A novel marine bacterial strain, PV-4(T), isolated from a microbial mat located at a hydrothermal vent of Loihi Seamount in the Pacific Ocean, has been characterized. This micro-organism is orangey in colour, Gram-negative, polarly flagellated, facultatively anaerobic and psychrotolerant (temperature range, 0-42 degrees C). No growth was observed with nitrate, nitrite, DMSO or thiosulfate as the electron acceptor and lactate as the electron donor. The major fatty acid detected in strain PV-4(T) was iso-C(15 : 0). Strain PV-4(T) had ubiquinones consisting mainly of Q-7 and Q-8, and possessed menaquinone MK-7. The DNA G+C content of the strain was 53.8 mol% and the genome size was about 4.5 Mbp. Phylogenetic analyses based on 16S rRNA gene sequences placed PV-4(T) within the genus Shewanella. PV-4(T) exhibited 16S rRNA gene sequence similarity levels of 99.6 and 97.5 %, respectively, with respect to the type strains of Shewanella aquimarina and Shewanella marisflavi. DNA from strain PV-4(T) showed low mean levels of relatedness to the DNAs of S. aquimarina (50.5 %) and S. marisflavi (8.5 %). On the basis of phylogenetic and phenotypic characteristics, the bacterium was classified in the genus Shewanella within a distinct novel species, for which the name Shewanella loihica sp. nov. is proposed. The type strain is PV-4(T) (=ATCC BAA-1088(T)=DSM 17748(T)).

This study was aimed at assessing the potential use of electrospun fibers as drug delivery vehicles with focus on the different diameters and drug contents to control drug release and polymer fiber degradation. A drug-loaded solvent-casting polymer film was made with an average thickness of 100 microm for comparative purposes. DSC analysis indicated that electrospun fibers had a lower T(g) but higher transition enthalpy than solvent-casting polymer film due to the inner stress and high degree of alignment and orientation of polymer chains caused by the electrospinning process. Inoculation of paracetanol led to a further slight decrease in the T(g) and transition enthalpy. An in vitro drug release study showed that a pronounced burst release or steady release phase was initially observed followed by a plateau or gradual release during the rest time. Fibers with a larger diameter exhibited a longer period of nearly zero order release, and higher drug encapsulation led to a more significant burst release after incubation. In vitro degradation showed that the smaller diameter and higher drug entrapment led to more significant changes of morphologies. The electrospun fiber mat showed almost no molecular weight reduction, but mass loss was observed for fibers with small and medium size, which was characterized with surface erosion and inconsistent with the ordinarily polymer degrading form. Further wetting behavior analysis showed that the high water repellent property of electrospun fibers led to much slower water penetration into the fiber mat, which may contribute to the degradation profiles of surface erosion. The specific degradation profile and adjustable drug release behaviors by variation of fiber characteristics made the electrospun nonwoven mat a potential drug delivery system rather than polymer films and particles.

OBJECTIVE

Rehabilitation research for individuals with chronic stroke is increasingly addressing spatiotemporal asymmetries (STA). Understanding the reliability of STA between sessions is essential for determining whether treatment responses exceed day-to-day variation. The purpose of this study was to determine the minimal detectable change (MDC) and test-retest reliability in STA and gait speed measures for individuals with chronic stroke.

METHODS

Twenty-six individuals with chronic (>6 months) stroke participated in 2 visits. At each visit, participants walked across a 14-ft (4.3-m) GAITRite mat at comfortable gait speed (CGS) and fast gait speed (FGS). Spatiotemporal asymmetries ratios (paretic/nonparetic limb) were calculated for step length, stance time, and swing time. Intraclass correlation coefficients (ICC, 2,1) were calculated to determine reliability between sessions. Minimal detectable change values were calculated to determine the smallest change that is considered "real."

RESULTS

Spatiotemporal asymmetry measures were consistent between sessions, with ICCs ranging from 0.93 to 0.98. Asymmetry ratio MDC values were calculated for step length (CGS: 0.15; FGS: 0.19), swing time (CGS: 0.26; FGS: 0.20), and stance time (CGS: 0.09; FGS: 0.10). Gait speed MDC was 0.20 m/s and 0.22 m/s, respectively, for the CGS and FGS walking conditions.

CONCLUSIONS

There were considerable differences among MDC values for the various STA measures, suggesting that larger changes will be necessary to show improvement for certain measures (eg, swing time asymmetry). These data will assist with setting clinical goals for patients with chronic stroke and will be useful for evaluating interventions designed to minimize temporal and spatial interlimb asymmetries during walking.

BACKGROUND

We address some critical but unknown parameters of individuals and populations of Norway rats (Rattus norvegicus) that influence leptospiral infection, maintenance and spirochetal loads shed in urine, which contaminates the environment ultimately leading to human infection.

RESULTS

Our study, conducted in Salvador, Brazil, established the average load of leptospires in positive kidneys to be 5.9 x 10(6) per mL (range 3.1-8.2 x10(6)) genome equivalents (GEq), similar to the 6.1 x 10(6) per ml (range 2.2-9.4 x10(6)) average obtained from paired urines, with a significant positive correlation (R2=0.78) between the two. Based on bivariate and multivariate modeling, we found with both kidney and urine samples that leptospiral loads increased with the age of rats (based on the index of body length to mass), MAT titer and the presence of wounding/scars, and varied with site of capture. Some associations were modified by sex but trends were apparent. Combining with data on the demographic properties and prevalence of leptospiral carriage in rat populations in Salvador, we estimated that daily leptospiral loads shed in the urine of a population of 82 individuals exceeded 9.1 x 10(10) leptospires.

CONCLUSIONS

These factors directly influence the risk of leptospiral acquisition among humans and provide essential epidemiological information linking properties of rat populations with risk of human infection.

Nonalcoholic fatty liver is involved in the development of nonalcoholic steatohepatitis and chronic liver injury. Impairment of hepatic transsulfuration reactions is suggested to be critically linked with alcoholic liver injury, but its role in nonalcoholic fatty liver remains unknown. We examined the early changes in sulfur-amino acid metabolism and their implication in nonalcoholic fatty liver disease (NAFLD). Male rats were provided with a standard liquid diet or a high-fat liquid diet (HF) for 3 wk. An additional group of rats received the HF diet supplemented with betaine (1%). HF diet intake elevated hepatic triglyceride and serum tumor necrosis factor-alpha (TNFalpha) concentrations. Antioxidant capacity of liver cytosol against hydroxyl and peroxyl radicals was reduced significantly. Hepatic S-adenosylmethionine (SAM) and glutathione (GSH) decreased, but hypotaurine and taurine concentrations increased. Methionine adenosyltransferase (MAT) activity, not its concentration, was depressed, whereas both activity and concentration of cysteine dioxygenase and GSH S-transferase were elevated. Betaine supplementation of the HF diet inhibited hepatic fat accumulation and serum TNFalpha elevation. The decrease in cytosolic antioxidant capacity was also prevented. MAT activity and its concentration were induced significantly. Hepatic SAM and GSH increased and elevation of hypotaurine and taurine was depressed. The results indicate that the metabolism of S-containing substances is significantly disturbed by the HF diet, suggesting a causal role of impairment of hepatic transsulfuration reactions in NAFLD. Betaine supplementation protects the liver from nonalcoholic steatosis and oxidative stress most probably via its effects on the transsulfuration reactions.

Living marine stromatolites at Highborne Cay, Bahamas, are formed by microbial mat communities that facilitate precipitation of calcium carbonate and bind and trap small carbonate sand grains. This process results in a laminated structure similar to the layering observed in ancient stromatolites. In the modern marine system at Highborne Cay, lamination, lithification and stromatolite formation are associated with cycling between three types of microbial communities at the stromatolite surface (Types 1, 2 and 3, which range from a leathery microbial mat to microbially fused sediment). Examination of 923 universal small-subunit rRNA gene sequences from these communities reveals that taxonomic richness increases during transition from Type 1 to Type 3 communities, supporting a previous model that proposed that the three communities represent different stages of mat development. The phylogenetic composition also changes significantly between these community types and these community changes occur in concert with variation in biogeochemical rates. The dominant bacterial groups detected in the stromatolites include Alphaproteobacteria, Planctomycetes, Cyanobacteria and Bacteroidetes. In addition, the stromatolite communities were found to contain novel cyanobacteria that may be uniquely associated with modern marine stromatolites. The implications of these findings are discussed in the context of current models for stromatolite formation.

Electrospinning has recently been developed as an efficient technique to develop polymeric nanofibres. Various synthetic and natural biodegradable polymers have been electrospun into fibres with diameters in the nanometre range (< 1 microm). The fibre diameter, structure and physical properties of the nanofibre matrices can be effectively tuned by controlling various parameters that affect the electrospinning process. The dimension and structure of electrospun polymeric nanofibre mats resembles mostly the collagen phase of natural extracellular matrix. This, combined with excellent physical properties such as high surface area, high porosity, interconnective pores of the nanofibre matrices and appropriate mechanical properties, well-controlled degradation rates and biocompatibility of the base polymer, make biodegradable polymeric nanofibre matrices ideal candidates for developing scaffolds for tissue engineering. This article reviews the recent advances in the development of synthetic biodegradable nanofibre-based matrices as scaffolds for tissue engineering.

Electrospun nanocomposite scaffolds were fabricated by encapsulating multi-walled carbon nanotubes (MWNT) in poly (lactic acid) (PLA) nanofibers. Scanning electron microscopy (SEM) confirmed the fabrication of nanofibers, and transmission electron microscopy identified the alignment and dispersion of MWNT along the axis of the fibers. Tensile testing showed an increase in the tensile modulus for a MWNT loading of 0.25 wt% compared with electrospun nanofibrous mats without MWNT reinforcement. Conductivity measurements indicated that the confined geometry of the fibrous system requires only minute doping to obtain significant enhancements at 0.32 wt%. Adipose-derived human mesenchymal stem cells (hMSCs) were seeded on electrospun scaffolds containing 1 wt% MWNT and 0 wt% MWNT, to determine the efficacy of the scaffolds for cell growth, and the effect of MWNT on hMSC viability and proliferation over two weeks in culture. Staining for live and dead cells and DNA quantification indicated that the hMSCs were alive and proliferating through day 14. SEM images of hMSCs at 14 days showed morphological differences, with hMSCs on PLA well spread and hMSCs on PLA with 1% MWNT closely packed and longitudinally aligned.

Intrasubject variation in bioavailability (rate and extent) and disposition of furosemide 40 mg was investigated using a repeated, randomized, double-blind cross-over study in 8 healthy subjects. Two generic tablet formulations (Lasix and Furix) and intravenous furosemide were compared on 6 separate days. Extensive intrasubject variability after oral administration was observed in AUC, mean absorption time (MAT) and urinary excretion. The variability (error variance) within the dosage forms was as large as that between the two generics. These variations most probably depended on the absorption process, since the repeated i.v. doses showed only marginal intrasubject variability. Absolute bioavailability was 56% for Lasix and 55% for Furix (AUC). The range was 20 to 84% between individuals and the maximal range within one individual was 20 to 61%. Confidence interval and Bayesian analysis showed a high probability of non-equivalence not only between but also within the generics when the separate cross-over experiments were analyzed (8 observations). When extending the analysis to 16 observations, bioequivalence was demonstrated for the two generic tablets. Rate of absorption, quantified as MAT, was 128 min for Lasix and 98 min for Furix (16 observations). Since MAT was significantly longer (p less than 0.001) than the mean residence time after the i.v. dose (57 min), absorption was evidently the rate-limiting step in the overall kinetics of oral furosemide. Intraindividual variation in absorption is a confounding factor in bioavailability studies of furosemide using limited numbers of subjects. This is important to consider when designing and evaluating bioavailability studies for drugs showing these variations.

BACKGROUND

Current estimates suggest an HIV prevalence of 42% among people who inject drugs (PWIDs) in Dar es Salaam, while HIV prevalence is estimated to be 8.8% among the general population in the city. To address the HIV epidemic in this population, the government of Tanzania began establishing HIV prevention, treatment and care services including outreach and medication assisted treatment (MAT) for PWIDs in 2010. We assessed gender inequities in utilization of outreach and MAT services and evaluated differences in HIV risk behaviors between female and male PWIDs.

METHODS

Routine outreach data between December 2010 to mid-August 2012 and baseline data on clients enrolling in methadone from February 2011 to August 2012 were utilized. Binomial regression was used to estimate adjusted relative risk estimates comparing females to males.

RESULTS

From December 2010 to August 2012, 8,578 contacts were made to drug users; among them 1,898 were injectors. A total of 453 injectors were eligible and referred to MAT, of which, 443 enrolled in treatment. However, regarding total outreach contacts, outreach to PWID, referral to MAT and enrollment in MAT, 8% or less of drug users accessing services were women. In contrast, weighted estimations from surveys suggest that 34% of PWIDs are female, and this approximation is similar to recent population size estimations. Overall, 43% of traditional outreach workers conducting outreach with drug users were female. Though reporting higher levels of condom usage, female PWID were more likely to report multiple sex partners, anal sex, commercial sex work and struggle under a higher burden of addiction, mental disorders and abuse.

CONCLUSIONS

Services have not been mobilized adequately to address the clear needs of females who inject drugs. A clear and urgent need exists for women-centered strategies that effectively engage female PWID into HIV prevention services.

Kombucha is a sweetened tea beverage that, as a consequence of fermentation, contains ethanol, carbon dioxide, a high concentration of acid (gluconic, acetic and lactic) as well as a number of other metabolites and is thought to contain a number of health-promoting components. The sucrose-tea solution is fermented by a symbiosis of bacteria and yeast embedded within a cellulosic pellicle, which forms a floating mat in the tea, and generates a new layer with each successful fermentation. The specific identity of the microbial populations present has been the focus of attention but, to date, the majority of studies have relied on culture-based analyses. To gain a more comprehensive insight into the kombucha microbiota we have carried out the first culture-independent, high-throughput sequencing analysis of the bacterial and fungal populations of 5 distinct pellicles as well as the resultant fermented kombucha at two time points. Following the analysis it was established that the major bacterial genus present was Gluconacetobacter, present at >85% in most samples, with only trace populations of Acetobacter detected (<2%). A prominent Lactobacillus population was also identified (up to 30%), with a number of sub-dominant genera, not previously associated with kombucha, also being revealed. The yeast populations were found to be dominated by Zygosaccharomyces at >95% in the fermented beverage, with a greater fungal diversity present in the cellulosic pellicle, including numerous species not identified in kombucha previously. Ultimately, this study represents the most accurate description of the microbiology of kombucha to date.

A new vaccine, 4CMenB, is composed of surface proteins of Neisseria meningitidis and is aimed to target serogroup B (MenB) isolates. The vaccine components are present in meningococcal isolates of other serogroups allowing potential use against meningococcal isolates belonging to non-B serogroups. Isolates of serogroup X (MenX) have been emerged in countries of the African meningitis belt. 4CMenB may offer a vaccine strategy against these isolates as there is no available capsule-based vaccine against MenX. We used the Meningococcal Antigen Typing System (MATS) to determine presence, diversity and levels of expression of 4CMenB antigens among 9 MenX isolates from several African countries in order to estimate the potential coverage of MenX by the 4CMenB vaccine. We performed bactericidal assays against these isolates, using pooled sera from 4CMenB-vaccinated infants, adolescents and adults. The African MenX isolates belonged to the same genotype but showed variation in the vaccine antigens. MATS data and bactericidal assays suggest coverage of the 9 African MenX isolates by 4CMenB but not of two unrelated MenX isolates from France. 4CMenB vaccine can be considered for further investigation to control MenX outbreaks in Africa.

Falls are common in patients with cerebellar ataxia (CA). Identification of gait variables associated with a higher risk of falls allows us to detect fallers and initiate protective procedures early. Gait variability, which is increased in CA patients, is a good predictor of falls in elderly subjects and patients with neurodegenerative diseases. The relationship between gait variability and fall risk in patients with different cerebellar disorders was systematically investigated. A total of 48 patients with different cerebellar ataxia entities [adult-onset cerebellar atrophy (SAOA) (n = 23), unknown entity (n = 7), vascular (n = 5), post-cerebellitis (n = 6), congenital (n = 2), Louis-Bar syndrome (n = 2), ethyltoxic (n = 2) posttraumatic (n = 1)] were examined using a GAITRite® sensor mat. Spatial and temporal variability parameters were used for ANOVA testing and logistic regression models with categorized fall events as dependent variables. Gait variability in the fore-aft direction showed significant differences between the fall groups (p < 0.05-0.01). Model effects were highest for walking with slow speed (correct prediction 0.50-0.72). The speed-dependent integral of gait variability markers showed a higher discriminatory power (correct prediction 0.74-0.94). Gait variability is linked to the fall risk of patients with CA, slow walking and temporal gait variability being most relevant. The use of speed-dependent integrals of gait variability improves the accuracy of fall prediction. To predict fall risks in cerebellar ataxia, gait variability measurements made during slow walking should be included in a gait analysis procedure. The effects of speed-adjusted physiotherapeutic interventions have to be further investigated.

Comparative serological examination of 300 serum samples from sheep slaughtered in the main abattoir in Cairo, Egypt revealed a higher prevalence of toxoplasmosis (43.7%) with the modified agglutination test (MAT), followed by the enzyme linked immune-sorbant assay (ELISA) (41.7%) and the indirect fluorescent antibody test (IFAT) (37%), while the lowest prevalence was detected with the dye test (DT) (34%). When the data from the first three serological tests were compared with that of the DT test, which was used as a reference test for toxoplasmosis, MAT had the highest sensitivity (96%), followed by ELISA (90.1%) and IFAT, which demonstrated the lowest sensitivity (80.4%). Conversely, IFAT had the highest specificity (91.4%), followed by MAT (88.9%) and ELISA (85.9%).

Remnant ice shelves along the northern coast of Ellesmere Island, Nunavut, Canada ( approximately 83 degrees N) provide a habitat for cryo-tolerant microbial mat communities. Bioassays of bacterial and primary production were undertaken to quantify the short-term physiological response of the mats to changes in key variables that characterize this cryo-ecosystem (salinity, irradiance and temperature). The heterotrophic versus autotrophic community responses to these stressors differed markedly. The heterotrophic bacteria were extremophilic and specifically adapted to ambient conditions on the ice shelf, whereas the autotrophic community had broader tolerance ranges and optima outside the ambient range. This latter, extremotrophic response may be partly due to a diverse suite of pigments including oligosaccharide mycosporine-like amino acids, scytonemins, carotenoids, phycobiliproteins and chlorophylls that absorb from the near UV-B to red wavelengths. These pigments provide a comprehensive broadband strategy for coping with the multiple stressors of high irradiance, variable salinity and low temperatures in this extreme cryo-environment.

OBJECTIVE

The safety, tolerability, and pharmacokinetics of intravenous *i.v.) montelukast sodium (Singulair, MK-0476), and the oral bioavailability of montelukast sodium in healthy males and healthy females were studied.

METHODS

This was a two-part study. Part I was a four-period study in males of rising i.v. doses of montelukast sodium (3, 9, and 18 mg) administered as 15-minute constant-rate i.v. infusions (Periods 1-3), followed by a 10-mg oral tablet dose of montelukast sodium (Period 4) under fasting conditions. Part II was a four-period study in females of i.v. montelukast sodium (9 mg) infused over 15 and 5 minutes (Periods 5 and 6, respectively) or injected as a bolus over 2 minutes (Period 7), followed by a 10-mg oral tablet dose of montelukast sodium (Period 8). Plasma samples were collected and analyzed by HPLC.

RESULTS

In males (N = 6), as the i.v. dose of montelukast sodium increased from 3 to 18 mg, the area under the plasma concentration-time curve of montelukast sodium from time 0 to infinity (AUC) increased proportionately. The mean values of plasma clearance (CL), steady-state volume of distribution (Vss), plasma terminal half-life (t1/12), and mean residence time in the body (MRTi.v.) of montelukast sodium were 45.5 ml/min, 10.5 1, 5.1 hr, and 3.9 hr, respectively, and remained essentially constant over the i.v. dosage range. Following oral administration of a 10-mg tablet of montelukast sodium, the AUC, maximum plasma concentration (Cmax), time when Cmax occurred (Tmax), apparent t1/12, mean absorption time (MAT), and bioavailability (F) of montelukast sodium averaged 2441 ng.hr/ml, 385 ng/ml. 3.7 hr, 4.9 hr, 3.4 hr, and 66%, respectively. Following i.v. administration of 9 mg of montelukast sodium to females (N = 6), the values of CL, Vss, t1/2, and MRT i.v. averaged 47.6 ml/min, 9.6 1, 4.5 hr, and 3.6 hr, respectively. Following oral administration of a 10-mg tablet to females, the mean AUC, Cmax, Tmax, apparent t1/2, MAT and F were 2270 ng.hr/ml, 350 ng/ml, 3.3 hr, 4.4 hr, 2.6 hr, and 58%, respectively. These parameter values were similar to or slightly smaller than those in healthy males receiving the same i.v. and oral doses.

CONCLUSIONS

The disposition kinetics of montelukast sodium were linear. Gender had little or no effect on the kinetics of montelukast sodium. Safety results from this study indicate that intravenous doses of montelukast sodium from 3 to 18 mg and a 10-mg oral dose are well tolerated.

The purpose of this study was to compare kinematic gait parameters measured with an instrumented walkway system (GAITRite(R)) and a video-based system (peak performance motus 3.1(R)). Subjects walked across a GAITRite mat with embedded pressure sensors. Reflective markers were attached to subjects' shoes and video capture was simultaneously performed during each trial. Video data were then digitized manually using peak software. Correlation coefficients for all parameters measured with both systems were high >>/=0.94). Significant differences between systems were found with analysis of variance (ANOVA) for two parameters, step length and stride velocity (P=0.003, 0.0002). The results of this study indicate that the instrumented walkway gave comparable results for temporal parameters but further investigation is needed to evaluate the fidelity of its spatial performance.

The IME1 gene of Saccharomyces cerevisiae is required for initiation of meiosis. Transcription of IME1 is detected under conditions which are known to induce initiation of meiosis, namely starvation for nitrogen and glucose, and the presence of MATa1 and MAT alpha 2 gene products. In this paper we show that IME1 is also subject to translational regulation. Translation of IME1 mRNA is achieved either upon nitrogen starvation, or upon G1 arrest. In the presence of nutrients, constitutively elevated transcription of IME1 is also sufficient for the translation of IME1 RNA. Four different conditions were found to cause expression of Ime1 protein in vegetative cultures: elevated transcription levels due to the presence of IME1 on a multicopy plasmid; elevated transcription provided by a Gal-IME1 construct; G1 arrest due to alpha-factor treatment; G1 arrest following mild heat-shock treatment of cdc28 diploids. Using these conditions, we obtained evidence that starvation is required not only for transcription and efficient translation of IME1, but also for either the activation of Ime1 protein or for the induction/activation of another factor that, either alone or in combination with Ime1, induces meiosis.

The highly dynamic nature of voltage-gated Na+ channel (VGSC) expression and its controlling mechanism(s) are not well understood. In this study, we investigated the possible involvement of nerve growth factor (NGF) in regulating VGSC activity in the strongly metastatic Mat-LyLu cell model of rat prostate cancer (PCa). NGF increased peak VGSC current density in a time- and dose-dependent manner. NGF also shifted voltage to peak and the half-activation voltage to more positive potentials, and produced currents with faster kinetics of activation; sensitivity to the VGSC blocker tetrodotoxin (TTX) was not affected. The NGF-induced increase in peak VGSC current density was suppressed by both the pan-trk antagonist K252a, and the protein kinase A (PKA) inhibitor KT5720. NGF did not affect the Nav1.7 mRNA level, but the total VGSC alpha-subunit protein level was upregulated. NGF potentiated the cells' migration in Transwell assays, and this was not affected by TTX. We concluded that NGF upregulated functional VGSC expression in Mat-LyLu cells, with PKA as a signaling intermediate, but enhancement of migration by NGF was independent of VGSC activity.

Coccidioides species, the fungi responsible for the valley fever disease, are known to reproduce asexually through the production of arthroconidia that are the infectious propagules. The possible role of sexual reproduction in the survival and dispersal of these pathogens is unexplored. To determine the potential for mating of Coccidioides, we analyzed genome sequences and identified mating type loci characteristic of heterothallic ascomycetes. Coccidioides strains contain either a MATMATMAT loci of most ascomycetes. Along with their cDNA structures, we determined that all genes in the MAT loci are transcribed. Two genes frequently found in common sequences flanking MAT idiomorphs, APN2 and COX13, are within the MAT loci in Coccidioides, but the MATMATMAT loci occurred prior to the separation of Coccidioides from Uncinocarpus reesii. An analysis of 436 Coccidioides isolates from patients and the environment indicates that in both Coccidioides immitis and C. posadasii, there is a 1:1 distribution of MAT loci, as would be expected for sexually reproducing species. In addition, an analysis of isolates obtained from 11 soil samples demonstrated that at three sampling sites, strains of both mating types were present, indicating that compatible strains were in close proximity in the environment.

The purpose of this study was to examine the influence of anticipation of pain, sensory perception of pain and pain-related fear and disability beliefs on the gait characteristics of patients with chronic low back pain (CLBP). Thirty-one individuals with CLBP (16 men and 15 women) and 24 healthy individuals (11 men and 13 women) between 20 to 56 years of age participated in this study. Anticipated pain and the pain actually felt were measured with two separate visual analogue scales before and after preferred and fast walking performances. Pain-related fear and disability beliefs were measured with the Fear-Avoidance Belief Questionnaire (FABQ) and the Disability Belief Questionnaire (DBQ). Spatial and temporal walking parameters were measured at preferred and fast walking performances using a computerized gait mat. Analysis of variance demonstrated significant differences between patients and healthy individuals in step length, single support time and walking velocity (P<0.05). Within the CLBP group, stepwise regression analysis showed that FABQ (physical activity) and anticipated pain were the strongest predictors of velocity deficits in preferred and fast walking respectively. Anticipation and fear of pain accounted significantly for the velocity deficits in walking. Standard clinical gait assessments must incorporate psychological measures of pain experience.

Tree growth and survival were assessed in 283 populations of Scots pine (Pinus sylvestris L.) originating from a broad geographic range and grown at 90 common-garden experimental sites across Europe, and in 101 populations grown at 14 sites in North America. Growth and survival were analysed in response to climatic transfer distance, the difference in mean annual temperature (MAT) between the site and the population origin. Differences among populations at each site, and across sites for regional groups of populations, were related to climate transfer distance, but in opposite ways in the northern vs. southern parts of the species range. Climate transfers equivalent to warming by 1-4 degrees C markedly increased the survival of populations in northern Europe >>or= 62 degrees N, < 2 degrees C MAT) and modestly increased height growth>>or= 57 degrees N but decreased survival at < 62 degrees N and modestly decreased height growth at < 54 degrees N latitude in Europe. Thus, even modest climate warming will likely influence Scots pine survival and growth, but in distinct ways in different parts of the species range.

Prostate carcinoma is one of the most common malignancies affecting males, resulting in a high rate of morbidity and mortality. This hormone-dependent malignancy is characteristically associated with a high incidence of osteoblastic skeletal lesions. However, osteolytic lesions invariably accompany blastic ones. In the current study, we assessed the role of parathyroid hormone-related peptide (PTHRP), a potent bone-resorbing agent, in contributing to bone breakdown and prostatic skeletal metastasis using a syngeneic rat prostate cancer model. The full-length cDNA encoding rat PTHRP was subcloned as a Hind III insert in the sense orientation into the mammalian expression vector pRc-CMV to generate the expression vector pRc-PTHRP-S. Both control and experimental plasmids were stably transfected into low PTHRP-producing Dunning R3227, Mat Ly Lu rat prostate cancer cells. Following antibiotic selection, monoclonal cell lines expressing the highest amount of PTHRP mRNA and immunoreactive PTHRP were selected as experimental tumor cells for further analysis. Increased PTHRP production by these cells had no significant effect in vitro on the invasive capacity of these cells. Control and experimental cells were inoculated s.c. into the right flank or by the intracardiac (i.c.) route into the left ventricle of inbred male Copenhagen rats. No skeletal metastases occurred after s.c. injection with either cells. In contrast, i.c. inoculation led to lumbar vertebra metastasis and consequent hind-limb paralysis. Furthermore, histological examination of skeletal metastases in experimental animals showed a marked increase in osteoclastic activity. Our results demonstrate that PTHRP can increase osteoclastic osteolysis in the presence of focal osseous prostate cancer metastases and may contribute to the lytic lesions which generally accompany osteoblastic lesions in prostate cancer.

Biodegradation inhibition of a benchmark chemical, 2,4-dichloro-phenoxyacetic acid methyl ester (2,4-DME), was used to quantify the inhibitory effects of heavy metals on aerobic microbial degradation rates of organic chemicals. This procedure used lake sediments and aufwuchs (floating mats) collected in the field or from laboratory microcosms. Effects of CuCl2, HgCl2, ZnCl2, Cd(NO3)2, and Cr(NO3)3 at initial concentrations ranging from 0.3 microM to 73 mM (approximately 0.1 to 10,000 mg liter-1) were investigated. In general, such metallic compounds appeared to be considerably more inhibitory to the biodegradation of an organic chemical than high concentrations of microbially toxic organics studied previously. Effects of various metal concentrations were evaluated based on the following: (i) estimated MICs, (ii) concentrations that caused a significant effect on biodegradation parameters (both a greater than 10% decrease in Vmax and a greater than 10% increase in t1/2 for 2,4-DME degradation), and (iii) concentrations that caused biodegradation half-life doublings (HLDs). The MICs of metals in sediment were lowest for Zn2+ (0.10 microM) and highest for Cd2+ and Cu2+ (0.9 and 1.2 microM, respectively). The MICs of metals in aufwuchs were lowest for Hg2+ (0.01 microM), intermediate for Cu2+ and Zn2+ (0.42 and 0.62 microM, respectively), and highest for Cr3+ and Cd2+ (3.4 and 5.6 microM, respectively). Compared with Cu2+ on aufwuchs, 70 times more Zn2+, 250 times more Cr3+, and 1,000 times more Cd2+ was required to significantly affect aufwuchs biodegradation rate parameters and coefficients (Vmax and t1/2). Aufwuchs was significantly affected by the lowest Hg2+ concentration tested (5 microM).(ABSTRACT TRUNCATED AT 250 WORDS)