Enzymatic redox reactions of the explosive 4,6-dinitrobenzofuroxan (DNBF): implications for its toxic action

With an aim to understand the toxicity mechanisms of the explosive 4,6-dinitro- benzofuroxan (DNBF), we studied its single-electron reduction by NADPH:cytochrome P450 reductase and ferredoxin:NADP(+) reductase, and two- electron reduction by DT-diaphorase and Enterobacter cloacae nitroreductase. The enzymatic reactivities of DNBF and another explosive 2,4,6-trinitrotoluene (TNT) were similar, except for the much lower reactivity of DNBF towards nitroreductase. DNBF was less cytotoxic in FLK cells than TNT. However, their action shared the same mechanisms, oxidative stress and activation by DT-diaphorase. The lower cytotoxicity of DNBF may be explained by the negative electrostatic charge of its adduct with water which may impede cellular membrane penetration, and by the formation of its less reactive adducts with intracellular reduced glutathione.     

Human toxocariasis: a seroepidemiological survey in schoolchildren of Sorocaba, Brazil

A seroepidemiological survey for toxocariasis, among 180 schoolchildren of the public schools of Sorocaba City, state of S?o Paulo, Brazil, was carried out from August 2000 to July 2001. ELISA test was performed using excretory and secretory antigens for the detection of IgG anti-Toxocara antibodies. Information regarding the children was obtained from the parents or legal guardians. The results showed that the mean age was 5.4 +/- 1.4 years, the infection coefficient (IC) was 38.3 and the infection risk was higher among the children living in the city outskirts (IC = 47.4) where the socioeconomic conditions were worse than in the central region of the city (IC = 11.1). There was an association between higher frequency of seroreactivity in the ELISA test and the condition of living in a house with a yard and/or unpaved street. The same was observed in relation to a history of enteroparasitism. There was also an association between a seronegative ELISA test and previous treatment of pet dogs and/or cats with vermifuge. Based on these results, the authors propose that public health programs should include anthelmintic for dogs and cats during the antirabies vaccination campaigns, in order to diminish environmental contamination with Toxocara spp. eggs and consequently human infection.     

Pregnancy rates and metabolic profiles in cattle treated with propylene glycol prior to embryo transfer

The objective of this study was to determine the effect of a sustained propylene glycol administration to recipients of frozen/thawed in vivo derived bovine embryos. Heifers were treated with oral propylene glycol for the last 20 days before embryo transfer (n = 142), and untreated as controls (n = 133). Progesterone, insulin, insulin-like growth factor-I, glucose, urea and triglyceride were analysed in blood on Day 0 and Day 7 of the estrous cycle corresponding to embryo transfer. The heifers were selected as recipients when showing progesterone levels <2.0 ng/ml (Day 0) and >2.5 ng/ml (Day 7), according to corpus luteum quality on Day 7 by technicians unaware of animals treated. Within treated animals, significantly more recipients were selected, and increased progesterone, corpus luteum quality, pregnancy and calving rates were recorded. Day 7 progesterone concentrations were higher in heifers treated and transferred. Propylene glycol increased insulin and insulin-like-growth factor-I, but glucose, urea and triglyceride did not vary. Furthermore, insulin-like-growth factor-I, glucose and triglyceride increased at estrous time, but urea decreased and insulin remained unaltered. Together with the sustained gain in pregnancy rates throughout the experiment (2 years), other evidences suggested that the observed effects did not rely on nutritional deficiency. Thus, propylene glycol improved pregnancy rates after embryo-transfer, and progesterone, insulin and insulin-like-growth factor-I are probably involved in this effect.     

Selectively increasing inducible heat shock protein 70 via TAT-protein transduction protects neurons from nitrosative stress and excitotoxicity

Induction of heat shock protein 70 (Hsp70) via sublethal stress protects neurons from subsequent lethal injuries. Here we show that specific and efficient intracellular transduction of Hsp70 can be achieved utilizing an 11 amino acid leading sequence from human immunodeficiency virus (TAT-Hsp70) in primary neuronal cultures. Western blot and immunohistochemistry demonstrated intracellular accumulation of Hsp70 in insoluble protein fractions and mitochondrial compartments. We then examined the effects of Hsp70 overexpression using TAT-Hsp70 in models of nitrosative and excitotoxic neuronal death in vitro. Neurons were pre-incubated with 300 nM TAT-Hsp 70 overnight, then exposed to either peroxynitrite (ONOO-) or glutamate. TAT-Hsp70 maintained cellular respiration, inhibited extracellular lactate dehydrogenase release, and/or reduced cell death assessed by flow cytometry vs. vehicle, wild-type Hsp70, and TAT-beta-galactosidase controls. Hsp70 transduction using a TAT fusion protein is an effective method to selectively increase Hsp70 in neurons and is sufficient to provide neuroprotection from nitrosative stress and excitotoxicity. Further study is needed to confirm whether TAT-Hsp70 is protective in in vivo models of brain injury.     

One step closer to fixing association studies: evidence for age- and gender-specific allele frequency variations and deviations from Hardy-Weinberg expectations in controls

Association studies are the most powerful method available for identifying modest gene effects in complex disorders, but they often produce inconsistent results. With the rapidly growing SNP databases, haplotype maps and high throughput genotyping, the use of association studies is expected to increase; therefore, it is critical and timely that the problems with study design are identified and fixed. We questioned if unrecognized allele and genotype frequency variations in controls could be responsible for some of the inconsistent association findings. We performed a population genetic study of apolipoprotein E (APOE) and cytochrome P450 2D6 (CYP2D6) in 1,748 individuals ranging in age from newborns to centenarians. Although APOE and CYP2D6 are two of the most commonly used candidate genes, this is the first study to examine age- and gender-specific frequency distributions over the entire age spectrum, using a large, ethnically and geographically uniform population. We found significant, previously unrecognized variations in APOE allele frequencies, and deviations from Hardy-Weinberg expectations in CYP2D6 genotype frequencies starting at birth. The allele frequency variations within controls were larger than some reported case-control differences. We demonstrate that unrecognized frequency fluctuations in controls are a serious and potentially common confounder whose impact on association studies has not been appreciated, and one that can be addressed with proper study design. We recommend that population genetic studies be performed on commonly used candidate markers and that rigorous standards be applied for case-control matching.     

Land degradation affects shrub growth responses to precipitation in a semiarid rangeland of north‐eastern Patagonia (Argentina)

Arid land degradation diminishes the proportion of precipitation conducted to infiltration and increases the proportion lost to run‐off and evaporation. Consequently, we expect that the effects of annual precipitation on shrub growth vary with land degradation as a result of changes in soil available water. Chuquiraga avellanedae is the dominant shrub and the main indicator of land degradation in semiarid rangelands of north‐eastern Patagonia. We chose two communities with a different degree of land degradation: an herbaceous steppe with shrubs (HSS) and a degraded shrub steppe (SS). Vegetative growth of C. avellanedae was determined nondestructively using a double‐sampling approach. Soil water content was estimated for the two communities using a soil water balance model. Linear regressions were used to evaluate the relationships between shrub growth and (i) annual precipitation and (ii) mean available water during the period of high vegetative growth in the soil layer that each plant community concentrates their roots. In SS, with elevated clay content, there were more roots of C. avellanedae in the upper layers of soil while in HSS, with coarse‐textured soil, C. avellanedae had more roots in deeper layers. Vegetative growth of C. avellanedae, both in HSS and SS communities, was positively related to annual precipitation but, for a given precipitation, C. avellanedae presented higher vegetative growth in HSS than in SS. We also found a positive relationship between vegetative growth and soil available water, and this relationship did not differ between communities. SS presented lower water availability because of lower infiltration rates. Our results showed that, irrespective of the degree of land degradation, plants respond directly to water content of the soil layers where most roots are present at a specific window of time.     

Blue Carbon Storage in Tropical Seagrass Meadows Relates to Carbonate Stock Dynamics,Plant–Sediment Processes,and Landscape Context: Insights from the Western Indian Ocean

Globally, seagrass ecosystems are considered major blue carbon sinks and thus indirect contributors to climate change mitigation. Quantitative estimates and multi-scale appraisals of sources that underlie long-term storage of sedimentary carbon are vital for understanding coastal carbon dynamics. Across a tropical–subtropical coastal continuum in the Western Indian Ocean, we estimated organic (C_org) and inorganic (C_carb) carbon stocks in seagrass sediment. Quantified levels and variability of the two carbon stocks were evaluated with regard to the relative importance of environmental attributes in terms of plant–sediment properties and landscape configuration. The explored seagrass habitats encompassed low to moderate levels of sedimentary C_org (ranging from 0.20 to 1.44% on average depending on species- and site-specific variability) but higher than unvegetated areas (ranging from 0.09 to 0.33% depending on site-specific variability), suggesting that some of the seagrass areas (at tropical Zanzibar in particular) are potentially important as carbon sinks. The amount of sedimentary inorganic carbon as carbonate (C_carb) clearly corresponded to C_org levels, and as carbonates may represent a carbon source, this could diminish the strength of seagrass sediments as carbon sinks in the region. Partial least squares modelling indicated that variations in sedimentary C_org and C_carb stocks in seagrass habitats were primarily predicted by sediment density (indicating a negative relationship with the content of carbon stocks) and landscape configuration (indicating a positive effect of seagrass meadow area, relative to the area of other major coastal habitats, on carbon stocks), while seagrass structural complexity also contributed, though to a lesser extent, to model performance. The findings suggest that accurate carbon sink assessments require an understanding of plant–sediment processes as well as better knowledge of how sedimentary carbon dynamics are driven by cross-habitat links and sink–source relationships in a scale-dependent landscape context, which should be a priority for carbon sink conservation.     

Although Abundant in Tumor Tissue,Mast Cells Have No Effect on Immunological Micro-milieu or Growth of HPV-Induced or Transplanted Tumors

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