Effects of alkaline-electrolyzed and hydrogen-rich water,in a high-fat-diet nonalcoholic fatty liver disease mouse model

AIM To identify the effect of hydrogen-rich water(HRW) and electrolyzed-alkaline water(EAW) on high-fat-induced non-alcoholic fatty acid disease in mice.METHODS Mice were divided into four groups:(1) Regular diet(RD)/regular water(RW);(2) high-fat diet(HFD)/RW;(3) RD/EAW; and(4) HFD/EAW. Weight and body composition were measured. After twelve weeks, animals were sacrificed, and livers were processed for histology and reverse-transcriptase polymerase chain reaction. A similar experiment was performed using HRW to determine the influence and importance of molecular hydrogen(H2) in EAW. Finally, we compared the response of hepatocytes isolated from mice drinking HRW or RW to palmitate overload.RESULTS EAW had several properties important to the study:(1) pH = 11;(2) oxidation-reduction potential of-495 mV; and(3) H2 = 0.2 mg/L. However, in contrast to other studies, there were no differences between the groups drinking EAW or RW in either the RD or HFD groups. We hypothesized that the null result was due to low H2 concentrations. Therefore, we evaluated the effects of RW and low and high HRW concentrations(L-HRW = 0.3 mg H2/L and H-HRW = 0.8 mg H2/L, respectively) in mice fed an HFD. Compared to RW and L-HRW, H-HRW resulted in a lower increase in fat mass(46% vs 61%), an increase in lean body mass(42% vs 28%), and a decrease in hepatic lipid accumulation(P < 0.01). Lastly, exposure of hepatocytes isolated from mice drinking H-HRW to palmitate overload demonstrated a protective effect from H2 by reducing hepatocyte lipid accumulation in comparison to mice drinking regular water.CONCLUSION H2 is the therapeutic agent in electrolyzed-alkaline water and attenuates HFD-induced nonalcoholic fatty liver disease in mice.

The Habitats, Burrowing Behavior, Physiology Adaptation and Life Cycle of Spadefoot Toads (<i>Pelobates syriacus, Boettge</i>r, 1869) at the Southern Limit of Its Distribution in Israel

The present study describes the habitats, life cycle, larvae growth, burrowing behavior and terrestrial adaptation of Spadefoot toads in Israel based on observations and data collected during more than 30 years in northern Israel. The distribution area in Israel is from the north in the Upper Galilee and Golan Heights (annual rainfall range of 500 - 1000 mm) to the southern coastal plain (annual rainfall of about 250 mm). Among the 51 different breeding places of amphibians, only ponds where water was available for a few months were used by Spadefoot toads and metamorphosed populations were found around these ponds. The larvae underwent metamorphosis during the summer and autumn in northern Israel, and during the spring in central and southern Israel. A negative correlation exists between the percentage of toads burrowing and soil moisture levels, with greater burrowing behavior occurring under dry conditions. The plasma concentration increased during burrowing by electrolytes and urea accumulations. The burrowing behavior helped Spadefoot toads survive in this area at the southern border of its distribution.

Sex-Based Variation of Gene Expression in the Gonads and Fins of Russian Sturgeon (<i>Acipenser gueldenstaedtii</i>)

Russian sturgeon (Acipenser gueldenstaedtii) is a primitive freshwater fish and a source of black caviar. The genes involved in sexual determination and differentiation are still unknown and there are no molecular markers for sex identification in this species. Studying the variation of the sex-based differences in genomic sequences and in gene expression in the sturgeon may lead to markers of sex in early stages of development and advances in aquaculture, as well as provide novel insights about the evolution of reproduction, sex determination, and sexual differentiation mechanisms in vertebrates. Previous studies by our and other groups have identified differentially expressed genes in the gonads of adult female and male sturgeon. The current study aimed to test whether these genes were also differentially expressed in non-gonadal tissue, namely fins. We measured by qRT-PCR the mRNA levels of 29 known and novel sex-related genes in the gonads and fins of males (4 years old) and females (7 years old;sexual maturation is earlier in males than in females). Six genes (ATP6, IGFRM, LIA1A, S1A, NPL1A, GAPDH and SOX9) showed higher expression in female fins. However, only ATP6 mRNA levels differed in fins of males and females of the same age (4 years old). These findings underscore the impracticality of sex identification based on gene expression in non-gonadal tissue and the need for genetic sex markers in the Russian sturgeon.

The Impact of Nitrogen and Phosphorus Dynamics on the Kinneret Phytoplankton: II: Chlorophyta, Cyanophyta, Diatoms and Peridinium

Lake Kinneret long-term data of the epilimnetic concentrations (ppm) and loads (tones) of the total Nitrogen (TN), total Phosphorus (TP), total inorganic Nitrogen (TIN), total Dissolved Phosphorus (TDP), Phytoplankton groups’ biomass, water level (WL) and Jordan River Discharge were analyzed. Previously collected data compiled aimed at an insight into the causative background for the modification of Phytoplankton community change. The study was carried out by searching for relations between algal groups’ densities and nutrient conditions in the Epilimnion by the use of statistical methods (Simple and Fractional Polynomial Regressions). The study is aimed at analyzing the relations between algal biomass and nutrient contents. It was found that Nitrogen decline and slight increase of phosphorus were followed by Peridinium (Photo 1)?decline and biomass increase of non-peridinium algae. It is suggested that nitrogen supply for algal growth is mostly from external sources, and the reduction of nitrogen in the epilimnion was caused by external removal. Contrary to nitrogen, phosphorus sourcing is only partly external (dust deposition, drainage basin) and mostly internal through double channels: Microbial mineralization of bottom sediments and Peridinium cysts mediation. The resulted complexity of the Kinneret ecosystem structure is nitrogen limitation, and enhancement of Non-peridinium algal growth, mostly Cyanobacteria.

The Impact of Nitrogen and Phosphorus Dynamics on the Kinneret Phytoplankton: I: Cyanophytes-Peridinium Alternate

During 1970-2001, several ecological changes were documented in the Lake Kinneret Ecosystem: Decline of total Nitrogen (TN) and increase of total Phosphorus (TP) loads with a corresponding decline of TN/TP ratio. The phytoplankton assemblages indicated consequence Increase of Cyanobacteria accompanied by elevation of the biomass of Chlorophyta and Diatoms. The Kinneret ecosystem functioning represented two superimposed events: the decline of TN enhanced Cyanobacteria and elevation of TP that was reflected by an increase of Chloropytes and Diatoms. Removal of Nitrogen was achieved but P load remained high enough to enhance Chlorophyta and Diatoms. Consequently, a top priority of future management perspective should be reduction of Phosphorus inputs.

Pathogenicity Assay for <i>Cochliobolus heterostrophus</i>G-Protein and MAPK Signaling Deficiency Strains

Cochliobolus heterostrophus is an agriculturally important and emerging model pathogen for studying the signaling hierarchies' role during the maize host colonization. In particular, G-protein and MAPK-linked pathways are playing a major role during pathogenesis. Although gene disruption studies are an efficient way of identifying the role of these cascades, differentiating between the mutant strains’ virulence ability may become an intricate task. For example, in C. heterostrophus, mutants in a G-protein α subunit gene, cga1, are defective in mating and appressorium formation, but unlike mutants in homologous genes in other fungal pathogens, the cga1 mutants remained highly virulent to corn under some host physiological conditions. Here, we used the cga1 strain as a model for developing an in vivo sensitive and accurate pathogenicity assay. A detailed and well controlled analysis of wild type (WT) and cga1 pathogenic behavior revealed that detached leaves are significantly more vulnerable to the disease than intact ones. In intact leaves, cga1 mutants were less infective of maize under most conditions. This difference was maximized when the first seedling leaf was chosen for inoculation and when the infected leaves, with spores or mycelia fragments droplets, were incubated for a period of four days. This optimal condition set enabled us to classify the C. heterostrophus G-protein signaling mutants deficient in α, β or both subunits in order of decreasing virulence: WT > cga1> cgb1> cga1 cgb1. The method presented proved to be accurate and sensitive enough to identify even slight variations in virulence. Moreover, it could be modified for use in studies of other foliar phytoparasitic fungi.

Gene Expression Modulation of Two Biosynthesis Pathways via Signal Transduction in <i>Cochliobolus heterostrophus</i>

G-protein-linked pathways have evolved to allow responses to extracellular agonists (hormones, neurotransmitters, odors, chemoattractants, light and nutrients) in eukaryotic cells, ranging from simpler systems, including yeasts, filamentous fungi and slime molds, to more complex organisms, such as mammals. Although the role of G-protein and mitogen-activated protein kinase (MAPK) in filamentous fungi has been studied for over a decade, downstream elements are less known, and the study of target genes has evolved mainly in recent years. Here, we examined the involvement of G-protein subunits and MAPK in controlling the expression of two distinct target genes. These genes were selected from an array database according to their unique expression profile and the role of closely related genes found in other Ascomycetes. One of these genes is BPH, which encodes the enzyme responsible for cytochrome P450-dependent benzoate hydroxylation in microsomes. The other gene is CIPA, which encodes isoflavone reductase (IfR), an enzyme involved in the synthesis of phytoalexin, which catalyzes an intermediate step in pisatin biosynthesis. The expression profile of these two genes was determined in a series of signaling deficiency mutants that were grown on different media using a DNA microarray. Comparison of the expression profile in the two wild type strains and mutants deficient in the G-protein α or β subunits or in MAPK, revealed a unique control mechanism for the BPH and CIPA genes. The two genes are highly expressed during the infection of the host plant leaves and may associate with the fungal response to the host. Signaling via G-protein or MAPK was shown to be related to cascades that altered the expression of these genes in response to the growth condition. This work demonstrates that signal transduction pathways are controlling genes that, although sharing an environmental dependent response, participate in distinct biosynthesis pathways. Moreover, the transcriptional profile may point to distinct and shared roles of the signaling components.

Production and Purification of Cutinase from <i>Fusarium oxysporum</i>Using Modified Growth Media and a Specific Cutinase Substrate

Cutinases are hydrolytic enzymes used by phytopathogenic fungi to gain entry into plants by breaking down the cuticular barrier of higher plants. Cutinase displayed hydrolytic activity not only towards cutin, the main component of the plant cuticle, but also towards a variety of soluble synthetic esters, insoluble triglycerides and polyesters. Therefore, cutinase was evaluated for use in the chemical, food and cotton bio-scouring industry and for synthetic fibers modification. This research examined the production and purification of extracellular cutinase from the phytopathogenic fungus Fusarium oxysporum. The addition of apple cutin or its hydrolysate to the fungus growth medium resulted in an enhanced secretion of cutinase into the extracellular fluid. Testing 1-hexadecanol as an alternative to natural cutin to induce cutinase production resulted in a high process yield under modified growth conditions. Cutinase enzyme production was followed by an optimized purification method for enzyme preparation using high-performance liquid chromatography and high-specificity 4-nitrophenyl (16-methyl sulfide ester) hexadecanoate (pNMSEH) cutinase substrate. Electrophoresis sodiumdodecyl sulfate-polyacrylamide and isoelectric focusing gels enabled the final separation and identification of the protein. The purified cutinase had an approximate molecular weight of 20 kDa and an isoelectric point of 4.7. The method presented here could be modified and used for cutinase production and purification in other microorganisms that exhibit cutinolytic activity.

Oxygen sufficiency controls TOP mRNA translation via the TSC-Rheb-mTOR pathway in a 4E-BP-independent manner

遇到 hypoxic 应力的房间由 downregulating 保存资源和精力蛋白质合成。这里我们 demonstratethat 在这回答的一机制是最高的 mRNAs 的翻译压抑编码 translationalapparatus 的部件。规定的这个模式包含 TSC 和 Rheb，当 Rheb 的 TSC1 或 TSC2 或 overexpression 的大美人在剥夺氧的房间救了最高的 mRNA 翻译。导致压力的翻译 repressionof 这些 mRNAs 密切与 4E-BP 的 hypophosphorylated 状态相关，一个翻译抑压者。然而， seriesof 4E-BP loss-of-function 和 gain-of-function 实验在氧或生长因素剥夺下面反驳在 phosphorylation statusof 4E-BP 和最高的 mRNAs 的翻译压抑之间的一种 cause-and-effect 关系。而且，血缺氧的 repressiveeffect 类似于由 Torin 1，而是更多 pronouncedthan 猛禽或 rictor 大美人由 mTOR 活动的有效抑制到达了那。同样，猛禽或 rictor 的缺乏它温和地 downregulated TOPmRNAs 的基础翻译效率，没能压制最高的 mRNAs 的调停氧的翻译激活。最后， TIA-1 和 TIAR co 击倒，二 RNA 有约束力的蛋白质以前在氨基的酸饥饿的房间在最高的 mRNAs 的翻译压抑含有， failedto 在另外的压力条件下面减轻最高的 mRNA 翻译。因此，近似翻译管理者 ofTOP mRNAs 的性质留下逃犯。