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PubMed1975

Plasma proteins in canine gastric lymph.

Because it has not been possible to collect gastric lymph by cannulation of lymph vessels, the methods of renal micropuncture were adapted to collection and analysis of 0.5- to 3-mu samples obtained by puncturing lymph vessels on or immediately adjacent to the ventral surface of the dog's stomach. Samples of gastric lymph collected immediately after completion of surgical exposure of the stomach were subjected to acrylamide gel electrophoresis, and they were found to show all the protein bands exhibited by simultaneously collected samples of plasma. Quantitative analysis of similar samples by cellulose acetate electrophoresis gave the following ratios of protein concentration in gastric lymph to plasma: total protein, 0.51; albumin, 0.68; total globulins, 0.42; alpha1-globulin, 0.49; alpha2-globulin, 0.62; beta-globulin, 0.35; fibrinogen, 0.39; and gamma-globulin, 0.46. When samples were collected over 4 hr the concentrations of all proteins in lymph increased, and an argument is presented that this was not the result of increasing small pore size as the result of injury. When the mucosa was irrigated with solutions of dithiothreitol, a substance causing plasma shedding, the concentrations of proteins in gastric lymph over a 4-hr period were similar to those found in control experiments. Gastric lymph mixed with 100 mN HCl was found to have proteolytic activity.

Alpha-Globulins↗
PubMedDate not supplied

Improved method for assay of estradiol and progesterone receptors with special reference to breast cancer.

Isoelectric focusing of estradiol receptor (ER) and multiple point dextran-coated charcoal (DCC) technique for progesterone receptor (PgR) were adapted for assay of both receptors in the same cytosol sample. Technical improvements and increased recovery of receptors were achieved. 1.0 fmol/ml cytosol of ER may be detected; sensitivity of PgR was 5-10 fmol/ml cytosol. Quantitation of ER was made in 414 (95%) out of 434 consecutive breast cancers. Inclusion of isoelectric focusing records showing only qualitative indication for presence of ER resulted in 98% ER containing tumours, supporting the aspect that strictly ER negative tumours are a rarity. Implications for therapy in disseminated disease are discussed.

Animals↗
PubMed1980

Automated "Gamma-flo" radioimmunoassay of urinary cyclic AMP.

The "Gammaflow" automated assay concept of Brooker et al. (Science 194:270, 1976) has been adapted to the assay for urinary cyclic AMP in patients' samples on the now commercially available Squibb Gamma-FloTM system, a totally automated continuous-flow immunoassay instrument. The instrument aspirates the unknown sample, combines it with radioligand and specific antiserum, incubates the mixture, separates antibody-bound radioligand from free radioligand, counts the radioactivity of the bound ligand, computes the standard curve, and determines subsequent unknown samples by comparison to the standard curve. The urinary cyclic AMP assay we report here is rapid, accurate, and concentrations of cyclic AMP as low as 15 nmol/L can be detected. The assays, which can be done at 60 determinations per hour, are accurate at all dilutions of urine tested and inter-assay (day-to-day) CVs were 3.9% at the midpoint of the standard curve (B/B0 = 0.49).

Autoanalysis↗
PubMed1976

Kinetic determination of hexosaminidases for Tay-Sachs heterozygote screening.

We describe a method for continuous kinetic measurement of hexosaminidase activity, and have applied it to detection of heterozygotes for Tay-Sachs gene. In contrast to existing single-point methods, a ph of 4.5, which is optimal for hexosaminidase activity on the substrate (4-methylumbelliferyl-N-acetyl-beta-d-glucosaminide) is maintained while the increase in fluorescence produced by 4-methylumbelliferone, the reaction product, is being determined. Under the conditions described the reaction follows zero-order kinetics, and activity is linearly related to serum dilution. There is a fairly narrow but practicable range of optimal substrate concentration. The presence of substrate in concentrations exceeding 1 mmol/liter results in progressive quenching of fluorescence and a decrease in apparent enzyme activity. The procedure is standardized with 4-methylumbelliferone in a solution of the substrate. This method is adaptable to use with automated multiple-point discrete-sample devices and especially with the fluorometric kinetic analyzers now being developed.

Clinical Enzyme Tests↗
PubMed1978

Nucleic acids quantitation in polyacrylamide disc gels with a modified double beam spectrophotometer.

Quantitation of RNA and protein bands separated in polyacrylamide disc gel electrophoresis is time consuming and not reliable when performed after staining processes. The best theoretical approach is the use of absorption spectra of the samples. The present paper reports the construction of an efficient and inexpensive adapter to perform direct scanning of 100 mm gels in a Unicam SP-800 double beam spectrophotometer. Conditions for gel casting and electrophoresis were established for RNA. A linear response between 0.25 micrograms and 15 micrograms per individual peak was found with a quantitative limit of detection of 0.25 micrograms. Spectral analysis of each band can be performed allowing identification of samples as nucleotides and detection of contaminants.

Electrophoresis, Polyacrylamide Gel↗
PubMed1997

Tissue culture adaptation of natural isolates of simian virus 40: changes occur in viral regulatory region but not in carboxy-terminal domain of large T-antigen.

The regulatory region of natural isolates of simian virus 40 (SV40) is different from that of laboratory-adapted strains of the virus. The latter have a nucleotide sequence duplication within the enhancer region which varies slightly with each strain, whereas the duplication is lacking in fresh isolates of SV40, which contain an 'archetypal' regulatory region. Many isolates also display nucleotide differences in the DNA encoding the carboxy terminus of large tumour antigen (T-ag). To determine whether genetic changes in these two regions of the SV40 genome were detectable during laboratory adaptation and long-term passage, low-passage virus stocks of two laboratory strains which had detailed passage histories spanning more than 25 years (Baylor strain and VA45-54) were analysed using PCR, cloning and sequencing assays. Both laboratory and archetypal regulatory regions were present in low-passage stocks. Following duplication in the regulatory region, no additional changes were detectable. The variable region at the T-ag carboxy terminus did not undergo any change with tissue culture passage and may serve as a useful site for taxonomic classification of different strains of SV40. Cloned genomes containing single or duplicated enhancers derived from both SV40 strains were viable in CV-1 cells. Attempts to induce regulatory region duplications by 14 serial passages of SV40 archetypal strains in monkey cells were not successful. The results are compatible with tissue culture adaptation of SV40, reflecting either selection of a rare variant pre-existing in the original sample or generation of a rare regulatory region duplication in infected cells.

Adaptation, Physiological↗
PubMed2003

Psychological responses to episodic chest pain.

Patients with chronic musculo-skeletal pain have been profiled as "dysfunctional", "interpersonally distressed" or "adaptive copers". The relevance of these for episodic visceral pain is unknown. Our aim was to replicate conceptually the taxonomy in patients with episodic visceral pain. Patients with chest pain and gastro-esophageal reflux disease (GERD; n=25), coronary artery (CAD; n=20), or with chest pain but without either reflux or coronary artery disease (non-cardiac chest pain--NCCP; n=23) were assessed using several standard affective and cognitive measures relevant to pain. Differences between the diagnostic groups were explored. K-means cluster analysis broadly replicated the three groups found in previous research but the "interpersonally distressed" group had few members. An additional cluster analysis suggested a more parsimonious solution for the sample was a two-cluster one, which approximated to the "adaptive coper" and "dysfunctional" profiles. Membership of both the three- and two-cluster profiles was not associated with membership of specific diagnostic category.

Adaptation, Psychological↗
PubMed1986

Reports of paternal alcoholism: reliability across siblings.

Recent studies of the correlates of having an alcoholic relative have relied heavily upon study participants' reports of the drinking behavior of their parents and other near relatives. These investigations have rarely attempted to validate the reports of familial alcohol problems or report on the reliability of information across other family members. The present study examined the reliability of reports of parental drinking behavior in a sample of 88 sibling pairs on versions of the Short Michigan Alcoholism Screening Test adapted so that subjects endorsed items pertaining to their father's (F-SMAST) or mother's (M-SMAST) drinking behavior. Because of the relative infrequency of offspring reports of maternal problem drinking in this sample, only data from the F-SMAST were suited for analysis. Siblings' reports of specific paternal drinking consequences showed relatively low agreement on items requiring inference (e.g., the presence or absence of guilt) and relatively high agreement on items referring to specific behavioral acts or consequences (e.g., seeking help, being arrested, driving under the influence). Global judgments of paternal alcoholism and overall scores on the F-SMAST showed adequate levels of reliability. It is concluded that, although it is always desirable to validate offsprings' reports of relatives' drinking, such reports tend to agree with those of other siblings, and the use of these reports is reasonable when validation is impossible or unfeasible.

Adult↗
PubMed2006

Almost complete colour constancy achieved with full-field adaptation.

A successive asymmetric colour-matching task was used to study the changes in colour appearance of simulated Munsell samples. Colour shifts were induced with two Planckian illuminants, standard illuminant A (u'=0.256, v'=0.524) and illuminant S (u'=0.174, v'=0.392). Measurements were conducted with a 20 degrees field and a 120 degrees field. Adaptation period varied from 1 to 30s with the smaller field and from 1 to 60s with the larger field. Colour shifts were specified in terms of a modified Brunswik ratio (BR). Higher values of BR were associated with longer adaptation periods but only when the larger background was used. Supplementary experiments showed that the changes in colour appearance were related to a slight shift in the perceived colour of the background. The timing of the colour shifts are modelled in terms of cone opponent responses. High values of BR correspond to almost complete von Kries adaptation in all three cone types.

Adaptation, Ocular↗
PubMed2001

Evaluation of a new immunoturbidimetric test (Liatest antithrombin III) for determination of antithrombin antigen.

Antithrombin (AT) is a glycoprotein mainly synthesised in the liver, and is one of the most important inhibitors of coagulation. Although AT activity determination is more important in clinical practice and anti-IIa and anti-Xa chromogenic tests are widely available, sometimes AT antigen measurement is preferred. Immunoradiometric and enzyme linked immunoabsorbent assay (ELISA) tests have been the only methods available so far for AT antigen determination, but both of them are relatively complicated, time consuming, and the ELISA test cannot be used for single sample assessment. A new immunoturbidimetric test (Liatest antithrombin III) is now available. We have adapted it for the Cobas Mira biomedical analyser. The intra- and inter-assay and total coefficients of variation for normal and low control plasma, as well as for normal pool plasma were low (< 3%). Blood samples from 32 patients with different clinical conditions were assessed both by the standard ELISA method and by the immunoturbidimetric method. The correlation coefficient was 0.95. According to our findings it seems that the immunoturbidimetric test (Liatest antithrombin III) for determination of AT antigen automated for the Cobas Mira biochemical analyser is easy to perform, time sparing and precise, with good comparability to other assays. It is suitable for measuring of AT antigen in routine laboratory work.

Antigens↗
PubMed2000

Italian norms for the Harvard Group Scale of Hypnotic Susceptibility, Form A.

Norms for an Italian translation of the Harvard Group Scale of Hypnotic Susceptibility, Form A (HGSHS:A) by Shor and Orne (1962) are presented. Subjects recruited from 1986 to 1989 were pooled, resulting in a sample of 376 participants (297 women and 79 men). The normative data were generally congruent with earlier normative studies in score distribution, item difficulty levels, and reliability. Women had significantly higher hypnotizability scores and item pass rates than men. The reliability scores of the Italian adaptation of the HGSHS:A were the same as a previously reported Danish sample and higher than a German sample, but lower than those of the Australian, Canadian, and original American samples. These results suggest that the Italian version of the HGSHS:A is an efficient tool for initial hypnotizability screening in an Italian context.

Adult↗
PubMed1997

Quality of life among patients with glaucoma in Sweden.

PURPOSE: To assess the health-related quality of life (HRQOL) in an unselected sample of patients with glaucoma. METHODS: The HRQOL was studied in a cross-sectional survey in 270 patients from two departments of ophthalmology in different parts of Sweden by using the Swedish Health-Related Quality of Life Survey, a generic HRQOL-instrument adapted from the Medical Outcomes Study. RESULTS: Compared to a random Standard Population Sample, the glaucoma patients showed lower mean score only in the scale of 'Pain' (p < 0.05, effect size 0.17), but higher mean score in 'Sexual functioning' (p < 0.01, effect size 0.35). Significant factors correlated to a lower score on HRQOL-scales were decreased visual acuity and visual field, and comorbidity of cardiovascular and non-vascular systemic diseases. Regarding the topical medications, beta-blockers with or without miotic agents had no negative impact on the HRQOL. CONCLUSION: Our findings indicate that health-related quality of life in glaucoma patients in general is good, especially when vision is intact, and that topical beta-blockers do not have any negative influence.

Aged↗
PubMed2005

Structural and functional differences in isoforms of mouse major urinary proteins: a male-specific protein that preferentially binds a male pheromone.

The MUPs (major urinary proteins) of the house mouse, Mus domesticus, are lipocalins that bind and slowly release male-specific pheromones in deposited scent marks. However, females also express these proteins, consistent with a second role in encoding individual signatures in scent marks. We have purified and characterized an atypical MUP from the urine of male C57BL/6J inbred mice, which is responsible for the binding of most of the male pheromone, 2-sec-butyl-4,5-dihydrothiazole, and which is also responsible for the slow release of this pheromone from scent marks. This protein is absent from the urine of female mice of the same strain. The protein has been characterized by MS, leading to unequivocal identification as a previously uncharacterized gene product, providing compelling evidence for the expression of this gene in liver and manifestation in urine. These properties contrast strongly with those of the other MUPs in the same urine sample, and suggest that the requirement to manifest a male-specific pheromone has been met by evolution of a cognate protein specifically adapted to the binding and release of this ligand. This atypical MUP is also present in a random sample of wild-caught male mice, confirming that this protein is not specific to the inbred mouse strain but is present in natural populations also.

Amino Acid Sequence↗
PubMed2005

The detection and characterisation of Dichelobacter nodosus from cases of ovine footrot in England and Wales.

Footrot, caused by the strictly anaerobic bacterium Dichelobacter nodosus, is the most common cause of lameness in sheep in Great Britain but problems exist in association with its diagnosis and control. The fastidious nature of D. nodosus means that complex media and several weeks are required for characterisation. An alternative method to simplify and enhance the detection of D. nodosus in clinical samples is therefore highly desirable. In terms of control, anecdotal evidence from the farming community suggests that the commercially available vaccine, based on Australian isolates of D. nodosus, is not widely employed in this country due to its perceived inefficacy. Seven hundred and six isolates, collected from outbreaks in England and Wales, were therefore used to investigate these issues. A 16S rRNA PCR was adapted to detect D. nodosus in clinical material within 1 day of sampling; a 15% increase in detection compared with culture and less than 1% false negatives were achieved. This represents a major advance in the rapid diagnosis of footrot and will be of great value to practitioners and diagnostic laboratories. Bacterial virulence was tested using protease thermostability and zymogram assays, whilst serogrouping was performed by slide agglutination. All isolates demonstrated virulence patterns previously recorded in Australia and all nine serogroups of D. nodosus (A-I) were represented. Serogroup H was predominant. There was, therefore, no evidence for the presence of novel strains of D. nodosus compared with Australia suggesting the need for further investigation into farmers' views on the use of the commercial vaccine in Great Britain.

Animals↗
PubMed2006

Clinical relevance of Aspergillus isolation from respiratory tract samples in critically ill patients.

INTRODUCTION: The diagnosis of invasive pulmonary aspergillosis, according to the criteria as defined by the European Organisation for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG), is difficult to establish in critically ill patients. The aim of this study is to address the clinical significance of isolation of Aspergillus spp. from lower respiratory tract samples in critically ill patients on the basis of medical and radiological files using an adapted diagnostic algorithm to discriminate proven and probable invasive pulmonary aspergillosis from Aspergillus colonisation. METHODS: Using a historical cohort (January 1997 to December 2003), all critically ill patients with respiratory tract samples positive for Aspergillus were studied. In comparison to the EORTC/MSG criteria, a different appreciation was given to radiological features and microbiological data, including semiquantitative cultures and direct microscopic examination of broncho-alveolar lavage samples. RESULTS: Over a 7 year period, 172 patients were identified with a positive culture. Of these, 83 patients were classified as invasive aspergillosis. In 50 of these patients (60%), no high risk predisposing conditions (neutropenia, hematologic cancer and stem cell or bone marrow transplantation) were found. Typical radiological imaging (halo and air-crescent sign) occurred in only 5% of patients. In 26 patients, histological examination either by ante-mortem lung biopsy (n = 10) or necropsy (n = 16) was performed, allowing a rough estimation of the predictive value of the diagnostic algorithm. In all patients with histology, all cases of clinical probable pulmonary aspergillosis were confirmed (n = 17). Conversely, all cases classified as colonisation had negative histology (n = 9). CONCLUSION: A respiratory tract sample positive for Aspergillus spp. in the critically ill should always prompt further diagnostic assessment, even in the absence of the typical hematological and immunological host risk factors. In a minority of patients, the value of the clinical diagnostic algorithm was confirmed by histological findings, supporting its predictive value. The proposed diagnostic algorithm needs prospective validation.

Aged↗
PubMed1982

Temperament questionnaires in clinical research.

Parental questionnaires concerning the nine temperament variables studied in the New York longitudinal study have been developed for the measurement of children's temperament at six, 12 and 24 months of age. Retest reliability for the questionnaires was found to be generally satisfactory and at a level similar to that found in other temperament studies. The temperament characteristics of a large standardization sample, studied longitudinally over the three ages, showed little relation to the children's gender, birth order or social class. Factor analyses of the variables showed that two of the patterns were identical at 12 and 24 months of age: one pattern consisted of mood, approach, adaptability and distractibility, and the other of activity, approach and intensity. The standardization sample showed about the same rates of the 'difficult' and the 'slow-to-warm-up' temperament types, as found in early samples. The stability of the subjects' temperament scores over time was low to moderate, and a majority of the subjects changed their temperament type over the 18 months covered by the longitudinal study. The use of temperament questionnaires in clinical research is illustrated in three current studies.

Child, Preschool↗
PubMed1997

Social self-efficacy and short-term variability in social relationships: the MacArthur successful aging studies.

Dealing with others entails both stability and short-term variability of the functions and outcomes of social relationships. The authors argue that patterns of short-term intraindividual variability in social relationships and self-efficacy beliefs contribute interpretable information about social adaptation. On the basis of 23 repeated weekly measurements of a sample of 32 participants ages 56 to 88 years, the authors examined the extent to which fluctuations in perceived relational outcomes are related to fluctuations of social self-efficacy. Results showed that individuals differ systematically in respect to the extent to which they experience and display fluctuations in self-efficacy and availability of social relationships. Moreover, when individuals perceive others to be available across time, social self-efficacy beliefs are stronger and fluctuate less across time.

Adaptation, Psychological↗
PubMed2002

Psychological functioning in boys of short stature: effects of different levels of growth hormone secretion.

AIM: To examine the relationship between growth hormone (GH) and psychological functioning, especially self-perception and well-being, in 60 prepubertal boys of short stature with a wide range of GH levels. METHODS: A comparison was made of the well-being and self-perception of children with GH insufficiency, children with idiopathic short stature (ISS), a normative sample and healthy boys with normal stature. RESULTS: Children with GH insufficiency had a more negative perception of their own physical appearance than the normative sample. They perceived themselves as more alert but also more inhibited than both the children with ISS and the healthy boys with normal stature. In comparison with the healthy boys with normal stature they perceived themselves as having more stability. The parents of the boys with GH insufficiency also perceived their children as being more stable compared with how the parents of boys with ISS perceived their children. To elucidate the effects of GH on psychological functioning a multiple regression analysis was performed. CONCLUSION: The lower the levels of GH the more inhibited were the boys of short stature, as perceived both by themselves and by their parents. The boys with GH insufficiency had a more negative perception of their physical appearance than the normative sample.

Adaptation, Psychological↗

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