Two novel toxin variants revealed by whole-genome sequencing of 175 Clostridium botulinum type E strains.

We sequenced 175 Clostridium botulinum type E strains isolated from food, clinical, and environmental sources from northern Canada and analyzed their botulinum neurotoxin (bont) coding sequences (CDSs). In addition to bont/E1 and bont/E3 variant types, neurotoxin sequence analysis identified two novel BoNT type E variants termed E10 and E11. Strains producing type E10 were found along the eastern coastlines of Hudson Bay and the shores of Ungava Bay, while strains producing type E11 were only found in the Koksoak River region of Nunavik. Strains producing BoNT/E3 were widespread throughout northern Canada, with the exception of the coast of eastern Hudson Bay.

Localization of c-Myb and induction of apoptosis by antisense oligonucleotide c-Myb after angioplasty of porcine coronary arteries.

Previous studies have shown that inhibition of the proto-oncogene c-myb inhibits neointimal formation in various animal models. However, the temporal and spatial expression of c-Myb in the vessel wall after injury is not known, and the mechanism of action of antisense oligonucleotide (AS-ODN-c-myb) inhibition remains unclear. One potential effect of cell cycle dysregulation by inhibition of c-myb is an increase in the rates of apoptosis. In this study, c-Myb expression after percutaneous transluminal coronary angioplasty (PTCA) injury and induction of apoptosis after AS-ODN-c-myb treatment were determined. Immunohistochemistry and cellular phenotyping were used to localize c-Myb expression in porcine coronary arteries at various time intervals after PTCA. In vitro, the effects of AS-ODN-c-myb on the apoptosis of porcine vascular smooth muscle cells (PVSMCs) and endothelial cells were determined by using a cell-death ELISA and time-lapse video microscopy. In vivo, local delivery of AS-ODN-c-myb was performed after PTCA of pig coronary arteries, and apoptosis was quantified at 6 hours. c-Myb is induced in pig coronary arteries after angioplasty, with maximal expression in inflammatory cells at 18 hours and in vascular smooth muscle cells at 3 to 7 days. In vitro, AS-ODN-c-myb enhanced PVSMCs (6.8+/-0.8% [P=<0.001] versus 0.5% serum) but not endothelial cell apoptosis (1.4+/-0.5% [P=NS] versus 0.5% serum). In vivo, 6 hours after porcine coronary angioplasty and delivery of AS-ODN-c-myb, the proportion of apoptotic cells within the media was 4.2+/-0.8% (PTCA alone), 2.3+/-0.2% (PTCA+vehicle), and 9.0+/-1.1% (PTCA+AS-ODN-c-myb; P<0.05 versus PTCA alone and P<0.01 versus PTCA+saline). c-Myb is expressed after PTCA of pig coronary arteries, and AS-ODN-c-myb induces apoptosis of PVSMCs in vitro and medial cells in vivo.

Effect of epinephrine on muscle glycogenolysis during exercise in trained men.

To test the hypothesis that an elevation in circulating epinephrine increases intramuscular glycogen utilization, six endurance-trained men performed two 40-min cycling trials at 71 +/- 2% of peak oxygen uptake in 20-22 degrees C conditions. On the first occasion, subjects were infused with saline throughout exercise (Con). One week later, after determination of plasma epinephrine levels in Con, subjects performed the second trial (Epi) with an epinephrine infusion, which resulted in a twofold higher (P < 0.01) plasma epinephrine concentration in Epi compared with Con. Although oxygen uptake was not different when the two trials were compared, respiratory exchange ratio was higher throughout exercise in Epi compared with Con (0.93 +/- 0.01 vs. 0.89 +/- 0.01; P < 0.05). Muscle glycogen concentration was not different when the trials were compared preexercise, but the postexercise value was lower (P < 0.01) in Epi compared with Con. Thus net muscle glycogen utilization was greater during exercise with epinephrine infusion (224 +/- 37 vs. 303 +/- 30 mmol/kg for Con and Epi, respectively; P < 0.01). In addition, both muscle and plasma lactate and plasma glucose concentrations were higher (P < 0.05) in Epi compared with Con. These data indicate that intramuscular glycogen utilization, glycolysis, and carbohydrate oxidation are augmented by elevated epinephrine during submaximal exercise in trained men.

Effect of CHO ingestion on exercise metabolism and performance in different ambient temperatures.

Two series of experiments were conducted to examine the effect of ingesting beverages with differing carbohydrate (CHO) concentrations and osmolalities on metabolism and performance during prolonged exercise in different environmental conditions. In series 1, 12 subjects performed three cycling exercise trials to fatigue at 70% VO2peak in either 33 degrees C (N = 6) (HT1) or 5 degrees C (N = 6) (CT). Subjects ingested either a 14% CHO solution (osmolality = 390 mosmol.1(-1) (HCHO); a 7% CHO solution (330 mosmol.1(-1) (NCHO) or a placebo (90 mosmol.1(-1) (CON1). In series 2, six subjects performed the same three trials at 33 degrees C (HT2), while ingesting either NCHO, a 4.2% CHO solution (240 mosmol.1(-1) (LCHO) or a placebo) (240 mosmol.1(-1) (CON2). Plasma glucose was higher (P < 0.05) in HCHO than NCHO, which in turn was higher (P < 0.05) than CON1 in both CT and HT1. Plasma glucose was lower (P < 0.05) in CON2 compared with NCHO and LCHO in HT2. The fall in plasma volume was greater (P < 0.05) in HCHO than other trials in both CT and HT1 but was not different when comparing the three trials in HT2. Exercise time was not different when comparing the trials in either HT1 or HT2 but was longer (P < 0.05) in NCHO compared with HCHO, which, in turn, was longer (P < 0.05) than CON1 in CT. These data demonstrate that, during prolonged exercise in the heat, fatigue is related to factors other than CHO availability. In addition, during exercise in 5 degrees C a 7% CHO solution is more beneficial for exercise performance than a 14% CHO solution.

Effect of Maillard reaction products on ruminal and fecal acid-resistant E. coli, total coliforms, VFA profiles, and pH in steers.

Six ruminally cannulated Angus-cross steers (362 kg) were used in a replicated 3 x 3 Latin square design to determine effects of supplementing Maillard reaction products (MRP) on acid-resistant E. coli and coliform populations. Steers were fed roughage-based diets supplemented (DM basis) with either 10% soybean meal (SBM), 10% nonenzymatically browned SBM (NESBM), or 10% SBM top-dressed with 45 g of a lysine-dextrose Maillard reaction product (LD-MRP). Equal weights of dextrose, lysine hydrochloride, and deionized water were refluxed to produce the LD-MRP. The NESBM was manufactured by treating SBM with invertase enzyme, followed by heating to induce nonenzymatic browning. Steers were allowed slightly less than ad libitum access to diets fed twice daily and were adapted to their respective treatments within 10 d. On d 11, ruminal and fecal samples were collected at 0, 2, 4, 6, 8, and 12 h after feeding from each of the steers and transported to the laboratory for microbial analysis. Ruminal samples and feces were analyzed for pH and VFA, and both ruminal fluid and feces were tested for acid-resistant E. coli and total coliforms by incubating samples in tryptic soy broth adjusted to pH 2, 4, and 7. Ruminal pH and total VFA concentrations did not differ among treatments. The molar proportion of ruminal acetate was higher (P < 0.05) for steers receiving NESBM than for steers receiving SBM and LD-MRP. At pH 4, steers that received NESBM had lower (P < 0.05) ruminal populations of E. coli and total coliforms than steers that received SBM. No differences were observed for ruminal E. coli and total coliforms at pH 2 and 7. Fecal pH was lower (P < 0.05) for steers fed NESBM than for steers fed SBM or LD-MRP. Molar proportions of fecal acetate were lower (P < 0.05) and proportions of butyrate and isovalerate were higher (P < 0.05) for steers fed NESBM compared with steers fed SBM. Fecal E. coli at pH 4 was lower (P < 0.05) for steers fed NESBM than for steers fed LD-MRP. Fecal E. coli and total coliforms at pH 2 and 7 did not differ among treatments. Dietary MRP had limited effectiveness at decreasing acid-resistant coliforms in the rumen and feces of cattle. Acid resistance in coliforms may depend on protein availability.

Evaluation of finishing performance, carcass characteristics, acid-resistant E. coli and total coliforms from steers fed combinations of wet corn gluten feed and steam-flaked corn.

Crossbred beef steers (n = 615) were used in a 152-d experiment to compare steam-flaked corn (SFC) diets containing 0, 30, or 60% wet corn gluten feed (WCGF). On d 114 to 118, ruminal and fecal samples were collected from 180 steers and analyzed for pH, VFA, and total and acid-resistant Escherichia coli and coliforms. Acid resistance of E. coli and coliform populations was determined by exposure of the samples for 1 h in pH 2, 4, and 7 citric acid/sodium phosphate buffers. Increasing levels of WCGF linearly decreased total ruminal VFA (P = 0.01) and total fecal VFA (P = 0.06), but linearly increased ruminal and fecal acetate:propionate (P < 0.01) ratio and ruminal and fecal pH (P < 0.05). Feeding increasing WCGF levels resulted in a quadratic response (P < 0.05) with respect to numbers of ruminal E. coli and total coliform populations resistant to pH 4 exposure. Steers fed 30% WCGF had higher (0.7 log units) ruminal E. coli and total coliforms after exposure at pH 4 compared to steers fed 0 or 60% WCGF. Populations of E. coli and total coliforms at pH 2 and 7 were similar for all dietary treatments. Dietary WCGF linearly increased DMI (P = 0.07) and liver abscesses (P = 0.03) and linearly decreased dietary NEg (P = 0.02). Average daily gain and feed efficiencies were greatest when steers were offered 30% WCGF (quadratic, P < 0.05). Dietary manipulations that reduce acid concentrations may not correspond to changes in acid resistance of E. coli and total coliform populations detected in the gastrointestinal tracts of cattle. Moderate levels of WCGF complement SFC finishing diets.

Evaluating the effect of glove coating on the shear bond strength of porcelain laminate veneers.

Gloves have become a standard tool in dentistry due to infection control concerns. But the starch powder with which many of these gloves are coated may contaminate surfaces on contact, creating potential problems during adhesive procedures such as the placement of porcelain veneers. The authors studied the powder's effect on the strength of the shear bond of etched porcelain to etched human tooth enamel and compared it to the effect of a powder-free polymer glove coating. While the starch coating led to a statistically significant decrease in shear bond strength, the powder-free coating produced no significant decrease in strength.

Negative effects of powdered latex gloves in clinical dentistry.

With the advent of CDCs "universal precautions" and OSHA's bloodborne pathogen standard, the number of health-care workers wearing latex gloves has increased dramatically in the law few years. Reports of irritation, contact, and allergic reactions to powdered latex gloves in both medicine and dentistry are on the rise as well. However, other problems related to powdered gloves and their effect on clinical dentistry are just beginning to be addressed. One recent study demonstrated that the starch used as a lubricant on latex rubber gloves significantly reduced the shear bond gloves significantly reduced the shear bond strength of etched porcelain to etched enamel with a resin adhesive system by nearly 40%. Question also arise as to the proper handling of dental implants and the effect that powdered gloves may have on their success based on the negative effects of post-surgical foreign body granulomas reported in the medical community.

Naturalistic assessment of children's compliance to teachers' requests and consequences for compliance.

Naturally occurring levels of teacher commands, child compliance to those commands, and positive and negative teacher feedback were studied in 19 teachers and 130 children in kindergarten through third grade. Seventy-five of the children had been identified as "making a good social adjustment" to school (high-rated) and 55 children were identified as "not making a good social adjustment" to school (low-rated). Results of intensive observation over a 4-wk period showed that: (a) individual teachers differed significantly in their overall use of commands; however, they did not differentially respond to high- versus low-rated children; (b) high-rated children were more likely to comply with commands than were low-rated children; (c) although the overall level of positive social consequences was extremely low, there was some indication that high-rated children were more likely to receive positive feedback for compliance than were low-rated children; (d) low-rated children received significantly more positive feedback than high-rated children for noncompliance; (e) teachers gave negative feedback for noncompliance at an equal level to both groups of children; and (f) although repeated teacher commands following noncompliance were equal across groups, low-rated children were exposed to significantly higher levels of repeated commands following compliance than were high-rated youngsters.

Clinical versatility of new compomer restorative technology.

The introduction of restorative materials known as compomers is revolutionizing the field of restorative dentistry. This versatile material combines the most positive attributes of both glass ionomers and composite resins, such as continuous fluoride release, excellent adhesion, ease in placement, exceptional shade blending, and high polishability. This article examines the advantages of the compomer (Compoglass, Ivoclar Vivadent, Amherst, NY) in relation to those of other restorative materials, details the indications for its use, and provides a procedural analysis of its application through a comprehensive description of a case report.

Embolization of the ovarian veins as a treatment for patients with chronic pelvic pain caused by pelvic venous incompetence (pelvic congestion syndrome).

Ovarian and pelvic (internal iliac) varices have long been recognized as a source of chronic pelvic pain in women. The technique of transcatheter embolotherapy for ovarian and pelvic varices requires selective catheterization of the ovarian and internal iliac veins, followed by contrast venography and embolization. The long-term effects of treatment are the subject of ongoing investigation. This article provides a concise background on ovarian and pelvic varices and reviews the recently published literature on their embolization for the treatment of pelvic venous incompetence (also known as pelvic congestion syndrome).

Between-site comparison of freshwater bacterioplankton by DNA hybridization.

Natural assemblages of freshwater bacterioplankton in enriched lowland rivers had greater cell-specific metabolic activity than those in gravel-pit ponds. Similarly, cell-specific activity and mean cell Size in calcareous headstreams tended to be greater than in intermittently-acid headstreams on millstone grit. DNA was extracted and purified from bacterioplankton assemblages, and between-site comparisons were made in terms of percentage similarity as indicated by DNA hybridization. Cluster analysis, using percentage-similarity matrices, placed bacterioplankton assemblages from different site types into distinct groups. This suggested that between-site physiological differences were related to intrinsically different bacterial composition rather than to different physiological response to different environmental conditions by essentially similar bacterial assemblages.

Effect of temperature on muscle metabolism during submaximal exercise in humans.

To study the effect of temperature on muscle metabolism during submaximal exercise, six endurance-trained men had one thigh warmed and the other cooled for 40 min prior to exercise using water-perfused cuffs. One cuff was perfused with water at 50-55 degrees C (HL) with the other being perfused with water at 0 degree C (CL). With the cuffs still in position, subjects performed cycling exercise for 20 min at a work load corresponding to 70% VO2,peak (where VO2,peak is peak pulmonary oxygen uptake) in comfortable ambient conditions (20-22 degrees C). Muscle biopsies were obtained prior to and following exercise and forearm venous blood was collected prior to and throughout the exercise period. Muscle temperature (Tmus) was not different prior to treatment, but treatment resulted in a large difference in pre-exercise Tmus (difference = 6.9 +/- 0.9 degrees C; P < 0.01). Although this difference was reduced following exercise; it was nonetheless significant (difference = 0.4 +/- 0.1 degree C; P < 0.05). Intramuscular [ATP] was not affected by either exercise or muscle temperature. [Phosphocreatine] decreased (P < 0.01) and [creatine] increased (P < 0.01) with exercise but were not different when comparing HL with CL. Muscle lactate concentration was not different prior to treatment nor following exercise when comparing HL with CL. Muscle glycogen concentration was not different when comparing the trials before treatment, but the post-exercise value was lower (P < 0.05) in HL compared with CL. Thus, net muscle glycogen use was greater during exercise with heating (208 +/- 23 vs. 118 +/- 22 mmol kg-1 for HL and CL, respectively; P < 0.05). These data demonstrate that muscle glycogen use is augmented by increases in intramuscular temperature despite no differences in high energy phosphagen metabolism being observed when comparing treatments. This suggests that the increase in carbohydrate utilization occurred as a direct effect of an elevated muscle temperature and was not secondary to allosteric activation of enzymes mediated by a reduced ATP content.

Variations in healthcare measures by insurance status for patients receiving ventilator support.

OBJECTIVE: To examine differences in healthcare delivery by expected health insurance status for hospitalized patients in diagnosis-related group (DRG) 475, respiratory system diagnoses requiring intubation and continuous ventilator support. DESIGN: A survey, derived from the Healthcare Cost and Utilization Project interstate database, of the care delivered to 21,149 adult patients in DRG 475 and hospitalized in one of 718 acute-care hospitals in nine states. Multivariate analysis was performed, controlling for demographic and hospital factors. RESULTS: Patients insured by health maintenance organizations (HMOs) had significantly lower rates of inpatient mortality (odds ratio [OR], 0.84; 95% confidence interval [CI95], 0.73-0.96), 14.3 more procedures performed (CI95, 11.5-17.2), 7.0% shorter hospitalizations (CI95, 12.5-1.6), and 5.2% higher charges (CI95, 0.4-10.0) than those with traditional private insurance. In addition, patients insured by Medicaid had 3.5% more procedures performed (CI95, 1.6-5.4), 10.4% longer lengths of hospitalization (CI95, 6.7-14.0), and 13.8% higher charges (CI95, 10.6-17.0) than those with traditional private insurance. Finally, the uninsured had significantly lower rates of inpatient mortality (OR, 0.87; CI95, 0.77-0.99), 8.5% more procedures performed (CI95, 6.0-11.1), 16.5% shorter hospitalizations (CI95, 21.5-11.6), and 13.4% lower charges (CI95, 17.8-9.0) than those with traditional private insurance. CONCLUSION: Variations in healthcare measures by insurance status for this DRG emphasize the importance of more careful analyses of insurance categories as a determinant of healthcare access and outcomes. Expected insurance status was an independent predictor of cost. Private insurance and HMO populations differed significantly in outcome and cannot be considered equivalent.