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Background: A convergence of studies has revealed sleep spindles to be associated with sleep-related cognitive processing and even with fundamental waking state capacities such as intelligence. However, some spindle characteristics, such as propagation direction and delay, may play a decisive role but are only infrequently investigated because of technical complexities., New Method: A new methodology for assessing sleep spindle propagation over the human scalp using noninvasive electroencephalography (EEG) is described. This approach is based on the alignment of time-frequency representations of spindle activity across recording channels., Results: This first of a two-part series concentrates on framing theoretical considerations related to EEG spindle propagation and on detailing the methodology. A short example application is provided that illustrates the repeatability of results obtained with the new propagation measure in a sample of 32 night recordings. A more comprehensive experimental investigation is presented in part two of the series., Comparison With Existing Method(s): Compared to existing methods, this approach is particularly well adapted for studying the propagation of sleep spindles because it estimates time delays rather than phase synchrony and it computes propagation properties for every individual spindle with windows adjusted to the specific spindle duration., Conclusions: The proposed methodology is effective in tracking the propagation of spindles across the scalp and may thus help in elucidating the temporal aspects of sleep spindle dynamics, as well as other transient EEG and MEG events. A software implementation (the Spyndle Python package) is provided as open source software., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: This communication is the second of a two-part series that describes and tests a new methodology for assessing the propagation of EEG sleep spindles. Whereas the first part describes the methodology in detail, this part proposes a thorough evaluation of the approach by applying it to a sample of laboratory sleep recordings., New Method: The tested methodology is based on the alignment of time-frequency representations of spindle activity across recording channels and is used for assessing sleep spindle propagation over the human scalp using noninvasive EEG., Results: Spindle propagation displays features that suggest wave displacements of global synaptic potential fields. Propagation patterns that are coherent (as opposed to random), laterally symmetrical, and highly repeatable within and between subjects were observed. Propagation was slower from posterior to anterior and from central to lateral brain regions than in the opposite directions. Propagation speeds varying between 2.3 and 7.0m/s were obtained. A distinct grouping of propagation properties was noted for a small cluster of frontal electrodes. No propagation between distantly separated scalp locations was observed. The values of spindle characteristics such as average frequency, RMS amplitude, frequency slope, and duration, depend largely on propagation direction but are only mildly correlated with propagation delay., Comparison With Existing Method(s): Results obtained are in line with many results published in the literature and offer new measures for describing sleep spindle behavior., Conclusions: Propagation properties provide new information about sleep spindle behavior and thus allow more precise automated assessments of spindle-related functions., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: Recurrent mutations in the Speckle-Type POZ Protein (SPOP) gene occur in up to 15% of prostate cancers. However, the frequency and features of cancers with these mutations across different populations is unknown., Objective: To investigate SPOP mutations across diverse cohorts and validate a series of assays employing high-resolution melting (HRM) analysis and Sanger sequencing for mutational analysis of formalin-fixed paraffin-embedded material., Design Setting and Participants: 720 prostate cancer samples from six international cohorts spanning Caucasian, African American, and Asian patients, including both prostate-specific antigen-screened and unscreened populations, were screened for their SPOP mutation status. Status of SPOP was correlated to molecular features (ERG rearrangement, PTEN deletion, and CHD1 deletion) as well as clinical and pathologic features., Results and Limitations: Overall frequency of SPOP mutations was 8.1% (4.6% to 14.4%), SPOP mutation was inversely associated with ERG rearrangement (P<.01), and SPOP mutant (SPOPmut) cancers had higher rates of CHD1 deletions (P<.01). There were no significant differences in biochemical recurrence in SPOPmut cancers. Limitations of this study include missing mutational data due to sample quality and lack of power to identify a difference in clinical outcomes., Conclusion: SPOP is mutated in 4.6% to 14.4% of patients with prostate cancer across different ethnic and demographic backgrounds. There was no significant association between SPOP mutations with ethnicity, clinical, or pathologic parameters. Mutual exclusivity of SPOP mutation with ERG rearrangement as well as a high association with CHD1 deletion reinforces SPOP mutation as defining a distinct molecular subclass of prostate cancer.

The generation of handwriting is a complex neuromotor skill requiring the interaction of many cognitive processes. It aims at producing a message to be imprinted as an ink trace left on a writing medium. The generated trajectory of the pen tip is made up of strokes superimposed over time. The Kinematic Theory of rapid human movements and its family of lognormal models provide analytical representations of these strokes, often considered as the basic unit of handwriting. This paradigm has not only been experimentally confirmed in numerous predictive and physiologically significant tests but it has also been shown to be the ideal mathematical description for the impulse response of a neuromuscular system. This latter demonstration suggests that the lognormality of the velocity patterns can be interpreted as reflecting the behavior of subjects who are in perfect control of their movements. To illustrate this interpretation, we present a short overview of the main concepts behind the Kinematic Theory and briefly describe how its models can be exploited, using various software tools, to investigate these ideal lognormal behaviors. We emphasize that the parameters extracted during various tasks can be used to analyze some underlying processes associated with their realization. To investigate the operational convergence hypothesis, we report on two original studies. First, we focus on the early steps of the motor learning process as seen as a converging behavior toward the production of more precise lognormal patterns as young children practicing handwriting start to become more fluent writers. Second, we illustrate how aging affects handwriting by pointing out the increasing departure from the ideal lognormal behavior as the control of the fine motricity begins to decline. Overall, the paper highlights this developmental process of merging toward a lognormal behavior with learning, mastering this behavior to succeed in performing a given task, and then gradually deviating from it with aging.

This paper reports the results of a model-based analysis of movements gathered in a 4×4 experimental design of speed/accuracy tradeoffs with variable target distances and width. Our study was performed on a large (120 participants) and varied sample (both genders, wide age range, various health conditions). The delta-lognormal equation was used for data modeling to investigate the interaction between the output of the agonist and the antagonist neuromuscular systems. Empirical observations show that the subjects must correlate more tightly the impulse commands sent to both neuromuscular systems in order to achieve good performances as the difficulty of the task increases whereas the correlation in the timing of the neuromuscular action co-varies with the size of the geometrical properties of the task. These new phenomena are discussed under the paradigm provided by the Kinematic Theory and new research hypotheses are proposed for further investigation of the speed/accuracy tradeoffs., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Analogues of glycolipids from Spingomonadacaece with O- and S- and SO2-linkages have been prepared using chelation induced anomerisation promoted by TiCl4. Included are examples of the anomerisation of intermediates with O- and S-glycosidic linkages as well as isomerisation of β-thioglycuronic acids (β-glycosyl thiols). The β-O-glucuronide and β-O-galacturonide precursors were efficiently prepared using benzoylated trichloroacetimidates. β-Glycosyl thiols were precursors to β-S-derivatives. Triazole containing mimics of the natural glycolipids were prepared using CuI promoted azide-alkyne cycloaddition reactions in THF. The glycolipid antigens are being evaluated currently for their effects on iNKT cells.

Developing strategies to maintain biodiversity requires baseline information on the current status of each individual species. The development of genetic techniques and their application to noninvasively collected samples have the potential to yield information on the structure of elusive animal populations and so are important tools in conservation management. Using DNA isolated from faecal samples can be challenging owing to low quantity and quality. This study, however, presents the development of novel real-time polymerase chain reaction assays using fluorescently labelled TaqMan(®) MGB probes enabling species and sex identification of Eurasian otter (Lutra lutra) spraints (faeces). These assays can also be used in determining an optimum microsatellite panel and can be employed as cost-saving screening tools for downstream genetic testing including microsatellite genotyping and haplotype analysis. The techniques are shown to work efficiently with L. lutra DNA isolated from tissue, hair, spraint, blood and anal jelly samples., (© 2013 John Wiley & Sons Ltd.)

This article presents an exploratory study investigating the possibility of predicting the time occurrence of a motor event related potential (ERP) from a kinematic analysis of human movements. Although the response-locked motor potential may link the ERP components to the recorded response, to our knowledge no previous attempt has been made to predict a priori (i.e. before any contact with the electroencephalographic data) the time occurrence of an ERP component based only on the modeling of an overt response. The proposed analysis relies on the delta-lognormal modeling of velocity, as proposed by the kinematic theory of rapid human movement used in several studies of motor control. Although some methodological aspects of this technique still need to be fine-tuned, the initial results showed that the model-based kinematic analysis allowed the prediction of the time occurrence of a motor command ERP in most participants in the experiment. The average map of the motor command ERPs showed that this signal was stronger in electrodes close to the contra-lateral motor area (Fz, FCz, FC1, and FC3). These results seem to support the claims made by the kinematic theory that a motor command is emitted at time t(0), the time reference parameter of the model. This article proposes a new time marker directly associated with a cerebral event (i.e. the emission of a motor command) that can be used for the development of new data analysis methodologies and for the elaboration of new experimental protocols based on ERP., (© 2012 Federation of European Neuroscience Societies and Blackwell Publishing Ltd.)

Fast reaching movements are an important component of our daily interaction with the world and are consequently under investigation in many fields of science and engineering. Today, useful models are available for such studies, with tools for solving the inverse dynamics problem involved by these analyses. These tools generally provide a set of model parameters that allows an accurate and locally optimal reconstruction of the original movements. Although the solutions that they generate may provide a data curve fitting that is sufficient for some pattern recognition applications, the best possible solution is often necessary in others, particularly those involving neuroscience and biomedical signal processing. To generate these solutions, we present a globally optimal parameter extractor for the delta-lognormal modeling of reaching movements based on the branch-and-bound strategy. This algorithm is used to test the impact of white noise on the delta-lognormal modeling of reaching movements and to benchmark the state-of-the-art locally optimal algorithm. Our study shows that, even with globally optimal solutions, parameter averaging is important for obtaining reliable figures. It concludes that physiologically derived rules are necessary, in addition to global optimality, to achieve meaningful ∆Λ extractions which can be used to investigate the control patterns of these movement primitives.

Objective: Spondylolisthesis is a pathological condition characterized by the slipping of a vertebral body, compared with the underlying one, following structural and/or degenerative changes of the spine. The purpose of this case series is to describe clinical presentations and the conservative physiotherapy management of 4 patients with low back pain and lumbar isthmic spondylolisthesis., Clinical Features: Four patients aged 25, 43, 36, and 50 years presented with low back pain of various duration. Radiographs confirmed the presence of lumbar isthmic spondylolisthesis. Outcome measures included numerical rating scale, disability outcome measure (Oswestry Disability Index), spinal instability tests (Prone Instability Test, Passive Lumbar Extension test), and muscle function tests (Aberrant Movement Patterns, Active Straight Leg Raising, Prone and Supine Bridge Tests)., Intervention and Outcomes: Treatment consisted of postural reeducation, stretching, and strengthening exercises. Over the course of individualized treatment, ranging from 8 to 10 treatment visits, outcomes improved for all 4 patients., Conclusion: This report describes varying clinical presentations and treatment of 4 patients with isthmic spondylolisthesis, suggesting that different pain generators could be managed by different conservative approaches.

Stopping the spread of the cholera epidemic in Haiti required engaging community health workers (CHWs) in prevention and treatment activities. The Centers for Disease Control and Prevention collaborated with the Haitian Ministry of Public Health and Population to develop CHW educational materials, train >1,100 CHWs, and evaluate training efforts.

The synthesis of S-glycosphingolipids based on uronic acids is described. These compounds are analogous to the highly immunostimulatory antigens isolated from the cell walls of bacteria of the Sphingomonas family. Key to the synthetic route is a stereoselective anomerization to give α-glycosyl thiol precursors. A route to a sphinganine precursor from pseudoephedrine glycinamide is also described., (© 2011 American Chemical Society)

In the context of the occidental population aging, new preventive approaches must be developed to reduce the disability related to brain stroke in the elderly. The stroke susceptibility assessment based on the analysis of human movements is one of the potential avenues needing investigation. As a first step in this direction, this paper reports results on the relationship linking the most important stroke risk factors to some characteristics of human movement. Various features were extracted using the Sigma-Lognormal model on 1440 stereotypical triangular movements performed by 120 subjects having different health conditions. These features were combined through a linear modeling to maximize the predictability of presence of stroke risk factors in the studied cohort. The receiver operating characteristic (ROC) curve and the area under this curve (AUC) were used to evaluate the clinical significance of this relationship. Using only the information derived from the movements, the six tested risk factors (cardiac problems, diabetes mellitus, hypercholesterolemia, hypertension, obesity, and cigarette smoking) can be predicted with an AUC ranging from .68 to .82., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Background: In behavioural economics, a 'nudge' describes configuration of a choice to encourage a certain action without taking away freedom of choice., Aim: To determine the impact of a 'nudge' strategy - prefilling either 3mL or 5mL syringes with midazolam - on endoscopic sedation practice., Methods: Consecutive patients undergoing sedation for EGD or colonoscopy were enrolled. On alternate weeks, midazolam was prefilled in either 3mL or 5mL syringes. Preprocedure sedation was administered by the endoscopist to achieve moderate conscious sedation; dosages were at the discretion of the endoscopist. Meperidine was not prefilled., Results: Overall, 120 patients received sedation for EGD [59 (5mL), 61 (3mL)] and 86 patients were sedated for colonoscopy [38 (5mL), 48 (3mL)]. For EGDs, average midazolam dose was significantly higher in the 5-mL group (5.2mg) vs. 3-mL group (3.3mg), (P<0.0001); for colonoscopies, average midazolam dose was also significantly higher in the 5-mL group (5.1mg) vs. 3-mL group (3.3mg), (P<0.0001). There was no significant difference in mean meperidine dose (42.1mg vs. 42.8mg, P=0.9) administered to both colonoscopy groups. No adverse sedation-related events occurred; no patient required reversal of sedation., Conclusions: These findings demonstrate that 'nudge' strategies may hold promise in modifying endoscopic sedation practice. Further research is required to explore the utility of 'nudges' in impacting other aspects of endoscopic practice., (© 2011 Blackwell Publishing Ltd.)

Objective: The aim of this retrospective study was to assess validity of the straight-leg raise (SLR) test using magnetic resonance imaging (MRI) results as a reference standard in a group of patients with L4-L5 and L5-S1 lumbar-herniated disks and sciatic pain. The relationship between diagnostic accuracy of this test, age classes, and grade of lumbar disk displacement was investigated., Methods: The charts of 2352 patients with sciatic pain with/without lumbar pain were examined. Results of the SLR were then compared with previous spinal MRI. A 2 × 2 contingency table was created, and analysis of sensitivity, specificity, positive and negative predictive values, diagnostic odds ratio, likelihood ratio (LR), and receiver operating characteristic (ROC) curve was carried out. Homogeneous age classes were created to compare them statistically., Results: Magnetic resonance imaging findings showed lumbar disk herniation (LDH) in 1305 patients. Of these subjects, 741 were positive on SLR testing. Sensitivity was 0.36, whereas specificity was 0.74. Positive and negative predictive values were 0.69 and 0.52, respectively. Positive LR was 1.38, and negative LR was 0.87. Diagnostic odds ratio was 1.59, and ROC analysis showed an area under the curve (AUC) of 0.596. The AUC decreased from 0.730 in the 16- to 25-year subgroup to 0.515 in the 76- to 85-year subgroup. Similar results were obtained in subjects with LDH and nerve root compression., Conclusions: Our results indicate low accuracy of the SLR in diagnosis of LDH if compared with MRI results. The discriminative power of the SLR seemed to decrease as age increased; thus, positive and negative results may be less conclusive in older patients., (Copyright © 2011 National University of Health Sciences. Published by Mosby, Inc. All rights reserved.)

The main goal of this work is to determine whether a computer mouse can be used as a low-cost device for the acquisition of two-dimensional human movement velocity signals in the context of psychophysical studies and biomedical applications. A comprehensive overview of the related literature is presented, and the problem of characterizing mouse movement acquisition is analyzed and discussed. Then, the quality of velocity signals acquired with this kind of device is measured on horizontal oscillatory movements by comparing the mouse data to the signals acquired simultaneously by a video motion tracking system and a digitizing tablet. A synthesis of the information gathered in this work indicates that the computer mouse can be used for the reliable acquisition of biosignals in the context of human movement studies, particularly for many applications dealing with the velocity of the end effector of the upper limb. This paper concludes by discussing the possibilities and limitations of such use.

In this paper, 14 healthy subjects in two age groups have produced rapid handwriting strokes with a direction reversal. The delta-lognormal model was used to obtain a detailed description of the velocity of these movements and of the neuromuscular synergy that produces them. This modeling also allowed the derivation of new hypothesis on the nature of the slowing effect due to aging (i.e., a direct effect or a coping strategy) and on its repartition on the different steps of the movement production (i.e., its preparation versus its execution). Our analysis revealed a substantial increase of neuromuscular response delays and a decrease of the command amplitudes with age. For the parameters that show a significant decrease in performance, the agonist and antagonist systems were affected similarly. In addition, we observed that the age has a proportional effect on the various time characteristics of the movements and that even in the case of a significant slowing down of the neuromuscular systems, the elderly can still achieve optimal movement responses, characterized by the use of a single delta-lognormal primitive. This performance might be related to the preservation of some movement timing properties and relationships between the agonist and the antagonist neuromuscular systems., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Nitriles are widespread in the environment as a result of biological and industrial activity. Nitrile hydratases catalyse the hydration of nitriles to the corresponding amide and are often associated with amidases, which catalyze the conversion of amides to the corresponding acids. Nitrile hydratases have potential as biocatalysts in bioremediation and biotransformation applications, and several successful examples demonstrate the advantages. In this work a real-time PCR assay was designed for the detection of Fe-type nitrile hydratase genes from environmental isolates purified from nitrile-enriched soils and seaweeds. Specific PCR primers were also designed for amplification and sequencing of the genes. Identical or highly homologous nitrile hydratase genes were detected from isolates of numerous genera from geographically diverse sites, as were numerous novel genes. The genes were also detected from isolates of genera not previously reported to harbour nitrile hydratases. The results provide further evidence that many bacteria have acquired the genes via horizontal gene transfer. The real-time PCR assay should prove useful in searching for nitrile hydratases that could have novel substrate specificities and therefore potential in industrial applications.

Aim: To enhance the information pertaining to the epidemiology of a collection of 378 Listeria spp. isolates obtained from several food-processing plants in Ireland over a 3-year period (2004-2007)., Methods and Results: The collection was characterized by pulsed-field gel electrophoresis (PFGE). The most prevalent pulse-type was PFGE profile I (n=14·5%) that consisted mainly of environmental Listeria spp. samples. Serotyping of 145 Listeria monocytogenes isolates was performed. The most common serovar was 1/2a and comprised 57·4% (n=77) of the L. monocytogenes collection. The other serovars were as follows: 4b (14·1%, n=19), 1/2b (9·7%, n=13), 4c (4·4%, n=6) and 1/2c (6·7%, n=9), respectively. Eleven isolates were identified as non-Listeria spp., the remaining ten L. monocytogenes isolates were nontypeable. The antimicrobial susceptibility testing revealed the antibiotic that isolates displayed the most resistance to was gentamicin (5%) followed by sulfamethoxazole-trimethoprim (2%), tetracycline and ciprofloxacin (1·5%)., Conclusions: The subtyping has indicated the diversity of the Listeria spp. The presence of serotype 1/2a, 1/2b and 4b in both raw and cooked ready-to-eat food products is a public health concern, as these serotypes are frequently associated with foodborne outbreaks and sporadic cases of human listeriosis. In addition, the emergence of antimicrobial-resistant L. monocytogenes isolates could have serious therapeutic consequences., Significance and Impact of Study: The molecular subtyping and the further characterization of these isolates may be valuable particularly in the context of a suspected common source outbreak in the future., (© 2010 The Authors. © 2010 The Society for Applied Microbiology.)

In this paper, the existence of oscillations for a class of recurrent neural networks with time delays between neural interconnections is investigated. By using the fixed point theory and Liapunov functional, we prove that a recurrent neural network might have a unique equilibrium point which is unstable. This particular type of instability, combined with the boundedness of the solutions of the system, will force the network to generate a permanent oscillation. Some necessary and sufficient conditions for these oscillations are obtained. Simple and practical criteria for fixing the range of parameters in this network are also derived. Typical simulation examples are presented.

The objective of the current study was to expand the knowledge of the role of acetogenic Bacteria in high rate anaerobic digesters. To this end, acetogens were enriched by supplying a variety of acetogenic growth supportive substrates to two laboratory scale high rate upflow anaerobic sludge bed (UASB) reactors operated at 37 degrees C (R1) and 55 degrees C (R2). The reactors were initially fed a glucose/acetate influent. Having achieved high operational performance and granular sludge development and activity, both reactors were changed to homoacetogenic bacterial substrates on day 373 of the trial. The reactors were initially fed with sodium vanillate as a sole substrate. Although % COD removal indicated that the 55 degrees C reactor out performed the 37 degrees C reactor, effluent acetate levels from R2 were generally higher than from R1, reaching values as high as 5023 mg l(-1). Homoacetogenic activity in both reactors was confirmed on day 419 by specific acetogenic activity (SAA) measurement, with higher values obtained for R2 than R1. Sodium formate was introduced as sole substrate to both reactors on day 464. It was found that formate supported acetogenic activity at both temperatures. By the end of the trial, no specific methanogenic activity (SMA) was observed against acetate and propionate indicating that the methane produced was solely by hydrogenotrophic Archaea. Higher SMA and SAA values against H(2)/CO(2) suggested development of a formate utilising acetogenic population growing in syntrophy with hydrogenotrophic methanogens. Throughout the formate trial, the mesophilic reactor performed better overall than the thermophilic reactor., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Objective: A malignant profile of early brain ischemia has been demonstrated in the Diffusion and Perfusion Imaging Evaluation for Understanding Stroke Evolution (DEFUSE) trial. Patients with a malignant profile had a low chance for an independent functional outcome despite thrombolysis within 3-6 h. We sought to determine whether CT angiography (CTA) could identify a malignant imaging profile within 3 h from symptom onset., Methods: We studied consecutive patients (04/02-09/07) with anterior circulation stroke who received CTA before intravenous thrombolysis within 3 h. We assessed the Alberta Stroke Program Early CT Score (ASPECTS) on CTA source images (CTASI). Intracranial thrombus burden on CTA was assessed with a novel 10-point clot burden score (CBS). We analyzed percentages independent (modified Rankin Scale score < or =2) and fatal outcome at 3 months and parenchymal hematoma rates across categorized combined CTASI-ASPECTS + CBS score groups where 20 is best and 0 is worst., Results: We identified 114 patients (median age 73 years [interquartile range 61-80], onset-to-tPA time 129 min [95-152]). Among 24 patients (21%) with extensive hypoattenuation on CTASI and extensive thrombus burden (combined score < or =10), only 4% (1/24) were functionally independent whereas mortality was 50% (12/24). In contrast, 57% (51/90) of patients with less affected scores (combined score 11-20) were functionally independent and mortality was 10% (9/90; p < 0.001). Parenchymal hematoma rates were 30% (7/23) vs. 8% (7/88), respectively (p = 0.008)., Conclusion: CTA identifies a large hyperacute stroke population with high mortality and low likelihood for independent functional outcome despite early thrombolysis., (Copyright 2010 S. Karger AG, Basel.)

Introduction: Abnormalities on acute magnetic resonance imaging predict outcome in minor stroke and transient ischaemic attack patients. We hypothesised that noncontrast computed tomography and computed tomography angiography findings in minor stroke and transient ischaemic attack patients would also predict functional outcome., Methods: We analysed consecutive patients with a transient ischaemic attack or a minor stroke with an National Institute of Health Stroke Scale or=2 ) at 90 days., Results: Among 457 patients, the median baseline National Institute of Health Stroke Scale score was 1. Median time from symptom onset to noncontrast computed tomography was 278 min (interquartile range 151-505) and median delay from noncontrast computed tomography to CT angiography was 3 min (interquartile range 0-13). At 90 days, 57 patients (12.5%) had a mRS >or=2. Clinical factors that were associated with functional impairment were age >or=60 years (RR 2.05 CI(95) 1.16-3.64) and baseline National Institute of Health Stroke Scale score >0 (RR 3.23 1.72-6.06). All the assessed computed tomography parameters (acute stroke on noncontrast computed tomography and intracranial or extracranial stenosis or occlusion) were individually predictive of functional impairment. A composite computed tomography imaging 'at risk' metric, defined by acute stroke on noncontrast computed tomography, Circle of Willis intracranial vessel occlusion or >or=50% stenosis, extracranial occlusion or >or=50% stenosis, was associated with poorer outcome (RR 2.92 CI(95) 1.81-4.71)., Conclusions: The presence of an acute stroke on noncontrast computed tomography or an intracranial or extracranial occlusion or stenosis was associated with an increased risk of functional impairment. Multi-modal computed tomography could be used to identify high-risk transient ischaemic attack or minor stroke patients.

Background: In patients with acute stroke receiving intravenous tissue plasminogen activator (tPA), we postulated that the presence of intracranial occlusion on CT angiography (CTA) modifies the benefit of thrombolysis., Materials and Methods: Using a retrospective cohort design, we identified patients with acute ischemic stroke in our CTA database between May 2002 and August 2007. All the patients had a CTA within 12 h of onset, a premorbid modified Rankin scale (mRS) < or = 1, and a baseline National Institute of Health Stroke Scale score(NIHSS)f > or = 6. The primary outcome was early effectiveness of tPA defined as an NIHSS score of < or = 2 at 24 h or a 4-point NIHSS improvement at 24 h. Secondary outcome included mRS < or = 1 at 90 days. The relationship between intracranial occlusion on CTA and benefit of tPA was assessed using a test for interaction., Results: A total of 287 patients met the criteria [occlusion present N =168; (98 with tPA; 70 without tPA) and occlusion absent N = 119; (52 with tPA; 67 without tPA)]. Those with intracranial occlusion were likely to have more severe strokes (NIHSS > or = 15; P < 0.001) and abnormal brain imaging (ASPECTS < or =7; P < 0.001). For outcome of 4-point NIHSS score improvement at 24 h, benefit from tPA was observed only among patients with a visible occlusion (absolute difference in favor of tPA: 20.4% vs. 0.7%; P = 0.06)., Conclusion: In patients with acute ischemic stroke, thrombolysis produced a better early clinical response among patients with intracranial occlusion, which needs to be confirmed in stroke thrombolysis trials.

Bacterial enzymes capable of nitrile hydrolysis have significant industrial potential. Microbacterium sp. AJ115, Rhodococcus erythropolis AJ270 and AJ300 were isolated from the same location in England and harbour identical nitrile hydratase/amidase gene clusters. Strain AJ270 has been well studied due to its nitrile hydratase and amidase activity. R. erythropolis ITCBP was isolated from Denmark and carries a very similar nitrile hydratase/amidase gene cluster. In this study, an identical nitrilase gene (nit1) was isolated from the four strains, and the nitrilase from strain AJ270 cloned and expressed in Escherichia coli. Analysis of the recombinant nitrilase has shown it to be functional with activity demonstrated towards phenylacetonitrile. A real-time PCR TaqMan assay was developed that allowed nit1 detection directly from soil enrichment cultures without DNA extraction, with nit1 detected in all samples tested. Real-time PCR screening of isolates from these soils resulted in the isolation of nit1 and also very similar nitrilase gene nit2 from a number of Burkholderia sp. The genes nit1 and nit2 have also been detected in many bacteria of different genera but are unstable in these isolates. It is likely that the genes were acquired by horizontal gene transfer and may be wide-spread in the environment.

The objective of this study was to examine the feasibility of using a two-step, fully biological and sustainable strategy for the treatment of carbohydrate rich wastes. The primary step in this strategy involves the application of thermostable enzymes produced by the thermophilic, aerobic fungus, Talaromyces emersonii, to carbohydrate wastes producing a liquid hydrolysate discharged at elevated temperatures. To assess the potential of thermophilic treatment of this hydrolysate, a comparative study of thermophilic and mesophilic digestion of four sugar rich thermozyme hydrolysate waste streams was conducted by operating two high rate upflow anaerobic hybrid reactors (UAHR) at 37 degrees C (R1) and 55 degrees C (R2). The operational performance of both reactors was monitored from start-up by assessing COD removal efficiencies, volatile fatty acid (VFA) discharge and % methane of the biogas produced. Rapid start-up of both R1 and R2 was achieved on an influent composed of the typical sugar components of the organic fraction of municipal solid waste (OFMSW). Both reactors were subsequently challenged in terms of volumetric loading rate (VLR) and it was found that a VLR of 9 gCOD l(-1)d(-1) at a hydraulic retention time (HRT) of 1 day severely affected the thermophilic reactor with instability characterised by a build up of volatile fatty acid (VFA) intermediates in the effluent. The influent to both reactors was changed to a simple glucose and sucrose-based influent supplied at a VLR of 4.5 gCOD l(-1)d(-1) and HRT of 2 days prior to the introduction of thermozyme hydrolysates. Four unique thermozyme hydrolysates were subsequently supplied to the reactors, each for a period of 10 HRTs. The applied hydrolysates were derived from apple pulp, bread, carob powder and cardboard, all of which were successfully and comparably converted by both reactors. The % total carbohydrate removal by both reactors was monitored during the application of the sugar rich thermozyme hydrolysates. This approach offers a sustainable technology for the treatment of carbohydrate rich wastes and highlights the potential of these wastes as substrates for the generation of second-generation biofuels.

Childhood obesity is increasingly prevalent in the community and is related to adverse cardiovascular outcomes during adulthood. In this study of healthy children, we evaluated the influence of adiposity and physical activity on carotid-femoral pulse wave velocity (PWV), an index of arterial stiffness and a marker of cardiovascular risk in adults. In 573 community-based children (mean age: 10.1+/-0.3 years; 51% boys), we measured body mass index and waist circumference. Percentage body fat was quantitated by dual-energy x-ray absorptiometry. Cardiorespiratory fitness (CRF) and physical activity levels were assessed using a 20-m shuttle run and 7-day pedometer count, respectively. PWV was estimated by applanation tonometry. In univariate analysis, PWV was positively correlated with body mass index (r=0.34), waist circumference (r=0.32), and percentage body fat (r=0.32; P<0.001 for all) and negatively correlated with CRF (r=-0.23; P<0.001) and pedometer count (r=-0.08; P=0.046). In separate multivariable linear regression models, body mass index, waist circumference, and percentage of body fat were independently and positively associated with PWV (P<0.01 for all) after adjusting for age, sex, systolic blood pressure, mean arterial pressure, heart rate, and CRF (P<0.01 for all). The influence of CRF on PWV was attenuated after adjusting for adiposity. In conclusion, increased body mass and adiposity and decreased CRF are associated with arterial stiffening in healthy prepubescent children.

DNA identification of non-invasive samples is a potentially useful tool for monitoring small mammal species. Here we describe a novel method for identifying five small mammal species: wood mouse, bank vole, common shrew, pygmy shrew and water shrew. Species-specific real-time polymerase chain reaction primers were designed to amplify fragments of the mitochondrial cytochrome b gene from hair and scat samples. We also amplified nuclear DNA from scats, demonstrating their potential as a source of DNA for population genetic studies., (© 2008 The Authors. Journal compilation © 2008 Blackwell Publishing Ltd.)

The time-courses of satellite cell (SC) activation and protein expression of hepatocyte growth factor (HGF), HGF activator (HGFA), HGFA inhibitor-1 (HAI-1), and HGFA inhibitor-2 (HAI-2) in human skeletal muscle, as well as serum HGF following a single bout of muscle lengthening contractions, were determined. Eight recreationally active participants were recruited for the study. Subjects performed 300 lengthening contractions involving the quadriceps femoris muscles of a single leg at a fixed velocity of 180 degrees/s. Percutaneous muscle biopsies were taken before (PRE) and at 4 h (T4), 24 h (T24), 72 h (T72), and 120 h (T120) following the exercise. The protocol resulted in an increase in the number of SCs [neural cell adhesion molecule (NCAM)-labeled cells] expressed relative to total myonuclei, at T24, compared with both PRE and T4 (P<0.05), and peaked at T72 (approximately 80% increase vs. PRE, P<0.05). HGF protein increased significantly in serum from baseline (PRE) to T4 (P<0.05). Active HGF protein was detected in skeletal muscle at rest [14.4+/-1.3 average integrated density value (IDV)/actin average IDV] and tended to increase at early time-points (P=0.12). HGFA protein increased significantly from PRE to T24 (P<0.05). HAI-1 protein increased significantly from PRE to T24 (P<0.05). HAI-2 (32 kDa) increased significantly from baseline (PRE) by T24 (P<0.05), and also by T72 and T120 (P<0.05). We conclude that a single bout of lengthening muscle contractions is sufficient to activate SCs, which may involve both a local and systemic HGF response to contraction-induced injury.

Background: In ischemic stroke, functional outcomes vary depending on site of intracranial occlusion. We tested the prognostic value of a semiquantitative computed tomography angiography-based clot burden score., Methods: Clot burden score allots major anterior circulation arteries 10 points for presence of contrast opacification on computed tomography angiography. Two points each are subtracted for thrombus preventing contrast opacification in the proximal M1, distal M1 or supraclinoid internal carotid artery and one point each for M2 branches, A1 and infraclinoid internal carotid artery. We retrospectively studied patients with disabling neurological deficits (National Institute of Health Stroke Scale score >or=5) and computed tomography angiography within 24-hours from symptom onset. We analyzed percentages independent functional outcome (modified Rankin Scale score

The genes encoding an enantioselective nitrile hydratase (NHase) from Rhodococcus erythropolis AJ270 have been cloned and an active NHase has been produced in Escherichia coli. Maximal activity was found when the genes encoding the alpha- and beta-subunits were transcribed as one unit and the gene encoding the P44k activator protein as a separate ORF on a single replicon. Addition of n-butyric acid and FeSO(4 )could improve NHase activity. Coexpression of the GroEL-GroES chaperone proteins increased activity in the absence of P44k protein but had no effect in the presence of P44k. The recombinant enzyme was highly enantioselective in the synthesis of S-(+)-3-benzoyloxy- 4-cyanobutyramide from the prochiral substrate 3-benzoyloxyglutaronitrile.

Safe drinking water and hygiene are essential to reducing Kenya's diarrhoeal disease burden. A school-based safe water and hygiene intervention in Kenya was evaluated to assess its impact on students' knowledge and parents' adoption of safe water and hygiene practices. We surveyed 390 students from nine schools and their parents at baseline and conducted a final evaluation of 363 students and their parents. From baseline to final evaluation, improvement was seen in students' knowledge of correct water treatment procedure (21-65%, P<0.01) and knowing when to wash their hands. At final evaluation, 14% of parents reported currently treating their water, compared with 6% at baseline (P<0.01). From 2004 to 2005, school absenteeism in the September-November term decreased in nine project schools by 35% and increased in nine neighbouring comparison schools by 5%. This novel programme shows promise for reducing school absenteeism and promoting water and hygiene interventions in the home.

The competitive and syntrophic interactions between different anaerobic bacterial trophic groups in sulphate limited expanded granular sludge bed (EGSB) reactors was investigated. The outcome of competition between the sulphate-reducing, methanogenic and syntrophic populations after development in reactors at varying influent COD/SO4 (2-) ratios was examined in batch activity tests with the inclusion of specific sulphate reducing bacteria (SRB) and methane producing archaea (MPA) inhibitors. SRB species could not out-compete MPA species for acetate at influent COD/SO4 (2-) ratios as low as 2. The SRB were seen to play a more significant role in the conversion of hydrogen but did not become completely dominant. HMPA were responsible for hydrogen utilization at an influent COD/SO4 (2-) ratio of 16, and were still dominant when the ratio was reduced to 4. It was only when the COD/SO4 (2-) ratio was reduced to 2 that the HSRB assumed a more influential role. SRB species were significant in the degradation of propionate at all COD/SO4 (2-) ratios applied. Sludge samples were analysed by scanning electron microscopy (SEM), granule size distribution and fluorescent in situ hybridization (FISH), combined with confocal laser scanning microscopy (CLSM), to monitor any changes in granule morphology under the various COD/SO4 (2-) ratios imposed during the reactor trial. In situ hybridization with domain- and species-specific oligonucleotide probes demonstrated a layered architecture with an outer layer harboring mainly Eubacterial cells and an inner layer dominated by Archaeal species.

The nitrile metabolising strains AJ270, AJ300 and AJ115 were isolated from the same location. The strains have very similar nitrile metabolising profiles. Sequencing of the 16S rRNA gene indicates that strains AJ270 and AJ300 are novel strains of Rhodococcus erythropolis while strain AJ115 is a novel Microbacterium strain very closely related to Microbacterium oxydans and Microbacterium liquefaciens. Analysis of the structure of the nitrile hydratase/amidase gene clusters in the three strains indicates that this region is identical in these strains and that this structure is different to other nitrile hydratase/amidase gene clusters. The major difference seen is the insertion of a complete copy of the insertion sequence IS1166 in the nhr2 gene. This copy of IS1166 generates a 10 bp direct duplication at the point of insertion and has one ORF encoding a protein of 434 amino acids, with 98% homology to the transposase of IS666 from Mycobacterium avium. A gene oxd, encoding aldoxime dehydratase is found upstream of the nitrile hydratase gene cluster and an open reading frame encoding a protein with homology to GlnQ type ABC transporters is found downstream of the nitrile hydratase/amidase genes. The identity of the nitrile hydratase/amidase gene clusters in the three strains suggests horizontal gene transfer of this region. Analysis of the strains for both linear and circular plasmids indicates that both are present in the strains but hybridisation studies indicate that the nitrile hydratase/amidase gene cluster is chromosomally located. The nitrile hydratase/amidase enzymes of strain AJ270 are inducible with acetonitrile or acetamide. Interestingly although a number of Fe-type nitrile hydratases have been shown to be photosensitive, the enzyme from strain AJ270 is not.

Microbacterium sp. AJ115 metabolises a wide range of nitriles using the two-step nitrile hydratase/amidase pathway. In this study, the amidase gene of Microbacterium sp. AJ115 has been inserted into the pCal-n-EK expression vector and expressed in Escherichia coli BL21(DE3)pLysS. The expressed protein is active in E. coli and expression of the amidase gene allows E. coli to grow on acetamide as sole carbon and/or nitrogen source. Expression of active amidase in E. coli was temperature dependent with high activity found when cultures were grown between 20 and 30 degrees C but no activity at 37 degrees C. On induction, the amidase represents 28% of the total soluble protein in E. coli. The expressed amidase has been purified in a single step from the crude lysate using the calmodulin-binding peptide (CBP) affinity tag. The V(max) and K(m) of the purified enzyme with acetamide (50 mM) were 4.4 micromol/min/mg protein and 4.5mM, respectively. The temperature optimum was found to be 50 degrees C. Purified enzyme demonstrated enantioselectivity with the ability to preferentially act on the S enantiomer of racemic (R,S)-2-phenylpropionamide. S-2-phenylpropionic acid is produced with an enantiomeric excess of >82% at 50% conversion of the parent amide.

The various problems associated with treating sulphate-containing wastewaters stem inherently from successful competitive interactions between sulphate reducing bacteria (SRB) and other bacteria involved in the process, resulting in the formation of H(2)S. Prevention of in-reactor sulphide generation by use of specific SRB inhibitors presents a potential solution. Nitrite has been reported to be a specific inhibitor of SRB but its possible toxicity to syntrophic and methanogenic members of the anaerobic consortium has not been investigated. In batch activity and toxicity tests, under both mesophilic and thermophilic conditions, nitrite, at concentrations of up to 150 mg L(-1), was found to be ineffective as a specific inhibitor of SRB, and was also shown to have an inhibitory effect on the activity of syntrophic and methane-producing bacteria in mesophilic and thermophilic digester sludge samples.

The present study investigated mesophilic anaerobic treatment of sulphate-containing wastewater in EGSB reactors and assessed the inclusion of nitrite in the reactor influent as a method for control of biological sulphate reduction. Two EGSB reactors, R1 and R2, were operated for a period of 581 days at varying volumetric loading rates, COD/SO4(2-) ratios and influent nitrite concentrations (R2 only). COD removal efficiencies of > 93% were achieved in both reactors at influent sulphate concentrations of up to 3,000 mg l(-1). A two-fold increase in the influent sulphate concentration, giving an influent COD/SO4(2-) ratio of 2, resulted in a reduction in reactor COD removal efficiency to 84% and 89%, in R1 and R2, respectively. Despite inclusion of nitrite in the R2 influent at concentrations up to 500 mg NO2-N l(-1), sulphate reduction proceeded similarly in R2 and R1, suggesting the ineffectiveness of nitrite as a potential inhibitor of SRB

Objective: To define the importance of nosocomial-acquired respiratory syncytial virus (RSV) infection in an area with a high prevalence of paediatric HIV-1 infection., Methods: A prospective study was performed that involved all children hospitalized to a general paediatric ward during the course of an RSV epidemic. These children were screened within 24 h of admission and subsequently at 3-4 day intervals for RSV infection using a direct immunofluorescence assay., Results: RSV was detected in 36 (11.8%) of the 305 children upon initial investigation. Fourteen (38.9%) of the 36 children with community-acquired RSV infection were HIV-1 infected. Repeat sampling for RSV infection was performed in 130 children who tested negative for RSV infection on initial screening and who were hospitalized for > or =3 days. Nosocomial acquisition of RSV occurred among 11.5% of these children, of whom 33% were HIV-1 infected. Fourteen (93.3%) of the children with nosocomial RSV acquisition were clinically symptomatic and treated for 'nosocomial-sepsis'. Furthermore, RSV was isolated from 54% of all children who were investigated for clinically diagnosed nosocomial sepsis. Two (13%) of 15 children with nosocomial RSV infection died; both had other underlying medical conditions., Conclusion: RSV is an important under-recognized cause of nosocomial infection and sepsis among children in this African country. Active interventions are warranted in addressing this problem, as has been successfully undertaken in developed countries.

The need to measure and evaluate orthotics and prosthetics (O&P) practice has received growing recognition in the past several years. Reliable and valid self-report instruments are needed that can help facilities evaluate patient outcomes. The objective of this project was to develop a set of self-report instruments that assess functional status, quality of life, and satisfaction with devices and services that can be used in an orthotics and prosthetics clinic. Selecting items from a variety of existing instruments, the authors developed and revised four instruments that differentiate patients with varying levels of lower limb function, quality of life, and satisfaction with devices and services. Evidence of construct validity is provided by hierarchies of item difficulty that are consistent with clinical experience. For example, with the lower limb function instrument, running one block was much more difficult than walking indoors. The instruments demonstrate adequate internal consistency (0.88 for lower limb function, 0.88 for quality of life, 0.74 for service satisfaction, 0.78 for device satisfaction). The next steps in their research programme are to evaluate sensitivity and construct validity. The Orthotics and Prosthetics Users' Survey (OPUS) is a promising self-report instrument which may, with further development, allow orthotic and prosthetic practitioners to evaluate the quality and effectiveness of their services as required by accreditation standards such as those of the American Board for Certification in Orthotics and Prosthetics that mandate quality assessment.

The filamentous fungus Fusarium lateritium produces cyanide hydratase when grown in the presence of cyanide. The cyanide hydratase protein produced at a high level in Escherichia coli shows a low but significant nitrilase activity with acetonitrile, propionitrile and benzonitrile. The nitrilase activity is sufficient for growth of the recombinant strain on acetonitrile, propionitrile or benzonitrile as the sole source of nitrogen. The recombinant enzyme shows highest nitrilase activity with benzonitrile. Site-directed mutagenesis of the F. lateritium cyanide hydratase gene indicates that mutations leading to a loss of cyanide hydratase activity also lead to a loss of nitrilase activity. This suggests that the active site for cyanide hydratase and nitrilase activity in the protein is the same. This is the first evidence of cyanide hydratase having nitrilase activity.

Chronic low-frequency stimulation has been used as a model for investigating responses of skeletal muscle fibres to enhanced neuromuscular activity under conditions of maximum activation. Fast-to-slow isoform shifting of markers of the sarcoplasmic reticulum and the contractile apparatus demonstrated successful fibre transitions prior to studying the effect of chronic electro-stimulation on the expression of the nicotinic acetylcholine receptor. Comparative immunoblotting revealed that the alpha- and delta-subunits of the receptor were increased in 10-78 day stimulated specimens, while an associated component of the surface utrophin-glycoprotein complex, beta-dystroglycan, was not drastically changed in stimulated fast skeletal muscle. Previous studies have shown that electro-stimulation induces degeneration of fast glycolytic fibres, trans-differentiation leading to fast-to-slow fibre transitions and activation of muscle precursor cells. In analogy, our results indicate a molecular modification of the central functional unit of the post-synaptic muscle surface within existing neuromuscular junctions and/or during remodelling of nerve-muscle contacts.

The enzymes nitrilase, cyanide dihydratase and cyanide hydratase are a group of closely related proteins. The proteins show significant similarities at the amino acid and protein structure level but the enzymes show many differences in catalytic capability. Nitrilases, while catalysing the hydration of nitrile to the corresponding acid, vary widely in substrate specificity. Cyanide dihydratase and cyanide hydratase use HCN as the only efficient substrate but produce acid and amide products, respectively. The similarities of all these enzymes at the amino acid level but the functional differences between them provide a rich source of material for the study of structure/function relationships in this biotechnologically important group of enzymes. This review provides an overview of current understanding of the genetics and biochemistry of this interesting group of enzymes.

African human immunodeficiency virus type 1 (HIV-1)-infected children were evaluated to define the burden of Pneumocystis carinii pneumonia (PCP) and its interaction with bacterial and viral pathogens. P. carinii was identified in 101 (43.7%) of 231 episodes of pneumonia among 185 HIV-1-infected children (median age, 4.5 months; range, 1.7-27.3 months). Receipt of trimethoprim-sulfamethoxazole (TMP-SMX) prophylaxis was not associated with a significant reduction (36%; 95% confidence interval [CI], -15.4% to 64.5%) in isolation of P. carinii among children considered to have received adequate prophylaxis (37.7% of children) compared with children who had never received any prophylaxis (48.5% of children). However, deaths among children with PCP who had been taking TMP-SMX prophylaxis were markedly reduced (98.6%; 95% CI, 89.1%-99.8%) compared with children who were not taking prophylaxis. Concurrent P. carinii infection was observed in 6 of 18, 11 of 26, and 4 of 6 HIV-1-infected children who had bacteremia, a respiratory virus isolated, or Mycobacterium species isolated, respectively.

Disruption of normal cystic fibrosis transmembrane conductance regulator- (CFTR)-mediated Cl(-) transport is associated with cystic fibrosis (CF). CFTR is also required for HCO(3)(-) transport in many tissues such as the lungs, gastro-intestinal tract, and pancreas, although the exact role CFTR plays is uncertain. Given the importance of CFTR in HCO(3)(-) transport by so many CF-affected organ systems, it is perhaps surprising that relatively little is known about the interactions of HCO(3)(-) ions with CFTR. We have used patch clamp recordings from native pancreatic duct cells to study HCO(3)(-) permeation and interaction with CFTR. Ion selectivity studies shows that CFTR is between 3-5 times more selective for Cl(-) over HCO(3)(-). In addition, extracellular HCO(3)(-) has a novel inhibitory effect on cAMP-stimulated CFTR currents carried by Cl(-). The block by HCO(3)(-) was rapid, relatively independent of voltage and occurred over the physiological range of HCO(3)(-) concentrations. These data show that luminal HCO(3)(-) acts as a potent regulator of CFTR, and suggests that inhibition involves an external anion-binding site on the channel. This work has implications not only for elucidating mechanisms of HCO(3)(-) transport in epithelia, but also for approaches used to treat CF.

The objective of this study was to determine the effect of high pressure (HP) on the inactivation of microbial contaminants in Cheddar cheese (Escherichia coli K-12, Staphylococcus aureus ATCC 6538, and Penicillium roqueforti IMI 297987). Initially, cheese slurries inoculated with E. coli, S. aureus, and P. roqueforti were used as a convenient means to define the effects of a range of pressures and temperatures on the viability of these microorganisms. Cheese slurries were subjected to pressures of 50 to 800 MPa for 20 min at temperatures of 10, 20, and 30 degrees C. At 400 MPa, the viability of P. roqueforti in cheese slurry decreased by >2-log-unit cycles at 10 degrees C and by 6-log-unit cycles at temperatures of 20 and 30 degrees C. S. aureus and E. coli were not detected after HP treatments in cheese slurry of >600 MPa at 20 degrees C and >400 MPa at 30 degrees C, respectively. In addition to cell death, the presence of sublethally injured cells in HP-treated slurries was demonstrated by differential plating using nonselective agar incorporating salt or glucose. Kinetic experiments of HP inactivation demonstrated that increasing the pressure from 300 to 400 MPa resulted in a higher degree of inactivation than increasing the pressurization time from 0 to 60 min, indicating a greater antimicrobial impact of pressure. Selected conditions were subsequently tested on Cheddar cheese by adding the isolates to cheese milk and pressure treating the resultant cheeses at 100 to 500 MPa for 20 min at 20 degrees C. The relative sensitivities of the isolates to HP in Cheddar cheese were similar to those observed in the cheese slurry, i.e., P. roqueforti was more sensitive than E. coli, which was more sensitive than S. aureus. The organisms were more sensitive to pressure in cheese than slurry, especially with E. coli. On comparison of the sensitivities of the microorganisms in a pH 5.3 phosphate buffer, cheese slurry, and Cheddar cheese, greatest sensitivity to HP was shown in the pH 5.3 phosphate buffer by S. aureus and P. roqueforti while greatest sensitivity to HP by E. coli was exhibited in Cheddar cheese. Therefore, the medium in which the microorganisms are treated is an important determinant of the level of inactivation observed.

Although primary genetic defects have been identified for some forms of inherited cardiomyopathy, it is not well understood how secondary abnormalities actually lead to muscle cell destruction. Since cardiomyopathies significantly influence morbidity and mortality rates world-wide, it is important to improve the differential diagnosis of these disorders and develop potential treatments for inherited diseases of the heart. Elucidation of the secondary molecular mechanisms underlying cardiac cell necrosis might help linking a specific mutation in a cardiac gene to acute heart failure. As disturbed Ca2+-homeostasis may contribute to heart failure, we have investigated the relative abundance and oligomeric status of the sarcoplasmic reticulum Ca2+-ATPase and phospholamban in various cardiomyopathies. These two proteins represent important factors in cardiac relaxation. The SERCA2 isoform of the Ca2+-ATPase represents a major Ca2+-removal system in cardiac muscle fibres and phospholamban is a regulator of Ca2+-pump activity. Although Ca2+-ATPase expression did not seem to be markedly altered, the comparative immunoblot analysis presented here clearly shows that phospholamban expression is increased in dilated cardiomyopathy, possibly explaining the decreased Ca2+-uptake in the disease. In contrast to the normal enzyme, the Ca2+-pump was demonstrated to exhibit an impairment of crosslinker-stabilized oligomerization in dilated cardiomyopathy. Since Ca2+-ATPase oligomerization is important for co-operative kinetics and protection against proteolytic degradation, the monomeric Ca2+-ATPase may trigger an abnormal contraction-relaxation cycle in dilated cardiomyopathy leading to heart failure.

We have been studying CFTR channels in guinea pig pancreatic duct cells and rather surprisingly found that luminal HCO3- had a pronounced inhibitory effect on cAMP-activated CFTR chloride currents. The block produced by HCO3- was rapid, voltage-independent and occurred over a physiological range of extracellular HCO3- concentrations. I- and ClO4- were also found to inhibit CFTR currents, but both were less effective than HCO3-. Although we have not identified how HCO3- is able to block CFTR our data suggests that an external anion-binding site on the channel itself is involved. Overall, our results show that luminal HCO3- acts as a potent inhibitor of CFTR channels (and by inference CFTR-mediated secretion), under normal physiological conditions. These data have implications not only for current models of pancreatic duct cell HCO3- transport, but also for other bicarbonate-secreting tissues, such as the liver, GI tract and lungs.

Background & Aims: Cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channels play an important role in HCO(3)(-) secretion by pancreatic duct cells (PDCs). Our aims were to characterize the CFTR conductance of guinea pig PDCs and to establish whether CFTR is regulated by HCO(3)(-)., Methods: PDCs were isolated from small intralobular and interlobular ducts, and their Cl(- )conductance was studied using the whole-cell patch clamp technique., Results: Activation of a typical CFTR conductance by adenosine 3',5'-cyclic monophosphate (cAMP) was observed in 114 of 204 cells (56%). A larger (10-fold), time- and voltage-dependent Cl(-) conductance was activated in 39 of 204 cells (19%). Secretin had a similar effect. Coexpression of both conductances in the same cell was observed, and both conductances had similar anion selectivity and pharmacology. Extracellular HCO(3)(-) caused a dose-dependent inhibition of both currents (K(i), approximately 7 mmol/L), which was independent of intracellular and extracellular pH, and the PCO(2) and CO(3)(2-) content of the bathing solutions., Conclusions: Two kinetically distinct Cl(-) conductances are activated by cAMP in guinea pig PDCs. Because these conductances are coexpressed and exhibit similar characteristics (anion selectivity, pharmacology, and HCO(3)(-) inhibition), we conclude that CFTR underlies them both. The inhibition of CFTR by HCO(3)(-) has implications for the current model of pancreatic ductal HCO(3)(-) secretion.