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Protein kinase C inhibitors potentiate angiotensin II-induced phosphoinositide hydrolysis and intracellular Ca2+ mobilization in renal mesangial cells.

Stimulation of mesangial cells with angiotensin II leads to rapid phosphoinositide hydrolysis and subsequent mobilization of intracellular Ca2+. Previous studies indicated that activation of protein kinase C (PKC) triggers a negative-feedback signal, which limits phosphoinositide turnover. By comparing the relative susceptibility of PKC isoenzymes to phorbol ester-induced down-regulation with the down-regulation of the functional cell response, i.e. feedback inhibition of inositol trisphosphate production, we inferred that PKC-alpha and PKC-delta are candidates for regulating phosphoinositide hydrolysis in mesangial cells. To test this hypothesis further, we examined the effects of inhibitors of PKC, that are reportedly not active on PKC-delta, on angiotensin II-stimulated phosphoinositide degradation and Ca2+ mobilization. Pretreatment of mesangial cells with the PKC inhibitors staurosporine and K252a potently augmented inositol trisphosphate and 1,2-diacylglycerol formation as well as Ca2+ mobilization in response to angiotensin II. These results suggest that PKC-alpha, but not PKC-delta, is the most likely candidate mediating feedback inhibition of angiotensin II-stimulated phosphoinositide turnover in mesangial cells.

Alkaloids↗

A critical evaluation of direct electrical protein detection methods.

During the last decennia many protein-related electrical phenomena have been studied and applied in a variety of measuring systems, from simple metal electrodes with adsorbed proteins to sophisticated systems with lipid bilayers. Many of the investigations concern the monitoring of immuno reactions. The basic underlying electrical effects of the observed phenomena are the protein modulated dielectric constant, conductivity, electrical potential, ion permeability and ion mobility. In this paper special attention is paid to the capacitive measurements with EIS systems as well as impedance and potential measurements with FET devices. The Donnan theory is treated and applied to the static ImmunoFET operation, explaining the relatively small effects which have been reported. Finally, an alternative approach is described in which the ImmunoFET is applied in a dynamic way, to circumvent the drawbacks of the static measurements.

Amino Acid Sequence↗

Rate of iodine volatilization and accumulation by filamentous fungi through laboratory cultures.

Five strains of basidiomycetes (Lentinula edodes, Coprinus phlyctidosporus, Hebeloma vinosophyllum, Pleurotus ostreatus and Agaricus bisporus), one strain of ascomycete (Hormoconis resinae) and six strains of imperfect fungi (Penicillium chrysogenum, Penicillium roquefortii, Cladosporium cladosporioides, Alternaria alternata, Aspergillus niger and Aspergillus oryzae) were cultured in a liquid medium containing a radioactive iodine tracer ((125)I), and were tested for their abilities to volatilize or accumulate iodine. Of the fungal strains tested, 11 strains volatilized a considerable amount of iodine, with L. edodes showing the highest volatilization rate of 3.4%. The volatile organic iodine species emitted from imperfect fungi cultures was identified as methyl iodide (CH(3)I). In contrast, six fungal strains in 12 strains accumulated a considerable amount of iodine from the medium with concentration factors of more than 1.0. Among these, Alt. alternata and Cl. cladosporioides accumulated more than 40% of the iodine in their hyphae, and showed high concentration factors of 22 and 18, respectively. These results suggest that filamentous fungi have a potential to influence the mobility and speciation of iodine by volatilization and accumulation. Considering their great biomass in soils, filamentous fungi may contribute to the global circulation of stable iodine and also the long-lived radioiodine, (129)I (half-life: 1.6 x 10(7)years), released from nuclear facilities into the environment.

Fungi↗

Heavy metals, polycyclic aromatic hydrocarbons and polychlorinated biphenyls in surface sediments of the Naples harbour (southern Italy).

The Naples's harbour is one of the largest and most important commercial and tourist port of the Mediterranean basin. It is located on the southeast coast of Italy and receives industrial and municipal wastewaters from the city of Naples. Due to its social and economic impact, a comprehensive assessment of levels and sources of contamination of bottom sediments in this area of the Mediterranean basin is essential to identify potential danger due to mobilization of contaminants produced by managing of the same sediments. In this study, superficial sediments collected from 189 sampling sites were analyzed for grain size, heavy metals (Cr, Cu, Ni, Pb, V, Zn, Co, Sn, Cd, Hg, As, Al and Fe), 16 priority polycyclic aromatic hydrocarbons (PAHs) and perylene and 38 individual polychlorinated biphenyl (PCB) congeners. Compared to the estimated local background, Cu, Zn, Pb, Cd, Sn and Hg show enrichment factors >3 and only Hg evidences a median value higher than the NOAA (effects range - median) guidelines. Principal component analysis allowed us to clearly discriminate two areas mainly affected by heavy metals contamination and influenced by different sources related to industrial, commercial and/or urban activities. Priority PAHs are predominantly represented by three-five-ring compounds with concentrations ranging between 9 and 31774 ng g(-1) and frequently higher than the NOAA ER-M index. A prevalent pyrolitic origin of PAH was assessed on the basis of the relative abundance of the different congeners and selected isomer ratios. The concentrations of PCBs, as sum of the 38 congeners, ranged from 1 to 899 ng g(-1), with a predominance of highly chlorinated (tetra- and penta-chlorobiphenyls) congeners. WHO-TEQ values, calculated for the PCDD-like PCB congeners, suggest a relatively high level of toxicity. Generally, the concentration of PAHs and PCBs were higher near the sites of intense industrial, shipping and/or commercial activities suggesting a direct influence of these sources on the pollutant distribution patterns.

Carbon↗

Use of electrokinetic measurements for characterization of columns used in capillary electrochromatography.

The separation mechanism in capillary electrochromatography (CEC) is a hybrid differential migration process, which entails the features of both high-performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE), i.e., chromatographic retention and electrophoretic migration. The focus of this paper is on the use of electrokinetic data, such as current, electroosmotic flow (EOF) and column efficiency measurements, that are readily available, for an improved understanding of CEC separations. A framework is presented here for the use of this data for evaluation of a variety of performance parameters including, conductivity ratio, interstitial EOF mobility, porosity, and zeta potential. This framework is applied for characterization of two monolithic columns with different chemistry that were manufactured in-house. The above-mentioned performance parameters were calculated for the two columns and it is found that the poly(VBC-EGDMA-SWNT) monolithic column with the GPTMS-PEI coating offers a significantly improved flow distribution in comparison to the poly(VBC-EGDMA) monolithic column. This observation is confirmed by performing separation of peptides on the two columns and height equivalent of a theoretical plate (HETP) measurements on the resulting peaks. It is shown that following our approach leads to an improved understanding of the separations achieved with the columns and to better column design.

Chromatography, Micellar Electrokinetic Capillary↗

Mechanisms of the inhibitory effects of amyloid beta-protein on synaptic plasticity.

Alzheimer's disease can be considered a protein misfolding disease. In particular, inappropriate processing of a proteolytic fragment of amyloid precursor protein, amyloid beta-protein (Abeta), in early stages of Alzheimer's disease may lead to stabilization of small oligomers that are highly mobile and have a potential to be extremely toxic assemblies. Recently, the importance of such soluble species of Abeta in triggering synaptic dysfunction, long before neuronal loss occurs, has become apparent. Animal models have revealed that plasticity of hippocampal excitatory synaptic transmission is relatively selectively vulnerable to Abeta both in vitro and in vivo. This review focuses on the mechanisms of Abeta inhibition of long-term potentiation at synapses in the rodent hippocampus from two complimentary perspectives. Firstly, we examine evidence that the synaptic activity of this peptide resides primarily in oligomeric rather than monomeric or fibrillar Abeta species. Secondly, the importance of different oxidative/nitrosative stress-linked cascades including JNK, p38 MAPK and NADPH oxidase/iNOS-generated reactive oxygen/nitrogen free radicals in mediating the inhibition of LTP by Abeta is emphasised. These mechanistic studies provide a plausible explanation for the sensitivity of hippocampus-dependent memory to impairment in the early preclinical stages of Alzheimer's disease.

Aged↗

A review and a framework of handheld computer adoption in healthcare.

Wide adoption of mobile computing technology can potentially improve information access, enhance workflow, and promote evidence-based practice to make informed and effective decisions at the point of care. Handheld computers or personal digital assistants (PDAs) offer portable and unobtrusive access to clinical data and relevant information at the point of care. This article reviews the literature on issues related to adoption of PDAs in health care and barriers to PDA adoption. Studies showed that PDAs were used widely in health care providers' practice, and the level of use is expected to rise rapidly. Most care providers found PDAs to be functional and useful in areas of documentation, medical reference, and access to patient data. Major barriers to adoption were identified as usability, security concerns, and lack of technical and organizational support. PDAs offer health care practitioners advantages to enhance their clinical practice. However, better designed PDA hardware and software applications, more institutional support, seamless integration of PDA technology with hospital information systems, and satisfactory security measures are necessary to increase acceptance and wide use of PDAs in healthcare.

Access to Information↗

Source, dispersion and combustion modelling of an accidental release of hydrogen in an urban environment.

Hydrogen is likely to be the most important future energy carrier, for many stationary and mobile applications, with the potential to make significant reductions in greenhouse gas emissions especially if renewable primary energy sources are used to produce the hydrogen. A safe transition to the use of hydrogen by members of the general public requires that the safety issues associated with hydrogen applications have to be investigated and fully understood. In order to assess the risks associated with hydrogen applications, its behaviour in realistic accident scenarios has to be predicted, allowing mitigating measures to be developed where necessary. A key factor in this process is predicting the release, dispersion and combustion of hydrogen in appropriate scenarios. This paper illustrates an application of CFD methods to the simulation of an actual hydrogen explosion. The explosion occurred on 3 March 1983 in a built up area of central Stockholm, Sweden, after the accidental release of approximately 13.5 kg of hydrogen from a rack of 18 interconnected 50 l industrial pressure vessels (200 bar working pressure) being transported by a delivery truck. Modelling of the source term, dispersion and combustion were undertaken separately using three different numerical tools, due to the differences in physics and scales between the different phenomena. Results from the dispersion calculations together with the official accident report were used to identify a possible ignition source and estimate the time at which ignition could have occurred. Ignition was estimated to occur 10s after the start of the release, coinciding with the time at which the maximum flammable hydrogen mass and cloud volume were found to occur (4.5 kg and 600 m(3), respectively). The subsequent simulation of the combustion adopts initial conditions for mean flow and turbulence from the dispersion simulations, and calculates the development of a fireball. This provides physical values, e.g. maximum overpressure and far-field overpressure that may be used as a comparison with the known accident details to give an indication of the validity of the models. The simulation results are consistent with both the reported near-field damage to buildings and persons and with the far-field damage to windows. The work was undertaken as part of the European Integrated Hydrogen Project-Phase 2 (EIHP2) with partial funding from the European Commission via the Fifth Framework Programme.

Accidents↗

From regionalization to homogenization: Nationwide metagenomic assessment of priority pathogens and the resistome in Polish hospital wastewater.

Hospital wastewater (HWW) is a critical hotspot for the dissemination of antibiotic resistance genes (ARGs) and pathogens. This study provides the first comprehensive metagenomic characterization of HWW across Poland, analyzing 64 medical facilities across two seasons via Nanopore long-read sequencing (total of 128 HWW samples). The HWW microbiome was mostly dominated by Proteobacteria, Bacteroidota, and Firmicutes. Multivariate analysis confirmed a significant seasonal shift in the resistome. Winter samples exhibited geographic regionalization, with localized hotspots of specific ARGs, including vancomycin resistance (operon van) and carbapenemase genes (blaOXA, blaNDM). Conversely, summer samples showed a significant trend toward nationwide homogenization, characterized by a uniform distribution of ESBL genes (blaTEM, blaCTX-M) and multidrug resistance (MDR) determinants, alongside the persistence of localized clinical hotspots. Klebsiella pneumoniae emerged as a central network hub, particularly in summer, showing strong correlations with ESBLs. Quantitative genomic co-occurrence analysis revealed a functional division within dominant taxa: while environmental species like Acinetobacter johnsonii comprised the general background microbiome, clinical pathogens such as Acinetobacter baumannii served as primary vectors, showing frequent associations with high-risk ARGs. Environmental and opportunistic bacteria, such as Aeromonas spp. and Citrobacter spp., were identified as putative 'bridge hosts' associated with mobile resistance determinants and potentially contributing to HGT. The findings indicate that seasonal factors, such as increased temperature and sub-inhibitory antibiotic concentrations, may contribute to the transition from regionalized to homogenized resistance profiles, demonstrating that background resistome convergence can coexist with point-source clinical outbreaks. This seasonal "blurring" of regional boundaries positions HWW as an active vector for large-scale antimicrobial resistance (AMR) dissemination. These results underscore the urgent need for nationwide metagenomic surveillance and advanced wastewater treatment strategies within the "One Health" framework to mitigate the environmental spread of WHO priority pathogens.

Acinetobacter baumannii↗

Why a mechanical disc?

Low back pain secondary to degenerative disc disease is an overwhelming and growing problem in the United States and Western countries. Most degenerative disc disease can be treated nonoperatively. There are, however, substantial numbers of patients who have not benefited from exhaustive nonoperative treatments and subsequently seek surgical solutions to their incapacitating back pain. Lumbar fusion for back pain and/or leg pain associated with degenerative disc disease is considered the gold standard by which other treatments are judged. A challenge to spinal fusion for degenerative disc disease is now being offered in the form of the artificial disc. The implantation of an artificial lumbar disc allows for maintenance or restoration of physiologic movement at affected segments. A major long-term complication of spinal fusion is degeneration of a disc adjacent to the fused segments. Theoretically, the maintenance of motion could minimize development of adjacent disc degeneration as seen with spinal fusion. It is interesting to note that fusion of the hip or knee is not considered a primary procedure, but fusion is a primary procedure for the lumbar spine. Four artificial lumbar discs are discussed in this article. Early results are promising in terms of clinical results and movement, but long-term follow-up clinical trials must be done in order to gain an accurate comparison with spinal fusion. Trials are currently ongoing. The clinical results up to now and the potential for maintaining lumbar mobility throughout life warrant continuation of this surgical procedure.

Arthroplasty, Replacement↗

Calcium dependence of Pi phosphorylation of sarcoplasmic reticulum Ca2+-ATPase at low water content: water dependence of the E2-->E1 conversion.

Enzymes entrapped in reverse micelles can be studied in low-water environments that have the potential of restricting conformational mobility in specific steps of the reaction cycle. Sarcoplasmic reticulum Ca2+-ATPase was incorporated into a reverse-micelle system (TPT) composed of toluene, phospholipids, Triton X-100 and varying amounts of water (0.5-7%, v/v). Phosphorylation of the Ca2+-ATPase by ATP required the presence of both water and Ca2+ in the micelles. No phosphoenzyme (EP) was detected in the presence of EGTA. Phosphorylation by Pi (inorganic phosphate) in the absence of Ca2+ was observed at water content below that necessary for phosphorylation by ATP. In contrast to what is observed in a totally aqueous medium, EP formed by Pi was partially resistant to dephosphorylation by Ca2+. However, the addition of non-radioactive Pi to the EP already formed caused a rapid decrease in radiolabelled enzymes, as expected for the isotopic dilution, indicating the existence of an equilibrium (E+Pi<-->EP). Phosphorylation by Pi also occurred in TPT containing millimolar Ca2+ concentrations in a range of water concentrations (2-5% v/v). The substrates p-nitrophenyl phosphate, acetyl phosphate, ATP and GTP increased the EP level under these conditions. These results suggest that: (1) the rate of conversion of the ATPase conformer E2 into E1 is greatly reduced at low water content, so that E2-->E1 becomes the rate-limiting step of the catalytic cycle; and (2) in media of low water content, Pi can phosphorylate both E1Ca and E2. Thus, the effect of enzyme hydration is complex and involves changes in the phosphorylation reaction at the catalytic site, in the equilibrium between E2 and E1 conformers, and in their specificity for substrates.

Adenosine Triphosphate↗

Elution mechanism in electrostatic ion chromatography with histidine as an isoelectric ampholytic mobile phase.

Electrostatic ion chromatography (EIC) using a zwitterionic stationary phase (formed by coating a C18 material with a hydrophobic zwitterionic surfactant) was studied with a mobile phase comprising an aqueous solution of histidine at the pH of its isoelectric point, together with non-suppressed conductometric detection. Anions and cations were found to be eluted as separate peaks, unlike the elution behaviour observed on the same system when pure water was used as mobile phase. An explanation was suggested in terms of protonation equilibria of the overall uncharged histidine to form small amounts of histidine cations and anions in the mobile phase which could act as counterions for analyte anions and cations. This suggestion was supported by measured pH changes occurring in the bands of eluted analyte anions (a decreased pH compared to the mobile phase) and cations (an increased pH compared to the mobile phase). The analytical potential of this type of EIC is discussed.

Chromatography, Liquid↗

Electrophoresis of concentrated mercury drops.

The electrophoretic behavior of concentrated monodispersed, positively charged mercury drops is investigated theoretically. The present study extends previous analyses by considering arbitrary surface potentials, double-layer polarization, and the interaction between adjacent double layers. The coupled equations describing the spatial variations in the flow field, the electric field, and the concentration field are solved by a pseudo-spectral method. For a low surface potential phi(r), the mobility increases monotonically with kappaalpha; kappa and alpha are respectively the reciprocal Debye length and the radius of a mercury drop. For medium and high phi(r), the mobility curve has a reflection point, which arises from the interaction of adjacent double layers, for kappaalpha. Also, if phi(r) is high, the mobility curve may exhibit a local minimum as kappaalpha varies. This phenomenon is pronounced if the concentration of the dispersed phase is high. If the double layer is thick, the mobility increases with phi(r), and the reverse is true if it is thin. We show that the higher the concentration of the dispersed phase the smaller the mobility, and as kappaalpha becomes large the mobility approaches a constant value, which is independent of the concentration of the dispersed phase. The mobility of mercury drops is larger than that of the corresponding rigid particles.

Journal Article↗

Spatial and temporal organic and heavy metal pollution at Mai Po Marshes Nature Reserve, Hong Kong.

An intensive monthly sampling of water and sediments from 12 sites over 8 months covering wet and dry seasons at Mai Po Marshes Nature Reserve was conducted during June 1997-February 1998. Major organic (C, N and P) and heavy metal pollutants (Cd, Cr, Cu, Ni, Pb, Zn) water and sediment samples were examined. The results showed that Mai Po Marshes were severely polluted by organic matter and heavy metals, and the water from Deep Bay appeared to be the source of pollution. Up to 13-55% chance that the sediments of Mai Po Marshes were classified as moderately to seriously metal contaminated materials, according to the guideline set by Hong Kong Government. Empirical models describing organic matter and heavy metal spatial and seasonal dynamics in the water and sediments were formulated, based on data analysis. During wet season (June-October), more than 58% variations of total P can be explained by ortho-P in water, while ammonia-N explained up to 90% variations of total Kjeldahl nitrogen in water. Throughout the whole sampling period (June-February), there were significant correlations (p<0.01) between total organic C in water. pH in the sediments and salinity in water appeared to be important factors determining heavy metal mobility in sediments, while potential metal release from the sediments is a concern when any oxidizing processes such as flooding or dredging are imposed on sediments.

Carbon↗

Synthetic zeolites as amendments for sewage sludge-based compost.

The effects of incorporating a synthetic zeolite (Zeolite P) in a range of concentrations (0.1-1.0 w:w) into an experimental horticultural compost, derived from sewage sludge, have been investigated. The impact of zeolite treatment on time-related changes of the labile zinc, copper, iron and manganese pools within the compost was compared to lime incorporation (5% w:w) and to a proprietary unamended peat-based compost. Addition of 0.5% and 1.0% zeolite significantly reduced labile zinc over a 90 day period. The highest zeolite treatment was more effective than liming; 0.5% zeolite was as effective as lime. Plant growth trials measuring transfer of metals to ryegrass (Lolium perenne L. cv Elka) in successive harvests demonstrated that both 1.0% zeolite and 5% lime treatment caused significant reduction in total metal transfer from soil-plant over a 116 day growth period. It is concluded that the use of synthetic zeolite as an amendment for compost of this type significantly reduces potential for soil metal mobility and soil-plant transfer.

Lolium↗

Differential effects of low molecular weight heparin and unfractionated heparin on circulating levels of antithrombin and tissue factor pathway inhibitor (TFPI): a possible mechanism for difference in therapeutic efficacy.

There is growing evidence on the superior efficacy and safety of low molecular weight heparins (LMWHs) over unfractionated heparin (UFH) for the treatment of both venous and arterial thromboembolism. Heparin exerts its function by potentiating antithrombin and by mobilizing tissue factor pathway inhibitor (TFPI) into the circulation. The present study was conducted to compare the effect of subcutaneous LMWH and infusion of UFH on these two anticoagulants in plasma. Eighteen healthy male volunteers were randomly allocated to therapy with continuous intravenous (iv) UFH (n=6) (initial infusion rate 450 IU/kg/day) or low molecular weight heparin (LMWH) (enoxaparin, 1.5 mg/kg/day) subcutaneously (sc) once daily for 72 hours. Free TFPI antigen, measured by a solid-phase two-site enzyme immunoassay, and antithrombin and protein C activities, measured by chromogenic assays, were assessed in plasma samples before, during, and after anticoagulant treatment. Infusion of UFH, but not subcutaneous LMWH, was found to attenuate the antithrombotic defense by a selective decrease of both circulating antithrombin (-21+/-7%, p<0.0001) and of free and endothelial-bound TFPI. The changes in antithrombin and TFPI by LMWH and UFH were statistically different between groups (p<0.001). The differential effect of UFH and LMWH on antithrombin and TFPI may explain the superior efficacy of subcutaneous LMWH compared with conventional intravenous UFH for treatment of both arterial and venous thrombosis.

Administration, Cutaneous↗

Head-only exposure to GSM 900-MHz electromagnetic fields does not alter rat's memory in spatial and non-spatial tasks.

Over the last decade, exposure to high frequency (2450 MHz) electromagnetic fields (EMFs) has been found to induce performance deficit in rodents in spatial memory tasks. As concern was expressed about potential biological effects of mobile communication microwaves, studies testing the effects of signals such as GSM were required. In a previous study, using head-only exposure to 900 MHz GSM EMF, we could not demonstrate any behavioural deficit in two simple learning tasks. The present study aimed at extending these results with more complex spatial learning tasks and a non-spatial task. In a first experiment, rats were trained in a radial-arm maze with a 10-s confinement between each visited arm. In a second experiment, a 15-min intra-trial delay was introduced after four visited arms. In a third experiment, non-spatial memory was tested in an object recognition task. In all experiments, performance of the head-only exposed rats (1 and 3.5 W/kg) was compared with that of sham and control rats. In the first experiment, a slightly improved performance was found after 3.5 W/kg exposure, a result that was not observed in the delay-task. In the third experiment, although some effects on exploratory activity were found, recognition memory was unaffected in exposed rats. Altogether, this set of experiments provides no evidence indicating that spatial and non-spatial memory can be affected by a 45-min head-only exposure to 900 MHz GSM EMF.

Animals↗

Solid phase speciation of arsenic by sequential extraction in standard reference materials and industrially contaminated soil samples.

Availability, mobility, (phyto)toxicity and potential risk of contaminants is strongly affected by the manner of appearance of elements, the so-called speciation. Operational fractionation methods like sequential extractions have been applied for a long time to determine the solid phase speciation of heavy metals since direct determination of specific chemical compounds can not always be easily achieved. The three-step sequential extraction scheme recommended by the BCR and two extraction schemes based on the phosphorus-like protocol proposed by Manful (1992, Occurrence and Ecochemical Behaviours of Arsenic in a Goldsmelter Impacted Area in Ghana, PhD dissertation, at the RUG) were applied to four standard reference materials (SRM) and to a batch of samples from industrially contaminated sites, heavily contaminated with arsenic and heavy metals. The SRM 2710 (Montana soil) was found to be the most useful reference material for metal (Mn, Cu, Zn, As, Cd and Pb) fractionation using the BCR sequential extraction procedure. Two sequential extraction schemes were developed and compared for arsenic with the aim to establish a better fractionation and recovery rate than the BCR-scheme for this element in the SRM samples. The major part of arsenic was released from the heavily contaminated samples after NaOH-extraction. Inferior extraction variability and recovery in the heavily contaminated samples compared to SRMs could be mainly contributed to subsample heterogeneity.

Arsenic↗