An investigation into secondary transfer-The transfer of textile fibres to seats.

Textile fibres can be transferred directly, person to person or person to scene (primary transfer), or indirectly via an intermediate (secondary transfer). In criminal cases involving a transfer of textile fibres, it is often the case that whilst the provenance of recovered 'crime significant' fibres is accepted by the defence, it is a particular activity leading to their transfer to a surface in question which is disputed. In such circumstances, transfer and persistence studies relating to fibres on a particular substrate in particular conditions assist in evaluating whether the presence of crime relevant fibres is more likely to have occurred by one particular activity compared to another. This study investigates the effect of a time delay between the primary transfer of fibres to a garment on the numbers of subsequently secondarily transferred fibres to a seat. Two donor garments composed of polyester and cotton fibres respectively were employed in this study and secondarily transferred to seats after time intervals of 0, 0.5, 2, 6 and 24h. The number of secondarily transferred fibres were recorded according to fibre type and time interval and compared against levels recorded at the primary transfer stage. The results showed that only a relatively small percentage of the original primary transfer is likely to be secondarily transferred and that the numbers found were inversely proportional to the time interval between the primary and secondary transfer. In addition, it was found that the secondary transfer of cotton fibres was an order of magnitude higher than for polyester.

Contamination with bacterial zoonotic pathogen genes in U.S. streams influenced by varying types of animal agriculture.

Animal waste, stream water, and streambed sediment from 19 small (<32km(2)) watersheds in 12U.S. states having either no major animal agriculture (control, n=4), or predominantly beef (n=4), dairy (n=3), swine (n=5), or poultry (n=3) were tested for: 1) cholesterol, coprostanol, estrone, and fecal indicator bacteria (FIB) concentrations, and 2) shiga-toxin producing and enterotoxigenic Escherichia coli, Salmonella, Campylobacter, and pathogenic and vancomycin-resistant enterococci by polymerase chain reaction (PCR) on enrichments, and/or direct quantitative PCR. Pathogen genes were most frequently detected in dairy wastes, followed by beef, swine and poultry wastes in that order; there was only one detection of an animal-source-specific pathogen gene (stx1) in any water or sediment sample in any control watershed. Post-rainfall pathogen gene numbers in stream water were significantly correlated with FIB, cholesterol and coprostanol concentrations, and were most highly correlated in dairy watershed samples collected from 3 different states. Although collected across multiple states and ecoregions, animal-waste gene profiles were distinctive via discriminant analysis. Stream water gene profiles could also be discriminated by the watershed animal type. Although pathogen genes were not abundant in stream water or streambed samples, PCR on enrichments indicated that many genes were from viable organisms, including several (shiga-toxin producing or enterotoxigenic E. coli, Salmonella, vancomycin-resistant enterococci) that could potentially affect either human or animal health. Pathogen gene numbers and types in stream water samples were influenced most by animal type, by local factors such as whether animals had stream access, and by the amount of local rainfall, and not by studied watershed soil or physical characteristics. Our results indicated that stream water in small agricultural U.S. watersheds was susceptible to pathogen gene inputs under typical agricultural practices and environmental conditions. Pathogen gene profiles may offer the potential to address both source of, and risks associated with, fecal pollution.

Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) at ambient freshwater beaches.

Methicillin-resistant Staphylococcus aureus (MRSA) are a threat to human health worldwide, and although detected at marine beaches, they have been largely unstudied at freshwater beaches. Genes indicating S. aureus (SA; femA) and methicillin resistance (mecA) were detected at 11 and 12 of 13 US Great Lakes beaches and in 18% or 27% of 287 recreational water samples, respectively. Eight beaches had mecA+femA (potential MRSA) detections. During an intensive study, higher bather numbers, staphylococci concentrations, and femA detections were found in samples collected after noon than before noon. Local population density, beach cloud cover, and beach wave height were significantly correlated with SA or MRSA detection frequency. The Panton-Valentine leukocidin gene, associated with community-acquired MRSA, was detected in 12 out of 27 potential MRSA samples. The femA gene was detected less frequently at beaches that met US enterococci criteria or EU enterococci 'excellent' recreational water quality, but was not related to Escherichia coli-defined criteria. Escherichia coli is often the only indicator used to determine water quality at US beaches, given the economic and healthcare burden that can be associated with infections caused by SA and MRSA, monitoring of recreational waters for non-fecal bacteria such as staphylococci and/or SA may be warranted.

Spatial, Temporal, and Matrix Variability of Clostridium botulinum Type E Toxin Gene Distribution at Great Lakes Beaches.

Clostridium botulinum type E toxin is responsible for extensive mortality of birds and fish in the Great Lakes. The C. botulinum bontE gene that produces the type E toxin was amplified with quantitative PCR from 150 sloughed algal samples (primarily Cladophora species) collected during summer 2012 from 10 Great Lakes beaches in five states; concurrently, 74 sediment and 37 water samples from four sites were also analyzed. The bontE gene concentration in algae was significantly higher than in water and sediment (P < 0.05), suggesting that algal mats provide a better microenvironment for C. botulinum. The bontE gene was detected most frequently in algae at Jeorse Park and Portage Lake Front beaches (Lake Michigan) and Bay City State Recreation Area beach on Saginaw Bay (Lake Huron), where 77, 100, and 83% of these algal samples contained the bontE gene, respectively. The highest concentration of bontE was detected at Bay City (1.98 × 10(5) gene copies/ml of algae or 5.21 × 10(6) g [dry weight]). This study revealed that the bontE gene is abundant in the Great Lakes but that it has spatial, temporal, and matrix variability. Further, embayed beaches, low wave height, low wind velocity, and greater average water temperature enhance the bontE occurrence.

Genes Indicative of Zoonotic and Swine Pathogens Are Persistent in Stream Water and Sediment following a Swine Manure Spill.

Manure spills into streams are relatively frequent, but no studies have characterized stream contamination with zoonotic and veterinary pathogens, or fecal chemicals, following a spill. We tested stream water and sediment over 25 days and downstream for 7.6 km for the following: fecal indicator bacteria (FIB), the fecal indicator chemicals cholesterol and coprostanol, 20 genes for zoonotic and swine-specific bacterial pathogens by presence/absence PCR for viable cells, one swine-specific Escherichia coli toxin gene (STII gene) by quantitative PCR (qPCR), and nine human and animal viruses by qPCR or reverse transcription-qPCR. Twelve days postspill, and 4.2 km downstream, water concentrations of FIB, cholesterol, and coprostanol were 1 to 2 orders of magnitude greater than those detected before, or above, the spill, and genes indicating viable zoonotic or swine-infectious Escherichia coli were detected in water or sediment. STII gene levels increased from undetectable before or above the spill to 10(5) copies/100 ml of water 12 days postspill. Thirteen of 14 water (8/9 sediment) samples had viable STII-carrying cells postspill. Eighteen days postspill, porcine adenovirus and teschovirus were detected 5.6 km downstream. FIB concentrations (per gram [wet weight]) in sediment were greater than in water, and sediment was a continuous reservoir of genes and chemicals postspill. Constituent concentrations were much lower, and detections less frequent, in a runoff event (200 days postspill) following manure application, although the swine-associated STII and stx2e genes were detected. Manure spills are an underappreciated pathway for livestock-derived contaminants to enter streams, with persistent environmental outcomes and the potential for human and veterinary health consequences.

The prevalence of two 'commonly' encountered synthetic target fibres within a large urban environment.

A target fibre study was carried out to assess the random prevalence of two ostensibly commonly encountered synthetic fibre types; black acrylic and blue polyester. The study was performed in an environment which maximised the number of random contacts between textile garments in the population and specific surfaces, namely; seating relating to buses, public houses and cinemas found within a large urban conurbation. Surface debris tapings were collected from samples of bus seats (30), pub seats (54) and cinema seats (53). Using low power stereomicroscopy, a total of 114 and 68 fibres, superficially similar to the respective black acrylic and blue polyester target fibres, were recovered from these tapings. The full range of comparative microscopical and instrumental analysis used in operational forensic laboratories was performed on the recovered fibres. No matches were found with either of the target fibres. These findings are in accordance with similar studies which show that the probability of an 'adventitious' match with a particular fibre type/colour combination is extremely low. In addition, the findings demonstrate that the current techniques and instrumentation employed by operational forensic laboratories are fit for purpose. Importantly, the findings demonstrate that databases and surveys (e.g. fibre population studies) which do not consider the analytical/comparison processes, must not be used in isolation when evaluating fibre evidence at source level.

Bacterial pathogen gene abundance and relation to recreational water quality at seven Great Lakes beaches.

Quantitative assessment of bacterial pathogens, their geographic variability, and distribution in various matrices at Great Lakes beaches are limited. Quantitative PCR (qPCR) was used to test for genes from E. coli O157:H7 (eaeO157), shiga-toxin producing E. coli (stx2), Campylobacter jejuni (mapA), Shigella spp. (ipaH), and a Salmonella enterica-specific (SE) DNA sequence at seven Great Lakes beaches, in algae, water, and sediment. Overall, detection frequencies were mapA>stx2>ipaH>SE>eaeO157. Results were highly variable among beaches and matrices; some correlations with environmental conditions were observed for mapA, stx2, and ipaH detections. Beach seasonal mean mapA abundance in water was correlated with beach seasonal mean log10 E. coli concentration. At one beach, stx2 gene abundance was positively correlated with concurrent daily E. coli concentrations. Concentration distributions for stx2, ipaH, and mapA within algae, sediment, and water were statistically different (Non-Detect and Data Analysis in R). Assuming 10, 50, or 100% of gene copies represented viable and presumably infective cells, a quantitative microbial risk assessment tool developed by Michigan State University indicated a moderate probability of illness for Campylobacter jejuni at the study beaches, especially where recreational water quality criteria were exceeded. Pathogen gene quantification may be useful for beach water quality management.

Geographic setting influences Great Lakes beach microbiological water quality.

Understanding of factors that influence Escherichia coli (EC) and enterococci (ENT) concentrations, pathogen occurrence, and microbial sources at Great Lakes beaches comes largely from individual beach studies. Using 12 representative beaches, we tested enrichment cultures from 273 beach water and 22 tributary samples for EC, ENT, and genes indicating the bacterial pathogens Shiga-toxin producing E. coli (STEC), Shigella spp. , Salmonella spp , Campylobacter jejuni/coli , and methicillin-resistant Staphylococcus aureus , and 108-145 samples for Bacteroides human, ruminant, and gull source-marker genes. EC/ENT temporal patterns, general Bacteroides concentration, and pathogen types and occurrence were regionally consistent (up to 40 km), but beach catchment variables (drains/creeks, impervious surface, urban land cover) influenced exceedances of EC/ENT standards and detections of Salmonella and STEC. Pathogen detections were more numerous when the EC/ENT Beach Action Value (but not when the Geometric Mean and Statistical Threshold Value) was exceeded. EC, ENT, and pathogens were not necessarily influenced by the same variables. Multiple Bacteroides sources, varying by date, occurred at every beach. Study of multiple beaches in different geographic settings provided new insights on the contrasting influences of regional and local variables, and a broader-scale perspective, on significance of EC/ENT exceedances, bacterial sources, and pathogen occurrence.

Widely tunable laser frequency offset lock with 30 GHz range and 5 THz offset.

We demonstrate a simple and versatile method to greatly extend the tuning range of optical frequency shifting devices, such as acousto-optic modulators (AOMs). We use this method to stabilize the frequency of a tunable narrow-band continuous-wave (CW) laser to a transmission maximum of an external Fabry-Perot interferometer (FPI) with a tunable frequency offset. This is achieved through a servo loop which contains an in-loop AOM for simple radiofrequency (RF) tuning of the optical frequency over the full 30 GHz mode-hop-free tuning range of the CW laser. By stabilizing the length of the FPI to a stabilized helium-neon (HeNe) laser (at 5 THz offset from the tunable laser) we simultaneously transfer the ~ 1 MHz absolute frequency stability of the HeNe laser to the entire 30 GHz range of the tunable laser. Thus, our method allows simple, wide-range, fast and reproducible optical frequency tuning and absolute optical frequency measurements through RF electronics, which is here demonstrated by repeatedly recording a 27-GHz-wide molecular iodine spectrum at scan rates up to 500 MHz/s. General technical aspects that determine the performance of the method are discussed in detail.

On the synthesis of DNA error correcting codes.

DNA error correcting codes over the edit metric consist of embeddable markers for sequencing projects that are tolerant of sequencing errors. When a genetic library has multiple sources for its sequences, use of embedded markers permit tracking of sequence origin. This study compares different methods for synthesizing DNA error correcting codes. A new code-finding technique called the salmon algorithm is introduced and used to improve the size of best known codes in five difficult cases of the problem, including the most studied case: length six, distance three codes. An updated table of the best known code sizes with 36 improved values, resulting from three different algorithms, is presented. Mathematical background results for the problem from multiple sources are summarized. A discussion of practical details that arise in application, including biological design and decoding, is also given in this study.

Effects on groundwater microbial communities of an engineered 30-day in situ exposure to the antibiotic sulfamethoxazole.

Effects upon microbial communities from environmental exposure to concentrations of antibiotics in the µg L(-1) range remain poorly understood. Microbial communities from an oligotrophic aquifer (estimated doubling rates of only once per week) that were previously acclimated (AC) or unacclimated (UAC) to historical sulfamethoxazole (SMX) contamination, and a laboratory-grown Pseudomonas stutzeri strain, were exposed to 240-520 µg L(-1) SMX for 30 days in situ using filter chambers allowing exposure to ambient groundwater, but not to ambient microorganisms. SMX-exposed UAC bacterial communities displayed the greatest mortality and impairment (viable stain assays), the greatest change in sensitivity to SMX (dose-response assays), and the greatest change in community composition (Terminal Restriction Fragment Length Polymorphism; T-RFLP). The sul1 gene, encoding resistance to SMX at clinically relevant levels, and an element of Class I integrons, was not detected in any community. Changes in microbial community structure and SMX resistance over a short experimental period in previously nonexposed, slow-growing aquifer communities suggest concentrations of antibiotics 2-3 orders of magnitude less than those used in clinical applications may influence ecological function through changes in community composition, and could promote antibiotic resistance through selection of naturally resistant bacteria.

Gene and antigen markers of shiga-toxin producing E. coli from Michigan and Indiana river water: occurrence and relation to recreational water quality criteria.

The relation of bacterial pathogen occurrence to fecal indicator bacteria (FIB) concentrations used for recreational water quality criteria (RWQC) is poorly understood. This study determined the occurrence of Shiga-toxin producing Escherichia coli (STEC) markers and their relation to FIB concentrations in Michigan and Indiana river water. Using 67 fecal coliform (FC) bacteria cultures from 41 river sites in multiple watersheds, we evaluated the occurrence of five STEC markers: the Escherichia coli (EC) O157 antigen and gene, and the STEC virulence genes eaeA, stx1, and stx2. Simple isolations from selected FC cultures yielded viable EC O157. By both antigen and gene assays, EC O157 was detected in a greater proportion of samples exceeding rather than meeting FC RWQC (P < 0.05), but was unrelated to EC and enterococci RWQC. The occurrence of all other STEC markers was unrelated to any FIB RWQC. The eaeA, stx2, and stx1 genes were found in 93.3, 13.3, and in 46.7% of samples meeting FC RWQC and in 91.7, 0.0, and 37.5% of samples meeting the EC RWQC. Although not statistically significant, the percentage of samples positive for each STEC marker except stx1 was lower in samples that met, as opposed to exceeded, FIB RWQC. Viable STEC were common members of the FC communities in river water throughout southern Michigan and northern Indiana, regardless of FIB RWQC. Our study indicates that further information on the occurrence of pathogens in recreational waters, and research on alternative indicators of their occurrence, may help inform water-resource management and public health decision-making.

Comparing wastewater chemicals, indicator bacteria concentrations, and bacterial pathogen genes as fecal pollution indicators.

The objective of this study was to compare fecal indicator bacteria (FIB) (fecal coliforms, Escherichia coli [EC], and enterococci [ENT]) concentrations with a wide array of typical organic wastewater chemicals and selected bacterial genes as indicators of fecal pollution in water samples collected at or near 18 surface water drinking water intakes. Genes tested included esp (indicating human-pathogenic ENT) and nine genes associated with various animal sources of shiga-toxin-producing EC (STEC). Fecal pollution was indicated by genes and/or chemicals for 14 of the 18 tested samples, with little relation to FIB standards. Of 13 samples with <50 EC 100 mL(-1), human pharmaceuticals or chemical indicators of wastewater treatment plant effluent occurred in six, veterinary antibiotics were detected in three, and stx1 or stx2 genes (indicating varying animal sources of STEC) were detected in eight. Only the EC eaeA gene was positively correlated with FIB concentrations. Human-source fecal pollution was indicated by the esp gene and the human pharmaceutical carbamazepine in one of the nine samples that met all FIB recreational water quality standards. Escherichia coli rfbO157 and stx2c genes, which are typically associated with cattle sources and are of potential human health significance, were detected in one sample in the absence of tested chemicals. Chemical and gene-based indicators of fecal contamination may be present even when FIB standards are met, and some may, unlike FIB, indicate potential sources. Application of multiple water quality indicators with variable environmental persistence and fate may yield greater confidence in fecal pollution assessment and may inform remediation decisions.

Influence of groundwater recharge and well characteristics on dissolved arsenic concentrations in southeastern Michigan groundwater.

Arsenic concentrations exceeding 10 microg/l, the United States maximum contaminant level and the World Health Organization guideline value, are frequently reported in groundwater from bedrock and unconsolidated aquifers of southeastern Michigan. Although arsenic-bearing minerals (including arsenian pyrite and oxide/hydroxide phases) have been identified in Marshall Sandstone bedrock of the Mississippian aquifer system and in tills of the unconsolidated aquifer system, mechanisms responsible for arsenic mobilization and subsequent transport in groundwater are equivocal. Recent evidence has begun to suggest that groundwater recharge and characteristics of well construction may affect arsenic mobilization and transport. Therefore, we investigated the relationship between dissolved arsenic concentrations, reported groundwater recharge rates, well construction characteristics, and geology in unconsolidated and bedrock aquifers. Results of multiple linear regression analyses indicate that arsenic contamination is more prevalent in bedrock wells that are cased in proximity to the bedrock-unconsolidated interface; no other factors were associated with arsenic contamination in water drawn from bedrock or unconsolidated aquifers. Conditions appropriate for arsenic mobilization may be found along the bedrock-unconsolidated interface, including changes in reduction/oxidation potential and enhanced biogeochemical activity because of differences between geologic strata. These results are valuable for understanding arsenic mobilization and guiding well construction practices in southeastern Michigan, and may also provide insights for other regions faced with groundwater arsenic contamination.

Spatial and temporal changes in microbial community structure associated with recharge-influenced chemical gradients in a contaminated aquifer.

In a contaminated water-table aquifer, we related microbial community structure on aquifer sediments to gradients in 24 geochemical and contaminant variables at five depths, under three recharge conditions. Community amplified ribsosomal DNA restriction analysis (ARDRA) using universal 16S rDNA primers and denaturing gradient gel electrophoresis (DGGE) using bacterial 16S rDNA primers indicated: (i). communities in the anoxic, contaminated central zone were similar regardless of recharge; (ii). after recharge, communities at greatest depth were similar to those in uncontaminated zones; and (iii). after extended lack of recharge, communities at upper and lower aquifer margins differed from communities at the same depths on other dates. General aquifer geochemistry was as important as contaminant or terminal electron accepting process (TEAP) chemistry in discriminant analysis of community groups. The Shannon index of diversity (H) and the evenness index (E), based on DGGE operational taxonomic units (OTUs), were statistically different across community groups and aquifer depths. Archaea or sulphate-reducing bacteria 16S rRNA abundance was not clearly correlated with TEAP chemistry indicative of methanogenesis or sulphate reduction. Eukarya rRNA abundance varied by depth and date from 0 to 13% of the microbial community. This contaminated aquifer is a dynamic ecosystem, with complex interactions between physical, chemical and biotic components, which should be considered in the interpretation of aquifer geochemistry and in the development of conceptual or predictive models for natural attenuation or remediation.

Surgical site infections following pediatric liver transplantation: risks and costs.

PURPOSE: Infectious complications following orthotopic liver transplantation (OLT) represent a significant cause of morbidity and mortality in both adults and children. In adults, surgical site infections complicating OLT have been shown to significantly increase resource utilization, but their impact in children has not been studied. In this study we identify risk factors for surgical site infections in children undergoing primary OLT for end-stage liver disease and estimate their impact on patient survival, graft survival, length of stay, and charges. METHODS: All pediatric liver transplants (n = 77) less than 16 years of age from the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Liver Transplantation Database were included in the analysis. Surgical site infections (n = 25) were defined as wound infections, abdominal abscesses, and bacterial or fungal infections of the liver, intestine, or peritoneum during the initial transplant admission. Risk of infection was estimated using logistic regression, survival rates were estimated using the Kaplan-Meier method, and length of stay and charges were compared using Student's t-test. Multivariate analysis of charges was performed using linear regression. RESULTS: Of the 77 patients, 25 (32.5%) developed a surgical site infection. Several factors were associated with increased risk of infections, including a leak at the biliary anastomosis (odds ratio [OR] 115, P = 0.003), preoperative white blood cell count (OR = 1.28, P = 0.009), surgery > 7 h (OR = 15.0, P = 0.011), HLA mismatches (OR = 6.0, P = 0.03), and female gender (OR = 8.0, P = 0.038). Surgical site infections did not significantly decrease either patient survival or graft survival, and increased hospital stay by an average of 21 days (P = 0.14). After controlling for other factors, patients who developed surgical site infections incurred on average $132,507 (P = 0.03) more in charges than patients who did not develop infections. CONCLUSIONS: Surgical site infections in pediatric patients following liver transplantation are significantly influenced by surgical technique and endogenous patient characteristics. Though survival outcomes are not different, the development of such infections has significant implications for resource utilization in the care of these patients.

Escherichia coli and enterococci at beaches in the Grand Traverse Bay, Lake Michigan: sources, characteristics, and environmental pathways.

This study quantified Escherichia coli (EC) and enterococci (ENT) in beach waters and dominant source materials, correlated these with ambient conditions, and determined selected EC genotypes and ENT phenotypes. Bathing-water ENT criteria were exceeded more frequently than EC criteria, providing conflicting interpretations of water quality. Dominant sources of EC and ENT were bird feces (10(8)/d/bird), storm drains (10(7)/d), and river water (10(11)/d); beach sands, shallow groundwater and detritus were additional sources. Beach-water EC genotypes and ENT phenotypes formed clusters with those from all source types, reflecting diffuse inputs. Some ENT isolates had phenotypes similar to those of human pathogens and/or exhibited high-level resistance to human-use antibiotics. EC and ENT concentrations were influenced by collection time and wind direction. There was a 48-72-h lag between rainfall and elevated EC concentrations at three southern shoreline beaches, but no such lag at western and eastern shoreline beaches, reflecting the influence of beach orientation with respect to cyclic (3-5 d) summer weather patterns. In addition to local contamination sources and processes, conceptual or predictive models of Great Lakes beach water quality should consider regional weather patterns, lake hydrodynamics, and the influence of monitoring method variables (time of day, frequency).

Identifying relationships between baseflow geochemistry and land use with synoptic sampling and R-mode factor analysis.

The relationship between land use and stream chemistry is often explored through synoptic sampling of rivers at baseflow conditions. However, baseflow chemistry is likely to vary temporally and spatially with land use. The purpose of our study is to examine the usefulness of the synoptic sampling approach for identifying the relationship between complex land use configurations and stream water quality. This study compares biogeochemical data from three synoptic sampling events representing the temporal variability of baseflow chemistry and land use using R-mode factor analysis. Separate R-mode factor analyses of the data from individual sampling events yielded only two consistent factors. Agricultural activity was associated with elevated levels of Ca2+, Mg2+, alkalinity, and frequently K+, SO4(2-), and NO3-. Urban areas were associated with higher concentrations of Na+, K+, and Cl-. Other retained factors were not consistent among sampling events, and some factors were difficult to interpret in the context of biogeochemical sources and processes. When all data were combined, further associations were revealed such as an inverse relationship between the proportion of wetlands and stream nitrate concentrations. We also found that barren lands were associated with elevated sulfate levels. This research suggests that an individual sampling event is unlikely to characterize adequately the complex processes controlling interactions between land use and stream chemistry. Combining data collected over two years during three synoptic sampling events appears to enhance our ability to understand processes linking stream chemistry and land use.

Impaired system A amino acid transport mimics the catabolic effects of acid in L6 cells.

BACKGROUND: Metabolic acidaemia stimulates protein catabolism in skeletal muscle cells, leading to muscle wasting. As this occurs without decreasing cytosolic pH, the initial signal is unclear. A possible explanation is that extracellular pH acts on solute transporters at the cell surface, inhibiting nutrient influx. DESIGN: Influx through glucose and Pi transporters and System A amino acid transporters into L6 skeletal muscle cells was assessed using 3H-2-deoxyglucose (2-DG), 33Pi and 14C-methylaminoisobutyrate (MeAIB), respectively. Protein degradation (PD) was assessed from 14C efflux from cells prelabelled with 14C-Phe. Branched-chain amino acids and Phe were assayed by selective fluorimetric assays. RESULTS: While acid (pH 7.1) had little immediate effect on 2-DG or 33Pi influx, exposure to pH 7.1 rapidly inhibited MeAIB influx. To determine whether System A inhibition was sufficient to trigger PD, it was blocked at pH 7.5 by a saturating dose (10 mmol L(-1)) of nonmetabolisable substrate (MeAIB). Like acid, this increased PD and decreased total protein. It also mimicked the decreases in protein synthesis, DNA synthesis, glucose transport and glycolysis, and depletion of branched-chain amino acids and Phe, which are induced in L6 by acid. The onset of inhibition of PD by an extracellular Gln load was retarded at pH 7.1, and stimulation of PD by acid was negligible if PD had already been stimulated by Gln depletion. The stimulatory effect of MeAIB on PD was selectively blunted by an excess of Gln, whereas the inhibitory effect of Gln on PD was blocked by excess MeAIB. CONCLUSIONS: The similarity of changes in response to MeAIB and acid implies that these share a common intracellular signalling pathway triggered by inhibition of System A. Even though System A is only a minor contributor to total Gln influx in L6 cells, it is suggested that blockade of System A with acid or MeAIB induces a catabolic state by denying Gln access to a key intracellular regulatory site.

Evaluating behavior of oxygen, nitrate, and sulfate during recharge and quantifying reduction rates in a contaminated aquifer.

This study evaluates the biogeochemical changes that occur when recharge water comes in contact with a reduced aquifer. It specifically addresses (1) which reactions occur in situ, (2) the order in which these reactions will occur if terminal electron acceptors (TEAs) are introduced simultaneously, (3) the rates of these reactions, and (4) the roles of the aqueous and solid-phase portions of the aquifer. Recharge events of waters containing various combinations of O2, NO3, and SO4 were simulated at a shallow sandy aquifer contaminated with waste fuels and chlorinated solvents using modified push-pull tests to quantify rates. In situ rate constants for aerobic respiration (14.4 day(-1)), denitrification (5.04-7.44 day(-1)), and sulfate reduction (4.32-6.48 day(-1)) were estimated. Results show that when introduced together, NO3 and SO4 can be consumed simultaneously at similar rates. To distinguish the role of aqueous phase from that of the solid phase of the aquifer, groundwater was extracted, amended with NO3 and SO4, and monitored overtime. Results indicate that neither NO3 nor SO4 was reduced during the course of the aqueous-phase study, suggesting that NO3 and SO4 can behave conservatively in highly reduced water. It is clear that sediments and their associated microbial communities are important in driving redox reactions.