Detection, quantification, and pharmacokinetics of furosemide and its effects on urinary specific gravity following IV administration to horses

Furosemide is a potent loop diuretic used for the prevention of exercise-induced pulmonary hemorrhage in horses. This drug may interfere with the detection of other substances by reducing urinary concentrations, so its use is strictly regulated. The regulation of furosemide in many racing jurisdictions is based on paired limits of urinary SG (<1.010) and serum furosemide concentrations (>100 ng/ml). To validate this regulatory mechanism, a liquid chromatography/mass spectrometry/mass spectrometry method employing a solid-phase extraction procedure and furosemide-d5 as an internal standard was developed. The method was used to determine the pharmacokinetic parameters of furosemide in equine serum samples and its effects on urinary SG after IV administration (250 mg) to 10 horses. Pharmacokinetic analysis showed that serum concentrations of furosemide were well described by a two-compartmental open model. Based on results in this study, it is very unlikely for horses to have serum furosemide concentrations greater than 100 ng/ml or urine SG less than 1.010 at 4 hours after administration (250 mg IV). However, it should be remembered that urine SG is a highly variable measurement in horses, and even without furosemide administration, some horses might naturally have urine SG values less than 1.010.     

Characterization and Purification of Glutathione S-Transferase from the Liver and Gill Tissues of Ağrı Balık Lake Trout Salmo trutta labrax and the Effects of Heavy Metal Ions on Its Activity

Glutathione S-transferase (GST) from the liver and gill tissues of A?r? Bal?k Lake Trout (also known as Black Sea Trout) Salmo trutta labrax was characterized and purified, and the toxic effects of some heavy metal ions on the enzyme's activity were analyzed. Liver GST was purified 930 times, resulting in 56% yield using glutathione–agarose affinity chromatography and a specific activity of 60.87 endotoxin units (EU)/mg protein. Using the same procedure, gill GST was purified 576 times, resulting in a 60% yield and specific activity of 46.8 EU/mg protein. The purity check of the purified enzymes was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Optimal pH, ionic strength, and stable pH were found for each tissue, and separate K_M and V_max values were determined for reduced glutathione and 2,4-dinitrochlorobenzene substrates. Heavy metal ions that have toxic effects on living organisms and are known to contribute to environmental pollution were selected, and their in vitro effects on enzyme activity were analyzed. The IC50 values and K_i constants of those metal ions showing an inhibitory effect on GST activity were determined.

Received November 24, 2014; accepted March 11, 2015     

Determination of Heat Requirements and Effective Heat Summations of Some Pomegranate Cultivars Grown in Southern Anatolia

Temperature (high and low) is one of the most limiting environmental factors for growth and production of fruits in different parts of the world. For temperate-zone fruit species, a certain total heat requirements are needed in order to obtain ripe fruits. The aim of this study was to determine heat requirements of three commercial pomegranate cultivars (‘Suruc’, ‘Katirbasi’ and ‘Hicaznar’) grown in Southern Anatolia region. The cultivars showed a narrower range of heat requirements for flowering (growing degree hours, GDH), i.e. ‘Suruc’ (25,000), ‘Katirbasi’ (25,270), ‘Hicaznar’ (28,000) and a wide range of heat requirements for fruit ripening, i.e. ‘Suruc’ (73,670), ‘Katirbasi’ (74,105) and ‘Hicaznar’ (88,052) GDH. Under the Sanliurfa province conditions, two years average of effective heat summation requirement from bud swelling to 50 % flowering stage for ‘Suruc’, ‘Katirbasi’ and ‘Hicaznar’ cultivars are calculated as 643, 655 and 718 growing degree days (GDD) and from bud swelling to harvest are 2734, 2802 and 3289 GDD. It can be concluded that the effective heat summation of Sanliurfa province is enough for commercial growing for all pomegranate cultivars.     

Purification of glucose-6-phosphate dehydrogenase and glutathione reductase enzymes from the gill tissue of Lake Van fish and analyzing the effects of some chalcone derivatives on enzyme activities

Glucose-6-phosphate dehydrogenase (G6PD) and glutathione reductase (GR) are metabolically quite important enzymes. Within this study, these two enzymes were purified for the first time from the gills of Lake Van fish. In the purifying process, ammonium sulfate precipitation and 2′,5′-ADP Sepharose 4B affinity column chromatography techniques for glucose-6-phosphate dehydrogenase, temperature degradation and 2′,5′-ADP Sepharose 4B affinity column chromatography for glutathione reductase enzyme were used. The control of the enzyme purity and determination of molecular weight were done with sodium dodecyl sulfate polyacrylamide gel electrophoresis. K _M and V _max values were determined with Lineweaver–Burk plot. Besides, the effects of some chalcone derivatives on the purified enzymes were analyzed. For the ones showing inhibition effect, % activity–[I] figures were drawn and IC₅₀ values were determined. K _i value was calculated by using Cheng–Prusoff equation.     

Acaricidal effects of the essential oil of Origanum minutiflorum (Lamiaceae) against Rhipicephalus turanicus (Acari: Ixodidae)

The acaricidal effects of the volatile essential oil Origanum minutiflorum O. Schwarz & P.H. Davis (Lamiaceae) against adult Rhipicephalus turanicus was evaluated at a variety of concentrations and exposure times. Generally tick mortality increased with concentration and exposure. Ticks exposed to vapors from cotton wicks containing at least 10 μl/L resulted in complete (100%) mortality at 120 min. The major constituent of essential oil obtained from the plant material of O. minutiflorum was carvacrol.     

Genetic variation and environmental stability of grain mineral nutrient concentrations in <Emphasis Type="Italic">Triticum dicoccoides</Emphasis> under five environments

Nineteen wild emmer wheat [Triticum turgidum ssp. dicoccoides (Körn.) Thell.] genotypes were evaluated for the grain concentrations of phosphorous (P), potassium (K), sulfur (S), magnesium (Mg), calcium (Ca), zinc (Zn), manganese (Mn), iron (Fe) and cooper (Cu) under five different environments in Turkey and Israel. Each mineral nutrient has been investigated for the (1) genotype by environment (G × E) interactions, (2) genotype stability, (3) correlation among minerals and (4) mineral stability. Among the macronutrients analyzed, grain concentrations of Ca (range 338–2,034 mg kg^?1) and S (range 0.18–0.43%) showed the largest variation. In the case of micronutrients, the largest variation was observed in the grain Mn concentration (range 13–87 mg kg^?1). Grain concentrations of Fe and Zn also showed important variation (range 27–86 and 39–115 mg kg^?1, respectively). Accessions with higher nutrient concentrations (especially Zn and Fe) had also greater grain weight, suggesting that higher grain Zn and Fe concentrations are not necessarily related to small grain size or weight. Analysis of variance showed that environment was the most important source of variation for K, S, Ca, Fe, Mn and Zn, explaining between 44 and 78% of the total variation and G × E explained between 20 and 40% of the total variation in all the minerals, except for S and Zn where its effect accounted for less than 16%. Genotype was the most important source of variation for Cu (explaining 38% of the total variation). However, genotype effect was also important for Mg, Mn, Zn and S. Sulfur and Zn showed the largest heritability values (77 and 72%, respectively). Iron exhibited low heritability and high ratio value between the G × E and genotype variance components \( \left( {\sigma_{\text{GE}}^{2} /\sigma_{G}^{2} } \right) \), suggesting that specific adaptation for this mineral could be positively exploited. The wild emmer germplasm tested in the current study revealed some outstanding accessions (such as MM 5/4 and 24-39) in terms of grain Zn and Fe concentrations and environmental stability that can be used as potential donors to enhance grain micronutrient concentrations in wheats.     

Comparative study on the characteristics of knitted fabrics made of vortex-spun viscose yarns

Murata vortex spinning system is based on the air jet spinning system. The vast majority of previous works deal with the properties of vortex spun (VS) yarn and the spinning system. In this study, we investigated knitted fabrics from VS yarn in comparison with fabrics from ring (RS), compact (CS) and open-end rotor (OES) spun yarns made from viscose. The effect of yarn spinning system on dimensional and physical properties of knitted fabrics was explained with specific attention to fabrics from VS yarn. Shrinkage of fabrics from VS yarn has the lowest at widthwise direction, while having the highest at lengthwise direction. It is shown that the order of fabric spirality and twist liveliness for yarns from different spinning systems are quite similar. However, relation between loop shape factor and angle of spirality is inconsistent. Angle of spirality of fabrics from VS yarn is higher than fabrics from OES yarn, but lower than that of others. The bursting strength of fabrics from VS yarn is lower than that of those from RS and CS yarns and higher than that of those from OES yarn. From this study, it is also evident that fabrics from VS yarn have the lowest pilling tendency and highest resistance to abrasion.     

Localization profiles of natriuretic peptides in hearts of pre-hibernating and hibernating Anatolian ground squirrels (Spermophilus xanthoprymnus)

The Anatolian ground squirrel (Spermophilus xanthoprymnus) is a typical example of true mammalian hibernators. In order to adapt to extreme external and internal environments during hibernation, they lower their body temperatures, heart rates and oxygen consumption; however, pathological events such as ischemia and ventricular fibrillation do not occur in their cardiovascular systems. During the hibernation, maintenance of cardiac function is very important for survival of ground squirrels. Natriuretic peptides (NPs) are key factors in the regulation of cardiovascular hemostasis. Since NPs’ role on the protection of heart during hibernation are less clear, the aim of this study was to investigate dynamic changes in NPs content in the cardiac chambers and to reveal the possible role of NPs on establishing cardiac function in ground squirrel during hibernation using immunohistochemistry. The immunohistochemical results indicate that cardiac NP expressions in atrial and ventricular cardiomyocytes were different from each other and were sex-independent. ANP and BNP were expressed in a chamber-dependent manner in female and male squirrel hearts. Furthermore, cardiac NPs expression levels in hibernation period were lower than those at the pre-hibernation period. During prehibernation period, ANP, BNP and CNP were expressed in the white and beige adipocytes of epicardial adipose tissue (EAT); while during hibernation period, the brown adipocytes of EAT were positive for BNP and CNP. These data suggest that the hibernation-dependent reduction in levels of NPs, particularly ANP, in cardiac chambers and EAT may be associated with low heart rate and oxygen consumption during hibernation. However, further studies are needed to better delineate the roles of NPs during the hibernation.