Clinical features of Clostridium difficile infection and molecular characterization of the isolated strains in a cohort of Danish hospitalized patients.

The purpose of this study was to compare clinical features of Clostridium difficile infection (CDI) to toxin gene profiles of the strains isolated from Danish hospitalized patients. C. difficile isolates were characterized by PCR based molecular typing methods including toxin gene profiling and analysis of deletions and truncating mutations in the toxin regulating gene tcdC. Clinical features were obtained by questionnaire. Thirty percent of the CDI cases were classified as community-acquired. Infection by C. difficile with genes encoding both toxin A, toxin B and the binary toxin was significantly associated with hospital-acquired/healthcare-associated CDI compared to community-acquired CDI. Significantly higher leukocyte counts and more severe clinical manifestations were observed in patients infected by C. difficile containing genes also encoding the binary toxin together with toxin A and B compared to patients infected by C. difficile harbouring only toxin A and B. In conclusion, infection by C. difficile harbouring genes encoding both toxin A, toxin B and the binary toxin were associated with hospital acquisition, higher leukocyte counts and severe clinical disease.

Spontaneous fermentation of traditional sago starch in Papua New Guinea.

Sago starch is an important dietary carbohydrate in lowland Papua New Guinea (PNG). An investigation was conducted to determine whether microbes play a role in its preservation using traditional methods. In 12 stored sago samples collected from PNG villages, lactic acid bacteria (LAB) were present (> or = 3.6 x 10(4)cfu/g) and pH ranged from 6.8 to 4.2. Acetic and propionic acids were detected in all samples, while butyric, lactic and valeric acids were present in six or more. In freshly prepared sago, held in sealed containers in the laboratory at 30 degrees C, spontaneous fermentation by endogenous microflora of sago starch was observed. This was evident by increasing concentrations of acetic, butyric and lactic acids over 4 weeks, and pH reducing from 4.9 to 3.1: both LAB and yeasts were involved. Survival of potential bacterial pathogens was monitored by seeding sago starch with approximately 10(4)/g of selected organisms. Numbers of Bacillus cereus, Listeria monocytogenes and Staphylococcus aureus fell to < 30/g within 7 days. Salmonella sp. was present only in low numbers after 7 days (< 36/g), but Escherichia coli was still detectable after three weeks (> 10(2)/g). Fermentation appeared to increase the storability and safety of the product.

Sequential coexpression of the multidrug resistance genes MRP and mdr1 and their products in VP-16 (etoposide)-selected H69 small cell lung cancer cells.

Resistance to drugs included in the multidrug-resistance phenotype has been attributed to overexpression of either mdr1 or MRP genes and their products in numerous cell lines, while coexpression, to our knowledge, has not previously been reported in the same cells. Human small cell lung cancer H69/VP cells were developed by continuous incubation in increasing doses of VP-16. In reverse transcription-PCR assays we found over-expression of both mdr1 and multidrug-resistance protein (MRP) genes, and immunoblots showed both elevated P-glycoprotein and MRP in H69/VP cells. Double immunocytochemical staining demonstrated the expression of both MRP and P-glycoprotein in the same cells, indicating that the observations do not result from the selection of two independent clones. Examination of early passages of H69/VP cells showed that overexpression of MRP mRNA occurred prior to mdr1. Thus, cell lines and clinical samples in the future should be tested for both mdr1/P-glycoprotein and MRP since a positive result for one of the phenotypes does not preclude the existence of the other.

Executive impairment among the functionally dependent: comparisons between schizophrenic and elderly subjects.

OBJECTIVE: Executive deficits have traditionally been associated with frontal lobe brain damage. They are relevant to a variety of disabling mental conditions, including schizophrenia and Alzheimer's disease. To measure these deficits, the authors developed the Executive Interview, a 25-item, 15-minute interview. It has been validated among elderly subjects across a wide range of functional impairment. METHODS: Forty young, chronically ill schizophrenic residents of a state mental health facility and 104 elderly residents, representing three levels of care, of a comprehensive retirement community were tested with the Executive Interview and the Mini-Mental State. RESULTS: When age, gender, education, and number of prescribed medications were controlled, cognitive impairment on the Executive Interview and Mini-Mental State rose with level of care. The Executive Interview alone discriminated between subjects at each level of care, and it was more sensitive to cognitive impairment than the Mini-Mental State. Executive Interview scores correlated the strongest with level of care. Mini-Mental State scores, number of prescribed medications, and age also correlated significantly. Schizophrenic patients showed as much executive impairment on the Executive Interview as elderly subjects at the same level of care despite significant differences in age, sex, and neuroleptic use. Executive Interview and Mini-Mental State scores were highly correlated among the elderly but less so among the schizophrenic patients. Cross-group differences were also found in the pattern of failure on selected Executive Interview items despite similar total Executive Interview scores. CONCLUSIONS: Increasing executive dyscontrol is associated with the need for increasing levels of care and supervision. This finding is neither age nor disease specific. Cross-group differences on selected Executive Interview items suggest the existence of disease-specific patterns of failure. Their recognition could prove useful in the identification of anatomically or pathophysiologically distinct subgroups among patients with executive dyscontrol.

Performance of whole blood IFN-gamma test for tuberculosis diagnosis based on PPD or the specific antigens ESAT-6 and CFP-10.

OBJECTIVE: To evaluate the QuantiFERON-TB test in BCG-vaccinated, non-BCG-vaccinated and tuberculosis (TB) patient donor groups, and to compare its diagnostic performance with that of a blood test based on the Mycobacterium tuberculosis specific antigens ESAT-6 and CFP-10. DESIGN: Analysis of the IFN-gamma responses of whole blood cells from BCG-vaccinated or non-BCG-vaccinated donors or patients with tuberculosis, stimulated with PPD, ESAT-6 or CFP-10 antigens, and evaluation of the specificity and sensitivity of the test. RESULTS: None of the non-vaccinated donors showed positive responses to M. tuberculosis-PPD, ESAT-6 or CFP-10. In BCG-vaccinated donors, 9/19 (47%) donors responded to the QuantiFERON-TB test based on M. tuberculosis-PPD, whereas 2/19 (10.5%) responded to either ESAT-6 or CFP-10. Comparable levels of sensitivity were obtained with the QuantiFERON-TB test based on M. tuberculosis-PPD (79%) and ESAT-6 or CFP-10 antigens (72%). CONCLUSION: Our results demonstrate that the whole blood test based on M. tuberculosis-PPD did not efficiently distinguish BCG-vaccinated donors from individuals with disease due to M. tuberculosis. The introduction of new recombinant antigens specific for M. tuberculosis, such as ESAT-6 or CFP-10, should increase the specificity of the whole blood test and enable discrimination between TB infection, atypical mycobacterial reactivity and reactivity due to BCG vaccination. Such a test would provide a quantum improvement over the current practice of using the tuberculin skin test for TB control and elimination.

Psychiatric length of stay determinants in a military medical center.

In this 2-year prospective study, 1040 consecutive admissions to an adult inpatient psychiatric unit of the United States Air Force's largest tertiary care medical center were assessed for length of stay (LOS) determinants. Twenty-one demographic, clinical, and diagnostic variables were examined for their effect on LOS. The median LOS at our medical center was 9 days and was similar to the national median LOS of 10 days for psychiatric units in general hospitals. We found several clinical, nonclinical, and diagnostic variables to be independent predictors of LOS, accounting for 31% of the variance, but that much of the variance remained unaccounted for. Primary DSM-III-R discharge diagnoses were helpful in distinguishing a nonoverlapping, short-stay group from a long-stay group. HCFA Diagnostic Related Group (DRG) estimates for LOS were assessed for their ability to predict LOS in our institution. With the exception of substance abuse/dependence disorders, DRGs consistently underestimated LOS. Previous studies using the coefficient of variation (COV) have demonstrated the inability of DRGs to distinguish homogeneous diagnostic groups. However, in this study, COV was helpful in differentiating a majority of DRGs by LOS despite the general underestimation of LOS by DRG. These results continue to support the inadequacy of the DRGs in determining equitable reimbursement and the difficulties in predicting psychiatric LOS despite the inclusion of clinical and nonclinical variables.

Selective interactions of concanavalin A at lipid membranes on the surface of an optical fiber.

An optical configuration was developed for sampling fluorescence coupled into an optical fiber from evanescent wave excitation of fluorescent materials at a lipid membrane on a quartz fiber surface. Selective interactions of pyrene-labelled concanavalin A located on a phosphatidyl choline-cholesterol lipid membrane with fluorescein isothiocyanate-labelled dextran in bulk aqueous solution were monitored by the intrinsic fluorescence sensing configuration. Monosialoganglioside, G(M1), was employed as a receptor in a phospholipid membrane on an optical fiber for selective measurement of pyrene-labelled concanavalin A in solution. Quantitative measurement was hindered by non-selective adsorption of cancanavalin A, but the potential for use of a lipid membrane in a fluorometric biosensor was established.

Inclusive K(S);(0)K(S);(0) resonance production in ep collisions at HERA.

Inclusive K_{S};{0}K_{S};{0} production in ep collisions at the DESY ep collider HERA was studied with the ZEUS detector using an integrated luminosity of 0.5 fb;{-1}. Enhancements in the mass spectrum were observed and are attributed to the production of f_{2}(1270)/a_{2};{0}(1320), f_{2};{'}(1525) and f_{0}(1710). Masses and widths were obtained using a fit which takes into account theoretical predictions based on SU(3) symmetry arguments, and are consistent with the Particle Data Group values. The f_{0}(1710) state, which has a mass consistent with a glueball candidate, was observed with a statistical significance of 5 standard deviations. However, if this state is the same as that seen in gammagamma-->K_{S};{0}K_{S};{0}, it is unlikely to be a pure glueball state.

Measurements of the exclusive decays of the upsilon(5S) to meson final states and improved B(s)* mass measurement.

Using 420 pb(-1) of data collected on the upsilon(5S) resonance with the CLEO III detector, we reconstruct B mesons in 25 exclusive decay channels to measure or set upper limits on the decay rate of upsilon(5S) into B meson final states. We measure the inclusive B cross section to be sigma(upsilon(5S) --> BB(X)) = (0.177 +/- 0.030 +/- 0.016) nb and make the first measurements of the production rates of sigma(upsilon(5S) --> B*B*) = (0.131 +/- 0.025 +/- 0.014) nb and sigma(upsilon(5S) --> BB*) = (0.043 +/- 0.016 +/- 0.006) nb, respectively. We set 90% confidence level limits of sigma(upsilon(5S) -->BB) < 0.038 nb, sigma(upsilon(5S) --> B(*)B(*)pi) < 0.055 nb and sigma(upsilon(5S) --> BBpipi) < 0.024 nb. We also extract the most precise value of the B(s)* mass to date, M(B(s)*) = (5411.7 +/- 1.6 +/- 0.6) MeV/c2.

Charmonium Decays of Y(4260), psi(4160), and psi(4040).

Using data collected with the CLEO detector operating at the CESR e+e- collider at sqrt[s]=3.97-4.26 GeV, we investigate 15 charmonium decay modes of the psi(4040), psi(4160), and Y(4260) resonances. We confirm, at 11 sigma significance, the BABAR Y(4260)-->pi+pi- J/psi discovery, make the first observation of Y(4260)--> pi(0)pi(0) J/psi (5.1 sigma), and find the first evidence for Y(4260)-->K+K- J/psi(3.7 sigma). We measure e+e- cross sections at sqrt[s]=4.26 GeV as sigma(pi+pi- J/psi)=58(+12)(-10)+/-4 pb, sigma(pi(0)pi(0) J/psi)=23(+12)(-8)+/-1 pb, and sigma(K+K- J/psi)=9(+9)(-5)+/-1 pb, in which the uncertainties are statistical and systematic, respectively. Upper limits are placed on other decay rates from all three resonances.

Proton gradient-driven import of the 16 kDa oxygen-evolving complex protein as the full precursor protein by isolated thylakoids.

The biogenesis of the lumenal 16 kDa protein of the photosynthetic oxygen-evolving complex was analysed using an assay for the import of proteins by isolated thylakoids. The precursor protein is imported with high efficiency in the light in both the presence and absence of stromal extract. Import is almost completely blocked in the dark or if the uncoupler nigericin is present in the light. The data indicate that transport across the thylakoid membrane is driven by a proton motive force in which the proton gradient is the dominant component, and that the full precursor protein can be transported across the thylakoid membrane without prior cleavage by the stromal processing peptidase.

The delta pH-driven, ATP-independent protein translocation mechanism in the chloroplast thylakoid membrane. Kinetics and energetics.

Previous studies have shown that proteins are transported across the chloroplast thylakoid membrane by two very different mechanisms, one of which requires stromal factors and ATP, whereas the other mechanism is ATP independent but completely reliant on the thylakoidal delta pH. We have examined the role of the delta pH in the latter mechanism by simultaneously monitoring the magnitude of delta pH (by 9-aminoacridine fluorescence quenching) and the rate of import of the 23-kDa photosystem II protein into isolated pea thylakoids. We show that protein import can take place, at low but significant rates, at very low values of delta pH (in the region of 1.2-1.4), and that plots of the rate of protein import against proton concentration gradient are probably hyperbolic in nature. There is no evidence for a threshold level of delta pH which is required to drive translocation of the 23-kDa protein. Addition of uncouplers midway during import incubations results in a rapid and complete inhibition of translocation, showing that the continuous presence of the delta pH is required for translocation to take place. During import into intact chloroplasts, the intermediate-size 23-kDa protein substrate for the thylakoidal protein transport machinery is found only in the stromal fraction at all values of delta pH, suggesting that the initial interaction with the machinery is relatively weak, reversible and delta pH-independent. We therefore propose that the delta pH is required for both the initiation and completion of translocation; these roles are in marked contrast to the roles of protonmotive force in mitochondrial and sec-dependent bacterial protein transport.

Effects of regular handling and blood sampling by wing vein puncture on the performance of broilers and pullets.

1. Broilers and pullets were regularly handled or bled by wing vein puncture over periods of 5 and 31 weeks respectively. 2. Neither procedure had any effect on body weight, food consumption nor on egg production, egg weight, shell quality or the percentage of non-marketable eggs.

Meningiomas, dicentric chromosomes, gliomas, and telomerase activity.

Lack of telomere maintenance during cell replication leads to telomere erosion and loss of function. This can result in telomere associations which probably cause the dicentric chromosomes seen in some tumour cells. One mechanism of telomere maintenance in dividing cells is the action of telomerase, a ribonucleoprotein enzyme that adds TTAGGG repeats onto telomeres and compensates for their shortening during cell division. Over 90 per cent of extracranial malignant neoplasms have been found to have telomerase activity. This study sought to determine if there was a relationship between absence of telomerase activity and presence of dicentric chromosomes in meningiomas and to what extent the other main group of central nervous system tumours, the gliomas, expressed telomerase activity. Telomerase activity was measured on 25 meningiomas and 29 gliomas. Four of the meningiomas were atypical variants and 11 were positive for dicentric chromosomes. Twenty-five of 29 gliomas were glioblastoma multiforme tumours. Measures were taken to ensure absence of false positives due to primer-dimer interaction and false negatives due to protein degradation or the presence of Taq polymerase inhibitors. All 25 meningiomas and the four low-grade gliomas (WHO grade II) were telomerase activity-negative. Seven (28 per cent) of the 25 glioblastoma multiforme tumours showed telomerase activity. The absence of telomerase activity in meningiomas and the high frequency of telomere associations support the hypothesis that these tumours are benign, transformed but pre-crisis. The relatively low frequency of telomerase activity in the malignant glioblastoma multiforme suggests that most of these tumours may have other mechanisms of telomere maintenance and that the potentially therapeutic telomerase inhibitors will not be of great value in the future management of the majority of patients suffering from these tumours.