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PubMed2003

Impact of introducing quality control/quality assurance (QC/QA) guidelines in respiratory specimen processing.

OBJECTIVES: To assess the impact of the introduction of a new quality control/quality assurance (QA/QC) protocol on the processing and reporting of respiratory specimens. METHODS: After implementation of guidelines for processing respiratory specimens, an investigation was carried out over a six-month period on 200 specimens, 105 sputa, and 95 deep tracheal aspirates (DTAs), assessed blindly by two independent investigators. Data regarding disagreement were arranged into two subgroups. A minor disagreement was defined as a difference in the two assessments of < 10 or 10-25 for white blood cell (WBC) or squamous epithelial cell (SEC) counts. A major disagreement was defined as one assessor reporting < 10 and the other > 25 for either WBC count or SEC count, or one assessor reporting the specimen as non-assessable, or both assessors having a minor disagreement in both the WBC count and the SEC count. RESULTS: Agreement was obtained on 111 samples. For 45 specimens, a major disagreement was documented, and in 44 cases, a minor disagreement was recorded, WBC being the most common cause of divergence. Data for sputa and DTAs were examined separately: of 45 major disagreements, 64.4% were observed for DTAs, while minor disagreements were recorded mostly for sputa. The role of the settings in which samples were taken in affecting quality was studied. Among the 105 sputa, 27 were from Community Health Centers (CHCs) and 78 from hospitalized patients. Agreement between the two observers was obtained in 48.1% of CHC cases versus 60.2% of hospital samples. To investigate how many of the rejected samples presented WBC > 25 suggestive of infection, we looked at the 107 samples rejected during the six-month period, grouped according to the suspected diagnosis. The highest number of rejected samples falls in the category of unrelated (non-respiratory) diagnosis, and clinical suspicion is not helpful in Gram stain interpretation. The annual saving (not culturing, not testing, and not treating) derived from this simple QC procedure totals about 5000 Euro. CONCLUSIONS: Standardization in microscopic screening of respiratory samples is difficult to achieve. Criteria for rejection must be adapted to local conditions after discussion with clinicians to increase their compliance with the newly introduced guidelines and to avoid sending unnecessary specimens. The effects on patient management and cost control are significant.

Guidelines as Topic↗

PubMedDate not supplied

Red cell phenotyping using hexadimethrine bromide (Polybrene) in a microplate system.

To provide rapidly phenotyped units of blood, we adapted the hexadimethrine bromide (Polybrene) technique to microplate technology. Pilot samples from 282 donor units were phenotyped for antigens in the Rh, Kidd, Kell, Duffy, and Ss systems with a standard tube-testing method and a Polybrene-microplate (P-MP) technique. Diluted antisera and a 1 percent red cell suspension were used to give P-MP reactions that were accurate and easy to interpret. One microplate, containing 96 tests, was prepared and read within 15 minutes for P-MP tests yielding direct agglutination (Rh, Jkb, Fya, Fyb), or within 19 minutes for P-MP tests requiring an antiglobulin phase (Jka, K, S, s). No false-positive results were found. No false-negative reactions were found in typing for Rh, K, S, s, or Jka antigens. Phenotyping for Jkb and Duffy antigens gave a false-negative rate of less than 0.015. Considerable savings in reagents were obtained through microplate miniaturization and through the enhancement of apparent antibody avidity in the Polybrene/low ionic medium which permitted dilution of reagent antisera. The P-MP technique affords a rapid, accurate, simple, and inexpensive means of phenotyping large numbers of donor units.

Blood Group Antigens↗

PubMed2001

Information needs of patients with cancer: results from a large study in UK cancer centres.

As part of a multi-centred study evaluating a communication skills training model for clinicians, we collected information preferences using an adaptation of Cassileth's Information Needs questionnaire from a heterogeneous sample of 2331 patients. Results showed that 87% (2027) wanted all possible information, both good and bad news and 98% (2203) preferred to know whether or not their illness was cancer. Cross tabulation of responses revealed no significant differences in information preferences for tumour site or treatment aims but did show an effect of age and sex. The few 58/440 (13.2%) patients who stated that in general they preferred to leave disclosure of details up to the doctor, tended to be older patients more than 70 years of age (chi square = 26.01, df = 2, P< 0.0001), although paradoxically they still wanted to know certain specific details. In comparison to men women preferred to know the specific name of the illness (chi square = 4.9, df = 1, P< 0.02) and what were all the possible treatments (chi square = 8.26, df = 1, P< 0.004). The results from this very large sample provide conclusive evidence that the vast majority of patients with cancer want a great deal of specific information concerning their illness and treatment. Failure to disclose such information on the grounds that significant numbers of patients prefer not to know is untenable.

Adult↗

PubMed2000

Rhythms of pituitary-adrenal activity during sleep in patients with Cushing's disease.

Previous studies have indicated a dependence of nocturnal pituitary-adrenal secretory activity on central nervous sleep processes in healthy humans: Under normal physiological conditions the release of ACTH/cortisol is inhibited during early sleep and becomes entrained to periods of NonREM sleep during late sleep. Here, we compared nocturnal dynamics in plasma concentrations of ACTH/cortisol in 7 patients with Cushing's disease with those of 7 healthy controls matched in age and sex with the patients. The patients in part were repeatedly tested. The total of 13 nights is composed of 7 nights of hyperpulsatile secretion pattern (5 patients) and 6 nights from hypopulsatile secretion pattern (4 patients). After an adaptation night polysomnographic sleep recordings were obtained and blood was sampled every 15 min between 23.00 and 7.00 h. Controls displayed the typical minimum in ACTH/cortisol concentrations during the early part of the night and maximum concentrations during the late part of the night, whereas ACTH/cortisol levels of Cushing patients indicated a relatively constant elevated pituitary-adrenal activity throughout the night, lacking any circadian variation. Autocorrelation functions revealed the presence of cortisol secretory rhythms with a similar period length in healthy controls (155.6+/-17.4 min) and patients with a hyperpulsatile pattern (142.4+/-6.6 min). In patients displaying hypopulsatility, no significant rhythmicity was observed. However, regardless of the type of secretory pulsatility, adrenal secretory activity started predominantly during periods of NonREM sleep (p<0.01) in healthy controls as well as in patients with Cushing's disease. This data indicates that the normal nocturnal circadian oscillation of pituitary-adrenal activity is absent in Cushing patients, whereas a link between pituitary-adrenal activity and ultradian rhythms of sleep appears to be preserved.

Activity Cycles↗

PubMed2004

[The Osnabruck Work Capabilities Profile (O-AFP) for persons with psychiatric illness: concept, development, and testing in schizophrenic patients].

The goal of this paper is presentation of an inventory for the assessment of basic capabilities underlying work and occupational behaviour of psychiatric patients in the course of their treatment and rehabilitation. The "Osnabruck Work Capabilities Profile" (O-AFP) is an instrument filled in by staff, which consists of three scales comprising ten items each, measuring "Learning Ability", "Social Communication Ability" and "Adaptation". In the study presented, these scales were confirmed by factor analyses of data from 194 schizophrenic patients. Test analyses yielded good item and scale characteristics. The only exception to this are elevated item difficulty scores of the third scale (adaptation). This was attributed to specific effects of selecting the sample of patients. Furthermore, low or insignificant correlations with the patients' symptom scores indicate the discriminant validity of the scales. It is concluded that the O-AFP is well suited for application with different forms of occupational therapy.

Adult↗

PubMed1973

Intervent dilution chromatography: concept for separation of strongly interacting macromolecules.

Intervent dilution chromatography separates interacting macromolecules by subjecting them to a dynamic environment in which the association constant is continuously varied. The dynamic environment is produced by using the sieving properties of a gel to repeatedly propel the molecular complex across an intervent boundary. Behind the boundary, at high intervent concentrations, the complex dissociates; ahead of the boundary, the component molecules are separated by adsorption processes. By selective adjustment of the intervent composition of the sample and the conditions of column equilibration, the process is adapted to a particular need. This report describes the chromatographic concept and shows how parameters are adjusted to obtain the desired separation. In this study a particularly difficult separation, i.e., the separation of ribosomal proteins from ribosomal RNA, is chosen to illustrate the power of the procedure.

Chromatography, Gel↗

PubMed1993

Gene replacement in Halobacterium halobium and expression of bacteriorhodopsin mutants.

A gene replacement method has been developed to express bacteriorhodopsin mutants in the archaeon Halobacterium halobium. Selectable plasmids carrying the bacterioopsin gene (bop) were integrated at the chromosomal bop locus of H. halobium. Under nonselective conditions, recombinants were isolated that had lost the integrated plasmid and retained a single chromosomal copy of the bop gene. This approach was used to construct a bop deletion strain. By using this strain, recombinants were obtained that express wild-type bacteriorhodopsin and mutants known to be defective in proton translocation. The expressed proteins were purified in a membrane fraction similar to purple membrane and were characterized in this form. UV/visible spectra of dark- and light-adapted bacteriorhodopsin from wild-type and Asp-96 mutants were identical to those of purple membrane. Arg-82, Asp-85, and Asp-212 mutants had 10- to 50-nm red shifts in their absorption maxima and showed altered light adaptation. The proton translocation activity of the wild-type samples and purple membrane was comparable, whereas the mutants had 0-60% of wild-type activity. These results support earlier studies of proton translocation mutants expressed in Escherichia coli.

Amino Acid Sequence↗

PubMed2001

Effect of starch application into the proximal duodenum of ruminants on starch digestibility in the small and total intestine.

Four Slovakian Black-and-white bulls (LW 410 +/- 12 kg; Exp. 1) and four Slovakian Black-and-white non lactating dairy cows (LW 475 +/- 14 kg; Exp. 2) with permanent ruminal cannulas, duodenal T-cannulas and ileal re-entrant cannulas were used in a 4 x 4 Latin square design to determine the postruminal capacity of starch digestion. In Exp. 1 bulls received 5.4 kg DM from corn silage and 3.6 kg DM from alfalfa hay, in Exp. 2 cows consumed only 2.1 kg DM corn silage and 1.9 kg DM alfalfa hay. Additionally, either 750 or 1500 g (Exp. 1) or resp. 1000 or 2000 g (Exp. 2) gelatinized corn or wheat starch per animal and day were applied as pulse doses or as infusion into the proximal duodenum. In both experiments the duodenal and ileal nutrient flow, as well as the faecal excretion without starch application, were measured in a pre-period. After starting starch application ileal digesta and faeces were sampled over 120 h after 9 or 23 days of adaptation respectively. Cr2O3 was used as a flow marker. It was shown, that the capacity of starch utilisation in the small intestine was limited. The effect of different doses of bypass-starch was more pronounced than the effect of different starch sources. Starch digestibility decreased with increasing amounts of starch in the intestine (Exp. 1: corn starch: from 74.3 to 68.0%, P < 0.001; wheat starch: from 76.7 to 67.4%, P < 0.001; Exp. 2: corn starch: from 71.4 to 50.3%. P < 0.001; wheat starch: from 73.8 to 53.1%, P < 0.001). Corn starch was 0.6 to 2.4% units (P < 0.05) and 2.4 to 2.8% units (P < 0.001) less digested than wheat starch in Exp. 1 and Exp. 2, respectively.

Animal Feed↗

PubMed2002

Reassessing the possibility of life on venus: proposal for an astrobiology mission.

With their similar size, chemical composition, and distance from the Sun, Venus and Earth may have shared a similar early history. Though surface conditions on Venus are now too extreme for life as we know it, it likely had abundant water and favorable conditions for life when the Sun was fainter early in the Solar System. Given the persistence of life under stabilizing selection in static environments, it is possible that life could exist in restricted environmental niches, where it may have retreated after conditions on the surface became untenable. High-pressure subsurface habitats with water in the supercritical liquid state could be a potential refugium, as could be the zone of dense cloud cover where thermoacidophilic life might have retreated. Technology based on the Stardust Mission to collect comet particles could readily be adapted for a pass through the appropriate cloud layer for sample collection and return to Earth.

Earth, Planet↗

PubMed2006

A screening system for detecting genetic risk markers of type 1 diabetes in dried blood spots.

BACKGROUND: Certain alleles among the genes that code for the human leukocyte antigens (HLA) confer susceptibility or resistance to the development of autoimmunity that causes type 1 diabetes (T1D). A number of ongoing diabetes research studies analyze dried blood spots (DBS) from newborn infants for HLA-D alleles to identify higher-risk children as early as possible. A commercially available assay to detect such alleles has recently become available using a dissociation- enhanced lanthanide fluorescence system found in many newborn screening laboratories. METHODS: We adapted the system for use with DBS and improved the sample set-up for greater efficiency. We also developed an independent system for data analysis based on a spreadsheet program. These modifications were applied to HLA-DQB1 gene locus (DQB) analysis of 117 newborn DBS, and the results we obtained were compared with independent reference values. RESULTS: Our assay modifications and independent data analysis improved sample throughput and result tabulation. DQB results from the modified assay were consistent with the reference values in all but one sample, which showed a partial match. CONCLUSIONS: The modifications described here make this commercially available assay more suitable for high-throughput applications such as newborn screening. Our results show that this system allows highly accurate detection of DQB alleles that influence T1D risk.

Blood Chemical Analysis↗

PubMed2000

Adaptation of FACE methodology for microanalysis of total hyaluronan and chondroitin sulfate composition from cartilage.

Protocols for analyzing the fine structure of hyaluronan and chondroitin sulfate using fluorophore-assisted carbohydrate electrophoresis of 2-aminoacridone-derivatized hyaluronidase/chondroitinase digestion products were adapted for direct analysis of previously characterized cartilage-derived samples. The chondroitin sulfate disaccharide compositions for fetal and 68 year human aggrecan from FACE analyses were DeltaDi4S (50%), DeltaDi6S (43%), and DeltaDi0S (7%); and DeltaDi4S (3%), DeltaDi6S (96%), and DeltaDi0S (1%), respectively. The nonreducing terminal structures included predominantly 4S-galNAc with minor amounts of 6S-galNAc and Di6S for the fetal aggrecan sample and, in addition, included 4,6S-galNAc in the 68 year aggrecan sample. FACE analysis of a proteinase K digest of rat chondrosarcoma tissue gave an internal disaccharide composition for its chondroitin sulfate chains of DeltaDi0S (7%) and DeltaDi4S (93%) with no DeltaDi6S and DeltaDi4, 6S detected, while DeltaDiHA from hyaluronan was 5% of the total. Analysis of nonreducing terminal structures indicated the presence of 4S-galNAc (51%), galNAc (27%), and Di4S (22%) with no 4,6S-galNAc or Di6S detected. Unexpectedly, FACE analysis detected putative linkage oligosaccharide structures from the chondroitin sulfate chains including both unsulfated (85%) and 4-sulfated (15%) linkage oligosaccharides. Finally, the number averaged chain length estimated from the ratio of the molar fluorescence of the Deltadisaccharides to that of the nonreducing termini or the linkage oligosaccharide structures was calculated as approximately 16 kDa. A tissue glucose concentration of 0.72 g/l was also measured. These results for both samples as determined by FACE analysis were similar to results previously reported, using more labor and time intensive procedures, validating the FACE protocols.

Aged↗

PubMed2002

Use of the regression analysis technique to determine the true phosphorus digestibility and the endogenous phosphorus output associated with corn in growing pigs.

The objectives of this study were to determine true phosphorus (P) digestibility and the endogenous P output associated with corn for growing pigs using the regression analysis technique. Four barrows, average initial body weight of 25 kg, were fitted with a T-cannula and fed four diets according to a 4 x 4 Latin square design. Four cornstarch-based diets, containing four levels of P at 0.7, 1.5, 2.2 and 2.8 g/kg dry matter intake (DMI), were formulated from corn. Each experimental period comprised 8 d with 4-d adaptation and 4-d collection of ileal digesta and fecal samples. The apparent ileal and fecal P digestibility values in corn were affected (P < 0.05) by P contents in the assay diets. The apparent ileal and fecal P digestibility values increased from -51.0 to 33.3% and from -41.4 to 39.1%, respectively, as P content increased from 0.7 to 2.8 g/kg DMI. Linear relationships (P < 0.05), expressed as g/kg DMI, between the apparent ileal and fecal digestible P and the total intake of dietary P, suggested that true P digestibility and the endogenous P outputs associated with corn can be determined by the regression analysis technique. There were no differences (P > 0.05) in true P digestibility values (54.0 +/- 6.5 vs. 59.8 +/- 8.5%) and the endogenous P outputs (0.693 +/- 0.128 vs. 0.670 +/- 0.160 g/kg DMI) between the ileal and the fecal determinations. The endogenous fecal P output represented 12.3% and 25.8% of the daily total and available P requirements in growing pigs recommended by the National Research Council in 1998. The present literature data of apparent digestibility and availability underestimate the true digestive utilization of P in corn for growing pigs by approximately 35%. Current diet formulation on the bases of total, apparent P digestibility and availability values in corn inevitably leads to P overfeeding and excessive P excretion in pigs.

Animals↗

PubMed2000

An improved design for vacuum tube drains.

Mini vacuum drains can be helpful in reconstructive surgery. The drains may be assembled using a scalp needle and a vacuum tube. The addition of the adapter that is used with the vacuum tubes for blood sampling improves the ease of use and practicality of these drains and increases their safety.

Equipment Design↗

PubMed1993

Mother-adolescent agreement on the symptoms and diagnoses of adolescent depression and conduct disorders.

OBJECTIVE: To determine rates and correlates of mother-adolescent agreement on the symptoms and diagnoses of depression and conduct disorders. METHOD: The Schedule for Affective Disorders and Schizophrenia in School Age Children was administered to a community sample of 460 adolescents and their mothers. Race, gender, family adaptability and cohesion, and living arrangement were evaluated as predictors of agreement. RESULTS: Mother-adolescent agreement was low (kappa < 0.40) for all diagnoses and symptoms with these exceptions: feeling tired (kappa = 0.40), inattention (kappa = 0.50), vandalism (kappa = 0.50), and chronic violation of rules (kappa = 0.72). Cohesion and adaptability were associated with mother-adolescent agreement on the symptoms and diagnosis of depression. This association was not demonstrated as consistently for agreement on the symptoms and diagnosis of conduct disorders. CONCLUSION: Study findings underscore the necessity of collecting data from both a parent and the adolescent when assessing an adolescent's psychological status.

Adolescent↗

PubMed2001

Predicting dimensions of personality disorder from domains and facets of the Five-Factor Model.

We compared the utility of several trait models for describing personality disorder in a heterogeneous clinical sample (N = 94). Participants completed the Schedule for Nonadaptive and Adaptive Personality (SNAP; Clark, 1993b), a self-report measure that assesses traits relevant to personality disorder, and two measures of the Five-Factor Model: the Revised NEO Personality Inventory (NEO-PI-R; Costa and McCrae, 1992) and the Big Five Inventory (BFI; John, Donahue, & Kentle, 1991). Regression analyses indicated substantial overlap between the SNAP scales and the NEO-PI-R facets. In addition, use of the NEO-PI-R facets afforded substantial improvement over the Five-Factor Model domains in predicting interview-based ratings of DSM-IV personality disorder (American Psychiatric Association, 1994), such that the NEO facets and the SNAP scales demonstrated roughly equivalent levels of predictive power. Results support assessment of the full range of NEO-PI-R facets over the Five-Factor Model domains for both research and clinical use.

Adaptation, Psychological↗

PubMed1986

Enzyme-linked immunofiltration assay (ELIFA) for the detection of specific antibodies (IgG-IgM-IgA-IgE) in farmer's lung disease.

Until now the detection of chymotrypsic or polysaccharidic arcs among precipitating systems indicated a positive diagnosis of Farmer's lung disease (FLD). It was shown that the occurrence of chymotrypsic activity is not absolutely correlated with the presence of clinical disease. The presence of different classes of specific precipitating antibodies was looked for and correlated with clinical symptoms in an attempt to find additional positive criteria for the diagnosis of FLD. The enzyme-linked immunofiltration assay (ELIFA) was adapted to detect the different specific antibody classes. 1400 serum samples were studied from 1350 farmers exposed to mouldy hay. Of 1358 samples examined from asymptomatic subjects tested as part of a systematic screening of the profession, 1356 had either no precipitins or have precipitins detected solely against non-chymotrypsic and non-polysaccharidic antigens. Ten specimens with precipitins joined the study. Only two serum samples from two symptomless individuals showed positive serological criteria for FLD as evidenced by the presence of a chymotrypsic band. The remaining forty-two serum samples from twenty-four patients with clinical signs of extrinsic allergic alveolitis had serological criteria for FLD as either chymotrypsic or polysaccharidic arcs. Only specific IgG in asymptomatic subjects was demonstrated, and when a supplementary class of specific antibody (IgM, IgA or IgE) was detected, there was clinical evidence of FLD in all except one case. Thus ELIFA could prove to be a useful tool for the positive diagnosis and prognosis of patients with FLD. In addition this method allows the recognition of a particular functional antigenic fraction of M. faeni by all classes of specific antibodies between different serum samples.

Antibodies, Bacterial↗

PubMed1989

"Reverse" DNA hybridization method for the rapid identification of subgingival microorganisms.

A "reverse" hybridization method is described, in which whole chromosomal DNA was extracted from 10-20 colonies of "unknown" strains in pure culture and labelled with digoxigenin by a random primer technique. DNA probes were prepared from a total of 23 strains and hybridized with targets containing 100 ng purified, denatured DNA from 38 reference strains fixed to nitrocellulose. 21/23 digoxigenin-labelled DNA probes successfully detected all members of the homologous species present on filters. Probes to Fusobacterium nucleatum strains 364 and MG detected 3/4 and 1/4 members of this species, respectively; 13/23 probes were 100% specific, but cross reactions between 10 probes and DNA targets from closely related, heterologous species occurred in 15/834 possible instances. False-positive reactions that occurred between closely related species were, however, easily distinguished and did not prevent the accurate identification of probe strains. Digoxigenin-labelled probes were capable of detecting 100 pg of homologous DNA. The reverse hybridization procedure allows identification or grouping of a large number of isolates within 3 days and provides a more economical means of characterizing subgingival isolates than predominant cultivable techniques and conventional phenotypic testing. This method could be adapted for the direct identification of microorganisms in subgingival plaque samples.

Bacteria↗

PubMed2006

Beliefs about using consumer consultants in inpatient psychiatric units.

A key recommendation of consumer organizations and governments has been the employment of consumer consultants in inpatient psychiatric facilities, but the attitudes of mental health clinicians towards this measure remain inconsistent. The aims of this study were to examine mental health clinicians' attitudes about the role of mental health consumer consultants in inpatient psychiatric units, and to ascertain if participants' age, type of inpatient unit, or grade of staff influenced their attitudes towards consultants. The Consumer Participation and Consultant Questionnaire was used, which was adapted from the Mental Health Consumer Participation Questionnaire. A convenience sample of 47 mental health professionals from two adult inpatient psychiatric units located in a large Australian public general hospital participated in the study. The findings, overall, showed that participants supported the inclusion of consumer consultants in psychiatric units in areas that indirectly impinged on their current roles. Age, level of nurses, and place of employment did not affect their beliefs, but type of occupation was influential. Nurses were less supportive of aspects of consumer consultants' roles that overlapped with the traditional roles of the nurse. The findings have implications for clinical practice, education, and further research, and these are discussed.

Adult↗