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PubMed2005

Determination of endogenous glycosaminoglycans derived disaccharides in human plasma by HPLC: validation and application in a clinical study.

SB-424323 is a new, orally active anti-thrombotic agent presently in phase-II clinical development, with limited hemorrhagic risk and a unique mechanism of action involving the induction of glycosaminoglycans (GAGs) biosynthesis. The objective of the present study was to develop a simple and rapid high performance liquid chromatography (HPLC) method for determination of endogenous GAGs derived disaccharides in plasma samples from a phase-II clinical study of SB-424323. Sample preparation was a simple heat treatment of the diluted plasma followed by digestion of endogenous GAGs with chondroitinase ABC to yield unsaturated disaccharides, 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-D-galactose (DeltaDi-0S), 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose (DeltaDi-4S), and 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose (DeltaDi-6S). These disaccharides were recovered and purified using centrifugal filtration through a filter with 3000 molecular weight cut-off along with externally added internal standard 2-acetamido-2-deoxy-3-O-(2-O-sulfo-beta-D-gluco-4-enepyranosyluronic acid)-D-galactose (DeltaDi-UA2S). A gradient reverse phase HPLC separation was developed on a Waters Symmetry C(18) column (4.6 mm x 150 mm, 5 microm) with a gradient mobile phase system consisting of 0.8 mM tetrabutylammonium hydrogen sulfate and 2mM sodium chloride and acetonitrile at a flow rate of 1.0 mL/min. The eluate was monitored with an ultraviolet detector set at 230 nm. Plasma standard curves were linear (r(2)> or =0.994) in the concentration range 1.0-20 microg/mL with a lower limit of quantification (LLOQ) of 1.0 microg/mL for each of the disaccharide. The mean measured quality control (QC) concentrations for the disaccharides deviated from the nominal concentrations in the range of -8.92 to 5.61% and -16.3 to 16.7%, for inter and intra-day, respectively. The inter and intra-day precision in the measurement of QC samples, were in the range of 3.21 to 18.2% relative standard deviation (R.S.D.) and 0.32 to 20.9% R.S.D., respectively. The inter and intra-day precision in the measurement of endogenous GAGs derived disaccharides in human control plasma, were in the range of 5.8 to 15.9% R.S.D. and 1.17 to 7.74% R.S.D., respectively. Stability of the processed samples was confirmed up to 48 h in the auto-sampler. The method is simple, reliable, and easily adaptable to analysis of large number of samples under logistics of a clinical study. The present method has been used to investigate the GAGs levels in the plasma of patients in a phase II clinical study of SB-424323.

Adolescent↗
PubMed1996

Antigen sampling across epithelial barriers and induction of mucosal immune responses.

Epithelial barriers on mucosal surfaces at different sites in the body differ dramatically in their cellular organization, and antigen sampling strategies at diverse mucosal sites are adapted accordingly. In stratified and pseudostratified epithelia, dendritic cells migrate to the outer limit of the epithelium, where they sample antigens for subsequent presentation in local or distant organized lymphoid tissues. In simple epithelia, specialized epithelial M cells (a phenotype that occurs only in the epithelium over organized lymphoid follicles) deliver samples of foreign material by transepithelial transport from the lumen to organized lymphoid tissues within the mucosa. Certain pathogens exploit the M cell transport process to cross the epithelial barrier and invade the mucosa. Here we review the features of M cells that determine antigen and pathogen adherence and transport into mucosal lymphoid tissues.

Animals↗
PubMed2005

Health related quality of life of adolescents in Vietnam: cross-cultural adaptation and validation of the Adolescent Duke Health Profile.

Quality of life (QOL) of adolescents has been scarcely documented in a general population sample. The study was aimed at translating and adapting the Adolescent Duke Health Profile to Vietnamese, validating the questionnaire, determining reference value, and identifying determinants of poor QOL. Following a cross-cultural methodology, the Adolescent Duke Health Profile (ADHP) had content adapted to Vietnamese by alteration of 2 out of 17 items. Test-retest validity was checked in 408 adolescents. Construct validity and internal consistency were assessed in a 1408 probability sample of adolescents in Vietnam, and determinants were analysed by multiple linear regression. The ADHP showed satisfactory internal consistency (Cronbach's alpha=0.87-0.92) and satisfactory construct validity in relation with drug abuse or not, or with parent situation living in couple or alone. Test-retest reliability was acceptable (ICC=0.7-0.8) and major determinants were age, sex, education, chronic disease, alcohol and drug use. This works provide a validated, simple health related QOL scale suited to adolescent population with reference values. Taking into account determinants identified will help program prevention and intervention health policy and to evaluate the effect of these actions.

Adolescent↗
PubMedDate not supplied

Factors associated with breast self-examination among jordanian women.

The purpose of this cross-sectional, correlational, descriptive study was to investigate factors and beliefs that may be related to the practice of breast self-examination (BSE) among a group of Jordanian women. The sample consisted of 519 women from two major universities in Jordan. About 36% of the sample were university employees and 64% were graduate and undergraduate students. Stratified random sampling was used to enroll the undergraduate students while graduate students and employees were selected by convenience sampling. The study instrument was an adapted version of Champion's Revised Health Belief Model Scale (CRHBMS). The results were analyzed using a chi-square test and a stepwise multiple regression. The main findings indicate that although the majority of the sample population (67%) had heard/read about BSE, only a quarter of them reported that they had ever practiced BSE in the previous 12 months, and only 7% had performed it on a regular monthly basis. Confidence, motivation, susceptibility, and fewer barriers were variables that showed a positive association with BSE practice in the previous year, while benefits, susceptibility, and motivation influenced the intention to perform BSE in the future. Women's age, level of education, having heard or read about breast tumors, and personal history of breast tumors were also found to be significant predictors of BSE practice.

Adolescent↗
PubMed2006

Evaluation of two different models to predict BRCA1 and BRCA2 mutations in a cohort of Danish hereditary breast and/or ovarian cancer families.

To meet the increasing demand for BRCA1 and BRCA2 mutation analysis, a robust system for selecting families who have a higher chance of a mutation has become important. Several models have been developed to help predict which samples are more likely to be mutation positive than others. We have undertaken a complete BRCA1 and BRCA2 mutation analysis in 267 Danish families with high-risk family history. We found deleterious mutations in 28% (76) of the families, 68% (52) of those in BRCA1 and 32% (24) in BRCA2. We compared our results with two popular manual models developed to estimate the chance of a positive result. One is the recently published Manchester model and the other is the Frank 2 model updated by Myriad Genetic Laboratories, Inc. Neither of the models would have suggested screening all mutation-positive samples. The Manchester model would have suggested screening 124 of the families in the cohort, thereby detecting 54 of 76 mutations (sensitivity 71%; specificity 63%), whereas the Frank 2/Myriad model would have found 60 of 76 mutations by screening 169 samples if a 10% likelihood was adapted (sensitivity 79%; specificity 43%). The updated Manchester model suggested screening 172 families whereby 64 mutations would have been detected (sensitivity 84%; specificity 44%). We conclude that although both models would have reduced the number of samples screened significantly, up to 28% of the mutations would not have been found by applying these models to this Danish cohort of families. This raises the question whether models designed for specific populations can be used in a wider setting.

BRCA1 Protein↗
PubMed1996

Behavioral outcomes after pediatric closed head injury: relationships with age, severity, and lesion size.

This study investigated the behavioral outcomes and adaptive functioning of 138 children with mild to severe closed head injury in the 6- to 16-year age range. Each child was evaluated with the Personality Inventory for Children-Revised. A subset of this sample (n = 77) received the Vineland Adaptive Behavior Scales. Results revealed little evidence for group differences based on severity of closed head injury on scales associated with psychopathology on the Personality Inventory for Children-Revised. However, children with severe closed head injury were viewed as experiencing more difficulties than children with mild-moderate closed head injury on those components of the Personality Inventory for Children-Revised most closely associated with cognitive functions. In addition, on the Vineland Adaptive Behavior Scales, severely injured children had lower scores on the Communication and Socialization scales than children with mild-moderate injury. Relationships between the size of frontal and extrafrontal lesions from concurrent magnetic resonance imaging and behavioral outcomes were not apparent. This study suggests that outcome measures assessing adaptive behavior and cognitive functions are more sensitive to severity of closed head injury than parent-based scales of internalizing and externalizing psychopathology.

Adolescent↗
PubMed2004

Newborn screening for congenital infectious diseases.

To estimate the prevalence of congenital toxoplasmosis, Chagas disease, cytomegalovirus, and rubella, blood samples on dried blood spot (DBS) from neonates (day 3-20 of life) were screened for immunoglobulin (Ig) M against Toxoplasma gondii, cytomegalovirus, rubella virus, and IgG against Trypanosoma cruzi by methods used for serum and adapted for use with DBS. Positive samples were further analyzed for IgM and IgG in serum from neonates and mothers. DBS samples from 364,130 neonates were tested for Toxoplasma gondii-specific IgM, and 15,873 neonates were also tested for IgM against cytomegalovirus and rubella virus and for Trypanosoma cruzi-specific IgG. A total of 195 were diagnosed with congenital toxoplasmosis, 16 with cytomegalovirus, and 11 with congenital rubella. One newborn had a confirmed result for Chagas disease, and 21 mothers had positive serum antibodies. These results suggest that infectious diseases should be considered for future inclusion in programs for newborn screening of metabolic diseases in disease-endemic areas.

Adult↗
PubMed1996

Multi-center European evaluation of HIV testing on serum and saliva samples.

OBJECTIVE: to evaluate the reliability of HIV antibody testing on saliva. DESIGN: matched serum and saliva samples were collected from both seronegative (n = 344) and seropositive (n = 125) individuals in five European countries. Duplicate saliva samples collected with Omni-Sal devices provided by Saliva Diagnostic System (SDS) were pooled before analysis. METHODS: all samples were analyzed by Recombinant HIV1 EIA Cambridge Bioscience and 2nd generation Abbott HIV 1&2 1A80. EIA procedures were adapted for saliva testing by modification of sample dilution and/or cut-off calculation. All saliva recording positive and/or doubtful EIA results were further analyzed by Western blot as a confirmatory method. RESULTS: EIA results obtained from sera analysis from both seropositives and seronegatives allowed for calculation of the tests' sensitivity (HIV1 Biotech: 99.2%-100%; Abbott: 100%) and specificity (both tests 100%). In the series of 125 saliva samples collected from seropositives, the EIA results were as follows: with Biotech (3 negative, 3 in the grey-zone and 119 reactive) and with Abbott (1 negative, 1 in the grey-zone and 123 reactive). One saliva sample found negative by both EIA tests, although fulfilling HIV1 WB criteria of positivity, was collected from an HIV2 infected person. Out of 125 saliva samples collected from seropositives, 121 produced positive Western Blot profiles, 4 were indeterminate and 1 was found negative whereas 125/125 sera were found positive. CONCLUSION: the reliability of HIV testing of saliva is dependent on the sensitivity of EIA tests and on the criteria used for the interpretation of Western blot tests as well. Although saliva testing offers numerous advantages for epidemiological purposes, it should not be recommended for diagnosis.

Blotting, Western↗
PubMed1987

[Comparative evaluation of 3 automated optical coagulation detection systems].

The Organon Teknika-General Diagnostics Coag A Mate X2 (CAM X2), Ortho Koagulab 40A, MLA Electra 700 (E 700), were simultaneously evaluated following the same protocol, on the same samples. Compared with the former photo-optical-based automated devices, these newer advanced instruments present a high throughput in fibrin endpoint detection, permitting fibrinogen determination with reliable results; they also present a higher degree of automatism, greater flexibility in use and easy mastering, allied to elaborated expression of results. Statistical comparison shows equivalent performance characteristics. Repetabilities are excellent for Prothrombin Time (CV: 1-2.5%) with 5 thromboplastins and for the prothrombin complex factors assays: they are also good for Activated Partial Thromboplastin Time (APTT) with 4 reagents (CV: 0.5 to 3%) provided that kaolin be not used on the KLO and E700; the same limitation exists for specific factors assays. Fibrinogen assays display good results (CV: 3-4% and 6-8%) respectively for times of 11 sec. (/=/ 2.4 g/l) and 23 sec. (/=/ 1 g/l). Standard linearities are generally excellent whatever the test. The correlation with results obtained on other devices is good (0.96 less than R less than 1). The most fully automated is the KLO with reliable system of sample delivery, automatic calibration and quality control function. The E700 is the slowest instrument, especially for APTT and the least adapted to large homogenous series, but its original loading system with binary coded cuvettes makes it interesting for emergencies and mixed tests. Controls and adjustments of the pumptubing are particularly required on the CAM X2 which is, on the other hand, the quickest instrument with its 4 photo-optical cells, well adapted to handling a large series of samples.

Autoanalysis↗
PubMed2001

Adaptation of a surface plasmon resonance biosensor with microfluidics for use with small sample volumes and long contact times.

The efficient delivery of sample to surface-immobilized sites is a key element in biosensing. For a surface plasmon resonance (SPR) biosensor, this has been addressed by constant flow through a microfluidic system with a sample injection loop (Sjölander, S.; Urbaniczky, C. Anal. Chem. 1991, 63, 2338-2345). The present study describes an alternative mode of sample delivery without constant unidirectional flow. It was implemented on a commercial Biacore X SPR biosensor equipped with a microfluidic cartridge, but with the fluidic handling performed by an externally computer-controlled syringe pump. We demonstrate that sample volumes as low as 2 microL can be reproducibly positioned to cover the sensor surfaces, manipulated in a serial fashion, efficiently mixed by applying an oscillatory flow pattern, and fully recovered. Compared to the traditional continuous unidirectional flow configuration, we found very similar kinetic responses at high analyte concentrations and slightly slower responses at low concentrations, most likely due to depletion of analyte from the small sample volumes due to surface binding. With the antibody-antigen systems tested, binding parameters were obtained that are generally within 10% of those from conventional experiments. In the new configuration, biosensor experiments can be conducted without the usual constraints in the surface contact time that are correlated with sample volume and mass transport rate. This can translate to improved detection limits for slow reactions and can facilitate kinetic and thermodynamic binding studies.

Biosensing Techniques↗
PubMed1999

Adaptation of corticosterone-but not beta-endorphin-secretion to repeated blood sampling in rats.

Effects of short-term repeated blood sampling on the secretion of corticosterone (CORT) and beta-endorphin (beta-END) were evaluated in male Wistar rats. Blood was drawn from the tail vein of conscious rats four times within 2 h both at the peak and trough period of the diurnal corticosterone secretion cycle. All rats were well accustomed to the procedure. The main findings were: (1) At both sampling intervals, CORT increased significantly in response to the first sampling and declined to baseline values in successive samples. (2) beta-END also increased significantly in response to the first sampling but remained elevated in successive samples. (3) Intensities of initial CORT and beta-END responses correlated positively with each other and with the baseline beta-END values. Feedback inhibition of CORT secretion with sustained elevation of beta-END titres suggests a moderate stress intensity of the repeated blood sampling procedures. In general, due to lack of short-term feedback inhibition, beta-END seems to reflect the effects of repeated administration of moderate intense stressors more closely than CORT.

Animal Welfare↗
PubMed1978

Assay of retinoids in biological samples by reverse-phase high-pressure liquid chromatography.

A separation procedure for retinoids based on reversephase high-pressure liquid chromatography with solvent mixtures of acetonitrile and water is described. The method may be applied to the screening of synthetic retinoids, which have potential for use in the prevention of cancer. It is easily adapted to a variety of biological samples and can be applied to other conventional retinoid assays in liver and plasma, detecting as little as 1 nmol retinyl esters and less than 0.3 nmol retinol per g tissue. The one-step chromatography results in separation and simultaneous determination of many of the synthetic retinoids and all of the natural retinoids, including the retinyl esters that are separated into their major fatty acid components. The method has been applied to the analysis of retinoid levels in the liver and intestine of vitamin A-deficient hamsters following a p.o. dose (0.5 mg/day for 2 days) of retinyl acetate or of a synthetic vitamin A analog and is predictive of the degree to which various synthetic retinoids can be converted to retinol and stored in the liver as retinyl esters. Because of its speed, excellent recoveries, and high resolution, the method offers significant advantages over previous, more lengthy procedures.

Animals↗
PubMed1983

Photoelectric signals from dried oriented purple membranes of Halobacterium halobium.

In dried oriented samples of purple membrane isolated from Halobacterium halobium, the photoelectric activity decreases and the light adaptation vanishes when the water content of the sample is lowered. In the photocycle the first steps of the proton movement were accelerated with decreasing humidity, while the last steps of the photocycle could not be observed. From the analysis of the photoelectric signal we conclude that at low humidities the protons move forward in the L decay and return to their original place during M decay.

Bacteriorhodopsins↗
PubMed2000

The multidimensional pain inventory profiles in patients with chronic cancer-related pain: an examination of generalizability.

This study examined the generalizability of the non-malignant pain patient profiles based on the Multidimensional Pain Inventory (MPI) to patients with cancer-related pain. Data were collected from 112 cancer patients. In total, 107/112 patients completed the MPI. Of the 96% of patients classified, only 60% were classified by the three main profiles. In this sample, there were 47.7% (n=51) Adaptive Copers, 9.3% (n=10) Dysfunctional, 2.8% (n=3) Interpersonally Distressed; 32.7% (n=35) Anomalous; 3.8% (n=4) Hybrid; and 3.8% (n=4) Unanalyzable. Because of the significantly lower pain severity, interference and affective distress scores, the Anomalous group could be considered Highly Adaptive. Given that 80% were classified as either Adaptive or Anomalous, these findings suggest that while the MPI-based profiles do apply, a two profile classification system may be more suitable for cancer patients than the usual three. In particular, the low proportion of patients classified as Interpersonally Distressed may reflect important differences in social support for cancer patients compared with non-cancer patients. Whereas the MPI-based profiles are consistent across non-malignant pain problems, it appears that the nature of cancer may affect the MPI-based profile classification system more than non-malignant pain problems do.

Adaptation, Psychological↗
PubMed2004

[Kell alloimmunization in pregnancy].

INTRODUCTION: Kell alloimmunization is a rare disease, although its incidence is the highest after after anti-D alloimmunization. METHODS: We report two recent cases and a review of the literature to describe practical management of Kell alloimmunization in pregnancy. DISCUSSION: When an immunization against the Kell antigen was diagnosed, amniocentesis was performed at 14 weeks gestation to determine the fetal blood group. If the fetus was Kell positive, a first fetal blood sample was drawn at 17 weeks gestation in case of fetal hydrops, and at 20 weeks without fetal hydrops. The diagnosis of anemia led to in utero transfusion. A second fetal blood sample was taken at 8 to 10 days, every two weeks during the second trimester and every three or four weeks during the third trimester. Fetal well-being was assessed with weekly sonography and rates of hemoglobin decline. These measures enable adapting the frequency of fetal blood sampling.

Adult↗
PubMed1982

Acculturation and stress in a low-income Puerto Rican community.

In this study, the acculturative stress experienced by Puerto Rican migrants to the urban U.S. is examined. Acculturative stress is defined as length of residence in a novel cultural environment while lacking psychosocial resources to support adaptation to that environment. In a sample of 67 patients seen in a public health nursing agency, it was found that acculturative stress was associated with poorer health status, more behavioral problems, and higher levels of ethnomedical belief. The implications of these results for research on stress and adaptation are discussed.

Acculturation↗
PubMedDate not supplied

Follow-up in patients with aspartylglucosaminuria. Part II. Adaptive skills.

Aspartylglucosaminuria is a lysosomal storage disorder inherited as an autosomal recessive trait. Progressive mental retardation is the main symptom, and the lifespan of the patient is abnormally short. The adaptive skills and maladaptive behaviour of 110 patients aged from 7 to 56 years were analysed using the Portage scale and the AAMD Adaptive Behaviour Scale Part two. The sample was divided into four groups; school-aged, young adults, adults and middle-aged. All patients needed help in the household and transactions outside the home. The school-aged turned out to be superior and middle-aged inferior to other groups concerning adaptive skills. The school-aged and young adults were quite independent in dressing, toileting, bathing and walking alone near the home. Out-of-home replacement became necessary in the majority in adulthood. Twenty-eight patients (25%) were regarded as having behavioural disturbances and the amount of personal disturbances increased significantly with age.

Activities of Daily Living↗
PubMedDate not supplied

[Method for the study of pulmonary function in laryngectomized patients].

OBJECTIVE: The aim of the study is to develop a instrumental method capable to achieve pulmonary air function test (spirometry) by assembling the spirometer mouthpiece to the laryngectomized patient's stoma. MATERIAL AND METHOD: Our study was carried out in 33 laryngectomized patients (all male). The spirometer tests were done with a Datospir 92 by Sibelmed equipment, which consists of a dry Fleish pneumotacographer with flow and volume chart register. We have made a stoma-spirometer adapter with a cardboard tube, an adhesive and silicone disc. RESULTS: The whole sample achieved excellent outcome with the stoma-spirometer adapter. Nor air leak neither high resistence were measured while spirometry was performed. CONCLUSIONS: We consider that the facts studied may enable us to add, pragmatically, new resources to the more effective understanding of the respiratory handicap in the laryngectomized population.

Aged↗