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Patterns of cortisol reactivity to laboratory stress.

Cortisol responses to a laboratory stress protocol were investigated in 82 male firefighters. Saliva samples were collected during an adaptation period beginning between 9 and 10 am, and then at the end of each of six 10-min trials (a mental arithmetic task, an inter-task recovery period, a speech task, and three recovery periods). Individual differences in the mean cortisol response to the stress tasks were characterized by variation in the direction of the response, as well as the size of the response. Neither pre-stress cortisol levels nor responses were correlated with cardiovascular and mood responses. Cortisol levels before stress task presentation were negatively correlated with recent stress severity. Larger mean cortisol responses were associated with lower reports of recent stress exposure, lower negative affect scores, and a coping style characterized less experience of anger, more control over anger expression, and a tendency to screen out threatening information in stressful situations. Thus, increased cortisol activity was associated with less recent stress exposure and a more adaptive behavioral style than for those whose cortisol levels fell or were largely unchanged in response to a laboratory stressor.

Adaptation, Physiological↗

Assessment of titrimetric and photometric methods for the determination of chymotrypsin catalytic activity in stool.

In this report a method for the spectrophotometric measurement of stool chymotrypsin (CT) is presented and compared to the more commonly used titrimetric method. Homogenisation of specimen and dissolution of CT is examined in detail. The advantages of the photometric method lie in the small size of sample required, and its easy adaptation to manual or automated analysis with a CT-specific amide substrate.

Catalysis↗

Influence of solute polarity in column-switching chromatography for the assay of drugs in plasma and urine.

The polarity of a drug is one of the most important parameters for the elaboration of switching systems. If the polarity of the drug is low or medium, "reversed-phase" chromatography is well adapted. The plasma or urine sample is diluted with water, centrifuged and injected first into a column of medium polarity (C2, CN or diol bonded phases). The compounds of interest are stopped on the top of the column and rinsed with water, then eluted and chromatographed on a C8 or C18 analytical column. A third column of still lower polarity can be added to improve the specificity of the system. In each successive step, the polarities of the mobile phases and columns should be decreased to reconcentrate the sample and reduce the band broadening that occurred in the previous step. Compounds of high polarity show almost no retention on reversed-phase columns, and normal-phase chromatography should be used. Aqueous solutions cannot be injected into polar bonded-phase columns as they lead to excessive band broadening. This problem can be solved by diluting plasma or urine with a large volume of a water-miscible organic solvent and injecting the clear supernatant. The compounds to be assayed are first reconcentrated on a polar column (NH2 or N(CH3)2 bonded phase) and then eluted. The selected "heart cut" of the eluate is chromatographed on another, more polar column. The influence of the polarity of drugs on the choice of switching systems is exemplified by assay methods for drugs of low, medium and high polarity.

Acetonitriles↗

A five-year clinical evaluation of Class II composite resin restorations.

OBJECTIVES: To study the clinical efficacy of posterior composite resin restorations placed in general practice after five years. METHODS: Two commercial composite resin systems were used. Three general practitioners placed the restorations at a Public Dental Health Service Office. The patients were not selected specifically for this study. Class II cavities were restored with Superlux Molar and P-50 APC composite systems on an alternate basis. At baseline, 63 restorations were placed in molars and premolars in 45 patients. For primary caries, generally a conservative cavity design was used (n = 23), while replacements of amalgams resulted in the larger conventional Class II design (n = 40). The restorations were assessed using a modified USPHS criteria. Wear measurements were determined by the Leinfelder method. Photographs and bite-wing radiographs were taken to supplement the clinical evaluation of colour match, marginal adaptation and recurrent caries. Saliva sampling was performed to determine the rate of secretion and the level of mutans streptococci and lactobacilli. RESULTS: At the five-year review 51 restorations were available for examination, of which nine restorations were rated clinically unacceptable. Including the failed restorations (n = 7), at the three-year review, a total of 16 restorations had failed (27.6%) over a five-year period. The most common reasons for failure were recurrent caries (n = 7) and marginal defects (n = 4). The mean wear of Superlux Molar was 167 microns and of P-50 APC 158 microns. Eight of the 11 patients with failed restorations due to caries and marginal defects had high counts of mutans streptococci at baseline. CONCLUSIONS: The failures in the present group of patients did not specifically relate to material, tooth type or cavity design. However, it is suggested that patient factors such as caries activity should be monitored and managed.

Adolescent↗

Simultaneous determination of norepinephrine, dopamine and serotonin in rat brain regions by ion-pair liquid chromatography on octyl silane columns and amperometric detection.

A highly sensitive and specific ion-pair liquid chromatography-electrochemical detection method is described for the simultaneous determination of rat brain regional norepinephrine, dopamine and serotonin levels using octylsilane (C8) columns. These amines were first isolated from tissue homogenates by adsorption on Amberlite CG-50 resin followed by separation on RP-8 columns with a mobile phase consisting of 0.05 M NaH2PO4 (pH 3.0) 0.02 mM EDTA, 1 mM heptanesulphonate-methanol (92:8, v/v) at 1.8 ml/min. Using 3,4-dihydroxybenzylamine as the internal standard, tissue recoveries (mean +/- S.D.) for norepinephrine, dopamine and serotonin were 87.5 +/- 2.6%, 61.8 +/- 10.5% and 72.9 +/- 7.5%, respectively. Assay sensitivities were sufficient for reliable quantitation of at least 200 pg of these compounds in a brain sample. The procedure is readily adaptable to determination of brain epinephrine and normetanephrine levels, as well. Finally, the reversed-phase system employed is highly flexible in that the same column and mobile phase conditions may be used for assay of biogenic amine metabolites.

Animals↗

The development of immunoassays to identify and quantify species source of gum arabic.

Gum arabic from Acacia senegal is commonly used as an additive in foodstuffs. Adulteration of gum arabic by other gums is a potential problem for reasons of safety and quality. This study aimed to develop and evaluate the use of enzyme-linked immunosorbent assays (ELISAs) for the detection of potential adulterants of gum arabic. Indirect competitive ELISAs (IC-ELISAs) were developed using the monoclonal antibodies SY CC7 (A. senegal), SY HH3 (Acacia seyal), and SY J1A1 (Combretum erythrophyllum). All IC-ELISAs had a working range of 0.005-10 mg/mL. The antibodies used were tested using the IC-ELISAs for cross-reactivity with other Acacia species and other gums. The antibodies were very specific for their respective antigens. Significant cross-reactivity was found for SY CC7 (between A. senegal and A. melliferae) and SY J1A1 (between C. erythrophyllum and A. seyal). The IC-ELISA was adapted further to test confectionery samples for the presence of gum arabic, which was successful, although recovery rates were reduced. Both IC- and plate trapped antigen ELISA (PTA-ELISA) formats were able to distinguish an adulterated sample of gum arabic when blended with either A. seyal or C. erythrophyllum. The PTA-ELISA was more sensitive for A. seyal than the IC-ELISA, but both were equally sensitive for C. erythrophyllum. The results suggest that the antibodies SY CC7, SY HH3, and SY J1A1 could be used in combination with each other for the detection of potential adulterants of A. senegal and the detection of gum arabic in foodstuffs.

Acacia↗

Determination of aluminum(III) in crystallized fruit samples using a multicommutated flow system.

A multicommutated flow system was developed for the determination of aluminum in crystallized fruit samples. Spectrophotometric determination is based on the reaction of aluminum with chrome azurol S. The binary sampling technique was implemented to improve mixing conditions and to minimize reagent consumption. Three different working zones were established (0.5-5.0, 5.0-25.0, and 10.0-100 ppm) using the zone sampling approach, allowing us to adapt the extent of the in-line dilution. The influence of the chemical and physical parameters on the performance of the system was studied. Detection limits of 0.1, 0.6, and 0.8 ppm were obtained for the lowest, the medium, and the highest dispersion system, respectively. The procedure was applied to the determination of aluminum in crystallized fruit extracts. The results were in agreement with those obtained by the reference flame atomic absorption procedure at a 95% confidence level. Repeatability (RSD) was better than 2.4% in all of the three application zones.

Aluminum↗

Measurement of dipolar couplings in a transducin peptide fragment weakly bound to oriented photo-activated rhodopsin.

Rhodopsin-containing disks, isolated from rod outer segments of bovine retina, align at high magnetic fields with their membrane normal parallel to the magnetic field. After light-activation of rhodopsin, transient binding of the C-terminal transducin undecapeptide, selectively labeled with 15N at Leu5 and Gly9, results in residual dipolar contributions to the 1J(NH) splittings for these two residues. Both residues show 1J(NH) splittings which are smaller than in the dark-adapted or rhodopsin-free sample, and return to their isotropic values at a rate determined by the decay of the meta II state of rhodopsin. The dipolar couplings indicate that in the bound state, N-H vectors of Leu5 and Gly9 make angles of 48+/-4 degrees and 40+/-8 degrees, respectively, with the disk normal. These 'transferred' dipolar couplings potentially offer a useful method for studying the conformation and orientation of flexible, low affinity ligands when bound to oriented integral membrane receptors.

Amino Acid Sequence↗

Adaptive test for testing the difference in survival distributions.

An adaptive test is proposed for the problem of testing the difference in survival distributions when the shape of the hazard ratio is unknown, hence the efficient test is unknown. The proposed adaptive test selects a test statistic from a finite set of the weighted logrank statistics T on the basis of the estimates of the efficiencies of the tests in T for given data. The efficiency estimator uses the length of the test based nonparametric confidence interval for the shift in a time transformed shift model. The suggested adaptive test is shown to be asymptotically efficient among the tests in T under the time transformed shift model and conditions commonly used in survival analysis. Simulations demonstrate that the adaptive test enjoys good small sample properties and in most situations is more powerful than the test using the maximum of the tests in T.

Computer Simulation↗

Prophylactic antibiotic treatment in patients with predicted severe acute pancreatitis: a placebo-controlled, double-blind trial.

BACKGROUND & AIMS: Antibiotic prophylaxis in necrotizing pancreatitis remains controversial. Until now, there have been no double-blind studies dealing with this topic. METHODS: A total sample size of 200 patients was calculated to demonstrate with a power of 90% that antibiotic prophylaxis reduces the proportion of patients with infected pancreatic necrosis from 40% placebo (PLA) to 20% ciprofloxacin/metronidazole (CIP/MET). One hundred fourteen patients with acute pancreatitis in combination with a serum C-reactive protein exceeding 150 mg/L and/or necrosis on contrast-enhanced CT scan were enrolled and received either intravenous CIP (2 x 400 mg/day) + MET (2 x 500 mg/day) or PLA. Study medication was discontinued and switched to open antibiotic treatment when infectious complications, multiple organ failure sepsis, or systemic inflammatory response syndrome (SIRS) occurred. After half of the planned sample size was recruited, an adaptive interim analysis was performed, and recruitment was stopped. RESULTS: Fifty-eight patients received CIP/MET and 56 patients PLA. Twenty-eight percent in the CIP/MET group required open antibiotic treatment vs. 46% with PLA. Twelve percent of the CIP/MET group developed infected pancreatic necrosis compared with 9% of the PLA group (P = 0.585). Mortality was 5% in the CIP/MET and 7% in the PLA group. In 76 patients with pancreatic necrosis on contrast-enhanced CT scan, no differences in the rate of infected pancreatic necrosis, systemic complications, or mortality were observed. CONCLUSIONS: This study detected no benefit of antibiotic prophylaxis with respect to the risk of developing infected pancreatic necrosis.

Adult↗

Quantum wave packet ab initio molecular dynamics: an approach to study quantum dynamics in large systems.

A methodology to efficiently conduct simultaneous dynamics of electrons and nuclei is presented. The approach involves quantum wave packet dynamics using an accurate banded, sparse and Toeplitz representation for the discrete free propagator, in conjunction with ab initio molecular dynamics treatment of the electronic and classical nuclear degree of freedom. The latter may be achieved either by using atom-centered density-matrix propagation or by using Born-Oppenheimer dynamics. The two components of the methodology, namely, quantum dynamics and ab initio molecular dynamics, are harnessed together using a time-dependent self-consistent field-like coupling procedure. The quantum wave packet dynamics is made computationally robust by using adaptive grids to achieve optimized sampling. One notable feature of the approach is that important quantum dynamical effects including zero-point effects, tunneling, as well as over-barrier reflections are treated accurately. The electronic degrees of freedom are simultaneously handled at accurate levels of density functional theory, including hybrid or gradient corrected approximations. Benchmark calculations are provided for proton transfer systems and the dynamics results are compared with exact calculations to determine the accuracy of the approach.

Journal Article↗

Linear-after-the-exponential (LATE)-PCR: an advanced method of asymmetric PCR and its uses in quantitative real-time analysis.

Conventional asymmetric PCR is inefficient and difficult to optimize because limiting the concentration of one primer lowers its melting temperature below the reaction annealing temperature. Linear-After-The-Exponential (LATE)-PCR describes a new paradigm for primer design that renders assays as efficient as symmetric PCR assays, regardless of primer ratio. LATE-PCR generates single-stranded products with predictable kinetics for many cycles beyond the exponential phase. LATE-PCR also introduces new probe design criteria that uncouple hybridization probe detection from primer annealing and extension, increase probe reliability, improve allele discrimination, and increase signal strength by 80-250% relative to symmetric PCR. These improvements in PCR are particularly useful for real-time quantitative analysis of target numbers in small samples. LATE-PCR is adaptable to high throughput applications in fields such as clinical diagnostics, biodefense, forensics, and DNA sequencing. We showcase LATE-PCR via amplification of the cystic fibrosis CFDelta508 allele and the Tay-Sachs disease TSD 1278 allele from single heterozygous cells.

Alleles↗

Attachment site(s) of retinal in bacteriorhodopsin.

After chemical reduction of the retinylidene-lysine Schiff base linkage in bacteriorhodopsin, the retinyl residue is covalently attached to Lys-216 (with a possible minor fraction on Lys-172) or to both Lys-216(172) and Lys-40/41. The linkage site (up to 100% on Lys-216; up to 70% on Lys-40/41) depends on whether the sample is reduced in the light or dark, whether the sample is light or dark adapted, and on temperature. Absorbance and circular dichroism spectra indicate that the retinyl residue is in its original binding site after reduction in the light. Thus, the different attachment sites may reflect changes that occur during the photoreaction cycle or during light/dark adaptation, or the reduction of accidental physiologically irrelevant Schiff base linkages to lysines close to the normal linkage in the structure of bacteriorhodopsin. In either case, the retinal does not leave its binding site. This last point severely limits the possible arrangements of the amino acid sequence in the bacteriorhodopsin tertiary structure and clearly distinguishes two models that are consistent with all criteria.

Bacteriorhodopsins↗

Appraisal of the MTT-based assay as a useful tool for predicting drug chemosensitivity in leukemia.

The MTT-based assay relies upon the cellular reduction of tetrazolium salts to their intensely colored formazans. The test is easy to perform in hematological malignancies and is adaptable for high throughput of samples, although there are some minor limitations in its application resulting from metabolic interference. This class of assays are highly accurate for predicting drug resistance, whereas their predictive value for drug sensitivity depends on the type of disease and drug or drug combination used. They have been found to predict clinical response to fludarabine FLD in B-CLL and were useful for predetermining clinical potential of a single drug or drug combination in AML patients. Extensive studies with ALL patients have supported their advantage for selecting effective drug treatment of the disease. To conclude, pretreatment chemosensitivity assays may help in the selection of chemotherapeutic drugs with the greatest likelihood for clinical effectiveness, and in the exclusion of uneffective therapy. This can lead to improved disease management, response, survival and use of financial resources.

Antineoplastic Agents↗

Subcutaneous tissue oxygen and carbon dioxide tensions during hyperbaric oxygenation: an experimental study in rats.

OBJECTIVE: To investigate the response of subcutaneous tissue oxygen (O2) and carbon dioxide (CO2) tensions to hyperbaric oxygenation. DESIGN: Experimental study. SETTING: University hospital, Finland. SUBJECTS: 10 Wistar rats. INTERVENTION: Subcutaneous tissue PO2 and PCO2 were directly measured with an implanted Silastic tube tonometer and capillary sampling technique while breathing air and exposed to hyperbaric oxygen (HBO) at 2.5 or 2.8 ATA pressure. Hyperbaric exposures were carried out in a large multiplace chamber pressurised with air. MAIN OUTCOME MEASURES: Subcutaneous tissue PO2 and PCO2. RESULTS: The mean subcutaneous PO2 rose from the baseline of 8 kPa (60 mmHg) to 16 kPa (112 mmHg) when rats breathed room air during pressurisation to 2.8 atm. When the rats breathed oxygen at 2.5 ATA the maximal mean tissue PO2 was four times higher than the mean starting value. During the HBO treatment at 2.8 ATA the tissue PO2 rose to a value about five times above baseline. The tissue PCO2 values almost doubled during the exposure to HBO at 2.5 ATA, probably because elimination of carbon dioxide was impaired. CONCLUSION: Measurements of tissue PO2 and PCO2 with an implanted Silastic tonometer and a capillary sampling technique can successfully be adapted to hyperbaric conditions. The method yielded reproducible results and is applicable to clinical use in hyperbaric medicine.

Animals↗

ADL dependence and perceived health among elderly deaf sign-language users: a comparison with a matched group of elderly hearing people.

The main purpose of the study was to investigate dependence in ADL and perceived health among elderly deaf sign-language users and to compare these findings with data from a matched hearing elderly population sample. Data on the deaf population were collected through assessments and structured interviews in sign language. Data on the hearing population were retrieved from the Swedish National Study and Ageing and Care (SNAC), while only a few variables from the questionnaire were used in the present study. Each deaf individual was matched by three hearing persons. Matching was performed according to gender, age, and living situation. The main finding is that elderly deaf sign-language users are more dependent in ADL but simultaneously report better health than their hearing counterparts. The differences in ADL dependence can be explained by factors linked to the communication problems experienced by deaf sign-language users, but also by cultural factors. The better health reported by the deaf sample may be attributed to adaptive mechanisms that deaf sign-language users have been claimed to develop. This research provides new information on a vulnerable group of elderly people, i.e. deaf sign-language users, that has previously been given little scientific attention.

Activities of Daily Living↗

Oral dydrogesterone treatment during the first trimester of pregnancy: the prevention of miscarriage study (PROMIS). A double-blind, prospectively randomized, placebo-controlled, parallel group trial.

OBJECTIVE: A planned study to test the hypothesis that dydrogesterone given during the first trimester of pregnancy can positively influence the immune system of women suffering from recurrent miscarriages and thereby lead to successful pregnancies. The primary objective is to show whether the administration of dydrogesterone during the first trimester of pregnancy increases the release of progesterone-induced blocking factor (PIBF) and causes a shift from Th-1 cytokines to Th-2 cytokines. Further objectives are an analysis of pregnancy outcome and to investigate the tolerability and safety of dydrogesterone use during pregnancy. METHODS: A prospectively randomized, placebo-controlled study of 20 mg dydrogesterone per day during the first trimester of pregnancy in women aged between 18 and 38 and with a history of idiopathic recurrent miscarriage. Start of treatment will be after ovulation. The study will be conducted in a two-stage adaptive design, starting with a sample size of 20 patients per treatment group. The concentrations of Th-1 and Th-2 cytokines in the serum and PIBF in the urine will be measured. Efficacy will be measured primarily by the ratio IFNgamma/IL-10. CONCLUSION: The results of this study will contribute to the discussion of a current concept of immunomodulation in pregnancy and potentially provide a treatment option for patients suffering from recurrent miscarriage.

Abortion, Habitual↗

An evaluation of a new desorption technique for air sampling sorbent tubes.

Air sampling sorbent tubes have been used for many years in determining concentrations of chemicals in the work environment. The conventional way to desorb analytes from the sorbents involves physical removal and transfer of the sorbent from the tube to a vial containing desorption solution. A new desorption technique, whole tube desorption--which involves adding the glass tube containing the sorbent to the vial containing the desorption solvents--has been evaluated. With this technique there is no transfer of sorbent out of the glass tube. Recovery efficiencies for several chemicals including ethylene oxide, methylene chloride and styrene were comparable to those obtained using traditional solvent desorption procedures. The utilization of this new desorption technique will involve a simple modification in the design of commercially available sorbent tubes so that the front and back sections can be physically separated. This new technique allows for a significant reduction in the time spent in the desorption of air sampling sorbent tubes and easier adaptation to laboratory robotic systems.

2,4-Dichlorophenoxyacetic Acid↗