Rapid Identification and Classification of Listeria spp. and Serotype Assignment of Listeria monocytogenes Using Fourier Transform-Infrared Spectroscopy and Artificial Neural Network Analysis.

The use of Fourier Transform-Infrared Spectroscopy (FT-IR) in conjunction with Artificial Neural Network software NeuroDeveloper™ was examined for the rapid identification and classification of Listeria species and serotyping of Listeria monocytogenes. A spectral library was created for 245 strains of Listeria spp. to give a biochemical fingerprint from which identification of unknown samples were made. This technology was able to accurately distinguish the Listeria species with 99.03% accuracy. Eleven serotypes of Listeria monocytogenes including 1/2a, 1/2b, and 4b were identified with 96.58% accuracy. In addition, motile and non-motile forms of Listeria were used to create a more robust model for identification. FT-IR coupled with NeuroDeveloper™ appear to be a more accurate and economic choice for rapid identification of pathogenic Listeria spp. than current methods.

Internal extracellular bacteria of Diaphorina citri Kuwayama (Hemiptera: Psyllidae), the Asian citrus psyllid.

The Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is an invasive insect pest that transmits Candidatus Liberibacter spp. This insect/pathogen system was first identified in North America in the early 2000's and has become the top threat to the citrus industry. Limited options for management of this problem exist; therefore, innovative pest management strategies are being developed. In this study, we describe the first step toward a paratransgenic approach (also referred to symbiotic control) for control of the insect vector or the pathogen. Culturable bacteria from the gut of Asian Citrus Psyllids were identified using standard culture techniques followed by sequencing of the cultured microorganisms. Further, 454 pyrosequencing of the gut was performed to audit bacterial presence in order to begin to identify any relationship between psyllid symbionts and C. Liberibacter spp.

Clustering of clinical and environmental Escherichia coli O104 isolates using the DiversiLab™ repetitive sequence-based PCR system.

The DiversiLab™ rep-PCR system was used to amplify DNA regions of 28 well-characterized Escherichia coli O104 strains to generate a digital DNA fingerprint profile for strain differentiation. E. coli O104 strains from human stools and other sources were examined. The results indicate that this system can cluster similar O104 strains rapidly.

Association and attraction of blueberry maggot fly Curran (Diptera: Tephritidae) to Pantoea (Enterobacter) agglomerans.

The attraction of washed, medium-free cells of Pantoea (Enterobacter) agglomerans to wild, adult Rhagoletis mendax Curran, the blueberry maggot fly, was evaluated in managed blueberry fields in Maine. Attraction was evaluated using Pherocon AM and Ladd traps, each tested with or without washed bacterial cells. Field studies showed significant increases in fly captures on the Pherocon AM traps. Apple volatiles odors on Ladd traps seemed to cancel the effects of bacterial odors. Aerobic heterotrophic bacteria were isolated and identified from alimentary organs within wild R. mendax. Isolates indentified included P. agglomerans. Blueberries collected in the field were surveyed for the presence of P. agglomerans and blueberries containing blueberry maggot larvae and noninfested blueberries were analyzed for amino acid content. Maggot-infested blueberry contained twice the amino acid nitrogen than that of noninfested blueberry. P. agglomerans, like with other pest tephritids, seems to be a cosmopolite with blueberry maggot.

Itch Severity Scale: a self-report instrument for the measurement of pruritus severity.

BACKGROUND: Assessing pruritus severity is difficult because of its subjective nature. A questionnaire that takes into account how the symptom is perceived by the patient may provide a more accurate representation of the pruritus. However, recently developed questionnaires do not specifically quantify severity of the symptom. OBJECTIVES: To develop a self-report questionnaire to measure pruritus severity and to provide initial evidence of its validity and reliability. METHODS: We modified a previously developed interview for the characterization and evaluation of pruritus, which was completed along with the RAND-36 Health Status Inventory and Dermatology Life Quality Index by patients with psoriasis-associated pruritus. Exploratory factor analysis, studies of internal consistency, and correlation analyses with health-related quality of life scores were used to help determine which components of the modified pruritus interview to include in the new questionnaire, the Itch Severity Scale (ISS). The ISS was then assessed for construct validity, internal consistency reliability and test-retest reliability. RESULTS: Seven of the initial 11 components of the modified pruritus interview were included in the ISS. ISS scores correlated moderately with physical (r=-0 x 483) and mental (r=-0 x 492) health composite scores of the RAND-36 and strongly with Dermatology Life Quality Index scores (r=0 x 628), evidence of construct validity. It had an internal consistency reliability of 0 x 80 and a test-retest reliability of 0 x 95. CONCLUSIONS: Based on this preliminary evidence of validity and reliability, this new seven-item ISS may be useful in comparing pruritus severity among different disease populations or in assessing pruritus treatment effectiveness.

Radiopaque alginate microcapsules for X-ray visualization and immunoprotection of cellular therapeutics.

Alginate-poly-L-lysine-alginate (APA) microcapsules have been explored as vehicles for therapeutic drug and cell delivery. The permselectivity of these capsules provides a unique means of controlled drug release and immunoisolation of encapsulated cells. Immunoisolation is especially attractive as it abrogates the need for chronic immunosuppressive therapy and opens up the possibility for the delivery of numerous cell sources including xenogeneic grafts. APA microcapsules containing cellular therapeutics have proven effective in the short-term treatment of a wide range of diseases requiring enzyme or endocrine replacement therapy, including type I diabetes. If these microcapsules could be noninvasively monitored with X-ray imaging modalities (i.e., fluoroscopy, CT, and digital subtraction angiography), questions such as the ideal transplantation site, the best means of delivery, and the long-term survival of grafts could be better addressed. We have developed two novel alginate-based radiopaque microcapsule formulations containing either barium sulfate (Ba X-Caps) or bismuth sulfate (Bi X-Caps). As compared to conventional, nonradiopaque APA capsules, Ba X-Caps and Bi X-Caps containing human cadaveric islets resulted in a decrease in cellular viability of less than 5% up to 14 days after encapsulation. Both radiopaque capsules were found to be permeable to lectins < or =75 kDa, but were impermeable to lectins > or =120 kDa, thus ensuring the blockage of the penetration of antibodies while allowing free diffusion of insulin and nutrients. The glucose-responsive insulin secretion of the radiopaque encapsulated human islets was found to be unaltered compared to that of unlabeled controls, with human C-peptide levels ranging from 3.21 to 2.87 (Ba X-Caps) and 3.23 to 2.87 (Bi X-Caps) ng/islet at 7 and 14 days postencapsulation, respectively. Using fluoroscopy, both Ba X-Caps and Bi X-Caps could be readily visualized as single radiopaque entities in vitro. Furthermore, following transplantation in vivo in mice and rabbits, single capsules could be identified with no significant change in contrast for at least 2 weeks. This study represents the first attempt at making radiopaque microcapsules for X-ray guided delivery and imaging of cellular therapeutics. While human cadaveric islets were used as a proof-of-principle, these radiopaque capsules may have wide ranging therapeutic applications for a variety of cell types.

The effects of vitamin C supplementation on symptoms of delayed onset muscle soreness.

AIM: The aim of this study was to compare the effects of 8 days of vitamin C (VC) supplementation on elbow flexor delayed onset muscle soreness (DOMS) to 8 days of placebo ingestion. METHODS: For 3 days prior to an exercise bout (2 x 20 eccentric elbow extensions), and for 5 days after, a treatment group ingested 3 x 1000 mg/day of VC versus 3 x 50 mg/day of glucose ingestion for the placebo group over the same time period. All subjects were prescreened via dietary recall to exclude any subjects with habitual VC consumption greater than 400 mg/day. Subject comprised 24 subjects (male and female) randomly divided into 2 groups of 12 subjects. Treatment group comprised 5 females and placebo group comprised 8 females. RESULTS: Data from a repeated measures ANOVA indicate that DOMS was successfully induced in both groups via significant time effects for strength loss (P = 0.0001), point tenderness (P = 0.0001), elbow flexor decreased range of motion (P = 0.013), and subjective pain (P = 0.0001). However, there were no significant between group differences in response to any of the aforementioned variables: strength loss (P = 0.202), point tenderness (P = 0.824), elbow flexor range of motion (P = 0.208), subjective pain (P = 0.342). CONCLUSIONS: The results of this study suggest that a VC supplementation protocol of 3 x 1000 mg/day for 8 days is ineffective in protecting against selected markers of DOMS.

Transformation of internal extracellular bacteria isolated from Rhagoletis completa cresson gut with enhanced green fluorescent protein.

We discovered Zeocintrade mark is an effective antibiotic against Enterobacter agglomerans and Klebsiella pneumoniae strains isolated from the walnut husk fly (Rhagoletis completa Cresson: Family Tephritidae) and that bleomycin resistance can be used as a selective marker in transforming plasmids. We transformed Ent. agglomerans and K. pneumoniae strains originally isolated from their close association with R. completa gut to produce enhanced green fluorescent protein, a variant of green fluorescent protein in the first demonstration of genetic transformation of internal extracellular bacteria isolated from a tephritid pest. We report methods for plasmid-mediated transformation of these bacteria, the expression of fluorescent marker protein from the transforming plasmids, and the stability of the transforming plasmid in the bacteria. We also discuss applications of this technology in the study of pest biology and control implementation.

Risk stratification after myocardial infarction using signal-averaged electrocardiographic criteria adjusted for sex, age, and myocardial infarction location.

BACKGROUND: The objectives were to investigate the factors influencing signal-averaged ECGs (SAECGs) recorded in patients after myocardial infarction (MI) and to develop criteria for predicting arrhythmic events (AEs) that account for these factors. METHODS AND RESULTS: SAECGs were recorded 5 to 15 days after MI in 2461 patients without bundle-branch block. The duration (QRSd), terminal potential (VRMS), and terminal duration (LAS) of the filtered QRS were measured. During follow-up (17 +/- 8 months), AEs (arrhythmic death; ventricular tachycardia, VT; ventricular fibrillation, VF) occurred in 80 patients (3.3%). Receiver operating characteristic curves showed that QRSd discriminated patients with all types of AEs, but VRMS and LAS discriminated only VT patients; QRSd minus LAS also discriminated AE patients. Sex, age, and MI location significantly affected the SAECG; survivors without VT or VF were divided into subgroups (2 sex x 4 age x 2 MI), and QRSd values exceeding the 70th percentile in each subgroup predicted AEs with a sensitivity of 65.4%. An unadjusted QRSd criterion showed the same overall sensitivity and specificity but with less uniform values for each subgroup. A Cox model was constructed by use of multiple prognostic indicators, and in rank order, QRSd, previous MI, and Killip class were predictive of AEs. CONCLUSIONS: SAECG adjustments for sex, age, and MI location did not improve sensitivity and specificity but produced a more uniform predictive performance. The proposed criteria are based only on QRSd, because late potentials (VRMS and LAS) did not discriminate patients with sudden death. Duration of high-level activity during QRS (QRSd-LAS) can predict AEs, suggesting that the arrhythmogenic substate involves a large mass of myocardium.

Myocardial infarction patients in the 1990s--their risk factors, stratification and survival in Canada: the Canadian Assessment of Myocardial Infarction (CAMI) Study.

OBJECTIVES: This study sought to evaluate the in-hospital and postdischarge mortality of patients with an acute myocardial infarction in the 1990s. BACKGROUND: The widespread implementation of therapeutic interventions that modify the natural history of coronary artery disease has led to changes in the profile and survival of patients with an acute myocardial infarction. Although data exist for selected subsets of patients with an acute myocardial infarction, at this time there is little recent prospective information on all patients presenting with an acute myocardial infarction, particularly for survival after hospital discharge. METHODS: All patients < or = 75 years old presenting with an acute myocardial infarction between July 1, 1990 and June 30, 1992 at nine Canadian hospitals were prospectively evaluated and followed up for 1 year. From November 1991, patients of all ages were included. In two centers, recruitment continued until December 31, 1992. A total of 3,178 patients were recruited. RESULTS: The in-hospital mortality rate of patients < or = 75 years old was 8.4%, and that at 1 year after hospital discharge was 5.3%. For patients of all ages recruited after November 1, 1991, the in-hospital mortality rate was 9.9% and 7.1% for 1 year after hospital discharge. For patients < or = 75 years old, age carried an independent in-hospital but no post discharge risk. Female patients had a twofold greater risk of dying in hospital. After hospital discharge, only 1.7% of patients < or = 75 years old and 1.9% of patients of all ages died of a presumed arrhythmic death. Premature ventricular contractions had no independent prognostic value. The relatively low in-hospital (5.3%) and postdischarge (6.1%) reinfarction rate may have contributed to improved survival. A greater reinfarction rate in patients >75 years old (17.4% vs. 9.6%, p < 0.001) may have contributed to their poorer outcome. CONCLUSIONS: One-year mortality after acute myocardial infarction continues to decrease, and changes in the prognostic value of traditional methods of risk stratification have occurred.

Multiple tendon rupture at unusual sites in rheumatoid arthritis.

A patient with seropositive nodular rheumatoid arthritis taking large doses of corticosteroids developed multiple tendon ruptures around weight bearing joints. The mechanisms underlying this rare complication are discussed.