population showed decreasing trend from closed canopy to disturbed sites (gap edge and gap center). The top soil was more appropriate to presence of earthworms although ecological groups have occupied different soil layers. As a conclusion, earthworms can be introduced as good bio-indicator of environmental changes that occur by disturbance.

Epigenetic silencing of gene expression has become an intensively studied topic regarding breast cancer during the last years. Previous investigations have shown that hypermethylation in gene promoters is an alternative to mutations cancer-related mechanism for inactivation of tumor suppressor gene function. The present study was designed for evaluation of promoter methylation in p53 and ATM tumor suppressor genes in Bulgarian patients with sporadic breast cancer. Sodium bisulfite conversion and methylation-specific PCR were performed on tumor DNA isolated from 55 patients with sporadic breast cancer. The data showed normal profile of methylation in the promoter region of p53 and ATM genes in all studied patients. Cases with hypermethylation were not observed. Our results did not support the involvement of p53 and ATM promoter hypermethylation in the neoplastic pathways of sporadic breast cancer. The lack of promoter hypermethylation in p53 and ATM is suggestive of a greater share of genetic aberrations in loss of tumor suppressor function of these genes and presumes the involvement of other epigenetic cancer mechanisms. To our knowledge this study provides the first data on promoter hypermethylation status of p53 and ATM genes in Bulgarian patients.

Itaconic acid (IA) is one of the hopeful substances within the cluster of organic acids. IA is used in artificial glass, bioactive compounds in pharmacy, medicine, agriculture, for the synthesis of fiber, resin, plastic, rubber, paints, surfactant, ion-exchange resins and lubricant. Most recurrently used microorganism for commercial production of IA is Aspergillus terreus. Some filamentous fungi belonging to Ustilaginales also produce IA. In the present work, an attempt was made to produce IA by Ustilago maydis employing Solid State Fermentation (SSF) from various agro wastes like ground nut shells, rice bran, rice husk, orange pulp, ground nut oil cake, orange pulp and sugarcane bagasse as carbon substrates, which were used after pretreatment. 10 g of each substrate was taken in a 500 ml conical flasks separately and supplemented with 20 mL nutrient solution containing glucose, at pH 3. One milliliter inoculum containing 1×107 spores was added and moisture was maintained at 60%. After incubation at 32°C for 5 days, the acid production was estimated by spectrophotometric method and by HPLC analysis. Interestingly, the yield of itaconic acid was promising with all the above substrates, where orange pulp, sugarcane bagasse and rice bran supported higher yields.

p27Kip1 has been hypothesized to play a major role in carcinogenesis. Most of the published data reported that loss of p27Kip1 expression was strongly associated with development and progression of tumour. The purpose of this study was to analyze p27Kip1 expression in normal, benign and malignant prostate cancer tissues and their association with the clinicopathological parameters. The expression of p27Kip1 was evaluated by an immunohistochemistry method. p27Kip1 expression was significantly higher in normal and benign prostate tissues (P<0.01). In contrast, some malignant tissues had no p27Kip1 expression and most had weak p27Kip1 expression. p27Kip1 expression was found to be decreased significantly with increasing Gleason scores (P=0.003). Most of prostate adenocarcinomas (PCa) with Gleason 8 and 9 showed loss of p27Kip1 expression. The expression was also positively correlated with prostate specific antigen level and age in PCa group (P=0.003 and 0.043 respectively), whereas no association was found between the p27Kip1 expression with tumour amount and age in benign prostatic hyperplasia group. This study suggests that loss of p27Kip1expression is essential during development and progression of prostate cancer.

Cancer of gallbladder is a hidden phenomenon and highly malignant with underprivileged diagnosis and poor survival. Study of cancer patterns amongst victims of Bhopal gas tragedy exposed to methyl isocyanate revealed higher incidence of gallbladder cancer that necessitated a more objective elucidation of the disease at its molecular level. Tissues of 92 cases of gallbladder cancer patients were taken in the study (31 men and 61 women, age range 16–85 years, mean age 45.8±1.50 years). Mutations of ATM, γ-H2AX and p53 were predominantly seen in the methyl isocyanate exposed cohort diagnosed with adenocarcinoma, with 61.4% (43/70), 54.3% (38/70) and 73% (51/70) respectively, involving infiltration into the papillary and mucinous region/cell types of the gallbladder. Out of these, the expression frequency of all the above three genes was higher in moderately differentiated adenocarcinoma in comparison to poorly and well-differentiated ones. The results of the present study support the hypothesis that ATM and p53 mutations provide fundamental genetic signatures influencing tumor behavior across patient subsets and invasiveness of the disease, while γ-H2AX is apparently an ordinary pathway involved in the genesis of tumors.

A refined and high efficiency protocol for in vitro regeneration of Commiphora wightii, a red-listed medicinal plant of medicinal importance, has been developed through optimized somatic embryogenesis pathway. Cultures from immature fruits were induced and proliferated on B5 medium supplemented with 2.26 µM 2,4-D. Embryogenic calli were obtained and then maintained for extended periods by alternately subculturing on modified MS medium supplemented with 1.11 µM BAP, 0.57 µM IBA and with 0.5% activated charcoal or without PGR every 3-4 weeks. Cyclic embryogenesis was obtained. Late torpedo and early cotyledonary stages somatic embryos were regularly harvested from PGR-free modified MS medium. It was found that percent moisture available in culture containers play a critical role in maturation and subsequent germination of somatic embryos of C. wighti. Maximum germination of more than 80% was achieved through media recycling. Somatic embryo derived plants (emblings) were acclimatized. After 5 months, acclimatized plants were out-planted in experimental field. These morphologically normal plants have been surviving for over 3 years. Molecular polymorphism was clearly evident when these plants were tested using RAPD primers, making the plants suitable for use in its species restoration program.

The Jamaican bitter yam (Dioscorea polygonoides) (ITIS) is known to possess potent antidiabetic and hypocholesterolemic properties and can therefore be exploited for associated nutraceutical/pharmaceutical purposes. It however possesses bioactive compounds known to promote organ damage when ingested in excess. This study investigates the effects of bitter yam consumption at a concentration of 5% on liver and kidney damage/function parameters. Normocholesterolemic mice fed bitter yam supplemented diets experienced significant increases in serum aspartate aminotransferase activity and bilirubin, magnesium and phosphorus concentrations. Significant increases were also observed in serum aspartate aminotransferase activity and blood urea nitrogen concentration of the genetically modified hypercholesterolemic mice fed supplemented diets. These results suggest mild kidney damage in both mice species and a significant increase in the rate of erythrocyte haemolysis in the normocholesterolemic mice.

The qualitative and quantitative composition of the essential oils obtained from male and female leaves of Juniperus thurifera L., (growing in Algeria) has been investigated for the first time. The essential oils were obtained by hydrodistillation (0.45% from female trees and 0.53% from male trees, v/w dried material) and analyzed by gas choromatography (GC) and gas choromatography-mass spectrometery (GC-MS). Seventy-seven compounds were identified, representing more than 97% of the oils. The major components were Sabinene, α-pinene and terpinene-4-ol. The concentrations of the oil constituents: α-thujene, α-pinene, α-phellandrene, p-cymene, linalyl acetate, Δ-amorphene, germacrene D-4-ol, and 4-epi-abietal were greater in the oil of the female tree than in the oil of the male tree. Conversely, the concentrations of α-terpinene, γ-terpinene, terpinene-4-ol, elemol, α-epi-cadinol and α-eudesmol were greater in the oil of the male tree than in the oil of the female tree. However, the concentration gradient trends for both female and male trees were similar for sabinene, myrcene, linalool, β-pinene, limonene, cis-sabinene hydrate terpinolene, α-terpineol. The antimicrobial activity of male and female J. thurifera essential oils was evaluated against 14 bacteria. The results showed a variable degree of antibacterial activity depending from the type of the oil (extracted from male or female trees). Essential oils of female trees were most effective.

Five Enterococcus isolates from some Nigerian traditional fermented foods were identified as Enterococcus gallinarum by using phenotypic and genotypic tests. Safety properties such as antibiotic susceptibility, virulence gene detection, haemolysin, gelatinase and bacteriocin production were determined using standard methods. There was no resistance to clinically relevant antibiotics. Virulence gene for collagen binding antigen and aggregation substance were detected in 60% of the E. gallinarum strains; while surface adhesin was detected in 20%, but none of the strains had cytolysin activator and gelatinase. Phenotype characterizations of the E. gallinarum isolates indicated that none of the isolates produced haemolysin and gelatinase. Enterococcus gallinarum C103 and U82 had no antimicrobial activity against all the selected bacteria pathogens while E. gallinarum W184, T71 and W21 were active against some of the indicator bacteria pathogens. Only E. gallinarum T71 and W21 showed broad spectra of antimicrobial activity. Combination of virulence factors did not appear in these food isolates. Therefore, these strains particularly the two strains with high spectra of antimicrobial activity could be exploited as functional starters in foods.