The endosomal trafficking regulator LITAF controls the cardiac Nav1.5 channel via the ubiquitin ligase NEDD4-2

The QT interval is a recording of cardiac electrical activity. Previous genome-wide association studies identified genetic variants that modify the QT interval upstream of LITAF (lipopolysaccharide-induced tumor necrosis factor-α factor), a protein encoding a regulator of endosomal trafficking. However, it was not clear how LITAF might impact cardiac excitation. We investigated the effect of LITAF on the voltage-gated sodium channel Nav1.5, which is critical for cardiac depolarization. We show that overexpressed LITAF resulted in a significant increase in the density of Nav1.5-generated voltage-gated sodium current I_Na and Nav1.5 surface protein levels in rabbit cardiomyocytes and in HEK cells stably expressing Nav1.5. Proximity ligation assays showed co-localization of endogenous LITAF and Nav1.5 in cardiomyocytes, whereas co-immunoprecipitations confirmed they are in the same complex when overexpressed in HEK cells. In vitro data suggest that LITAF interacts with the ubiquitin ligase NEDD4-2, a regulator of Nav1.5. LITAF overexpression down-regulated NEDD4-2 in cardiomyocytes and HEK cells. In HEK cells, LITAF increased ubiquitination and proteasomal degradation of co-expressed NEDD4-2 and significantly blunted the negative effect of NEDD4-2 on I_Na. We conclude that LITAF controls cardiac excitability by promoting degradation of NEDD4-2, which is essential for removal of surface Nav1.5. LITAF-knockout zebrafish showed increased variation in and a nonsignificant 15% prolongation of action potential duration. Computer simulations using a rabbit-cardiomyocyte model demonstrated that changes in Ca²⁺ and Na⁺ homeostasis are responsible for the surprisingly modest action potential duration shortening. These computational data thus corroborate findings from several genome-wide association studies that associated LITAF with QT interval variation.     

Purification and investigation of some kinetic properties of glucose-6-phosphate dehydrogenase from parsley (Petroselinum hortense) leaves

In this study, glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ oxidoreductase, EC 1.1.1.49; G6PD) was purified from parsley (Petroselinum hortense) leaves, and analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps: preparation of homogenate, ammonium sulfate fractionation, and DEAE-Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 8.79% and had a specific activity of 2.146 U (mg protein)(-1). The overall purification was about 58-fold. Temperature of +4 degrees C was maintained during the purification process. Enzyme activity was spectrophotometrically measured according to the Beutler method, at 340 nm. In order to control the purification of enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acrylamide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for enzyme. The molecular weight was found to be 77.6 kDa by Sephadex G-150 gel filtration chromatography. A protein band corresponding to a molecular weight of 79.3 kDa was obtained on SDS-polyacrylamide gel electrophoresis. For the enzymes, the stable pH, optimum pH, and optimum temperature were found to be 6.0, 8.0, and 60 degrees C, respectively. Moreover, KM and Vmax values for NADP+ and G6-P at optimum pH and 25 degrees C were determined by means of Lineweaver-Burk graphs. Additionally, effects of streptomycin sulfate and tetracycline antibiotics were investigated for the enzyme activity of glucose-6-phosphate dehydrogenase in vitro.     

A research on the microfungal flora of somegreenhouse soils in the vicinity of LapsekiÇanakkale,Turkey

Qualitative and quantitative distributions of microfungal flora of the some greenhouse soils in the vicinity of Lapseki, Canakkale were studied. A total of 25 soil samples were collected from 10 cm depth in five greenhouses. Using soil dilution technique, 128 different microfungal isolates were obtained Isolates collected belonged to Oomycetes (1), Zygomycetes (7), Ascomycetes (9) and 96 belonging to the Deuteromycetes Fifteen isolates were classified as Mycelia Sterilia. The genera of microfungi most abundant in the greenhouse soils were Aspergillus, Penicillium, Geomyces, Exophiala and Fusarium. Qualitative and quantitative distributions of the Aspergillus were higher than those of the other genera. It was found that the maximum value was in greenhouse 5 and the minimum value was in greenhouse 4. The solarization method was the most effective in reducing fungal numbers.     

The expression of HER-2/neu (c-erbB2), survivin and cycline D1 in serous ovarian neoplasms: their correlation with clinicopathological variables

Ovarian cancer is the most common cause of death among all gynecologic malignancies and a result of complex interaction of multiple oncogenes and tumor suppressor genes. The aim of this study was to evaluate expression of HER-2/neu (c-erbB2), survivin and cycline D1 biomarkers in serous ovarian neoplasms and their correlations with clinicopathological variables in serous ovarian cancers. We analyzed pathological specimens of 62 patients with benign (n = 25), borderline (n = 14) and malignant (n = 23) serous ovarian neoplasms. Immunohistochemical analysis was performed on formalin-fixed paraffin-embedded specimens. Significantly more immunoreactivity with HER-2/neu was detected in malignant tumors (100 %) compared to borderline (78.6 %) and benign tumors (48 %) (P < 0.01). Survivin expression was significantly higher in malignant tumors (91.3 %) than those found in borderline (71.4 %) and benign tumors (24 %) (P < 0.001). Similarly, higher cyclin D1 expression was observed in malignant tumors (95.6 %) compared to borderline (85.7 %) and benign tumors (48 %) (P < 0.001). Expression of all biomarkers analyzed significantly and gradually increased from benign to borderline and borderline to malignant serous tumors. In terms of clinicopathological variables, only tumor grade was associated with the expression of all biomarkers others exhibited different correlations in serous ovarian cancers. The expressions of HER-2/neu (c-erbB2), survivin and cycline D1 are positively correlated with the malignant potential of serous ovarian neoplasms.     

Interpretation of relevance of sodium-calcium exchange in action potential of diabetic rat heart by mathematical model

Sarcolemmal Na+-Ca2+ exchange plays a central role in ion transport of the myocardium and the current carried with it contributes to the late phase of the action potential (AP) besides the contribution of outward K+-currents. In this study, the mathematical model for AP of the diabetic rat ventricular myocytes [34] was modified and used for the diabetic rat papillary muscle. We used our experimentally measured values of two K+-currents; transient outward current, Ito and steady-state outward current, Iss, as well as L-type Ca2+-current, I(CaL), then compared with the simulated values. We have demonstrated that the prolongation in the AP of the papillary muscle of the diabetic rats are not due to the alteration of I(CaL) but mainly due to the inhibition of the K+-currents and also the Na+-Ca2+ exchanger current, I(Na-Ca). In combination with our experimental data on sodium-selenite-treated diabetic rats, our simulation results provide new information concerning plausible ionic mechanisms, and second a possible positive effect of selenium treatment on the altered I(Na-Ca) for the observed changes in the AP duration of streptozotocin-induced diabetic rat heart.     

Does diet variation determine the digestive tract length of Capoeta banarescui Turan,Kottelat, Ekmekci and Imamoglu, 2006?

This study tested the spatial variations in the digestive/intestine tract length of Capoeta banarescui, with regard to their diets in different habitats. Highly varied diets observed in a previous study within the same river system posed the question whether this flexibility is reflected in the digestive tract and intestine length of the species in the Ye?il?rmak River, Turkey. Totals of 382 specimens (standard length 4.6–19.1 cm) were captured by electro‐fishing along the river in September 2012 at 11 locations spanning elevations from 34 to 992 m. The stomach, intestine and total digestive tract lengths were measured, and stomach contents analysed from 196 specimens. For statistical analyses, the stomach, intestine and total digestive tract length were expressed as percentages of total weight and standard length. The data provided evidence that the digestive tract and intestine lengths varied significantly among locations in association with the diet. Fish having dominantly carnivorous diets (e.g. chironomid larvae/invertebrates) in two locations had significantly shorter intestines and digestive tracts than those with diets dominated by benthic algae and other plants. The data indicated that C. banarescui showed broad flexibility in their feeding habits. Feeding heavily on plant materials might lead to the development of longer digestive tracts, increasing the active surface area for digestion; alternatively, there may be less invested in development of the digestive tract when feeding primarily on carnivorous diets where the respective digestive enzymes are readily available. The data suggest that phenotypic plasticity in the digestive tract length of C. banarescui is associated more with the abundant protein‐rich carnivorous food sources in the studied habitats. Whether this digestive tract plasticity has a genetic background remains to be verified in future studies.     

Squalius semae,a new species of chub from the Euphrates River,Eastern Anatolia (Teleostei: Cyprinidae)

In a study of the fishes of the Euphrates River (Persian Gulf basin) three species of Squalius belonging to the cephalus-group were found: The Tohma population was identified as S. seyhanensis and the Merzimen and Hilvan populations as S. berak. The comparison of 28 metric and 5 meristic parameters and morphological characters showed that the populations of the northern Euphrates River drainages are distinct and belong to a hitherto unnamed species. We describe it here as Squalius semae sp. n.

http://www.zoobank.org/urn:lsid:zoobank.org:pub:D195B69C-C6CB-4DEB-B85D-5B695F917CE5     

Investigation of Heavy Metal Level and Mineral Nutrient Status in Widely Used Medicinal Plants’ Leaves in Turkey: Insights into Health Implications

The use of plants in treatments has been as old as humanity and it has preserved its popularity for centuries til now because of their availability, affordability and safeness. However, despite their widespread use, safety and quality issues have been major concerns in the world due to industrial- and anthropogenic-based heavy metal contamination risks. Thus, this study was attempted to analyze the heavy metal levels and mineral nutrient status of widely used medicinal plants in Turkey to have insights about their health implications on humans. The plant concentrations of B, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb and Zn were analyzed by ICP-OES in the leaves of 44 medical plants purchased from herbal markets of three different districts of Istanbul/Turkey. The measured lowest to highest concentrations were 0.065–79.749 mg kg⁻¹ B, 921.802–12,854.410 mg kg⁻¹ Ca, 0.020–0.558 mg kg⁻¹ Cd, 0.015–4.978 mg kg⁻¹ Cr, 0.042–8.489 mg kg⁻¹ Cu, 34.356–858.446 mg kg⁻¹ Fe, 791.323–15,569.349 mg kg⁻¹ K, 102.236–2837.836 mg kg⁻¹ Mg, 4.915–91.519 mg kg⁻¹ Mn, 10.224–3213.703 mg kg⁻¹ Na, 0.001–5.589 mg kg⁻¹ Ni, 0.003–3.636 mg kg⁻¹ Pb and 2.601–36.102 mg kg⁻¹ Zn. Those levels in plants were in acceptable limits though some elements in some plants have high limits which were not harmful. Variations (above acceptable limits) in element concentrations also indicated that these plants could be contaminated with other metals and that genetic variations may influence accumulation of these elements at different contents. Overall, analyzed medicinal plants are expected not to pose any serious threat to human health.

     

Zur genetik der 6-Phosphogluconatdehydrogenase (EC:1.1.1.44) bei Säugern

Zusammenfassung Das Enzym 6-PGD zeigt bei den verschiedensten Species einen genetisch gesteuerten Polymorphismus, der sich bei elektrophoretischer Darstellung in einer Variabilität des Isoenzymmusters äußert. Die Befunde bei den bislang untersuchten Säugern unter Einschluß des Menschen lassen sich formalgenetisch nach dem Modell: 1 Genlocus mit multipler Allelie interpretieren. Zur Prüfung der Frage, ob das Säugergenom einheitlich nur ein Gen für dieses Enzym enthält oder ob bei einzelnen Species eine Duplikation dieses Gens existiert, wurdene eine Anzahl Säuger verschiedener Ordnungen untersucht. In einer ersten Mitteilung wird hier über 6 verschiedene Species der Microtinae berichtet. Die Befunde sprechen durchweg für die Existenz von jeweils nur 1 Genlocus der 6-PGD. Bei Microtus oeconomus wurden 2 Allele an diesem locus nachgewiesen, bei Microtus ochrogaster 3 verschiedene Allele. Durch Familienuntersuchungen bei diesen beiden Species werden alle möglichen Kombinationen dieser Allele nachgewiesen.

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The genetics of 6-PGD (EC: 1.1.1.44) in various mammals I. Studies on 6 species of Microtinae, Rodentia. Isoenzyme polymorphism and family studies in Microtus oeconomus and Microtus ochrogaster

Summary The enzyme 6-PGD exhibits a genetically determined polymorphism which results in a variability of the isoenzyme pattern. In mammals so far examined the findings are to be interpreted according to the model of multiple alleles at one single gene locus. In order to test whether the mammalian genome is uniformly endowed with one single locus for this enzyme or whether a duplication of this locus is present in certain species, we examined various mammals of different orders. Our results in this first communication on 6 different Microtinae species are all in agreement with the assumption of a single 6-PGD locus. In Microtus oeconomus 2 alleles could be demonstrated and in Microtus ochrogaster 3 different alleles. Family studies performed with both species revealed the occurrence of all the expected combinations of these alleles.