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PubMed2003

RNA extraction and RT-PCR procedures adapted for the detection of enterovirus sequences from frozen and paraffin-embedded formalin-fixed spinal cord samples.

Methods for detecting enterovirus RNA in both paraffin-embedded, formalin-fixed and frozen spinal cord sections from amyotrophic lateral sclerosis (ALS) patients were established. A proteinase K digestion following the deparaffinization procedure was required for the fixed spinal cord sections, whereas only one step of crushing in phosphate buffered saline was necessary for the frozen samples prior to the extraction of the RNA. With an optimized reverse transcription and PCR procedure, enterovirus RNA could be detected from frozen and fixed archival spinal cord samples.

Amyotrophic Lateral Sclerosis↗
PubMed1992

Adaptation of a commercially available enzyme linked immunosorbent assay (ELISA) for the determination of thyroxine in canine plasma samples.

The purpose of the present study was to modify a previously evaluated enzyme linked immunosorbent assay (Larsson and Lumsden, 1980) for the determination of the thyroxine (T4) concentration in canine plasma samples. The test kit Enzymun-Test(Boehringer Mannheim) for determination of T4 concentrations in human plasma was used in connection with the analyzer system Enzymun-Test(Boehringer Mannheim) System ES-33. The assay protocol originally used by Larsson and Lumsden (1980) was modified by including an additional standard, prolongation of the incubation period for the immunoreaction and the enzymeassay, and reduction of the amount of T4 conjugate. The intra- and interassay coefficient of variations ranged from 3.8% to 12.5%. The detection limit of the assay was 3.9 nmol/l which is well below the T4 concentration observed in plasma from healthy dogs, and the T4 concentrations determined in plasma from healthy dogs before and 3 hours after administration of TRH (38.0 +/- 7.4 nmol/l and 51.8 +/- 8.5 nmol/l, respectively) were comparable to those reported in the literature.

Animals↗
PubMed2001

Adaptation of preventive interventions for a low-income, culturally diverse community.

OBJECTIVE: To describe essential elements in the adaptation of a prevention approach with a high-risk urban sample, chosen to contrast sharply with the primarily middle-class sample in which it had been originally tested. METHOD: Key elements of a preventive intervention for families with parental depression were adapted for use in the new context. RESULTS: A sequence of alliance-building events was implemented, involving engagement at three levels: community, caregivers, and family. The prevention approach was modified to include an expanded approach to defining depression and resilience; greater flexibility on the part of the clinician; more intensive engagement between clinician and family, with a focus on immediate daily concerns; as well as awareness of cultural issues and responsiveness to the subject's experience of violence. CONCLUSION: Core principles of helping family members to discuss the effects of depression and adversity on family life were affirmed.

Adolescent↗
PubMed2001

Contrast adaptation in human retina and cortex.

PURPOSE: Although cortical contrast adaptation has been extensively studied with both psychophysical and electrophysiological techniques, little is known about retinal contrast adaptation in humans. METHODS: Retinal and cortical long-term contrast adaptation was assessed with simultaneous measurement of pattern electroretinogram (PERG) and cortical visual evoked potentials (VEPs). This study involved three approaches: sampling of the contrast transfer function from 2.7% to 98% with adaptation to high (98%) and low (7.3%) contrasts, linearity of adaptation effects, and transfer of contrast adaptation between parallel and orthogonal grating orientations. RESULTS: Contrast adaptation affected retinal and cortical recordings quite differently. The VEP showed a sigmoid contrast transfer function, which was shifted toward higher contrasts (by a factor of 1.9), whereas amplitudes at higher test contrasts were enhanced to 127%. The PERG decreased in amplitude to approximately 90%, and the latency was significantly reduced by 4 to 6 msec (P < 0.05). All measured effects were linear with adaptation contrast. Orientation played no role in the PERG results, whereas the VEP was enhanced to 125% when tested parallel and to 150% when tested orthogonal to adaptation. CONCLUSIONS: VEP results confirm and extend previous findings and fit well with single-cell recordings. The PERG findings suggest that retinal contrast adaptation occurs and mainly operates in the temporal domain, comparable to rapid gain-control findings in cats and primates.

Adaptation, Ocular↗
PubMed1996

Analysis of respiratory water--a new method for evaluation of myocardial energy metabolism.

Aerobic ATP synthesis via oxidative phosphorylation causes a proportional production of respiratory water. Thus the amount of respiratory water produced at a given time should be a reliable measure of the current ATP demand of the mammalian myocardium. Respiratory water from isolated rabbit hearts was labeled by using the stable oxygen isotope 18O. The hearts were perfused according to the method of Langendorff (O. Langendorff. Pfluegers Arch. 61: 291-332, 1895) with 18O2-equilibrated Krebs-Henseleit solution. Control hearts were exclusively perfused with carbogen-equilibrated Krebs-Henseleit solution. Myocardial tissue was then lyophilized; the extracted water and samples from the coronary venous effluent were converted to CO2 by using the guanidine hydrochloride technique. The delta 18O values within the CO2 samples were determined by mass spectrometry and related to the standard mean ocean water (SMOW) scale. Compared with control hearts, the 18O-labeled hearts exhibited a significant increase of delta 18O values from tissue water (-47.50 +/- 0.64 vs. -40.35 +/- 2.05% SMOW; P < 0.05). The values were also significantly increased in the coronary venous effluent after a perfusion time of only 50 s (-47.50 +/- 0.64 vs. -43.66 +/- 0.91% SMOW; P < 0.05). Thus this first adaptation of the guanidine hydrochloride technique on microliter samples of myocardial tissue water and coronary venous effluent demonstrates that this method can be used to evaluate both respiratory activity and the kinetics of cardiac metabolic processes.

Adenosine Triphosphate↗
PubMed1998

Forced-choice staircases with fixed step sizes: asymptotic and small-sample properties.

Visual detection and discrimination thresholds are often measured using adaptive staircases, and most studies use transformed (or weighted) up/down methods with fixed step sizes--in the spirit of Wetherill and Levitt (Br J Mathemat Statist Psychol 1965;18:1-10) or Kaernbach (Percept Psychophys 1991;49:227-229)--instead of changing step size at each trial in accordance with best-placement rules--in the spirit of Watson and Pelli (Percept Psychophys 1983;47:87-91). It is generally assumed that a fixed-step-size (FSS) staircase converges on the stimulus level at which a correct response occurs with the probabilities derived by Wetherill and Levitt or Kaernbach, but this has never been proved rigorously. This work used simulation techniques to determine the asymptotic and small-sample convergence of FSS staircases as a function of such parameters as the up/down rule, the size of the steps up or down, the starting stimulus level, or the spread of the psychometric function. The results showed that the asymptotic convergence of FSS staircases depends much more on the sizes of the steps than it does on the up/down rule. Yet, if the size delta+ of a step up differs from the size delta- of a step down in a way that the ratio delta-/delta+ is constant at a specific value that changes with up/down rule, then convergence percent-correct is unaffected by the absolute sizes of the steps. For use with the popular one-, two-, three- and four-down/one-up rules, these ratios must respectively be set at 0.2845, 0.5488, 0.7393 and 0.8415, rendering staircases that converge on the 77.85%-, 80.35%-, 83.15%- and 85.84%-correct points. Wetherill and Levitt's transformed up/down rules--which require delta-/delta+ = 1--and the general version of Kaernbach's weighted up/down rule--which allows any delta-/delta+ ratio--fail to reach their presumed targets. The small-sample study showed that, even with the optimal settings, short FSS staircases (up to 20 reversals in length) are subject to some bias, and their precision is less than reasonable, but their characteristics improve when the size delta+ of a step up is larger than half the spread of the psychometric function. Practical recommendations are given for the design of efficient and trustworthy FSS staircases.

Contrast Sensitivity↗
PubMed2005

A plate assay for simultaneous screening of polysaccharide- and protein-degrading micro-organisms.

AIMS: To develop a plate assay for simultaneous screening of polysaccharide-degrading and protein-degrading micro-organisms. METHODS AND RESULTS: A plate assay, based on the visible solubilization of small substrate particles and the formation of haloes on Petri dishes, containing a mixture of diversely coloured insoluble polysaccharides and dye-labelled collagen as chromogenic substrates, was developed. This method was successfully applied for isolating the diverse polysaccharide- and/or protein-degrading bacteria from soil and sludge samples. Selected strains were identified using 16S rDNA partial sequencing; most of them belong to the genera Bacillus, Cellulomonas and Cellulosimicrobium. CONCLUSIONS: This novel approach provides unique and valuable information for direct primary screening when the target of selection is micro-organisms exhibiting protein-degrading activity, polysaccharide-degrading activity or a specific combination of them. SIGNIFICANCE AND IMPACT OF THE STUDY: This plate assay is convenient and easy to perform, rapid, and more adaptable for screening of a large number of samples, compared with other existing methods in the literature.

Actinomycetales↗
PubMed1980

Effects of adaptation on biodegradation rates in sediment/water cores from estuarine and freshwater environments.

Experiments were devised to determine whether exposure to xenobiotics would cause microbial populations to degrade the compounds more rapidly during subsequent exposures. Studies were done with water/sediment systems (ecocores) taken from a salt marsh and a river. Systems were tested for adaptation to the model compounds methyl parathion and p-nitrophenol. CO(2) released from radioactive parent compounds was used as a measure of mineralization. River populations preexposed to p-nitrophenol at concentrations as low as 60 mug/liter degraded the nitrophenol much faster than did control populations. River populations preexposed to methyl parathion also adapted to degrade the pesticide more rapidly, but higher concentrations were required. Salt marsh populations did not adapt to degrade methyl parathion. p-Nitrophenol-degrading bacteria were isolated from river samples but not from salt marsh samples. Numbers of nitrophenol-degrading bacteria increased 4 to 5 orders of magnitude during adaptation. Results indicate that the ability of populations to adapt depends on the presence of specific microorganisms. Biodegradation rates in laboratory systems can be affected by concentration and prior exposure; therefore, adaptation must be considered when such systems are used to predict the fate of xenobiotics in the environment.

Journal Article↗
PubMed1995

[Toxicity assessment of waste water samples with fish cell lines]

An in vitro cytotoxicity assay with fish cell lines was established in order to reduce the number of animals used in fish tests with the golden orfe (leuciscus idus melanotus). In three co-ordinated working laboratories the toxicity of seven chemicals was determined according to a standardized protocol using the permanent cell lines D11 and RTG-2. There was an almost linear correlation (r=0.99) between the IC50 values after neutral red uptake and crystal violet staining of the chemically treated RTG-2 cells. These cells were more sensitive than D11 cells (r=0.93) with regard to chemically induced cytotoxicity. The test protocol was adapted for the testing of waste water (ten samples). Two samples induced cytotoxic effects in RTG-2 cells while in none of the three laboratories the same samples were found cytotoxic in D11 cells. The data with the seven chemicals and the still limited number of waste water samples (10) lend strong support to the assumption that the fish cell line RTG-2 can be regarded as well suited for detecting toxic properties of waste water.

Journal Article↗
PubMed2000

Rapid recovery of photosystems on rewetting desiccation-tolerant mosses: chlorophyll fluorescence and inhibitor experiments.

In the mosses Racomitrium lanuginosum, Anomodon viticulosus and Rhytidiadelphus loreus, after a few days air dry, F:(v)/F:(m) reached, within the first minute of remoistening in the dark, two-thirds or more of the value attained after 40 min. A fast initial phase of recovery was completed within 10-20 min after which further change was slow. Initial recovery of Phi(PSII) in the light was somewhat slower, but was generally substantially complete within a similar time. Remoistening with 0.3 mM cycloheximide (CHX) or 3 mM dithiothreitol (DTT) made little difference to this short-term (40 min) recovery of either F:(v)/F:(m) or Phi(PSII); 3 mM chloramphenicol (CMP) had little effect on recovery of F:(v)/F:(m), but resulted in substantial (though not total) depression of Phi(PSII) and (14)CO(2) uptake. Effects of the protein-synthesis inhibitors and DTT were much more clearly apparent in longer-term experiments (>20 h) but only in the light. In the dark, the three inhibitors had at most only slight effects over periods of 60-100 h. In the light, CMP-treated samples of all three species showed a progressive decline of dark-adapted F:(v)/F:(m), falling to zero within 1-5 d (possibly due to blocking of the turnover of the D1 protein of PSII) and accelerated by DTT. CHX-treated samples showed a similar but slower decline. In the shade-adapted and relatively desiccation-sensitive Rhytidiadelphus loreus, slow recovery of F:(v)/F:(m) continued in the dark even in the presence of CMP and CHX for much of the 142 h of the experiment. The results indicate that in desiccation-tolerant bryophytes recovery of photosynthesis after periods of a few days air dry requires only limited chloroplast protein synthesis and is substantially independent of protein synthesis in the cytoplasm.

Adaptation, Physiological↗
PubMed2003

Adaptation to disability in chronic obstructive pulmonary disease: neglected relationships to older adults' perceptions of independence.

PURPOSE: This study extends understanding of the relationship between disability and independence in older adults with COPD. METHODS: An interviewer-administered questionnaire was used to examine disability, adaptation, perceptions of independence, and self-efficacy in a sample of 50 community-dwelling older adults. Odds ratios were used to investigate relationships between variables. RESULTS: Participants used a wide range of behavioural strategies to adapt to their disability including: limiting activities, optimizing performance (e.g. taking rests), compensating for lost function (e.g. using equipment), and obtaining help from others. The relative use of these adaptations varied across five domains of activity: personal care, in-home mobility, household activities, community mobility, and valued activities. Most participants felt very or extremely independent in all domains. In personal care, those who reported greater disability (O.R. = 0.26), more frequent attempts to optimize performance (O.R. = 0.57), or greater reliance on help from others (O.R. = 0.79) were significantly (p < 0.05) less likely to feel very or extremely independent. CONCLUSIONS: The relationship between disability and perceptions of independence depends on the nature of the activity and is influenced by factors that are amenable to study and intervention. In particular, the vast array of behavioural strategies available to older adults with COPD enables them to feel highly independent despite disability.

Activities of Daily Living↗
PubMedDate not supplied

[The experience of using the FTA-carriers for blood sample storage while conducting the molecular-genetic identification of unidentified killed persons--victims of military actions in the territory of the Chechen Republic].

The activity of adapting and introducing the FTA-carriers for the storage of blood samples of relatives of missing service men for the purpose of using them in the consequent identification measures in respect to non-identified killed persons has been conducted in the Russian Federation within the framework of identification of service men killed in the Chechen territory during the 1994-1996 armed conflict. The article generalizes the experience of practical usage of FTA-cards as an element in the technological sequence of the molecular-genetic identification expertise. Methods related with application of FTA-technologies were tested; the efficiency of polymerized chain reaction was evaluated by using the matrix of immobilized DNA for hypo variable segments HVS-1 and HVS-2 as well as for the D-loop of mitochondrial DNA and for chromosomal STR-locuses D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820.

Base Sequence↗
PubMed1985

A recommended procedure for ultrastructural immunohistochemistry on small human tissue samples.

Various fixation and staining procedures for the demonstration of surface and cytoplasmic antigens have been described. An immunostaining procedure was sought that would allow the demonstration of these antigens, especially in small human tissue samples at the ultrastructural level. A modification and adaptation of the technique of Eldred, Zucker, Karten, and Yazula (J Histochem Cytochem 31:285, 1983) was applied on several varieties of human tissue, including liver, skin, and lymphoid tissue, using monoclonal and polyclonal antibodies in an indirect peroxidase procedure. In this way a reliable and generally applicable procedure was developed that satisfied the following demands: Use of a universal fixative that allows preservation of the antigenicity of various antigens; Adequate penetration of the tissue by the immunological reagents; Optimal preservation of subcellular structures; and Possibility to store the tissue samples for considerable periods of time.

Antibodies, Monoclonal↗
PubMed1993

Adaptation of dental plaque to sorbitol after 3 months' exposure to chewing gum.

Five subjects used sorbitol-containing chewing gum for a period of 12 wk. Plaque was collected before and after the sorbitol exposure and also 12 wk after the termination of the exposure. The individual plaque samples were incubated with 14C-labeled sorbitol, and the medium was examined by HPLC. It was found that plaque samples from all subjects catabolized more sorbitol after the exposure. The flora adapted to sorbitol was persistent, and all the subjects had a higher capacity for metabolizing sorbitol even 12 wk after the end of the sorbitol exposure than before it. Formate, acetate, ethanol, and lactate were the major catabolites of sorbitol. Small amounts of succinate and propionate were found in some samples.

Acetates↗
PubMed2006

An overview of statistical approaches for adaptive designs and design modifications.

With adaptive design methods for clinical trials, design elements such as sample size or further interim sample sizes may be changed during the course of the trial depending on all previously collected data. The focus of the overview is on group sequential approaches where the types of adaptations need not be specified in advance. An overview of the different statistical approaches for adaptive design methods proposed in the literature is given, relations between these methods are described and some perspectives of application and for future research are discussed.

Clinical Trials as Topic↗
PubMed2002

Local adaptation in the Linum marginale-Melampsora lini host-pathogen interaction.

The potential for local adaptation between pathogens and their hosts has generated strong theoretical and empirical interest with evidence both for and against local adaptation reported for a range of systems. We use the Linum marginale-Melampsora lini plant-pathogen system and a hierarchical spatial structure to investigate patterns of local adaptation within a metapopulation characterised by epidemic dynamics and frequent extinction of pathogen populations. Based on large sample sizes and comprehensive cross-inoculation trials, our analyses demonstrate strong local adaptation by Melampsora to its host populations, with this effect being greatest at regional scales, as predicted from the broader spatial scales at which M. lini disperses relative to L. marginale. However, there was no consistent trend for more distant pathogen populations to perform more poorly. Our results further show how the coevolutionary interaction between hosts and pathogens can be influenced by local structure such that resistant hosts select for generally virulent pathogens, while susceptible hosts select for more avirulent pathogens. Empirically, local adaptation has generally been tested in two contrasting ways: (1) pathogen performance on sympatric versus allopatric hosts; and (2) sympatric versus allopatric pathogens on a given host population. In situations where no host population is more resistant or susceptible than others when averaged across pathogen populations (and likewise, no pathogen population is more virulent or avirulent than others), results from these tests should generally be congruent. We argue that this is unlikely to be the case in the metapopulation situations that predominate in natural host-pathogen interactions, thus requiring tests that control simultaneously for variation in plant and pathogen populations.

Adaptation, Physiological↗