Evaluation of Blood Parameters Alteration Following Low-dose Radiation Induced by Myocardial Perfusion Imaging.

INTRODUCTION: With increasing the usage of myocardial perfusion imaging (MPI) for the diagnosis of ischemic heart disease, we aimed to evaluate the side effects of low-dose radiation induced by this technique on blood elements, especially proteins and liver function factors. MATERIAL AND METHODS: 40 eligible patients (Mean age: 54.62±10.35, 22 female and 18 male), who had referred to the nuclear medicine department for MPI from May till August 2014, were enrolled in the study. A blood sample was taken from each patient just before and 24 hours after the injection of 740Mbq of Tecnetium-99m Methoxy isobutyl isonitrile (99mTc-MIBI) in the rest phase of the MPI in a reference medical laboratory; blood tests included total protein (TP), albumin (Alb), globulin (Glo), aspartate aminotransferase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), direct bilirubin (D.Bili), total bilirubin (T.Bili), serum iron (SI), total iron bounding capacity (TIBC), Albumin globulin ratioA/G ratio), and complete blood count (CBC). RESULTS: Injection of 740Mbq99mTc-MIBI caused a significant increase in serum levels of AST (p= 0.001), ALT (p= 0.001), SI (p= 0.030), TIBC (p= 0.003) and A/G Ratio (p= 0.020). However, following radiotracer injection, a significant decrease was noted in the serum levels of TP (p= 0.002), Alb (p= 0.014), Glo(p= 0.002), ALP (p= 0.001), D.Bili (p= 0.003) and T.Bili (p= 0.000). CONCLUSION: Due to increased usage of MPI, our data highlights the importance of monitoring the clinical and paraclinical effects of the procedure on vital organs and physiological pathways to reduce their adverse effects.

Genetic variation in the developmental regulation of cortical avpr1a among prairie voles.

Early experiences can have enduring impacts on brain and behavior, but the strength of these effects can be influenced by genetic variation. In principle, polymorphic CpGs (polyCpGs) may contribute to gene-by-environment interactions (G × E) by altering DNA methylation. In this study, we investigate the influence of polyCpGs on the development of vasopressin receptor 1a abundance in the retrosplenial cortex (RSC-V1aR) of prairie voles (Microtus ochrogaster). Two alternative alleles ('HI'/'LO') predict RSC avpr1a expression, V1aR abundance and sexual fidelity in adulthood; these alleles differ in the frequency of CpG sites and in methylation at a putative intron enhancer. We hypothesized that the elevated CpG abundance in the LO allele would make homozygous LO/LO voles more sensitive to developmental perturbations. We found that genotype differences in RSC-V1aR abundance emerged early in ontogeny and were accompanied by differences in methylation of the putative enhancer. As predicted, postnatal treatment with an oxytocin receptor antagonist (OTA) reduced RSC-V1aR abundance in LO/LO adults but not their HI/HI siblings. Similarly, methylation inhibition by zebularine increased RSC-V1aR in LO/LO adults, but not in HI/HI siblings. These data show a gene-by-environment interaction in RSC-V1aR. Surprisingly, however, neither OTA nor zebularine altered adult methylation of the intronic enhancer, suggesting that differences in sensitivity could not be explained by CpG density at the enhancer alone. Methylated DNA immunoprecipiation-sequencing showed additional differentially methylated regions between HI/HI and LO/LO voles. Future research should examine the role of these regions and other regulatory elements in the ontogeny of RSC-V1aR and its developmentally induced changes.

Synergistic Effects of NDRG2 Overexpression and Radiotherapy on Cell Death of Human Prostate LNCaP Cells.

BACKGROUND: Radiation therapy is among the most conventional cancer therapeutic modalities with effective local tumor control. However, due to the development of radio-resistance, tumor recurrence and metastasis often occur following radiation therapy. In recent years, combination of radiotherapy and gene therapy has been suggested to overcome this problem. The aim of the current study was to explore the potential synergistic effects of N-Myc Downstream-Regulated Gene 2 (NDRG2) overexpression, a newly identified candidate tumor suppressor gene, with radiotherapy against proliferation of prostate LNCaP cell line. MATERIALS AND METHODS: In this study, LNCaP cells were exposed to X-ray radiation in the presence or absence of NDRG2 overexpression using plasmid PSES- pAdenoVator-PSA-NDRG2-IRES-GFP. The effects of NDRG2 overexpression, X-ray radiation or combination of both on the cell proliferation and apoptosis of LNCaP cells were then analyzed using MTT assay and flow cytometery, respectively. RESULTS: Results of MTT assay showed that NDRG2 overexpression and X-ray radiation had a synergistic effect against proliferation of LNCaP cells. Moreover, NDRG2 overexpression increased apoptotic effect of X-ray radiation in LNCaP cells synergistically. CONCLUSION: Our findings suggested that NDRG2 overexpression in combination with radiotherapy may be an effective therapeutic option against prostate cancer.

Methylation of avpr1a in the cortex of wild prairie voles: effects of CpG position and polymorphism.

DNA methylation can cause stable changes in neuronal gene expression, but we know little about its role in individual differences in the wild. In this study, we focus on the vasopressin 1a receptor (avpr1a), a gene extensively implicated in vertebrate social behaviour, and explore natural variation in DNA methylation, genetic polymorphism and neuronal gene expression among 30 wild prairie voles (Microtus ochrogaster). Examination of CpG density across 8 kb of the locus revealed two distinct CpG islands overlapping promoter and first exon, characterized by few CpG polymorphisms. We used a targeted bisulfite sequencing approach to measure DNA methylation across approximately 3 kb of avpr1a in the retrosplenial cortex, a brain region implicated in male space use and sexual fidelity. We find dramatic variation in methylation across the avrp1a locus, with pronounced diversity near the exon-intron boundary and in a genetically variable putative enhancer within the intron. Among our wild voles, differences in cortical avpr1a expression correlate with DNA methylation in this putative enhancer, but not with the methylation status of the promoter. We also find an unusually high number of polymorphic CpG sites (polyCpGs) in this focal enhancer. One polyCpG within this enhancer (polyCpG 2170) may drive variation in expression either by disrupting transcription factor binding motifs or by changing local DNA methylation and chromatin silencing. Our results contradict some assumptions made within behavioural epigenetics, but are remarkably concordant with genome-wide studies of gene regulation.

Look Different: Effect of Radiation Hormesis on the Survival Rate of Immunosuppressed Mice.

BACKGROUND: Hormesis is defined as the bio-positive response of something which is bio-negative in high doses. In the present study, the effect of radiation hormesis was evaluated on the survival rate of immunosuppressed BALB/c mice by Cyclosporine A. MATERIAL AND METHODS: We used 75 consanguine, male, BALB/c mice in this experiment. The first group received Technetium-99m and the second group was placed on a sample radioactive soil of Ramsar region (800Bq) for 20 days. The third group was exposed to X-rays and the fourth group was placed on the radioactive soil and then injected Technetium-99m. The last group was the sham irradiated control group. Finally, 30mg Cyclosporine A as the immunosuppressive agent was orally administered to all mice 48 hours after receiving X-rays and Technetium-99m. The mean survival rate of mice in each group was estimated during time. RESULTS: A log rank test was run to determine if there were differences in the survival distribution for different groups and related treatments. According to the results, the survival rate of all pre-irradiated groups was more than the sham irradiated control group (p < .05). The highest survival time was related to the mice which were placed on the radioactive soil of Ramsar region for 20 days and then injected Technetium-99m. CONCLUSION: This study confirmed the presence of hormetic models and the enhancement of survival rate in immunosuppressed BALB/c mice as a consequence of low-dose irradiation. It is also revealed the positive synergetic radioadaptive response on survival rate of immunosuppressed animals.

Detection of t(9;22) b2a2 fusion transcript by flow cytometry.

INTRODUCTION: We assessed the feasibility of flow cytometry-fluorescent in situ hybridization technique in the detection of translocated mRNA in the cytoplasm of human peripheral blood nucleated cells. It is assumed that this assay can be applied as a diagnostic method in the detection of chromosomal translocation which commonly occurs in hematologic malignancies. METHODS: KCL-22 cell line and white blood cells from 21 CML patients were recruited in the study. Cells were isolated and fixed. After permeabilization, cells were resuspended in hybridization buffer and probes were added to the mixture. Subsequently, cells were washed and analyzed on the flow cytometer instrument. The flow cytometry results were compared with qRT-PCR and fluorescent microscope outcomes. RESULTS: Using the current principle, 97 ± 2.1% of the KCL-22 cells were labeled with b2a2 mRNA-specific probes. In addition, seven patients were recognized positive for t(9;22) b2a2. The percentage of cells containing abovementioned translocation in these patients was varied from 3.26% to 97.8%. There was no false-positive result in negative controls (K562 with BCR-ABL1 b3a2, NB4, and Jurkat cell lines) along with blood samples of normal controls. All the results obtained by flow-FISH were confirmed by qRT-PCR and fluorescent microscope. CONCLUSION: This strategy benefits from appropriate specificity, sensitivity, rapidity, and ability in the determination of malignant cell percentage. Therefore, it can improve traditional time-consuming and labor-intensive FISH method.

Effects of Hesperidin as a Radio-protector on Apoptosis in Rat Peripheral Blood Lymphocytes after Gamma Radiation.

INTRODUCTION: Hesperidin (HES), as the most abundant flavonoid existing in the citrus, is widely used by human daily. The radio-protective effects of Hesperidin have been confirmed in various measurement systems. This study aimed to evaluate the effects of Hesperidin on the changes in the apoptosis level and expression of apoptotic genes target (bax, bcl-2 and ration of bax/bcl-2) in the peripheral blood lymphocytes of male rats after gamma radiation. MATERIALS AND METHODS: 64 male rats were divided into eight groups: Control, HES (100 mg/kg b.w, orally, 7 days), whole body irradiation with 2 and 8Gy, pre-administrated with 50 and 100 mg/kg body weight of Hesperidin for 7 days before irradiation with 2 and 8 Gy. 24 hours after radiation, apoptotic lymphocytes were evaluated using PE Annexin V Apoptosis detection I kit and the levels of mRNA for bax and bcl-2 were evaluated by real time reverse transcription polymerase chain reaction. RESULTS: A significant reduction in apoptosis of the lymphocytes was demonstrated in group animals receiving 8 Gy compared to the group which received 2 Gy irradiation (p<0.0001). However, apoptosis significantly increased in group of rats who received Hesp before irradiation (p<0.05). The increase of apoptosis by Hesperidin administration can be attributed to the decreased expression of bax and significantly reduced expression of bcl-2 and finally increasing the ration of bax/bcl-2. CONCLUSION: The results suggest that administration of 50 and 100 mg/kg of Hesperidin induces apoptotic effects by changing expression level of bax, bcl-2 and also the ratio of bax/bcl2.

Construction of bacterial ghosts for transfer and expression of a chimeric hepatitis C virus gene in macrophages.

The bacterial ghost (BG) production is a field of biotechnology for applications in vaccine and drug delivery. We assessed the capacity of BG for delivery of a recombinant gene encoded for both cell mediated and antibody dependent epitopes of hepatitis C virus (HCV) into murine macrophages. Escherichia coli (E. coli) cells were transformed with the lysis plasmid (pHH43). To produce chimeric gene, NS3 (non-structural protein 3) and core regions of HCV genome were fused together by splicing by overlap extension (SOEing) PCR and were cloned into plasmid pEGFP-C1. Bacterial ghosts were loaded with recombinant pEGFP-C1 and then were transferred to murine macrophages (RAW 264.7). To investigate plasmid transfection and chimeric mRNA transcription, fluorescent microscopy and RT-PCR were used. In vitro studies indicated that bacterial ghosts loaded with pEGFP-C1 plasmid were efficiently taken up by murine macrophages and indicated a high transfection rate (62%), as shown by fluorescent microscopy. RT-PCR from extracted intracellular mRNAs for chimeric Core-NS3 gene showed a specific 607 bp fragment of the gene. The sequence analysis of purified PCR products demonstrated the expected unique mRNA sequence. We constructed a chimeric HCV gene containing both cell mediated and antibody dependent epitopes with a significant expression in murine macrophages delivered by bacterial ghost.

Biological control of Tiarosporella phaseolina the causal agent of charcoal rot of soybean.

Charcoal rot caused by Tiarosporella phaseolina (Tassi) Van der Aa is an important disease of soybean in Gorgan province of Iran. Experiments were carried out with 95 bactenal isolates that were collected from the rhizosphere of soybean plant. Among these bacteria only 50 isolates showed antagonistic effect on Tiarosporella phaseolina using dual culture test. Six highly effective bacteria were selected for subsequent studies. Based on biochemical physiological and morphological tests, isolates Pf-12 and Pf-63 were identified as Pseudomonas fluorescens, isolates B-13, B-42,B-126 and B-84 as Bacillus subtilis. The isolates of P. fluorescens produced antibiotics as well as volatile metabolites that inhibited mycelial growth of fungus. Bacillus subtilis isolates inhibited the fungal growth through volatile and non-volatile metabolites production. Only P. fluorescens isolates produced hydrogen cyanide. In greenhouse studies, the isolates B-13 and B-126 reduced 59% and 66% the intensity of charcoal rot of soybean respectively. The combinations of isolates B-13 and B-126 were also effective on reducing the intensity of disease.

Effect of tea dust residues to control root-knot nematode of tomato.

In this research, control of tomato root- knot nematode (Meloidogyne incognita) was conducted using tea dust residues at different rates. First, the species and race of nematode were identified by employing diagnostic keys. Then, with 5 replications in complete randomized design. Tea dust residues were used at 9 treatments (0, 5, 10, 15, 20, 25, 30, 35 and 40 g/kg of soil). Statistical analysis on mean treatments rates showed that treatment with 25 g/kg soil economically was effective in growth rates and reduction in gall index.

Study on interaction between root-knot nematode Meloidogyne javanica and wilt fungus Verticillium dahliae on olive seedlings in greenhouse.

Verticillium dahliae has been reported as a limiting factor in cotton, olive, potato and tomato fields from several countries in the world. Root-knot nematodes Meloidogyne javanica causes considerable damage to olive groves in olive growing areas. Since the presence of these two pathogens in olive trees and seedlings were confirmed in Golestan Province, this study was proposed to find the mode of their action and interaction with olive seedlings in greenhouse. The non-defoliant strain of the fungus (SS-4) was isolated from olive groves showing symptom in Golestan Province. M. javanica was also recovered from the infested olive seedlings. After species identification, it was reared on tomato seedlings var. Rutgers. The larvae were used as a source of inoculum. Conidia and microsclerotia of V. dahliae were used as a source of inoculum for pathogenesis in this study. Stem cuttings of olive cultivar Zard were transplanted in different sets of pots containing 720 ml. of sterilized loamy soil and sandy soil. Experiment was conducted in Completely Randomized Design with 6 treatments and 8 replicates including control, nematode alone, fungus alone, nematode and fungus simultaneously, nematode and fungus concomitantly, fungus two weeks prior to nematode, nematode and fungus concomitantly, nematode two weeks prior to fungus. Pots were inoculated with 1500 larvae of nematodes and 7200 microsclerotia of V. dahliae. Experiment was terminated after 9 months and following parameters were determined i.e. fresh weight of roots, number of galls and females, per root system and discoloration of leaf and root tissues. Presence of nematode prior to fungus caused reduction in colonization of fungus in the roots and the stems and vis presence of fungus prior to nematode caused reduction in number of galls produced by nematode. Sever symptom on aerial parts of plant was observed when both pathogens were inoculated simultaneously. However fresh weight of roots was reduced in all treatments. Galling and population density in root system were higher in pots filled with sandy soil than in loamy soil, but fungal colonization in roots and stems was much more in seedlings grown in loamy soil and diseases symptoms in aerial parts of plants were much severe in plants grown in loamy soil than in pots with sandy soil.

Interaction between root lesion nematode Pratylenchus vulnus and two species of Fusarium on growth and development of maple seedlings.

Producing healthy seedlings and distributing them to far areas, is one of the most important factor for developing forests. Because of easy planting, rapid growth and good wood quality, Acer velutinum is the most useful species, among the softwood and hardwood trees. Growth and development of the nurseries were effected by different pests and diseases, the nematodes are one of these agents. They are not actively surveyed in the forest nurseries in Iran. On a survey of maple seedling in the forest nurseries Pratylenchus vulnus, Fusarium solani and F. oxysporum were identified. The interaction between these two fungi and nematode were studied in randomized complete block design with four replications and six treatments. For mass production and multiplication of the nematodes, they were sterilized with Streptomycin Sulfate, they were reared on carrot disc culture. The nematodes were then added to each pot. The results showed a highly significant difference (p = 0.01) between different treatments. In the treatment with nematodes alone growth of seedlings was minimum and population density of nematodes in soil and root tissue was maximum. In this treatments 75% of seedlings were died and seedling average height were 2.25 cm. In the treatments with nematode and fungi together the population of nematodes were decreased and the growth of seedlings were increased, in comparison with nematode alone. F. oxysporum showed more antagonistic effect on nematodes than F. solani. In the nematode + F. oxysporum treatment, 25% of seedlings were died and seedling average height was 12.75 cm. In the nematode + F. solani treatment, 50% of seedlings were died and the seedling average height was 5.5 cm. This is the first report of pathogenecity of Pratylenchus vulnus and its interaction whit fungi on maple seedlings in Iran.

Barley leaf stripe disease.

Leaf stripe is one of the most important diseases of barley in Iran especially in Gorgan, Mazandran and near Tehran (Varamin). Most obvious symptoms of the disease are described. Long pale or yellow stripes become darker as the fungus sporulates on the leaf surface. Infected plants usually are stunted and produce sterile spikes, rarely a few seeds are produced. Infected spikes and late-forming tillers may produce fertile spikes. The fungus is seed brone and survives in the outer layers of infected seed. To study the seed-borne disease, we have used the different methods (ISTA). Coleoptiles of seedlings are infected by the fungus under cool, moist conditions, a soil temperature below 15 degrees C is necessary for seed infection. The fungus penetrates through coleoptiles and grows systemically within the plant, produces toxin and kills cells and discolors leaf tissue between veins, thus causing striped lesions. When conditions are wet or humid, spores are produced on the surface of leaves at above the time spikes of healthy plant. Morphological characteristics of the vegetative and reproductive structures of the fungus show that it is Drechslera graminea (Rabenh) Shoemaker.