A Regenerable Biosensing Platform for Bacterial Toxins.

Waterborne diarrheal diseases such as travelers' diarrhea and cholera remain a threat to public health in many countries. Rapid diagnosis of an infectious disease is critical in preventing the escalation of a disease outbreak into an epidemic. Many of the diagnostic tools for infectious diseases employed today are time-consuming and require specialized laboratory settings and trained personnel. There is hence a pressing need for fit-for-purpose point-of-care diagnostic tools with emphasis in sensitivity, specificity, portability, and low cost. We report work toward thermally reversible biosensors for detection of the carbohydrate-binding domain of the Escherichia coli heat-labile enterotoxin (LTB), a toxin produced by enterotoxigenic E. coli strains, which causes travelers' diarrhea. The biosensing platform is a hybrid of two materials, combining the optical properties of porous silicon (pSi) interferometric transducers and a thermoresponsive multivalent glycopolymer, to enable recognition of LTB. Analytical performance of our biosensors allows us to detect, using a label-free format, sub-micromolar concentrations of LTB in solution as low as 0.135 µM. Furthermore, our platform shows a temperature-mediated "catch-and-release" behavior, an exciting feature with potential for selective protein capture, multiple readouts, and regeneration of the sensor over consecutive cycles of use.

Fluorocarbon Plasma Gas Passivation Enhances Performance of Porous Silicon for Desorption/Ionization Mass Spectrometry.

Desorption/ionization on porous silicon mass spectrometry (DIOS-MS) is shown to be a powerful technique for the sensing of low-molecular-weight compounds, including drugs and their metabolites. Surface modification of DIOS surfaces is required to increase analytical performance and ensure stability. However, common wet chemical modification techniques use fluorosilanes, which are less suitable for high-throughput manufacturing and analytical repeatability. Here, we report an alternative, rapid functionalization technique for DIOS surfaces using plasma polymerization (ppDIOS). We demonstrate the detection of drugs, metabolites, pesticides, and doping agents, directly from biological matrices, with molecular confirmation performed using the fragmentation capabilities of a tandem MS instrument. Furthermore, the ppDIOS surfaces were found to be stable over a 162 day period with no loss of reproducibility and sensitivity. This alternative functionalization technique is cost-effective and amenable to upscaling, ensuring avenues for the high-throughput manufacture and detection of hundreds of analytes across various applications while still maintaining the gold-standard clinical technique using mass spectrometry.

Regulatory T cells and IgE expression in duodenal mucosa of Strongyloides stercoralis and human T lymphotropic virus type 1 co-infected patients.

BACKGROUND: Strongyloides stercoralis is an intestinal nematode unique in its ability to replicate in the human host, allowing ongoing cycles of autoinfection, persisting for decades within the same host. Although usually asymptomatic, overwhelming infections can occur in Strongyloides and HTLV-1 co-infected individuals (SS/HTLV-1). Regulatory T cells (Tregs) are able to blunt specific Th2 responses necessary to control the parasite. We previously reported that peripheral blood Tregs are increased in SS/HTLV-1 and correlate with low Th2 responses. We hypothesized that Tregs are also increased at the site of infection in duodenal mucosa. METHODS: Paraffin embedded duodenal biopsies were obtained from 10 SS/HTLV-1 patients, 3 controls with non-parasitic chronic duodenitis, and 2 healthy controls. Immunohistochemistry was performed using monoclonal antibodies against human CD3, CD8, IgE and FoxP3. The number of cells were counted using a conventional light microscope. The number of CD3+, CD8+, FoxP3+ and IgE positive cells per 0.35 mm2 was measured using ImagePro Plus software comparing areas adjacent or distant from parasite material. RESULTS: In patients with SS/HTLV-1, T lymphocyte counts and CD8+ cells were lower in areas adjacent to the parasite compared to non-adjacent areas (CD3+: adjacent: 6.5 [Interquartile range (IQR: 2.8-12.3)]; non-adjacent: 24.5 [IQR: 20.9-34.4]; Mann-Whitney p = 0.0003; CD8+: adjacent: 4.5 [IQR: 2.3-11.8]; non-adjacent: 21 [IQR: 15.3-42.9]; Mann-Whitney p = 0.0011). Tregs cells in the intestines (FoxP3+ expressing cells) were increased in patients with SS/HTLV-1 compared with patients with chronic duodenitis (SS/HTLV-1: 1.5 [IQR: 0.7-2.3]; duodenitis controls: 0 [range 0-0.7]; healthy controls: 0; Mann-Whitney p = 0.034). There was also a trend towards fewer eosinophils adjacent to the parasites. Among SS/HTLV-1 patients the number of IgE expressing cells was increased for in areas not adjacent to the parasite compared to non-adjacent areas (ANOVA, p = 0.001). CONCLUSIONS: Our data shows increased Treg cell numbers localized adjacent to the parasites in the duodenum SS/HTLV-1 patients. In addition, other T lymphocytes and IgE expressing cells were decreased adjacent to the parasites, suggesting an important role for Tregs in down-regulating local parasite effector responses.

Assessment of a multi-modal intervention for the prevention of catheter-associated urinary tract infections.

BACKGROUND: Catheter-associated urinary tract infections (CAUTIs) represent an important healthcare burden. AIM: To assess the effectiveness of an evidence-based multi-modal, multi-disciplinary intervention intended to improve outcomes by reducing the use of urinary catheters (UCs) and minimizing the incidence of CAUTIs in the internal medicine department of a university hospital. METHODS: A multi-modal intervention was developed, including training sessions, urinary catheterization reminders, surveillance systems, and mechanisms for staff feedback of results. The frequency of UC use and incidence of CAUTIs were recorded in three-month periods before (P1) and during the intervention (P2). FINDINGS: The catheterization rate decreased significantly during P2 [27.8% vs 16.9%; relative risk (RR): 0.61; 95% confidence interval (CI): 0.57-0.65]. We also observed a reduction in CAUTI risk (18.3 vs 9.8%; RR: 0.53; 95% CI: 0.30-0.93), a reduction in the CAUTI rate per 1000 patient-days [5.5 vs 2.8; incidence ratio (IR): 0.52; 95% CI: 0.28-0.94], and a non-significant decrease in the CAUTI rate per 1000 catheter-days (19.3 vs 16.9; IR: 0.85; 95% CI: 0.46-1.55). CONCLUSION: The multi-modal intervention was effective in reducing the catheterization rate and the frequency of CAUTIs.

Moderate consumption of red wine, but not gin, decreases erythrocyte superoxide dismutase activity: a randomised cross-over trial.

BACKGROUND AND AIMS: Several studies have shown that moderate alcohol consumption reduces the risk of coronary heart disease, a disease related to oxidative stress. However, the effects of different alcoholic beverages on antioxidant status are not fully known. Our aim was therefore to compare the effects of a moderate intake of an alcoholic beverage with high polyphenol content (red wine) and another without polyphenol content (gin) on plasma antioxidant vitamins, lipid profile and oxidability of low-density lipoprotein (LDL) particles. METHODS AND RESULTS: Forty healthy men (mean age, 38 years) were included in a randomised cross-over trial. After a 15-day washout period, subjects received 30 g/ethanol/d as either wine or gin for 28 days. Diet and exercise were monitored. Before and after each intervention, we measured serum vitamins, malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase activities, lipid profile, oxidized LDL and LDL resistance to ex-vivo oxidative stress. Compared to gin intervention, wine intake reduced plasma SOD activity [-8.1 U/gHb (95% confidence interval, CI, -138 to -25; P=0.009)] and MDA levels [-11.9 nmol/L (CI, -21.4 to-2.5; P=0.020)]. Lag phase time of LDL oxidation analysis also increased 11.0 min (CI, 1.2-20.8; P=0.032) after wine, compared to gin, whereas no differences were observed between the two interventions in oxidation rate of LDL particles. Peroxide concentration in LDL particles also decreased after wine [-0.18 nmol/mL (CI, -0.3 to-0.08;P=0.020)], as did plasma oxidized LDL concentrations [-11.0 U/L (CI,-17.3 to -6.1; P=0.009)]. CONCLUSION: Compared to gin, red wine intake has greater antioxidant effects, probably due to its high polyphenolic content.

Differential effects of moderate or heavy alcohol consumption on circulating adhesion molecule levels.

Epidemiological studies suggest that moderate but not heavy alcohol consumption provides protection against coronary heart disease. We assessed the relationship between alcohol consumption and serum levels of adhesion molecules involved in the pathogenesis of early atherosclerosis. One-hundred apparently healthy men with similar cardiovascular risk factors were divided into four groups according to ethanol intake. Moderate drinkers (20-40 g/day) showed lower serum intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) levels than abstainers (p < 0.05; both), as well as lower serum ICAM-1, VCAM-1 and E-selectin levels than heavy drinkers (p = 0.01; all). The latter also showed higher serum ICAM-1 and E-selectin levels than abstainers (p < 0.001; both). We conclude that moderate drinkers show a significant reduction of soluble endothelial adhesion molecule levels compared to abstainers and heavy drinkers, that may contribute to the protective effect of moderate alcohol consumption against atherosclerosis.

High ethanol intake and malnutrition in alcoholic cerebellar shrinkage.

To determine the influence of chronic ethanol intake and nutritional status on cerebellar shrinkage in alcoholism, we studied 12 undernourished patients with acute Wernicke's encephalopathy (WE), 12 undernourished and 24 well-nourished asymptomatic chronic alcoholics, and 24 age-matched well-nourished controls, using morphometric analysis of MRI scans with volumetry of the cerebellum. Alcoholics reported a mean daily intake of ethanol of 177+/-8 g over a period of 27+/-1 years. Most undernourished alcoholics and half of the well-nourished alcoholics, compared to one-tenth of the controls, showed a significant reduction in cerebellar volume (p< or =0.01, both). Alcoholics with cerebellar shrinkage (n=33) were older (p=0.05) and tended to report greater daily ethanol intake than alcoholics without cerebellar shrinkage (n=15), although not significantly so (p=0.09). Cerebellar volume correlated negatively with age in controls and asymptomatic alcoholics (r> or =0.52, p< or =0.01, both), with a significantly greater shrinkage for age in the latter (p=0.003). Logistic regression analysis showed that malnutrition (OR 6.6 [95%CI 1.7-25.6], p=0.005) and a daily ethanol intake of more than 140 g over ten years (OR 6.1 [95%CI 1.8-20.5], p=0.003) were independently associated with the development of cerebellar shrinkage.

Usefulness of CT and MR imaging in the diagnosis of acute Wernicke's encephalopathy.

OBJECTIVE: In this study, we analyzed the sensitivity and specificity of CT and MR imaging in the diagnosis of acute Wernicke's's encephalopathy. SUBJECTS AND METHODS: Three groups of subjects were studied: 15 patients with acute Wernicke's encephalopathy; 15 asymptomatic alcoholics; and 15 control subjects. Studies included clinical and laboratory examinations as well as CT and MR imaging of the brain. RESULTS: On CT scans, two patients with Wernicke's encephalopathy (13%) and no asymptomatic alcoholics showed low-density abnormalities in the paraventricular regions of the thalamus (p = .2414). On MR imaging, increased T2 signal of paraventricular regions of the thalamus was observed in seven patients (46%) with Wernicke's encephalopathy and one asymptomatic alcoholic (6%) (p < .01), and increased T2 signal of periaqueductal regions of the midbrain in six patients (40%) with Wernicke's encephalopathy and one asymptomatic alcoholic (6%) (p < .05). However, no significant differences were observed in the prevalence of mamillary body shrinkage between alcoholics with Wernicke's encephalopathy (six [40%]) and asymptomatic chronic alcoholics (four [27%]). The sensitivity of MR imaging in revealing evidence of this disease was 53% and the specificity, 93%. CONCLUSION: MR imaging is useful in confirming the diagnosis of acute Wernicke's encephalopathy. However, the absence of abnormalities on MR imaging does not exclude this diagnosis. CT proved not useful in the diagnosis of Wernicke's encephalopathy.

Ethanol acutely decreases calcium transients in cultured human myotubes.

Ethanol consumption frequently leads to a number of skeletal muscle disorders, including acute and chronic alcoholic myopathy. Ethanol has been found to interfere with signal transduction mechanisms in cardiac and smooth muscle cells. We studied the effects of ethanol on the intracellular calcium ([Ca2+]i) transients responsible for excitation-contraction coupling in human myotubes from chronic alcoholic patients and healthy controls. Cultured myotubes were loaded with the fluorescent Ca2+ indicator fura-2 and evaluated on a single-cell basis. Following electrical stimulation, ethanol caused a significant reversible dose-dependent reduction in [Ca2+]i transient amplitude, achieving a mean decrease of 36+/-5% at 300 mM ethanol (p < 0.01), without modifying the basal [Ca2+]i. This acute effect of ethanol was similar in myotubes obtained from chronic alcoholics and controls. Similarly, ethanol caused a dose-dependent reduction of [Ca2+]i transient amplitude in control samples when depolarization was elicited by 100 mM KCl (p < 0.01). Several potential mechanisms of ethanol action were studied in control muscle samples. Sarcolemmal Ca2+ entry was measured indirectly by monitoring Mn2+-quenching of intracellular fura-2 via the nitrendipine-sensitive Ca2+ channels during electrical pacing. Ethanol at doses of 100 mM and greater caused a dose-dependent reduction in the rate of quench (p < 0.01). In addition, the intracellular pool of Ca2+ releasable by caffeine was found to be reduced at 300 mM ethanol (p < 0.05). We conclude that ethanol reduces the [Ca2+]i transients underlying excitation-contraction coupling in human myotubes, and that this occurs to a similar extent in cells obtained from chronic alcoholics and controls. This acute effect of ethanol was primarily due to an inhibitory effect of ethanol on sarcolemmal Ca2+ influx via voltage-operated Ca2+ channels, although there may also be an effect on the Ca2+ sarcoplasmic reticulum loading state.

Activated lymphocytes (CD25+ CD69+ cells) and decreased CD19+ cells in well-nourished chronic alcoholics without ethanol-related diseases.

To assess lymphocyte subsets and expression of activation antigens in peripheral blood lymphocytes (PBLs) in chronic alcoholism, a cross-sectional study with 30 well-nourished chronic alcoholics and 30 controls was performed. Studies included detailed clinical and laboratory evaluation, nutritional status assessment, and determination of lymphocyte subpopulations, as well as activation antigens. A significant decrease of B cells (CD19+) was observed in chronic alcoholics, compared with controls (p < 0.001). A significant increase of PBLs expressing CD69 and CD25 (p < 0.01, both) in chronic alcoholics was also detected, whereas CD71 expression was unaffected. In addition, T lymphocytes expressing HLA-DR were significantly higher in chronic alcoholics than controls (p < 0.05). The serum level of soluble interleukin-2 receptor was also significantly higher in the alcoholic group, compared with controls (p = 0.04). Moreover, the estimated total lifetime dose of ethanol consumed correlated positively with the percentage of PBLs expressing CD25 (r = 0.48; p = 0.01) and negatively with PBLs expressing CD71 (r = -0.39; p = 0.04). By contrast, the changes were not related to age, nutritional status, or the presence of other ethanol-related diseases. In conclusion, chronic alcoholics present a significant decrease of B cells and an "incomplete activation state" of PBLs that depends on the dose of ethanol consumed.

Serum and muscle levels of alpha-tocopherol, ascorbic acid, and retinol are normal in chronic alcoholic myopathy.

Some authors have suggested a possible loss of antioxidant factors in alcoholic skeletal myopathy. To assess the relationship between ethanol consumption and serum and muscle levels of alpha-tocopherol, ascorbic acid, and retinol in chronic alcoholics with and without skeletal myopathy, a prospective cross-sectional study was performed in the Alcohol Unit of a 1000-bed university hospital. Twenty-five chronic male alcoholic patients (10 with skeletal myopathy) and 15 male controls of similar age were included. Evaluation of daily and lifetime ethanol consumption, assessment of anthropometric and protein nutritional parameters, and open biopsy of the left deltoid muscle were performed, as well as determinations of serum and muscle levels of retinol, alpha-tocopherol, and ascorbic acid by HPLC analysis. Ten of the 25 chronic alcoholic patients presented histological criteria of skeletal myopathy. Four alcoholics presented caloric malnutrition and three protein malnutrition. All of the muscle biopsies of the control group were entirely normal, as were their nutritional studies. The serum and muscular levels of alpha-tocopherol, ascorbic acid, and retinol were normal and were similar in both alcoholics and controls. Except for serum retinol, these values were also similar in alcoholic patients with or without skeletal myopathy. In the univariate analysis, we identified the total lifetime dose of ethanol (p < 0.003), the muscle arm area (p < 0.05), and serum levels of prealbumin (p < 0.03) and retinol-binding protein (p < 0.05) as factors influencing the development of alcoholic myopathy. However, in multivariate analysis, the total lifetime dose of ethanol was the only independent factor in relation to alcoholic myopathy (p < 0.003). Serum and muscle levels of the antioxidants alpha-tocopherol, ascorbic acid, and retinol do not influence the presence of skeletal myopathy in chronic alcoholic patients.

Brain imaging in alcoholism.

Copyright Rapid Science Ltd

Comparison of alcoholic cardiomyopathy in women versus men.

To compare the prevalence and cardiac status of male and female alcoholics with alcoholic cardiomyopathy during a 5-year period, all chronic alcoholics with dilated cardiomyopathy who had clinical symptoms of heart failure were included. Alcoholic cardiomyopathy was diagnosed in 10 chronic alcoholic women and in 26 men; the prevalence of alcoholic cardiomyopathy was similar in both sexes. No significant differences were observed in age, nutritional parameters, and clinical and radiologic data of heart failure between the 2 groups. Alcoholic women reported a significantly lower daily dose of ethanol (p = 0.002), a shorter duration of alcoholism (p = 0.017), and a lower total lifetime dose of ethanol consumption (p = 0.001), and had a lower New York Heart Association functional class than men. Women also had lesser ventricular dysfunction than men. In a multivariate analysis, left ventricular systolic dysfunction was related to the total lifetime dose of ethanol consumption (p <0.04), but not to gender. Finally, when patients were matched for left ventricular ejection fraction, women had consumed a lower total lifetime dose of ethanol than men (p <0.001). The prevalence of alcoholic women with dilated cardiomyopathy was found to be similar to that of alcoholic men, although women required a lower total lifetime dose of ethanol to develop the disease.

Testicular function in asymptomatic chronic alcoholics: relation to ethanol intake.

To evaluate the effect of ethanol on testicular function in chronic alcoholics without chronic liver disease, we studied 38 asymptomatic chronic alcoholics and 19 age-matched controls. Detailed clinical history, nutritional status, hormonal analysis, and seminal studies were conducted in each case and control. Alcoholic patients had an average of 39 +/- 2 years old (range: 26 to 60) and reported a daily ethanol consumption from 100 to 350 g (mean: 198 +/- 15) over a period of 18.0 +/- 1.2 years. Alcoholics exhibited a significant increase of the luteinizing hormone (p < 0.001) and a decrease of the Free Androgen Index, compared with controls (p < 0.05) that related significantly with the total lifetime dose of ethanol (p < 0.01, both). Seminal studies indicate that 39.4% of alcoholics had significantly reduced their spermatozoa count (p < 0.01), whereas significant morphological abnormalities were observed in 44.7% of the alcoholics (p < 0.01). Spermatozoa motility from alcoholics was also found to be altered in half of the patients (p < 0.01). A significant increase of serum luteinizing hormone, follicle-stimulating hormone, and sex hormone binding globulin levels, and a decrease of Free Androgen Index were observed in alcoholics with morphology and motility abnormalities (p < 0.05, all). In multivariate analysis, the only independent factor that determined the alterations in sperm (count, morphology abnormalities, and motility alterations) was the total lifetime of ethanol intake (p < 0.001, all). We conclude that alcoholics frequently develop a situation of primary hypogonadism related to a lifetime of ethanol consumption.

Chronic alcoholic myopathy: diagnostic clues and relationship with other ethanol-related diseases.

We report the clinical, laboratory, functional and histological features of 100 male alcoholic patients of whom 44 had chronic alcoholic myopathy (CAM). We evaluated the use of non-invasive tests in detecting CAM, and examined its relationship with other ethanol-related diseases such as cirrhosis and cardiomyopathy. Of the CAM patients, 24 (55%) presented clinical symptoms of myopathy, whereas proximal muscle atrophy was observed in 15 patients (35%). Thirty-seven (80%) had significantly decreased muscle strength by myometric measurement and 27 (60%) had abnormally increased serum muscle enzymes. In most of these patients, the myopathy was classified as mild. The most frequent histological findings were myocytolysis, fibre size variability and type II fibre atrophy. As there was a good correlation between clinical symptoms, decreased muscle strength on myometry and histological evidence of CAM, muscle biopsy may be avoidable in some of these patients. Cardiomyopathy and liver cirrhosis were more frequent in patients with CAM, and should be checked for in chronic alcoholics with skeletal myopathy.

[Endometriosis. Analysis of laparoscopic findings at the A.B.C. (American-British Cowdray) Hospital]. / Endometriosis. Análisis del hallazgo laparoscópico en el Hospital A.B.C.

Endometriosis approximately affects 10 to 15% of premenopausic women. Diagnosis mainly is made by laparoscopy. We revised all the laparoscopies which were made in labor service at the American British Cowdray Hospital from July 1st 1989 to June 30th 1994 in order to evaluate the incidence of endometriosis in the institution. They were made from 882 laparoscopies done, in 447 cases (50.68%) endometriosis was the main finding, this was similar to the reports published by other authors. The group of age mainly affected was from 20 to 29 years-old. Sterility was found in 53% of these patients, slightly above the results in other series. The 85% of the cases were in hospital for less than 24 hours. Complications were in 7% of them, none required a laparotomy, however.

Teratoma cervical del recién nacido: Tratamiento y táctica quirúrgica / Cervical teratoma in the neonate:Treatment and chirurgical approach

El teratoma cervical en el recién nacido es una entidad poco frecuente .Se presenta un paciente con una forma gigante del tumor y su esquema terapéutico.El diagnóstico precoz, la intubación orotraqueal, la cirugía adecuada sin demora y el seguimiento a largo plazo con determinación de la curva de alfa-fetoproteína, definen la sobrevida de estos pacientes

Nutritional status in chronically alcoholic men from the middle socioeconomic class and its relation to ethanol intake.

To determine the relationship between nutritional status and ethanol consumption, 250 chronically alcoholic men (mean age 41 +/- 11 years) entering an alcoholism treatment program were studied. A control group of 100 male volunteers (mean age 40 +/- 10 years) was also evaluated. Detailed clinical history, laboratory analysis and nutritional status assessment were carried out in each case and control. In addition, ethanol-related diseases such as liver disease, chronic pancreatitis, cardiomyopathy, myopathy and peripheral neuropathy were ruled out in all patients. The mean daily ethanol consumption of the alcoholics was 235 +/- 62 g over an average of 19 years. All of them belong to a very stable, middle-class working group of men. Only 25 (10%) alcoholics showed evidence of energy malnutrition, 15 (6%), of protein malnutrition, and 6 (2%) of both. In the multivariate analysis, the only independent factors for the development of malnutrition were the total lifetime dose of ethanol and calorie intake (ethanol excluded) (P < 0.01; both), as well as cirrhosis (P < 0.01) when protein malnutrition was considered. Alcoholic cirrhosis was diagnosed in 20 cases, skeletal myopathy in 117, dilated cardiomyopathy in 20 and peripheral neuropathy in 41. When patients with ethanol-related diseases were excluded, no significant differences in nutritional parameters were observed between chronic alcoholics and controls. We conclude that malnutrition is not as frequent as previously thought in middle socioeconomic class male alcoholics and its existence may be considered as another consequence of ethanol intake or secondary to the alcohol-related diseases.

[Uremic hemolytic syndrome induced by mitomycin C. Presentation of a series of 5 cases and review of the literature]. / Síndrome hemolítico urémico inducido por mitomicina C. Presentación de una serie de cinco casos y revisión de la literatura.

Five new cases of hemolytic uremic syndrome associated mitomycin C treatment in neoplastic patients are presented and clinical, biological, histological therapeutic and the evolution data of 92 other cases from the literature are analyzed. The results point out the high incidence of GI adenocarcinomas (65%), its rare appearance with total mitomycin doses below 50 mg/m2, its frequent clinical presentation with cardiovascular manifestations, the therapeutical difficulties and the possibility of a better prognosis with an early diagnosis.