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PubMed1991

[Negative schizophrenic symptoms and their detection].

The paper deals with the methodology and methods for the assessment of negative (or minus) symptoms of schizophrenia. Two approaches are distinguished: Rating scales and objective methods. Statistical analysis of symptom-ratings with comprehensive psychiatric assessment systems (eg Brief Psychiatric Rating Scale, Present State Examination) have supported the distinction between positive and negative symptoms. Therefore special scales have been constructed for the purpose of differentially assessing these symptoms, eg the Scale for the Assessment of Negative/Positive Symptoms (SANS, SAPS) or the Positive and Negative Syndrome Scale (PANSS). Relations to self-rating scales for subjectively experienced deficits and basic symptoms in schizophrenic patients are pointed out, eg Frankfurt Complaint Inventory (FBFB) or Scale for Emotional Blunting (SEB). Psychometric properties of the rating scales for negative symptoms have to be improved, especially their validity often is not shown. Objective--rater-independent--assessment methods for negative symptoms are rare and being experimentally developed. Some symptoms may be assessed by psychological tests as deficiencies in psychological functioning, other by laboratory based (automated) analysis of behaviour, eg speech or facial actions. These investigations increase the reliability of the symptom-assessments. The validation of the methods involves etiological, prognostic and treatment implications of the negative symptoms. Current results suggest, that the negative syndrome of schizophrenia is a heterogenous construct, which has further to be differentiated.

Affective Symptoms↗
PubMed1987

Determination of protein degradation rates using a rumen in vitro system containing inhibitors of microbial nitrogen metabolism.

1. A previously reported rumen in vitro system (Broderick, 1978) was modified to include chloramphenicol (CAP) with hydrazine sulphate (HS) to give quantitative recovery of protein breakdown products. Degradation rates were determined by regression v. time of log proportion remaining undegraded (computed by subtracting from added nitrogen the amount of N recovered as ammonia and amino acids). Concentrations of reagents giving optimal N recoveries and estimated degradation rates for casein and expeller soya-bean-meal (SBM) were: 1.0 mM-HS, 30 micrograms CAP/ml, 2.0 mM-mercaptoethanol, 3.3 mg maltose/ml, when protein was added at 0.125 mg N/ml. 2. Digestion of azo-casein and azo-albumin, solubilization of radioactivity from 14C-labelled casein, ovalbumin and bovine serum albumin (BSA), and hydrolysis of benzoyl-L-tyrosine p-nitroanilide and benzoyl-L-arginine p-nitroanilide were not significantly decreased by HS and CAP. This suggests that the inhibitors did not reduce microbial proteolysis. 3. Mean fractional degradation rates (/h) were: 0.395 casein, 0.135 BSA, 0.159 solvent-SBM, 0.045 expeller-SBM, 0.061 meat meal, 0.070 lucerne (Medicago sativa) hay. Extents of protein escape, estimated assuming rumen passage of 0.06/h, were 13, 28, 56 and 40% for casein, solvent-SBM, expeller-SBM and lucerne hay respectively. This method appears more reliable for assessing rumen degradability than buffer N solubility and protein digestibility with ficin protease. 4. Azo-dye treatment slowed the rate of casein degradation, measured by ammonia plus amino acid release, but did not alter digestion of BSA. 5. The validity of the inhibitor in vitro method for estimating protein degradability, as well as potential problems in its application, are discussed. The complete procedure may be limited to laboratories with automated analytical equipment, but a simplified version of the method may be more generally applicable.

Animals↗
PubMed1992

A test for the measurement of pulmonary diffusion capacity during high-intensity exercise.

The most commonly used technique for the measurement of pulmonary diffusing capacity (DL) is the single-breath hold technique requiring a 10-s breath-hold after the maximal inspiration of carbon monoxide (0.3% CO) and helium (10% He). To measure pulmonary diffusing capacity in our experiments, we had the added advantage of the use of the Gould Pulmonary Function Laboratory that automates the collection and recording of data and the calibration of equipment for each test. However, this technique, DL(CO), is difficult to use during exercise of moderate or elevated intensity because of the lengthy breath-hold. Thus, the purpose of the present study was to compare DL(CO) with 3-s and 5-s breath-holds to a 10-s breath-hold at rest and during moderate and intense exercise in 14 subjects. As expected, an increase in the DL(CO) was observed during moderate and intense exercise when compared to resting values (45.7 +/- 10.0 and 53.0 +/- 7.6 vs 32.1 +/- 7.7 ml CO min-1 mmHg-1). No difference was observed between values for DL(CO) measured at varying breath-hold times at rest (3 s: 32.9 +/- 7.4; 5 s: 32.0 +/- 7.5; 10 s: 31.4 +/- 8.2 ml CO min-1 mmHg-1) or during moderate exercise (3 s: 45.9 +/- 10.1; 5 s: 45.9 +/- 10.6; 10 s: 45.2 +/- 10.4 ml CO min-1 mmHg-1) or intense exercise (3 s: 52.1 +/- 8.3; 5 s: 54.3 +/- 9.3; 10 s: 52.6 +/- 5.2 ml CO min-1 mmHg-1). Reliability coefficients indicated that the use of a 3-s breath-hold was appropriate.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed2006

Development of an impregnated reagent and automation of solid-phase analytical derivatization for carbonyls: proof of principle.

This study undertakes reduction of scale and automation of a solid-phase analytical derivatization of carbonyls with 2,4-dinitrophenylhyrazine on a styrene-divinylbenzene resin (XAD-2). Three processes are tested. In the batch process, an aqueous phase consisting of 50 microL of sample and 150 microL of reagent solution is contacted with 6 mg XAD-2 by shaking. An impregnated reagent consisting of 2,4-dinitrophenylhydrazine hydrochloride (DNPH) deposited on XAD-2 enables two additional processes. In-vial derivatization with an impregnated reagent requires shaking 50 microL of sample with 6 mg of the impregnated reagent and reduced the reaction time from 10 to 5 min. The third process involves packing impregnated reagent a mini-column and flowing 50 microL of sample through under positive pressure supplied by a Harvard Pump. This reduces sample preparation time to 1 min. Studies are conducted with worst-case model analytes: butanone, 2-pentanone, and malonyldialdehyde. The carbonyl of the two ketones is hindered, and, thus, these two compounds react very slowly with DNPH in aqueous solution. Malonyldialdehyde is highly water soluble, and it does not react in aqueous phase but also would not sorb onto XAD-2 for reaction. Nevertheless, derivatization/extraction of all model compounds any of the three procedures result in reproducible and high yields.

Automation, Laboratory↗
PubMed2004

Applying apoB to the diagnosis and therapy of the atherogenic dyslipoproteinemias: a clinical diagnostic algorithm.

PURPOSE OF REVIEW: The first objective is to present the most recent evidence relating to the efficacy of apolipoprotein B as a diagnostic index of the risk of vascular disease and a therapeutic target for statin therapy. The second is to present a diagnostic algorithm for the apolipoprotein B100 dyslipidemias based on triglyceride and apoB. RECENT FINDINGS: The results from several recent prospective epidemiological studies demonstrate apoB to be superior to any of the cholesterol indices to estimate the risk of vascular disease. Similarly, the results of several of the major statin clinical trials demonstrate that apoB is a more adequate index of the adequacy of statin therapy than any of the cholesterol indices. Recent studies of lipoprotein subclass distribution in subjects with familial combined hyperlipidemia are reviewed. They demonstrate the limitations of the original lipid-based criteria and point to the necessity of using apoB as a fundamental diagnostic criterion for the disorder. A diagnostic algorithm for an apoB100 atherogenic dyslipoproteinemias is presented and the limitations of the lipid-based system described. SUMMARY: The evidence supporting the clinical use of apoB is solid, its measurement is standardized, and automated, inexpensive laboratory testing could easily be widely available. However, clinical benefit will only follow clinical application.

Algorithms↗
PubMed2004

The Pseudomonas aeruginosa PA01 gene collection.

Pseudomonas aeruginosa, a common inhabitant of soil and water, is an opportunistic pathogen of growing clinical relevance. Its genome, one of the largest among bacteria [5570 open reading frames (ORFs)] approaches that of simple eukaryotes. We have constructed a comprehensive gene collection for this organism utilizing the annotated genome of P. aeruginosa PA01 and a highly automated and laboratory information management system (LIMS)-supported production line. All the individual ORFs have been successfully PCR-amplified and cloned into a recombination-based cloning system. We have isolated and archived four independent isolates of each individual ORF. Full sequence analysis of the first isolate for one-third of the ORFs in the collection has been completed. We used two sets of genes from this repository for high-throughput expression and purification of recombinant proteins in different systems. The purified proteins have been used to set up biochemical and immunological assays directed towards characterization of histidine kinases and identification of bacterial proteins involved in the immune response of cystic fibrosis patients. This gene repository provides a powerful tool for proteome- and genome-scale research of this organism, and the strategies adopted to generate this repository serve as a model for building clone sets for other bacteria.

Bacterial Proteins↗
PubMed1977

Electrical impedance measurements in the reading and monitoring of broth dilution susceptibility tests.

Electrical impedance changes in the medium were studied during traditional broth dilution tests. Tests involved clinical isolates of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Enterobacter sp., Klebsiella, and enterococcus and ten antibiotics. Minimal inhibitory concentration (MIC) values from overnight visual readings were compared with MIC values determined from electrical impedance end points, using inocula of 10(6) organisms per ml. Ninety-three percent of the results were within one twofold dilution of each other. When the impedance end point was determined at 6 h, this correlation was lowered to 34%. By increasing the initial electrical impedance inoculum 10-fold, the correlation between the 6-h impedance MIC and the overnight visual MIC was improved to 74%. Ampicillin, tetracycline, and polymyxin E results accounted for most discrepancies. Continuous monitoring of impedance changes suggested that better correspondence could be obtained by adjusting the end point criteria for the 6-h impedance MIC with different antibiotics. Electrical impedance methods for reading bacterial end points in automated clinical laboratory instruments appear promising.

Culture Media↗
PubMed1996

Performance of fungal blood cultures by using the Isolator collection system: is it cost-effective?

Enhanced recovery of fungal isolates from blood by using the Isolator system has been reported previously. We examined bacterial and fungal blood cultures during a 14-month period to determine if this enhanced recovery required a separate fungal culture and to determine the differential utility between a fungal blood culture and a routine bacterial culture. During this period, 84 of 5,196 (1.6%) fungal blood cultures and 170 of 25,702 (0.6%) bacterial blood cultures were positive for yeast or filamentous fungi. Thirty-seven positive fungal cultures, simultaneously collected, had correspondingly positive bacterial cultures. An additional 15 positive fungal cultures yielded isolates that had either been previously recovered from a bacterial culture or were recovered from a bacterial culture collected within 48 h. Of the 32 unpaired fungal cultures remaining, 5 were Candida albicans whose unique isolation was believed to be the result of specimen sampling variance rather than any enhanced recovery characteristics of fungal culture methods. Examination of patient data relating to the 27 remaining isolates (24 patients episodes) showed that only five fungal blood cultures (0.096% of all collected) had any impact on patient therapy decisions, and one of these was judged to be the cause of unnecessary therapy. Our data suggest that separate fungal cultures of blood are not cost-effective for those laboratories using the Isolator for routine blood cultures and furthermore may not be cost-effective for laboratories using automated broth systems that are comparable to the Isolator in recovery of fungi.

Adult↗
PubMed1996

Vitek system antimicrobial susceptibility testing of O1, O139, and non-O1 Vibrio cholerae.

Vibrio cholerae causes epidemic diarrhea throughout the world. Fluid replacement is the primary therapy for cholera; however, high mortality rates often necessitate the use of antibiotics. V. cholerae, like most bacteria, has developed resistance to some antibiotics. In the early 1990s a new serotype strain, Bengal 0139, began a new wave of cholera epidemics. Bengal isolates showed unique trends in antimicrobial resistance. Many clinical laboratories use automated antibiotic susceptibility testing for V. cholerae. It is important to know if automated susceptibility test results for V. cholerae coincide with reported trends in antibiotic susceptibility. In the present study, we used the Vitek automated susceptibility system to determine the susceptibilities of 79 V. cholerae O1 isolates, 100 O139 isolates, and 112 non-O1 isolates. Vitek susceptibilities for V. cholerae showed a good correlation with preestablished epidemiological data. Although the new O139 serogroup showed a trend of increased resistance to trimethoprim-sulfamethoxazole and nitrofurantoin, it was more susceptible to ampicillin than previous serogroup O1 and non-O1 strains. Regardless of serogroup, > or = 98% of the V. cholerae isolates tested were susceptible to most antibiotics tested by us. It is important to continue susceptibility testing of all new isolates of V. cholerae because of emerging resistant strains. However, V. cholerae remains susceptible to most of the available antibiotics.

Cholera↗
PubMed2005

Reduced PCR sensitivity due to impaired DNA recovery with the MagNA Pure LC total nucleic acid isolation kit.

The increasing demand for molecular diagnostics in clinical microbiology laboratories necessitates automated sample processing. In the present study, we evaluated the performance of the MagNA Pure LC total nucleic acid isolation kit (M extraction) in comparison with the manual method (Si extraction) according to Boom et al. (R. Boom, C. J. A. Sol, M. M. M. Salimans, C. L. Jansen, P. M. Wertheim-van Dillen, and J. van der Noordaa, J. Clin. Microbiol. 28:495-503, 1990) for the detection of viral DNA by competitive quantitative PCR. Reconstruction experiments with HindIII-digested phage lambda DNA and HaeIII-digested phiX174 DNA showed that the recovery of DNA from phosphate-buffered saline, cerebrospinal fluid, EDTA-anticoagulated plasma, and EDTA-anticoagulated whole blood by M extraction is, on average, 6.6-fold lower compared to Si extraction. PCR signals of spiked PCR control DNAs for Epstein-Barr virus and varicella-zoster virus were also between 1.9- and 14.2-fold lower after M extraction compared to Si extraction, also suggesting impaired DNA recovery. M extraction of spiked cytomegalovirus strain AD 169 in whole blood showed a 5- to 10-fold reduction in PCR sensitivity compared to Si extraction. This reduction of PCR sensitivity was also observed when clinical whole blood samples were processed by M extraction. Before implementing M extraction, the clinical consequences of the reduced recovery should first be considered, especially when maximal sensitivity is required.

Cytomegalovirus↗
PubMed2003

Evaluation of abnormal liver function tests.

Interpretation of abnormalities in liver function tests is a common problem faced by clinicians. This has become more common with the introduction of automated routine laboratory testing. Not all persons with one or more abnormalities in these tests actually have liver disease. The various biochemical tests, their pathophysiology, and an approach to the interpretation of abnormal liver function tests are discussed in this review.

Alkaline Phosphatase↗
PubMed1991

A comparison of five devices for the bedside monitoring of heparin therapy.

Five instruments were tested for their capacity to monitor heparin therapy on whole blood at the bedside. The instruments were 512 Coagulation Monitor (Ciba-Corning), Thrombotrack (Nycomed), Automated Coagulation Timer (Hemotec), Hemochron-ACT and Hemochron-APTT (International Technidyne Corporation). Fifty subjects with various levels of heparinisation were tested on each instrument and were also assayed for antithrombin III, fibrinogen, haematocrit, platelet count and plasma heparin level. The results were compared with a reference APTT performed on the Automated Coagulation Laboratory 300R (Instrumentation Laboratories). The Hemochron-ACT correlated least well. The Hemotec and Thrombotrack were unsuitable in a clinical setting because of pipetting requirements, although the Thrombotrack did correlate well with the reference parameters. The 512 Coagulation Monitor was the simplest to use, but its maximum response corresponded to the midpoint of the reference APTT therapeutic range. The Hemochron-APTT was simple to use, had an adequate response range and correlated well with reference parameters.

Blood Coagulation Tests↗
PubMed2006

An automated method for rapid identification of putative gene family members in plants.

BACKGROUND: Gene duplication events have played a significant role in genome evolution, particularly in plants. Exhaustive searches for all members of a known gene family as well as the identification of new gene families has become increasingly important. Subfunctionalization via changes in regulatory sequences following duplication (adaptive selection) appears to be a common mechanism of evolution in plants and can be accompanied by purifying selection on the coding region. Such negative selection can be detected by a bias toward synonymous over nonsynonymous substitutions. However, the process of identifying this bias requires many steps usually employing several different software programs. We have simplified the process and significantly shortened the time required by condensing many steps into a few scripts or programs to rapidly identify putative gene family members beginning with a single query sequence. RESULTS: In this report we 1) describe the software tools (SimESTs, PCAT, and SCAT) developed to automate the gene family identification, 2) demonstrate the validity of the method by correctly identifying 3 of 4 PAL gene family members from Arabidopsis using EST data alone, 3) identify 2 to 6 CAD gene family members from Glycine max (previously unidentified), and 4) identify 2 members of a putative Glycine max gene family previously unidentified in any plant species. CONCLUSION: Gene families in plants, particularly that subset where purifying selection has occurred in the coding region, can be identified quickly and easily by integrating our software tools and commonly available contig assembly and ORF identification programs.

Arabidopsis↗