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PubMed1978

Comparison of WI-38, MRC-5, and IMR-90 cell strains for isolation of viruses from clinical specimens.

With the diminishing supply of the human fetal lung WI-38 cell strain, a replacement for viral isolation is needed. Two candidates are the human fetal lung strains MRC-5 and IMR-90. A comparison of WI-38, MRC-5, and IMR-90 was performed to evaluate efficiency and speed of viral isolation, clarity of cytophatic effect, and ease of growing the cells. The inocula were clinical specimens rather than tissue culture-adapted isolates. Frozen samples of 46 specimens that had previously yielded an isolate on WI-38 were thawed and inoculated onto WI-38, MRC-5, and IMR-90 cells. In addition, 95 freshly taken clinical specimens uf undetermined infectivity were inoculated onto the cell strains. Viral recovery rates were similar on all three strains, as were the appearance and speed of onset of the cytophatic effect. MRC-5 and WI-38 cells remained healthy until generation 36, whereas IMR-90 cells went into crisis by generation 20. The longer life span of the MRC-5 cells makes them more suitable than IMR-90 cells to replace the WI-38 strain for routine use in viral diagnosis.

Cell Line↗
PubMed2004

Correlation between plasma and urine phenylalanine concentrations.

In this pilot study, we show that plasma phenylalanine concentration can be predicted from urine concentration if the age of the patient is taken into consideration. This observation could open the way to a new monitoring of phenylketonuric patients in which painful frequent blood sampling, mandatory to adapt the low phenylalanine diet, could be mostly replaced by urinalysis. Compliance to treatment would be improved and hence also the ultimate mental development. Since this study was based on a small number of patients, validation of the model in a large multicentric survey is needed before it can be recommended.

Adolescent↗
PubMed1987

Extracellular accumulation of proline, serine and trehalose in the haemolymph of osmoconforming brackish-water mosquitoes.

Larvae of Culex tarsalis, a mosquito, are capable of surviving and developing in dilutions of sea water ranging from 0 mosmol l-1 to 700 mosmol l-1. In waters more dilute than 400 mosmol l-1, the larvae osmoregulate, whereas in those more concentrated than 400 mosmol l-1, the osmotic strength of the haemolymph parallels that of the medium, i.e. the larvae osmoconform. Over the full range of external concentrations tested, the larvae regulate the levels of Na+, K+, Mg2+, Ca2+ and Cl- in the haemolymph. Analyses of haemolymph samples from larvae adapted to media of 50 mosmol l-1 or 600 mosmol l-1 indicate that the increase in haemolymph osmotic concentration observed in media above 400 mosmol l-1 is due to the accumulation of organic compounds, particularly proline, serine and trehalose.

Amino Acids↗
PubMed2005

Multiplex approaches in protein microarray technology.

The success of genome sequencing projects has provided the basis for systematic analysis of protein function and has led to a shift from the description of single molecules to the characterization of complex samples. Such a task would not be possible without the provision of appropriate high-throughput technologies, such as protein microarray technology. In addition, the increasing number of samples necessitates the adaptation of such technologies to a multiplex format. This review will discuss protein microarray technology in the context of multiplex analysis and highlight its current prospects and limitations.

Animals↗
PubMed1985

Peripheral androgen levels in the male brush-tail possum (Trichosurus vulpecula).

Radioimmunoassays were established for the measurement of total androgens and the specific measurement of testosterone and 5 alpha-dihydrotestosterone in peripheral plasma of the brush-tail possum. Androgen concentrations were measured in blood collected from indwelling jugular cannulae to determine whether the normal pattern of androgen secretion in this species was episodic and to attempt to relate total androgen and the pattern of testosterone secretion to the changes previously reported in prostatic, but not epididymal, weight in the breeding season. Blood was collected from restrained animals at varying time-intervals during daylight hours and darkness. Despite an apparent good adaptation to the sampling procedure there was generally a progressive decline in plasma androgen level during the collection period. This was true for animals bled during or out of the breeding season. There was no significant seasonal effect on the androgen concentration in the initial blood sample. When less restraint was used, two of three animals showed fluctuations in androgen levels over the 7-h sampling period. Testosterone levels in blood obtained by cardiac puncture were four- to nine-fold higher than those of 5 alpha-dihydrotestosterone but levels of these androgens in samples obtained during the breeding season were not significantly different from those obtained out of season. The results do not argue for a pulsatile release of testosterone in the possum but do demonstrate a marked capacity for changes in the peripheral androgen concentration. There was a poor correlation between testosterone and 5 alpha-dihydrotestostosterone levels and prostatic weight.

Androgens↗
PubMed1994

Interleukin-1 beta concentration of gingival crevicular fluid.

The concentration of interleukin-1 beta is elevated in inflamed gingival tissue. Therefore a method for the measurement of interleukin-1 beta (Il-1 beta) in gingival crevicular fluid (GCF) using a commercially available Il-1 beta ELISA was developed. GCF was collected with periopaper strips and 4 protocols of sampling using filter paper strips were tested; the method with a recovery rate of 111.9% (SD: +/- 14.5%) was chosen for subsequent analysis of all samples. Il-1 beta concentration in GCF of periodontitis patients and a healthy control group was determined. Patients (n = 19, mean age: 29.3 years) had not been treated. The healthy control group (n = 14, mean age: 22.8 years) showed, after a hygiene regimen of 2 weeks, no clinical signs of gingival/periodontal inflammation. Probing depth, clinical attachment level, bleeding upon probing, and a modified plaque index were recorded. Il-1 beta could be detected in all GCF samples. The concentration ranged between 22.8 ng/ml and 150 ng/ml in the healthy control group and between 85.8 ng/ml and 882.2 ng/ml in the periodontitis patients. No sex-related differences were noted. According to our present results the determination of GCF Il-1 beta concentration is possible using commercially available test kits if the principle of sample preparation is adapted to the specific requirements of GCF analysis.

Adolescent↗
PubMed1983

Effect of energy concentration and source of nitrogen on numbers and types of rumen protozoa.

Two rumen-fistulated Holstein heifers were fed three protein-free semipurified diets: 70% roughage, 30% concentrate; 50% roughage, 50% concentrate; and 30% roughage, 70% concentrate. Wheat straw was the roughage, and the concentrate was a pelleted mixture of corn starch, dextrose, fat, urea, vitamins, and minerals. Each animal received the three dietary treatments in a randomized complete block design. On days 1, 3, and 5 following a 21-day adaptation, rumen fluid samples were taken before feeding and at 1, 2, 3, 4, and 5 h after feeding. Total protozoa concentrations for low, medium, and high concentrate diets were 1.5, 2.5, 4.1 X 10(5)/ml. Dasytricha concentrations were not affected by diet, but Entodinium, Diplodinium, Isotricha, Epidinium, and Ophryoscolex increased with increases of the concentrate proportion of the diets. Entodinium, the predominant protozoa with all diets, increased as concentrate proportion of the diet increased. In a second experiment the two heifers from Experiment 1 were fed semipurified diets that contained either urea or soybean meal as the nitrogen source. There were larger populations of Dasytricha and Isotricha when the diet contained soybean meal, and larger populations of Charon, Entodinium, and total protozoa when the diet contained urea. In both experiments, holotrichs were highest 1 to 2 h after feeding, and entodiniomorphs were highest before feeding and 4 to 6 h after feeding. An active protozoan population can exist in the rumen of cattle fed diets virtually devoid of natural protein, and protozoa are influenced by energy and nitrogen source.

Animals↗
PubMedDate not supplied

[Validation of a self-efficiency scale for the prevention of AIDS among adolescents].

OBJECTIVES: The objectives are obtain the internal consistency, validity and factorial structure of the Self-Efficacy for AIDS scale, which was developed in American population by Kasen, Vaughan and Walter (1992) and adapted in mexican samples for López and Padilla (1999). INSTRUMENTS: It is employed the Self-efficacy to Prevent the AIDS Scale of 27 items of López and Padilla (1999), together with closed questions Likert type, dicotomics and two opened questions on relevant aspects of the sexual conduct to validate the scale, under a format self-applied test of pencil and paper. MATERIAL AND METHODS: It is a correlacional study with a not experimental design of transverse type. It was accomplished with a simple random sample of 962 adolescents wich were taken of 60 public health centers of Monterrey. None went to consult by motives related to the AIDS. RESULTS: The scale was adjusted to a normal distribution (Zk-s(Normal) = 1.369; p = 0.047). It presents a mean of 95.14 and a standard deviation of 25.80. Its reliability, by the alpha of Cronbach, was 0.89. SEA-27 shows a structure of four independent factors that explain 58.47% of the total variance. CONCLUSIONS: Thus, it is concluded that SEA-27 is a reliable and valid scale.

Acquired Immunodeficiency Syndrome↗
PubMed1976

Simultaneous measurement of phenobarbital, diphenylhydantoin, and primidone in blood by high-pressure liquid chromatography.

We describe a sensitive, precise high-pressure liquid chromatographic method in which 5-(p-methylphenyl)-5-phenylhydantoin is used as the internal standard for simultaneous determination of diphenylhydantoin, phenobarbital, and primidone in whole blood and plasma. These anticonvulsant drugs are well separated from each other and from normal blood constituents in less than 10 min. The lower limit of detection for each drug is 100 ng for primidone, 200 ng for dilantin, and 300 ng for phenobarbital. The eluted drugs were detected by their absorption at 254 nm, and evaluated from their peak heights as compared to internal standard. The method was successfully adapted for pediatric samples (100 to 500 mul of whole blood or plasma). Fifty specimens were analyzed for phenobarbital and diphenylhydantoin and 25 specimens for primidone by a standard gas-chromatographic method and by our liquid-chromatographic method; the resulting correlation coefficient was greater than 0.98.

Chromatography, Gas↗
PubMedDate not supplied

[Anxiety and depression of cancer patients hospitalized and at home].

The aim of this study was to investigate the anxiety and depression of cancer patients hospitalized and at home. Using a descriptive, correlational and comparative design and the Roy Adaptation Model, a sample of 80 oncologic patients was studied. Several instruments were used to measure anxiety and depression (HADS), quality of life and symptoms (RSCL), sociodemographic factors, variables connected to the hospitalization, quality of the relationship with health practitioners, family members and friends and the degree of satisfaction for the received information and support. The examined variables were measured on the same patients at hospital and at home. About the 30% of the patients were anxious and depressed. Statistical analysis showed that while anxiety did not change from the hospital to home, depression increased soon after the discharge and decreased over time and after the increasing of the number of hospital access. Anxiety and depression were positively correlated to boredom during the hospitalization, physical symptoms, number of the patients children, and previous anxious and depressive problems. Anxiety and depression were negatively correlated to the ward comfort, the support of health practitioners, family members and friends and the satisfaction for the received information. Differences between this study and the international literature are discussed. Recommendations for the future research and nursing practice are given.

Adult↗
PubMed2004

Analysis of dyes in illicit pills (amphetamine and derivatives).

The determination of dyes present in illicit pills is shown to be useful and easy-to-get information in strategic and tactical drug intelligence. An analytical strategy including solid-phase extraction (SPE) thin-layer chromatography (TLC) and capillary zone electrophoresis equipped with a diode array detector (CZE-DAD) was developed to identify and quantify 14 hydrosoluble, acidic, synthetic food dyes allowed in the European Community. Indeed, these may be the most susceptible dyes to be found in illicit pills through their availability and easiness of use. The results show (1) that this analytical method is well adapted to small samples such as illicit pills, (2) that most dyes actually found belong to hydrosoluble, acidic, synthetic food dyes allowed in the European Community, and (3) that this evidence turns out to be important in drug intelligence and may be assessed into a Bayesian framework.

Amphetamines↗
PubMed2006

[Error permissibility of neural network used for renal corpuscle area enhancement].

In the automatic analysis system of the kidney-tissue image, boundary enhancement for glomerulus area is a vital step. Complex characteristics of kidney-tissue image leads to the difficulty in boundary features description. This paper suggests a kind of feature template under the special boundary definition. A nonlinear threshold surface is constructed by neural network, then the proper surface can be selected to enhance boundary with the influence of error permissibility being taken into account. Experimental results indicate that this learning method with error permissibility can enhance the boundary of glomerulus and suppress noises at the same time, so it can obtain good processed effects and have a fine performance highly adaptive to various sample images.

Humans↗
PubMed1988

High-density lipoprotein apolipoproteins in urine: II. Enzyme-linked immunoassay of apolipoprotein A-I.

We have developed a capture antibody, noncompetitive, enzyme-linked immunoassay for urinary apolipoprotein A-I (Apo A-I) in urine, with use of affinity-purified polyclonal antisera against Apo A-I. A 96-well microtiter plate format is used, with unconcentrated urine as sample and dilutions of serum or high-density lipoprotein (HDL) as standards. The intra- and interassay variation (CV) averaged 7.4% and 9.4%, respectively. The limit of detection is low (1.25 ng/L), and no cross-reactivity with Apo B, C, E, or A-II was detected. The mean (+/- SD) concentrations of Apo A-I in urine of patients with glomerular proteinuria were a thousandfold greater (38.4 +/- 23.1 mg/L) than in normal subjects (16.3 +/- 11.3 micrograms/L in men, 17.97 +/- 7.7 micrograms/L in women, a significant difference, P less than 0.001). Apo A-I measurements correlated very well (r = 0.92) with selectivity index assessment. The diurnal variation of the concentration of Apo A-I in urine appears to result from dilution related to fluid intake. This enzymatic method is easy to perform, can be used with large numbers of samples, and is adaptable for use in the routine clinical laboratory. The method holds promise for discriminating between normal and subclinical kidney disease populations by measuring the concentrations of urinary Apo A-I excreted on HDL particles.

Adolescent↗
PubMed1986

[Enzymuria].

Study of enzymuria rises more and more interest in human pathology as a diagnosis parameter of renal diseases or as an index of nephrotoxicity. In this two fields, the literature demonstrated the interest of enzymes of the brush border of proximal tubuli (alanine amino peptidase, gamma glutamyl transpeptidase, alkaline phosphatase) and of lysosomal enzymes (beta glucuronidase and N acetyl beta D glucosaminidase). The authors analyse the problems limiting the present use of these methods: incomplete knowledge of enzyme of the different parts of the renal tissue and of the mechanisms of enzymuria (choice of enzymes and time periods for sampling); analytical problems referring to the study of enzymes in a complex medium (treatment and storage of samples, choice of adapted methods for activity measurement); at last the way of expression the results is still to be defined.

Clinical Enzyme Tests↗
PubMed1982

[Randomization for controlled therapeutic trials].

The choice of an adequate randomization procedure, the organization of the randomization process and its correct implementation are questions that may influence the quality of a controlled therapeutic trial. Starting with a brief critical review of randomization procedures now available, this paper presents in greater detail one method which is thought to be optimal for clinic trials of typical sample sizes. The "adaptive randomization" procedure of Pocock and Simon [3] keeps all 2-8 factors of prognostic importance in satisfactory balance-as has been confirmed by trials in the past 4 years. An interactive computer program, RANDOM, is introduced, whereby easy and safe use of the procedure is enabled via a video terminal. Furthermore, standardized recording of the randomization process by a clerk for medical documentation in the central randomization office is described. The possibility of randomizing multi-factorial trials and the admissibility of eliminating already randomized patients from a trial are discussed. The working group on documentation and biometry of the 1st Department of Surgery, Vienna University Medical School, offers the program to any centres that may be interested and is also willing to undertake randomizations (by phone) for clinical trials within Austria.

Aged↗
PubMed1994

Revised estimates of diagnostic test sensitivity and specificity in suspected biliary tract disease.

BACKGROUND: The purpose of this study was to estimate the sensitivity and specificity of diagnostic tests for gallstones and acute cholecystitis. METHODS: All English-language articles published from 1966 through 1992 about tests used in the diagnosis of biliary tract disease were identified through MEDLINE. From 1614 titles, 666 abstracts were examined and 322 articles were read to identify 61 articles with information about sensitivity and specificity. Application of exclusion criteria based on clinical and methodologic criteria left 30 articles for analysis. Cluster-sampling methods were adapted to obtain combined estimates of sensitivities and specificities. Adjustments were made to estimates that were biased because the gold standard was applied preferentially to patients with positive test results. RESULTS: Ultrasound has the best unadjusted sensitivity (0.97; 95% confidence interval, 0.95 to 0.99) and specificity (0.95; 95% confidence interval, 0.88 to 1.00) for evaluating patients with suspected gallstones. Adjusted values are 0.84 (0.76 to 0.92) and 0.99 (0.97 to 1.00), respectively. Adjusted and unadjusted results for oral cholecystogram were lower. Radionuclide scanning has the best sensitivity (0.97; 95% confidence interval, 0.96 to 0.98) and specificity (0.90; 95% confidence interval, 0.86 to 0.95) for evaluating patients with suspected acute cholecystitis; test performance is unaffected by delayed imaging. Unadjusted sensitivity and specificity of ultrasound in evaluating patients with suspected acute cholecystitis are 0.94 (0.92 to 0.96) and 0.78 (0.61 to 0.96); adjusted values are 0.88 (0.74 to 1.00) and 0.80 (0.62 to 0.98). CONCLUSIONS: Ultrasound is superior to oral cholecystogram for diagnosing cholelithiasis, and radionuclide scanning is the test of choice for acute cholecystitis. However, sensitivities and specificities are somewhat lower than commonly reported. We recommend estimates that are midway between the adjusted and unadjusted values.

Acute Disease↗
PubMed1993

Irrigation with waste water: parasitological analysis of soil.

In third world countries, the agricultural use of treated wastewater represents an interesting solution. A recent report of W.H.O. shows the importance of strict parasitological criteria for such reuse. The aim of this paper is to study the conditions under which Ascaris eggs have been recovered from artificially contaminated soils (sandy, clay or loamy soil, mould garden). The eggs elution has been carried out from soil particles using various solutions (detergents, distilled water, formaldehyde, sodium hydroxide, sodium hypochlorite). The recovery percentage analysis showed the superiority of the sodium hypochlorite solution titrating 10 chlorometric degrees, whatever the soil type. The eggs concentration in the eluates has been carried out through the flotation technique by testing various reagents with densities ranging from 1.16 to 1.44. The zinc sulphate solution at 50%, 55% and 66% prove to be flotation agents that are well adapted to this sampling type. By carrying out the elution with a sodium hypochlorite solution titrating 10 chlorometric degrees and the concentration by flotation with a zinc sulphate solution at 55% the recovery percentages, which are independent of the parasitic load, vary from 66 to 78%.

Agriculture↗
PubMed1998

A method for examining differential mRNA expression along the crypt-villus axis of the human small intestine.

1. The aim of this study was to devise a method of segregating crypt and villus cell subpopulations from endoscopic human small intestinal biopsies which might be used to examine changes associated with functional differentiation at the molecular level. 2. Routine endoscopic biopsies from the human small intestine were subjected to a modified protocol of mechanical disruption and chelation to yield subpopulations of different cell types. The purity and character of the cell populations isolated was assessed by measuring enzyme activity and thymidine incorporation and by histology. A guanidinium isothiocyanate method was adapted for small samples to extract RNA from the isolated subpopulations, and probes for RNA with a known predilection for crypt and villus cells were used to further investigate the application and usefulness of the technique. 3. Sequential histological examination during the segregation protocol demonstrated that different cell types were removed serially from the biopsy samples. Cell-type enrichment of the segregated subpopulations was demonstrated by differential alkaline phosphatase activity and by differences in thymidine incorporation in the samples isolated. Sufficient quantities of RNA could be extracted from the segregated subpopulations for Northern blot analysis and the differential expression of mRNA for sucrase-isomaltase and transferrin receptor was demonstrated in the villus and crypt subpopulations respectively. 4. Messenger RNA can be successfully extracted from different cell types segregated from routine human endoscopic small intestinal biopsies. This technique should prove useful for investigating the mechanisms regulating the functional differentiation of epithelial cells in the small intestine and the regulatory mechanisms governing absorption of macromolecules.

Biopsy↗