Cloning, expression and characterization of monkey (Macaca fascicularis) CD137

CD137 plays an important role as a co-stimulatory molecule in activated T cells. Agonistic CD137 specific antibodies have been investigated as therapeutic agents to promote tumor-specific immune responses by direct activation of T cells. As part of the pre-clinical pharmacological evaluation of cynomolgus monkeys, monkey CD137 was cloned and characterized. The deduced amino acid sequence encoded a full-length gene of 254 amino acids 95% identical to human CD137. Sequence variants identified in monkey CD137 include four splice variants lacking the transmembrane domain. These variants were detectable in human including two previously unreported variants. Two missense single nucleotide polymorphisms were detected present in 42 and 50% of 36 monkeys tested. In both monkey and human, mRNA expression of full-length CD137 and splice variants were significantly increased in peripheral blood mononuclear cells (PBMCs) upon stimulation by anti-CD3 antibodies. Recombinant monkey CD137 protein was bound with high affinity by an agonistic anti-human CD137 antibody but not by an anti-mouse CD137 antibody. In summary, compared to human, monkey CD137 showed distinct extracellular domain amino acid sequence and sequence polymorphisms. Thus, antibodies directed against epitopes in this extracellular domain could have differences in pharmacologic activity between cynomolgus monkeys and human or across individual cynomolgus monkeys.     

Interaction of Crop Load and Irrigation on Yield,Fruit Size,Color and Stem-end Splitting Ratio of Apple c.v. ‘Gala,Galaxy’

In this study, the interaction between crop load and irrigation level on yield, fruit size, skin color and stem-end splitting fruit ratio in the apple cultivar ‘Gala, Galaxy’ grafted on rootstock M9 were investigated. Six irrigation programs were applied during the whole growth season: deficit irrigation (rates of 0.25 kc, 0.50 kc, 0.75 kc), full irrigation (rate of 1.00 kc), excess irrigation (rate of 1.25 kc) and non-irrigation (rates of 0.00 kc of “Class A” pan evaporation coefficient). Four crop loads in each irrigation application were performed by hand thinning after the June drop as a- a low crop load (3 fruits cm^?2 TCA), b- a medium crop load (5 fruits cm^?2 TCA), c- a heavy crop load (7 fruits cm^?2 TCA), and d- an un-thinned crop load (>?7 fruits cm^?2 TCA). The total tree yield increased with crop load and irrigation levels. Fruit size was significantly increased by the low crop load. Irrigation increased the fruit size compared to non-irrigation treatment. Further 0.75 kc, 1.00 kc and 1.25 kc irrigation treatments significantly increased the fruit length. Irrigation reduced the fruit flesh firmness. While the low crop load increased the skin red color, it decreased the fruit skin brightness. The yellowness of skin decreased with increasing in the irrigation amount. Irrigation reduced the skin brightness and yellowness, but it increased red color. Crop load and irrigation significantly affected the stem-end splitting fruit ratio. While the splitting fruit ratio increased with a decrease in the crop load, it decreased with an increase in irrigation amount, relatively. Consequently, the low and medium crop load treatments would be beneficial to increase the ratio of marketable fruits without any significant losses in yield for ‘Gala’ apple, especially under 0.75 kc deficit irrigation treatment.     

Simultaneous Determination of Phorate and Oxyfluorfen in Well Water Samples with High Accuracy by GC-MS After Binary Dispersive Liquid-Liquid Microextraction

The potential risk of pesticides to cause harm to humans and other organisms even at trace levels calls for sensitive and accurate analytical techniques for their simultaneous qualitative and quantitative determinations. In this study, a sensitive binary dispersive liquid-liquid microextraction (B-DLLME) strategy was developed for the simultaneous determination of phorate and oxyfluorfen by gas chromatography mass spectrometry after extraction/preconcentration from aqueous solution. An experimental design was used to optimize parameters of the B-DLLME method to obtain maximum outcome. Under the optimum conditions of B-DLLME, the limit of detection (LOD) for phorate and oxyfluorfen were found to be 0.41 μg L^?1 and 0.54 μg L^?1, respectively. The detection limits correlate to about 37 and 73 folds enhancement in detection powers when compared to direct GC-MS determination of phorate and oxyfluorfen, respectively. In order to find out the applicability of developed method to real samples, recovery tests were performed for 20 μg L^?1 of phorate and oxyfluorfen spiked in well water samples. Percent recovery values were found to be 94.5% for phorate and 101.9% for oxyfluorfen.     

Kinetic, Isotherm and Thermodynamic Analysis on Adsorption of Cr(VI) Ions from Aqueous Solutions by Synthesis and Characterization of Magnetic-Poly(divinylbenzene-vinylimidazole) Microbeads

The magnetic-poly(divinylbenzene-1-vinylimidazole) [m-poly(DVB-VIM)] microbeads (average diameter 53-212?μm) were synthesized and characterized; their use as adsorbent in removal of Cr(VI) ions from aqueous solutions was investigated. The m-poly(DVB-VIM) microbeads were prepared by copolymerizing of divinylbenzene (DVB) with 1-vinylimidazole (VIM). The m-poly(DVB-VIM) microbeads were characterized by N(2) adsorption/desorption isotherms, ESR, elemental analysis, scanning electron microscope (SEM) and swelling studies. At fixed solid/solution ratio the various factors affecting adsorption of Cr(VI) ions from aqueous solutions such as pH, initial concentration, contact time and temperature were analyzed. Langmuir, Freundlich and Dubinin-Radushkvich isotherms were used as the model adsorption equilibrium data. Langmuir isotherm model was the most adequate. The pseudo-first-order, pseudo-second-order, Ritch-second-order and intraparticle diffusion models were used to describe the adsorption kinetics. The apparent activation energy was found to be 5.024?kJ?mol(-1), which is characteristic of a chemically controlled reaction. The experimental data fitted to pseudo-second-order kinetic. The study of temperature effect was quantified by calculating various thermodynamic parameters such as Gibbs free energy, enthalpy and entropy changes. The thermodynamic parameters obtained indicated the endothermic nature of adsorption of Cr(VI) ions. Morever, after the use in adsorption, the m-poly(DVB-VIM) microbeads with paramagnetic property were separeted via the applied magnetic force. The magnetic beads could be desorbed up to about 97% by treating with 1.0?M NaOH. These features make the m-poly(DVB-VIM) microbeads a potential candidate for support of Cr(VI) ions removal under magnetic field.     

A Practical LC-MS/MS Method for the Detection of NDMA at Nanogram per Liter Concentrations in Multiple Water Matrices

N-nitrosodimethylamine (NDMA) is one of the most important disinfection by-products (DBPs) due to its carcinogenicity even at low concentrations which correspond to the levels occurring in drinking water and wastewater effluents. Therefore, NDMA is a candidate DBP that is expected to be regulated in the near future. However, the measurement of NDMA in the low nanogram per liter range is challenging because of the limitations of analytical techniques including both the sample preparation and the LC-MS/MS. Moreover, the accuracy of most of the current methods is only tested for drinking water and no information is present for other matrices. In this study, a combination of solid-phase extraction (SPE) and LC-MS/MS method that does not require high-resolution MS or advanced techniques for sample pretreatment is developed. Moreover, important factors that affect the optimization of the SPE method are provided to enable readers to optimize their own SPE procedures if necessary. The proposed method was validated for surface water, groundwater, and wastewater samples and the method quantification limit was 2?ng/L. In addition, the proposed method was used to determine the concentration of NDMA precursors measured as NDMA formation potential (NDMAFP) throughout a drinking water treatment plant at two different sampling periods. NDMAFP decreased by approximately 40?% in both samples. The concentrations ranged between 4 and 11.5?ng/L and the presence of these low concentrations underlines the need for an easy to use, yet sensitive method for the determination of NDMA in environmental matrices.     

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.

     

The distribution and immunolocalization of fibroblast growth factors (FGFs) in the rat oviduct during early pregnancy and the post-partum period

The mammalian oviduct provides a favourable environment for several reproductive processes, including ovum transport, sperm capacitation, fertilization and pre-implantation embryonic development. This environment is regulated by cyclic ovarian steroids, that is oestrogen, and growth factors. Fibroblast growth factors (FGFs) regulate the differentiation and growth of various cell types in the female genital tract. This study aimed to determine the localization of FGF1, FGF2, FGF receptor 1 (FGFR1) and 2 (FGFR2) in the rat oviduct, by immunohistochemistry, on day 5 of pregnancy and post-partum days 1, 3 and 5, and to demonstrate the possible functions of these proteins during early pregnancy and the post-partum period. On all examination days, cytoplasmic and nuclear FGF1 immunoreactivity was detected in the epithelium lining the infundibulum, ampulla and isthmus of the oviduct. Immunoreactivity was much stronger in the basal bodies of the cilia on the epithelium lining the infundibulum and ampulla. FGF1 immunoreactivity was also detected in stromal cells, myocytes and endothelial cells. Cytoplasmic FGF2 immunoreactivity was observed in the tunica muscularis, vascular myocytes and endothelial cells. While strong cytoplasmic FGF2 immunoreactions were observed in the stromal cells of the lamina propria, the luminal epithelium, some stromal cells and smooth muscle cells displayed a rather weak FGFR1 and FGFR2 immunoreactivity. Immunoreaction intensity did not differ between the periods examined. This study shows that FGF1, FGF2, FGFR1 and FGFR2 are produced by rat oviduct cells during pregnancy and the post-partum period, and reproductive physiology is regulated not only by hormonal mechanisms, but also by growth factors.     

Production and characterization of films from cotton stalk xylan

Composite film production based on cotton stalk xylan was studied, and the mechanical and physical properties of the films formed were investigated. Xylan and lignin were separated from cellulose by alkali extraction and, then, lignin was removed using ethanol washing. Self-supporting continuous films could not be produced using pure cotton stalk xylan. However, film formation was achieved using 8-14% (w/w) xylan without complete removal of lignin during xylan isolation. Keeping about 1% lignin in xylan (w/w) was determined to be sufficient for film formation. Films were produced by casting the film-forming solutions, followed by solvent evaporation in a temperature (20 degrees C) and relative humidity (40%) controlled environment. The elastic modulus and hypothetical coating strength of the films obtained by using 8% xylan were significantly different from the ones containing 10-14% xylan. The water vapor transfer rates (WVTR) decreased with increasing xylan concentration, which made the films thicker. The glycerol addition as an additional plasticizer resulting in more stretchable films having higher WVTR and lower water solubility values. As a result, film production was successfully achieved from xylan, which was extracted from an agricultural waste (cotton stalk), and the film-forming effect of lignin on pure xylan has been demonstrated.     

Distribution of hepatitis B virus (HBV) genotypes among HBV carriers in Isparta

BACKGROUND: The aim of this study was to investigate the prevalence of hepatitis B virus (HBV) genotypes in Isparta, Southwest of Turkey, as well as the clinical features and transmission route for patients with HBV infections. METHODS: Patients (n = 135) with HBV infection were included in the study. Epidemiological and clinical data were obtained. HBV genotypes were determined with a preS2 epitope ELISA kit. RESULTS: Although the HBV transmission route remained unidentified in 51.1% of the patients, blood contact was determined as the most common probable transmission route (38.5%). One hundred twenty-four (91.8%) of 135 samples, could be genotyped. One hundred fifteen (85.1%) were genotyped as type D/E, six (4.4%) were genotyped as type A, two (1.4%) were genotyped as type C, and one (0.7%) were genotyped as type F. CONCLUSION: Genotype D/E is determined as the predominant HBV genotype circulating in Isparta, Southwest of Turkey. No relationship between genotypes and disease severity and transmission route has been detected.     

Therapeutic effects of thymoquinone in doxorubicin-induced hepatotoxicity via oxidative stress,inflammation and apoptosis

Cancer is a lethal disease that is characterized by uncontrolled cell division and proliferation, and it results in death in many organisms. Doxorubicin (DOX) is a therapeutic agent used for treatment of many cancer types, but it induces serious hepatotoxicity. In this study, we aimed to determine possible hepato-therapeutic effects of thymoquinone (THQ) on DOX-induced hepatotoxicity in rats. Rats were divided into five groups (n = 8): Control, THQ (10 mg/kg/day/i.p for 14 days), Olive Oil (equal volume with THQ for 14 days), DOX (single dose, 15 mg/kg/i.p on 7th day) and DOX + THQ (10 mg/kg/day/i.p and DOX 15 mg/kg/i.p on 7th day). At the end of the experiment, liver tissues were extracted and evaluated histopathologically. eNOS, iNOS and Cas-3 immunostaining were performed to determine the expression levels. TUNEL method was used to determine apoptotic index. Furthermore, liver tissue total antioxidant status (TAS), total oxidant status (TOS), TNF-α and TGF-β levels were measured by ELISA assay. The DOX group showed histopathological deterioration compared to Control group. Moreover, apoptotic index, eNOS, iNOS and Cas-3 expressions increased in DOX group. While TAS level of the DOX group decreased, TOS level increased. TNF-α and TGF-β levels increased in DOX group. However, there was improvement in DOX + THQ group compared to DOX group. Moreover, apoptotic cell number, eNOS, iNOS and Cas-3 expressions decreased in DOX + THQ group compared to DOX group. We concluded that thymoquinone can be used as a phytotherapeutic for reducing DOX-induced liver damage.