The purpose of this study was to determine the cytotoxic effect on CaCo-2 intestinal cells of dialysates obtained from bacterial cultures of some enterobacterial opportunistic strains with different ...sources of isolation (food, stool culture, acute diarrhoea, urine culture), previously tested and selected for their intensive adherence and invasion capacity to the cellular substratum and also for their cytotoxic effect on cell monolayers. In this study the level of cytotoxicity was measured quantitatively by means of the MTT assay and qualitatively by transmission electron microscopy (TEM). The MTT method uses a tetrazolium salt for the quantitative spectrophotometric assay of CaCo-2 cells survival and proliferation rates in the presence of bacterial dialysates. This test detects the viable cells, which are able to reduce the tetrazolium salt and offers the advantages of a very simple, rapid and precise method. For TEM examination the ultrathin sections were prepared following the standard protocols. The most cytotoxic strains proved to be Citrobacter freundii 93 strain isolated from stool culture, and Enterobacter cloacae 43, isolated from food followed by E. coli 115 strain isolated from acute diarrhoea. These results correlate well with TEM results pointing out the cytotoxic effect of Enterobacter cloacae 43 strain and also its ability to induce attachment and to destroy the cell surface (A/E) of HEp-2 cells. Besides their great adherence and invasion capacity, the production and release of cytotoxic factors into the extracellular medium represent virulence factors in these strains. This could be responsible for the increase of the pathogenic potential of opportunistic bacteria and explain their implication in the etiology of severe infections and food-borne diseases. This study proved that the virulence of opportunistic pathogens is not correlated with the strain's origin, the most evident virulence features being exhibited by an Enterobacter cloacae strain isolated from food.
Detecting and understanding disturbance is a challenge in ecology that has grown more critical with global environmental change and the emergence of research on social–ecological systems. We identify ...three areas of research need: developing a flexible framework that incorporates feedback loops between social and ecological systems, anticipating whether a disturbance will change vulnerability to other environmental drivers, and incorporating changes in system sensitivity to disturbance in the face of global changes in environmental drivers. In the present article, we review how discoveries from the US Long Term Ecological Research (LTER) Network have influenced theoretical paradigms in disturbance ecology, and we refine a framework for describing social–ecological disturbance that addresses these three challenges. By operationalizing this framework for seven LTER sites spanning distinct biomes, we show how disturbance can maintain or alter ecosystem state, drive spatial patterns at landscape scales, influence social–ecological interactions, and cause divergent outcomes depending on other environmental changes.
Chromosome 13 is the largest acrocentric human chromosome. It carries genes involved in cancer including the breast cancer type 2 (BRCA2) and retinoblastoma (RB1) genes, is frequently rearranged in ...B-cell chronic lymphocytic leukaemia, and contains the DAOA locus associated with bipolar disorder and schizophrenia. We describe completion and analysis of 95.5 megabases (Mb) of sequence from chromosome 13, which contains 633 genes and 296 pseudogenes. We estimate that more than 95.4% of the protein-coding genes of this chromosome have been identified, on the basis of comparison with other vertebrate genome sequences. Additionally, 105 putative non-coding RNA genes were found. Chromosome 13 has one of the lowest gene densities (6.5 genes per Mb) among human chromosomes, and contains a central region of 38 Mb where the gene density drops to only 3.1 genes per Mb.
Measuring total body water in peritoneal dialysis patients using an ethanol dilution technique.
The accuracy with which total body water (TBW) is estimated is a direct determinant of the reliability ...of Kt/V urea measurements in peritoneal dialysis (PD) patients. Ethanol dilution has been previously shown to be a reliable measure of TBW. Advances in breath alcohol technology make this a feasible clinical tool.
We gave 19 fasting chronic PD patients 0.3 g/kg of ethanol (EtOH) orally on two separate occasions. Breath alcohol concentrations (BrACs), determined by dual-beam infrared analysis, were recorded at baseline and periodically thereafter until BrACs were less than 0.01%. The TBW was then determined by standard pharmacokinetic techniques.
TBW measurements were reproducible, with a mean between-run difference of -0.004 liter/kg (95% limits of agreement -0.040 to 0.032 by Bland-Altman). The Watson equations tended to underestimate TBW, with a mean difference EtOH-Watson) of +30. liters (SD 4.0 liters, P = 0.004) and a mean absolute difference of 4.1 liters (SD 2.7 liters, range -4.4 to 9.5 liters). Kt/V was calculated from dialysate and urline collection, using V as determined from TBW estimated from EtOH and Watson. The mean Kt/V(EtOH) was 2.31 (SD 0.50) compared with 2.46 (SD 0.52) using Watson. The mean absolute differences between the two Kt/V estimates was 0.26 (SD 0.20, range -0.87 to 0.57), with Kt/V overestimated by Watson in 14 patients. EtOH was well tolerated, and the procedure was completed in about four hours.
Measuring V by the BRAc technique does not require blood sampling, is reliable, and is reproducible. It is a potentially useful method for a periodic determination of volume that may allow for more accurate Kt/V measurement in PD patients.
Members of the Src family of kinases are abundant in platelets. Although their localization is known, their role(s) in platelet function are not well understood. Lyn is a Src-family kinase that ...participates in signal transduction pathways elicited by collagen-related peptide; it has also been implicated through biochemical studies in the regulation of von Willebrand factor signaling. Here, we provide evidence that Lyn plays a role in γ-thrombin activation of platelets. Unlike the wild-type platelets, platelets from Lyn-deficient mice do not undergo irreversible aggregation, produce thromboxane A2, or secrete adenosine diphosphate in response to submaximal γ-thrombin concentrations that cause secretion-dependent irreversible aggregation. Phosphorylation of Akt, a downstream effector of phosphatidylinositol 3-kinase, also requires a higher concentration of γ-thrombin in Lyn-deficient platelets than in wild-type platelets. These findings demonstrate that Lyn signaling is required for thrombin induction of secretion-dependent platelet aggregation. Specifically, Lyn is required under these conditions to enable thrombin-induced TxA2 production and adenosine diphosphate secretion, necessary steps in secretion-dependent platelet aggregation.
Tamoxifen is an estrogen receptor (ER) ligand with widespread use in clinical and basic research settings. Beyond its application in treating ER-positive cancer, tamoxifen has been co-opted into a ...powerful approach for temporal-specific genetic alteration. The use of tamoxifen-inducible Cre-recombinase mouse models to examine genetic, molecular, and cellular mechanisms of development and disease is now prevalent in biomedical research. Understanding off-target effects of tamoxifen will inform its use in both clinical and basic research applications. Here, we show that prenatal tamoxifen exposure can cause structural birth defects in the mouse. Administration of a single 200 mg/kg tamoxifen dose to pregnant wildtype C57BL/6J mice at gestational day 9.75 caused cleft palate and limb malformations in the fetuses, including posterior digit duplication, reduction, or fusion. These malformations were highly penetrant and consistent across independent chemical manufacturers. As opposed to 200 mg/kg, a single dose of 50 mg/kg tamoxifen at the same developmental stage did not result in overt structural malformations. Demonstrating that prenatal tamoxifen exposure at a specific time point causes dose-dependent developmental abnormalities, these findings argue for more considerate application of tamoxifen in Cre-inducible systems and further investigation of tamoxifen's mechanisms of action.
Temporary rivers and streams that naturally cease to flow and dry up can be found on every continent. Many other water courses that were once perennial now also have temporary flow regimes due to the ...effects of water extraction for human use or as a result of changes in land use and climate. The dry beds of these temporary rivers are an integral part of river landscapes. We discuss their importance in human culture and their unique diversity of aquatic, amphibious, and terrestrial biota. We also describe their role as seed and egg banks for aquatic biota, as dispersal corridors and temporal ecotones linking wet and dry phases, and as sites for the storage and processing of organic matter and nutrients. In light of these valuable functions, dry riverbeds need to be fully integrated into river management policies and monitoring programs. We also identify key knowledge gaps and suggest research questions concerning the values of dry riverbeds.
Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the ...transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.
ABSTRACT
To manage agroecosystems for multiple ecosystem services, we need to know whether the management of one service has positive, negative, or no effects on other services. We do not yet have ...data on the interactions between pollination and pest‐control services. However, we do have data on the distributions of pollinators and natural enemies in agroecosystems. Therefore, we compared these two groups of ecosystem service providers, to see if the management of farms and agricultural landscapes might have similar effects on the abundance and richness of both. In a meta‐analysis, we compared 46 studies that sampled bees, predatory beetles, parasitic wasps, and spiders in fields, orchards, or vineyards of food crops. These studies used the proximity or proportion of non‐crop or natural habitats in the landscapes surrounding these crops (a measure of landscape complexity), or the proximity or diversity of non‐crop plants in the margins of these crops (a measure of local complexity), to explain the abundance or richness of these beneficial arthropods. Compositional complexity at both landscape and local scales had positive effects on both pollinators and natural enemies, but different effects on different taxa. Effects on bees and spiders were significantly positive, but effects on parasitoids and predatory beetles (mostly Carabidae and Staphylinidae) were inconclusive. Landscape complexity had significantly stronger effects on bees than it did on predatory beetles and significantly stronger effects in non‐woody rather than in woody crops. Effects on richness were significantly stronger than effects on abundance, but possibly only for spiders. This abundance‐richness difference might be caused by differences between generalists and specialists, or between arthropods that depend on non‐crop habitats (ecotone species and dispersers) and those that do not (cultural species). We call this the ‘specialist‐generalist’ or ‘cultural difference’ mechanism. If complexity has stronger effects on richness than abundance, it might have stronger effects on the stability than the magnitude of these arthropod‐mediated ecosystem services. We conclude that some pollinators and natural enemies seem to have compatible responses to complexity, and it might be possible to manage agroecosystems for the benefit of both. However, too few studies have compared the two, and so we cannot yet conclude that there are no negative interactions between pollinators and natural enemies, and no trade‐offs between pollination and pest‐control services. Therefore, we suggest a framework for future research to bridge these gaps in our knowledge.
Centrally acting thyrotropin releasing hormone (TRH), independent of endocrine action, has been shown to regulate several metabolic and behavioral parameters in rats, including food intake and ...locomotor activity. The present study investigated and compared the effects of central TRH on feeding behavior in Siberian hamsters exposed to long (LP) or short (SP) photoperiods, which induce natural physiological states of obesity and leanness respectively. The effects of two TRH analogues, RX77368 (a metabolically stable TRH analogue) and TRH-Gly (an endogenous precursor to TRH with putative preferential action at the central TRH receptor, TRH-R2), were also investigated. All peptides were infused via the third ventricle (i.c.v.). Food intake was measured, and the proportion of time spent interacting with food, active or resting was scored. TRH (5 microg) significantly reduced food intake without producing associated changes in activity in hamsters maintained in both LP (p < 0.001) and SP (p < 0.05). A lower dose of TRH (0.5 microg) only decreased feeding significantly (p < 0.01) in hamsters exposed to SP, indicating that there may be an underlying difference in sensitivity to TRH depending on metabolic state. RX77368 (1 microg) produced substantial hypophagia (p < 0.001) and decreased the proportion of time spent interacting with food, but, unlike TRH, may produce this via an increase in locomotor activity. TRH-Gly (5 microg) produced a small decrease in food intake (p < 0.05), lasting for 6 h. We conclude that TRH and TRH analogues possess anorexigenic capacities in this species, with a likely site of action in the hypothalamus. Increased sensitivity to the hypophagic effects of central TRH may contribute to the long-term catabolic state induced by short photoperiods.
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