Use of a mammalian gonadotropin‐releasing hormone (GnRH) agonist to characterize pituitary GnRH receptors in white sturgeon (Acipenser transmontanus Richardson)

Aim of the present study was to characterize the pituitary GnRH receptor in white sturgeon, Acipenser transmontanus, using a superagonist analog of mammalian GnRH [D‐Ala⁶, des‐Gly¹⁰–Pro⁹]‐ethylamide and to investigate the possible effect of estradiol‐17β treatment on the concentration and affinity of the GnRH receptor in immature white sturgeon. The binding of ¹²⁵I‐GnRH‐A to sturgeon pituitary receptors was rapid and saturable at 4°C and 20°C. However, maximal binding at 20°C was almost two‐fold greater than the highest binding noted at 4°C. Specific binding of radioligand was directly related to the amount of tissue included in the assay system over the range of 5–20 mg fresh tissue equivalents per ml. The binding capacity of ¹²⁵I‐GnRH‐A with sturgeon pituitary tissue was much greater than radiolabeled GnRH. Administration of E₂ to immature sturgeon caused an almost two‐fold increase in GnRH‐A binding capacity (E₂ treated: Bmax = 2.87 fmoles 3 mg^?1 FTE; control: Bmax = 1.70 fmoles 3 mg^?1), and did not affect GnRH‐A binding affinity (E₂ treated: Ka = 0.13 × 10¹¹ m ^?1; control: Ka = 0.15 × 10¹¹ m ^?1). Overall, the study provides evidence that the GnRH analog is effective for characterizing the GnRH receptor in white sturgeon; however, more experimentation is necessary to determine whether E₂ administration to immature white sturgeon can increase the GnRH receptor capacity.     

Molecular detection of nematicidal crystalliferous Bacillus thuringiensis strains of Iran and evaluation of their toxicity on free-living and plant-parasitic nematodes

The characterization of nematode-effective strains and cry genes in the Iranian Bacillus thuringiensis (Bt) collection (70 isolates) is presented. Characterization was based on PCR analysis using 12 specific primers for cry5, cry6, cry12, cry13, cry14, and cry21 genes encoding proteins active against nematodes, crystal morphology, and protein band patterns as well as their nematicidal activity on root-knot nematode (Meloidogyne incognita) and two free-living nematodes (Chiloplacus tenuis and Acrobeloides enoplus). PCR results with primers for these genes showed that 22 isolates (31.5%) contain a minimum of one nematode-active cry gene. Strains containing the cry6 gene were the most abundant and represent 22.8% of the isolates. Bt strains harboring cry14 genes were also abundant (14.2%). cry21 and cry5 genes were less abundant, found in 4.2% and 2.8% of the strains, respectively. In total, six different nematode-active cry gene profiles were detected in this collection. Four isolates did not show the expected PCR product size for cry5, cry6, and cry21 genes; they might contain potentially novel insecticidal crystal protein genes. Twenty-two Bt isolates containing nematode-active cry genes were selected for preliminary bioassays on M. incognita. Based on these bioassays, four isolates were selected for detailed bioassays. Isolates YD5 and KON4 at 2 x 10(8) CFU/mL concentrations showed 77% and 81% toxicity on M. incognita, respectively. The free-living nematodes C. tenuis and A. enoplus were more susceptible and the highest mortality was observed within 48 h of incubation at all of the concentrations tested. Maximum mortality was recorded for isolates SN1 and KON4 at 2 x 10(8) CFU/mL concentrations and resulted in 68% and 77% adults deaths of C. tenuis and 68% and 72% for A. enoplus, respectively. Our results showed that PCR is a useful technique for toxicity prediction of nematicidal Bt isolates.     

Methylenetetrahydrofolate reductase gene polymorphism in diabetes and obesity

Methylenetetrahydrofolate reductase (MTHFR) polymorphism may play an important role in the pathophysiology of obesity and diabetes accompanied by obesity due to its influence on plasma homocysteine levels. There are significant and sometimes very strong relationship between levels of homocysteine and several multi-system diseases including CHD and CVA. To examine the association between MTHFR gene C677T polymorphism in diabetes and obesity with serum homocysteine levels. A total of 682 subjects were recruited in four groups (Normal, obese, diabetic and obese and diabetics). MTHFR gene C677T polymorphism was detected using PCR-RFLP technique. Serum homocysteine levels were measured using HPLC. There was a significant increase in the mean serum homocysteine levels in subjects carrying TT genotype (34.6 ± 26.5) compared to subjects carrying CC (15.1 ± 8) or CT genotype (16.4 ± 7.8) (P < 0.000). We found no significant differences for MTHFR allele and genotype frequencies between different groups. Our data have confirmed the association between serum homocysteine levels and MTHFR C677T genotype reported in other populations.     

Cytotoxicity of carboplatin on human glioblastoma cells is reduced by the concomitant exposure to an extremely low-frequency electromagnetic field (50 Hz, 70 G)

Glioblastoma multiforme (GBM) is a malignant brain cancer that causes high mortality in patients. GBM responds weakly to the common cancer treatments such as chemotherapy and radiotherapy and even surgery. Carboplatin is an alkylating agent widely used to treat cancer. However, resistance to this drug is a common problem in its use in cancer treatment. Concomitant exposure to extremely low-frequency electromagnetic fields (ELF-EMFs) and carboplatin is one unexplored possibility for overcoming this resistance. Indeed, many lines of evidence show that EMF affects cancer cells and drug action. In this study, we evaluated the effect of concomitant administration of carboplatin and EMF (50 Hz, 70 G) and also concomitant administration of carboplatin and static magnetic field (SMF) (70 G) on human glioma cell line (U-87). The results showed that cotreatment reduced the efficiency of carboplatin in U-87 cells, by decreasing caspase-3 in comparison to drug groups. Overall, EMF reduced the apoptotic effect of carboplatin, possibly through a redox regulation mechanism. Therefore, we have to avoid coadministration of magnetic field (MF) and carboplatin in tumor area, because the MF decreased the toxicity of the drug. However, further studies are needed to reveal the action mechanism of this combination therapeutic method.     

Variation in the Essential Oil Composition,Antioxidant Capacity,and Physiological Characteristics of Pelargonium graveolens L. Inoculated with Two Species of Mycorrhizal Fungi Under Water Deficit Conditions

In recent years, the application of arbuscular mycorrhizal fungi (AMF) has been considered to be an important strategy for improving crop yield and quality. In the present study, a factorial experiment based on a complete randomized design with two factors was performed to investigate the effect of AMF and water stress on the essential oil (EO) composition, antioxidant activity, and physiological and morphological characteristics of rose-scented geranium (Pelargonium graveolens L.). The factors included AMF inoculation (Rhizophagus intraradices, Funneliformis mosseae, and a mixture of both species) and irrigation levels [well-watered (WW), moderate drought stress (MDS), and severe drought stress (SDS)]. The main EO constituents were citronellol (31–37%) and geraniol (9–14%) in all treatments. Under water-stress conditions, some constituents increased, such as geraniol and geranyl formate, whereas others decreased, such as linalool, menthone and rose oxide. Overall, the highest amount of citronellol (37.3%) and geraniol (14.8%) was obtained in the plants inoculated with F. mosseae and R. intraradices under WW and MDS conditions, respectively. Antioxidant activity, total flavonoids, and phenolics were increased because of AMF inoculation, whereas a different trend was observed for the phenolic and flavonoid contents under water-stress conditions. Furthermore, water deficit elevated the amount of soluble carbohydrates as well as the proline content, whereas the amount of proline was lower in inoculated plants than in non-inoculated ones. All the growth parameters were improved in the AMF-inoculated plants compared to non-inoculated ones under different irrigation regimes. Drought conditions decreased the photosynthetic pigments and efficiency, whereas AMF plants ameliorated the adverse effect of drought conditions. In general, mycorrhizal inoculation resulted in an improvement in the growth parameters as well as the phytochemical and physiological characteristics of rose-scented geranium.

     

RANTES gene mRNA expression and its -403 G/A promoter polymorphism in coronary artery disease

Objective

Increased RANTES expression has been described to have a role in atherosclerosis plaque formation. Functional polymorphisms within RANTES promoter region have shown association with increased risk of coronary atherosclerosis (CAD). The aim of this study was to examine the RANTES mRNA expression in patients with CAD compared to patients without CAD and its association with RANTES − 403 G/A polymorphism in an Iranian population.

Methods

The study was performed on 319 patients who underwent coronary artery angiography and patients with > 50% stenosis in vessels considered as case groups (CAD+) N = 191 and normal vessels group as control (CAD−) N = 128. In each group 20 patients were examined for RANTES mRNA expression.RANTES mRNA expression was examined using quantitative real-time PCR. Genotyping of − 403 polymorphism was performed using PCR-RFLP technique.

Results

We found that RANTES mRNA expression was increased to 1.37 fold in CAD patients compared to the controls but the difference was not statistically significant. Also comparing the RANTES mRNA expression in patients with different RANTES − 403 G/A polymorphism showed that in patients carrying AA genotype RANTES mRNA expression was increased to 1.74 fold compared to patients carrying GG genotype and to 1.51 fold compared to patients carrying GA genotype. No significant difference for allele and genotype frequencies of RANTES − 403 polymorphism was found between cases and controls.

Conclusion

More studies on larger number of samples are required to further evaluate role of RANTES in pathogenesis of CAD.     

A novel multi-planar radiography method for three dimensional pose reconstruction of the patellofemoral and tibiofemoral joints after arthroplasty

Determining the 3D pose of the patella after total knee arthroplasty is challenging. The commonly used single-plane fluoroscopy is prone to large errors in the clinically relevant mediolateral direction. A conventional fixed bi-planar setup is limited in the minimum angular distance between the imaging planes necessary for visualizing the patellar component, and requires a highly flexible setup to adjust for the subject-specific geometries. As an alternative solution, this study investigated the use of a novel multi-planar imaging setup that consists of a C-arm tracked by an external optoelectric tracking system, to acquire calibrated radiographs from multiple orientations. To determine the accuracies, a knee prosthesis was implanted on artificial bones and imaged in simulated 'Supine' and 'Weightbearing' configurations. The results were compared with measures from a coordinate measuring machine as the ground-truth reference. The weightbearing configuration was the preferred imaging direction with RMS errors of 0.48 mm and 1.32 ° for mediolateral shift and tilt of the patella, respectively, the two most clinically relevant measures. The 'imaging accuracies' of the system, defined as the accuracies in 3D reconstruction of a cylindrical ball bearing phantom (so as to avoid the influence of the shape and orientation of the imaging object), showed an order of magnitude (11.5 times) reduction in the out-of-plane RMS errors in comparison to single-plane fluoroscopy. With this new method, complete 3D pose of the patellofemoral and tibiofemoral joints during quasi-static activities can be determined with a many-fold (up to 8 times) (3.4mm) improvement in the out-of-plane accuracies compared to a conventional single-plane fluoroscopy setup.