Diagnostic anoctamin-5 protein defect in patients with ANO5-mutated muscular dystrophy.

AIMS: Previously, detection of ANO5 protein has been complicated by unspecific antibodies, most of which have not identified the correct protein. The aims of the study were to specify ANO5 protein expression in human skeletal muscle, and to investigate if the ANO5 protein levels are affected by different ANO5 mutations in anoctaminopathy patients. METHODS: Four different antibodies were tested for ANO5 specificity. A sample preparation method compatible with membrane proteins, combined with tissue fractionation was used to determine ANO5 expression in cell cultures expressing ANO5, in normal muscles and eight patient biopsies with six different ANO5 mutations in homozygous or compound heterozygous states, and in other dystrophies. RESULTS: Only one specific monoclonal N-terminal ANO5 antibody was efficient in detecting the protein, showing that ANO5 is expressed as a single 107 kD polypeptide in human skeletal muscle. The truncating mutations c.191dupA and c.1261C>T were found to abolish ANO5 expression, whereas the studied point mutations had variable effects; however, all the ANO5 mutations resulted in clearly reduced ANO5 expression in the patient muscle membrane fraction. Attempts to detect ANO5 using immunohistochemistry were not yet successful. CONCLUSIONS: The data presented here indicate that the ANO5 protein expression is decreased in ANO5-mutated muscular dystrophy and that most of the non-truncating pathogenic ANO5 mutations likely destabilize the protein and cause its degradation. The method described here allows direct analysis of human ANO5 protein, which can be used in diagnostics, for evaluating the pathogenicity of the potentially harmful ANO5 variants of uncertain significance.

Online estimation of VFA, alkalinity and bicarbonate concentrations by electrical conductivity measurement during anaerobic fermentation.

This paper describes the use of electrical conductivity for measurement of volatile fatty acids (VFA), alkalinity and bicarbonate concentrations, during the anaerobic fermentation process. Two anaerobic continuous processes were studied: the first was a laboratory reactor for hydrogen production from molasses and the second was a pilot process for anaerobic digestion (AD) of vinasses producing methane. In the hydrogen production process, the total VFA concentration, but not bicarbonate concentration, was well estimated from the on-line electrical conductivity measurements with a simple linear regression model. In the methane production process, the bicarbonate concentration and the VFA concentration were well estimated from the simultaneous on-line measurements of pH and electrical conductivity by means of non-linear regression with neural network models. Moreover, the total alkalinity concentration was well estimated from electrical conductivity measurements with a simple linear regression model. This demonstrates the use of electrical conductivity for monitoring the AD processes.

Radicicol enhances the regeneration of skeletal muscle injured by crotoxin via decrease of NF-kB activation.

This study evaluated cellular and molecular effects of radicicol, a heat shock protein (HSP) inducer, on the regeneration of skeletal muscle injured by crotoxin, the main toxin isolated from Crotalus durissus terrificus venom. Regenerating muscles treated with radicicol had decreased NF-kB activation. Differentiating myoblasts treated with radicicol showed reduced number of NF-kB positive nuclei and increased fusion index. The results suggest that radicicol enhances regeneration of muscle by attenuating NF-kB activation and increasing myogenic differentiation.

Radicicol improves regeneration of skeletal muscle previously damaged by crotoxin in mice.

This work investigates the influence of heat shock proteins (HSPs) on necrosis and subsequent skeletal muscle regeneration induced by crotoxin (CTX), the major component of Crotalus durissus terrificus venom. Mice were treated with radicicol, a HSP inductor, followed by an intramuscular injection of CTX into the gastrocnemius muscle. Treated groups were sacrificed 1, 10 and 21 days after CTX injection. Muscle histological sections were stained with toluidine blue and assayed for acid phosphatase or immunostained with either neuronal cell adhesion molecule (NCAM) or neonatal myosin heavy chain (MHCn). Muscle samples were also submitted to Western blotting analysis. The results show that CTX alone and CTX combined with radicicol induced a similar degree of myofiber necrosis. CTX-injured muscles treated with radicicol had increased cross-sectional areas at 10 and 21 days post-lesion compared with untreated CTX-injured muscles. Additionally, radicicol significantly increased the number of NCAM-positive satellite cells in the gastrocnemius at one day post-CTX injury. CTX-injured muscles treated with radicicol contained more MHCn-positive regenerating myofibers compared with untreated CTX-injured muscles. These results suggest that HSPs contribute to the regeneration of myofibers damaged by CTX. Additionally, further studies should investigate the potential therapeutic effects of radicicol in skeletal muscles affected by Crotalus venom.

Recovery of deambulation after cardiothoracic surgery: a single center experience.

BACKGROUND: Surgical procedure and postoperative bed rest lead to musculoskeletal system alterations with a possibility of new walking dependence of patients who undergo cardiothoracic surgery, which is sometimes associated with prolonged hospitalization and increased health expenditure. AIM: The aim of this study was to assess the postoperative motor disability in inpatients admitted to the cardiothoracic surgical ward, and the results of customized rehabilitation in terms of recovery of postural changes and walking capacity with respect to the preoperative condition and destination after discharge. DESIGN: A prospective observational study was conducted. SETTING AND POPULATION: Four hundred seventeen inpatients, who had undergone cardiothoracic surgery, were enrolled between March 2011 and January 2012 in a Hospital Unit of Cardiothoracic Surgery. METHODS: A computerized system was used to collect data about ambulation at home, type and number of rehabilitation sessions proposed, ambulation at discharge, destination after discharge from ward of origin. All patients, who give their consent, undergone rehabilitative treatment on the ward of origin with an expert physiotherapist. RESULTS: Three hundred seventy-five inpatients were examined in Cardiac Surgery. One patient (0.26%) refused rehabilitative treatment. Two patients (0.53%) died. At the time of discharge 236 (74.45%) patients had recovered the ability to walk independently. After discharge 87.64% of patients was transferred to a specialist ward for intensive rehabilitation. Forty-two inpatients were enrolled in thoracic surgery. Two patients died whilst in hospital. At the time of discharge, 36 patients (94.73%) were able to walk independently. After discharge 80% of patients returned home. CONCLUSION: In our study, the application of an early and simple rehabilitation program on the ward of origin after surgery has made possible the recovery of ambulation of most inpatients who referred independence at home in a few days, limiting hospitalization and health expenditure. CLINICAL REHABILITATION IMPACT: Data about recovery of ambulation with respect to the preoperative condition and destination after discharge resume the importance of identifying motor impairment after surgery, in order to apply an adequate, early and feasible rehabilitation protocol to inpatients, limiting hospitalization and health expenditure.

Realistic dental care in jails and prisons: summary of proceedings.

As more knowledge becomes available, positive solutions to the complicated dental care problems that have emerged in this new field will be reached, and, eventually, health care practitioners will be able to fulfill the requirements of a complete health delivery system in our correctional institutions throughout the country.

Model based prediction of the clogging of an anaerobic fixed bed reactor.

This paper presents the use of nonlinear constrained optimization techniques in order to detect and evaluate the degree of clogging in an anaerobic fixed bed reactor. First, experimental results show that the validity of a mass balance model can degrade over the time. Using the available model of the process and nonlinear constrained optimization tools, it is established that these changes can be due to the decrease of the liquid volume into the reactor while the mean values of biomass concentrations increase, leading to the clogging of the reactor. These theoretical results are confirmed experimentally in evaluating the hydraulic retention time of the reactor using a tracer.

Evaluation of a four year experience with a fully instrumented anaerobic digestion process.

For several years, a 1 m3 fixed bed anaerobic digestion process has been operated for the treatment of distillery vinasses. This reactor has been fully instrumented with the following variables available on-line: pH, temperature, liquid and gas flow rates, gas composition (i.e., CH4, CO2 and H2), concentration of bicarbonate, chemical oxygen demand, total organic carbon, volatile fatty acids and partial and total alkalinity, these last four variables being measured twice by different techniques (i.e., using a TOC analyzer, a titrimetric sensor and an infrared spectrometer). The purpose of this paper is to compare the respective benefits of advanced instrumentation for the monitoring of wastewater treatment processes in general, and for anaerobic digestion in particular. It will also provide some statistical analysis of the time required to operate a fully instrumented wastewater treatment process. It is indeed well admitted in the literature that instrumentation is usually the main limitation step for using closed-loop control. However, it is our opinion that, in the near future, this situation will change. This point is discussed based on our four years practical experience.

On-line measurements of COD, TOC, VFA, total and partial alkalinity in anaerobic digestion processes using infra-red ectrometry.

This paper describes the use of a Fourier Transform Infra-Red (FT-IR) spectrometer as an on-line sensor to measure Chemical Oxygen Demand (COD), Total Organic Carbon (TOC), Volatile Fatty Acids (VFA), and Partial and Total Alkalinity (PA and TA) in anaerobic digestion processes for the treatment of industrial wastewaters. Comparison with manual off-line analysis and with an on-line industrial TOC analyser and an on-line titrimetric sensor (for the measurements of VFA, TA and PA) are provided to demonstrate the interest of spectral analysis in the mid infra-red domain for the monitoring of anaerobic digestion processes. In order to further illustrate the advantages of using such a technique, on-line measurements recorded during an accident of the pH regulation in the input (pH in the reactor went above 11 and biomass activity stopped) are shown. They demonstrate that, if carefully performed, the calibration can be extended outside its range while being still compatible with requirements of wastewater treatment processes.

Dynamic evaluation of a fixed bed anaerobic digestion process in response to organic overloads and toxicant shock loads.

This paper details a dynamic evaluation of a 1 m3 fixed bed anaerobic digestion reactor in response to organic overloads and toxicant shock loads. Raw industrial wine distillery wastewater was used as a reference substrate and several disturbances were applied to the process: (i) organic overloads with and without pH regulation in the feeding line, (ii) adding of ammonia in the input wastewater. The purpose of this study was to assess, using on-line instrumentation, the robustness of a fixed bed anaerobic digester. Anaerobic digestion processes have the reputation of being difficult to operate and prone to process instability due to external disturbances and the objective of this study was to demonstrate the possibility of such a reactor configuration for industrial use.

Automatic control of volatile fatty acids in anaerobic digestion using a fuzzy logic based approach.

A control law based on fuzzy logic was developed and validated for an anaerobic wastewater treatment process. The controlled variable was the concentration of volatile fatty acids (VFA) in the reactor and the manipulated variable was the input flow rate. In order to use it as the input of the fuzzy sets, the controlled variable was treated using an algorithm of interpolation, extrapolation and filtering. The treatment of VFA values attempted to anticipate the behaviour of the variable and to avoid the inherent delay of the response, associated to the time constant of the system. Furthermore, the controlled variable derivative was used as a second input of the fuzzy sets to increase or decrease the speed of the control action. The control law was applied to a 0.948 m3 fixed-bed anaerobic reactor treating raw and diluted (1:2) industrial distillery vinasses. The validation was performed establishing different transient states between different set points in the range of 0.8 and 1.8 g VFA/l and different concentrations of the influent. The control law proved to be reliable supplying an adequate control action in terms of amplitude and velocity to achieve the desired set point for different types of perturbation and control purposes.

MELD score and albumin replacement are related to higher costs during management of patients with refractory ascites.

BACKGROUND: Ascites is the most common complication of cirrhosis and indicates that the disease is at an advanced stage. In cirrhotic patients with refractory ascites, treatment is based on repeat paracentesis. The objective of this study is to evaluate the cost of paracentesis in cirrhotic patients and to determine the factors related to this cost. METHODS: This prospective study included all patients with cirrhosis who underwent paracentesis between March 2012 and March 2013 at the Outpatient Service of the Liver Transplantation Unit, Clinical Hospital, University of São Paulo School of Medicine. Microcost analysis was performed with individual tabbed data regarding the consumption of albumin and containers for ascites. The remaining cost components were drugs, materials used during the procedure, and human resources. Statistical analysis was performed using SPSS version 20. RESULTS: We conducted a total of 881 paracentesis procedures in a group of 155 patients that included 60.5% men and 39.5% women with a mean age of 57 years (range 20 to 80 years). Patients underwent an average of 5.3 paracentesis procedures per year (range 1 to 32). The total cost of all procedures was $193,126.60 and the most costly component was albumin ($87,162.10). The average cost per procedure was $219.50. The most frequent liver disease diagnoses were hepatitis C (24%) and alcoholic cirrhosis (24%). The majority of patients were on the liver transplant list (54.2%). Factors associated with higher costs in the period were a Model for End-Stage Liver Disease score higher than 24 (P = .001) and patients on the transplant waiting list (P = .042). CONCLUSIONS: Paracentesis in cirrhotic patients is a high-cost procedure in health care. The main factors related to cost are the volume of fluid drained due to the need for albumin replacement and the severity of liver disease that is related to the frequency of paracentesis.

Sensitivity to pH, product inhibition, and inhibition by NAD+ of 1,3-propanediol dehydrogenase purified from Enterobacter agglomerans CNCM 1210.

Because of its key role in the metabolism of glycerol during fermentation, 1,3-propanediol dehydrogenase (EC 1.1.1.202) of Enterobacter agglomerans CNCM 1210 was purified to homogeneity and studied with respect to its sensitivity to pH and to nucleotide and 1,3-propanediol concentrations. Enzyme activity was optimal at pH 7.8. The enzyme was competitively inhibited by NAD+ (Ki of 0.29 mM), and 1,3-propanediol exerted a strong inhibitory effect according to a mixed-type inhibition with a Ki of 13.7 mM and an a-factor of 9.0. It is proposed that these dehydrogenase properties be extended to the dehydrogenases of Citrobacter freundii and Klebsiella pneumoniae, which exhibited numerous similar physical properties.

Experimental evaluation of the effects of pravastatin on electrophysiological parameters of rat skeletal muscle.

The effects of daily chronic treatment for 6 months with pravastatin was evaluated on the performance of the skeletal muscle system of different rat groups. At all doses (0.1 mg/kg-20 mg/kg) the righting reflex and the electromyographic signals observed in vivo did not show any abnormality. At the end of the treatment the Extensor digitorum longus muscles were dissected from treated and control rats and their passive and active electrical parameters were analyzed in vitro by standard microelectrodes technique. Pravastatin did not modify the chloride conductance nor the excitability characteristics of the fibers. Chronic treatment with pravastatin does not produce any alteration of skeletal muscle function.

[Action of indoprofen on atrio-ventricular conduction. Preliminary note]. / Azione dell'indoprofene sulla conduzione atrio-ventricolare. Note preliminari.

The purpose of the present study was to evaluate the possible electrophysiological effects of Indoprophene, a new non steroidal analgesic anti-inflammatory agent, which might be employed in the treatment of chest-pain from myocardial ischemia. Ten patient, who were undergoing a electrophysiological study for diagnostic purposes, were given 400 mg of indoprophene by intravenous rapid bolus. The A-V conduction times were recorded in basic condition and 30 minutes after administration of the drug. The data we collected show that Indoprophene does not influences either normal or prolonged A-V conduction times. We conclude that the drug, as it has a good effect on pain and lacks haemodynamic consequences and effects on conduction and excitability, is recommendable for the treatment of chest-pain from myocardial ischemia.