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A modified dot-blot method of protein determination applied in the tannin-protein precipitation assay to facilitate the evaluation of tannin activity in animal feeds.

Tannins have received considerable attention from animal nutritionists as potential agents for modifying ruminal fermentation patterns, or for exploring new feed resources. This group of secondary plant compounds is defined by their ability to form complexes with proteins. A widely accepted method for assaying the biological activity of extracted tannins is the precipitation of bovine serum albumin. The protein carries a radioactive label (125I) to allow direct quantification from the precipitate. Tannin-protein complexes dissolve in sodium dodecylsulfate solution. A dot-blot assay for protein determination, which is based on the reversible binding of a fluorochrome, benzoxanthene yellow, to the protein spots and is not disturbed by the presence of detergents, can replace the radioactive method by a fluorimetric measurement. A novel alternative to the last part of the dot-blot assay is to scan the stained protein spots in situ using a video camera and computer image analysis. Several filter sets were tested and, within a concentration range of 0.1-2.0 mg protein/ml, each of them yielded results identical to the original method while the time required was only 30 % of the working time consumed by the original procedure. The modified dot-blot assay should be applicable to the evaluation of tannin activity in all shrub and tree foliages considered as animal feed.

Animal Feed↗

New methodologies for pharmacological treatment trials for alcohol dependence.

As the number of pharmacological agents for the treatment of alcoholism continues to proliferate, there is an increasing need to design studies that accurately measure the efficacy of these agents. Key study design issues include selecting subjects, calculating patient retention, determining medication compliance, maintaining adequate sample size to ensure that a clinically meaningful therapeutic effect will not be missed, and measuring the study outcome, including the amount of alcohol consumed and levels of craving. Because there is evidence that self-report of alcohol consumption and medication compliance can be unreliable in subjects, biological methods have been used. One type of direct measurement, carbohydrate-deficient transferrin levels, has been found to be a reasonably sensitive indicator of alcohol consumption that may provide meaningful outcome information. The ingestion of riboflavin with medication and its urinary measurement is an accepted method of compliance detection. Improved study design will lead to continued advances in pharmacotherapy for the treatment of patients with alcoholism.

Alcohol Deterrents↗

[Palliative treatment of malignant esophageal obstruction with a new endoscopically administered self-expanding nitinol stent: case report].

The endoscopic methods of palliative treatment in malignant esophageal stenoses caused by neoplasm which have been performed so far unfortunately do not provide permanently satisfying results. The implantation of self-expanding stents with the purpose of removing malignant esophageal obstructions has recently become the most acceptable method of treatment. By using that method it is possible to reduce or even completely remove difficulties in swallowing. In that way a significant improvement in life quality of patients with inoperable esophageal cancer is possible. The various types of metal endoprostheses used for the palliative treatment of esophageal malignant stenoses are described in this paper. Here is also presented a case of nitinol stent implantation in a 50 year old woman suffering from esophageal cancer. This type of therapeutic treatment has been applied in Croatia for first time.

Alloys↗

Comparison of indirect immunofluorescence and Western blot for detection of anti-human immunodeficiency virus antibodies.

There was 100% agreement between the results of indirect immunofluorescence (IF) and Western blot testing when these methods were used to detect antibodies to the human immunodeficiency virus in sera from 25 patients with acquired immune deficiency syndrome (AIDS), 20 patients with AIDS-related complex, 186 subjects at high risk for AIDS, and 40 healthy heterosexuals. However, there was only an 88.7% correlation between IF and Western blot results for 728 sera from blood and plasma donor centers that were selected on the basis of screening enzyme immunoassay reactivity. IF tests yielded nine false-negatives and were equivocal, yielding a nonspecific pattern of reactivity for both infected and uninfected cells for 73 of these specimens. The IF and Western blot methods were equal in performance for the detection of anti-human immunodeficiency virus antibodies in the high-risk and unselected low-risk groups, proving to be a practical approach for testing specimens from these subjects. However, the Western blot was the most acceptable method for the validation of specimens from groups at low risk for AIDS that were selected based on enzyme immunoassay reactivity.

AIDS-Related Complex↗

Noninvasive measurement of rate of left ventricular relaxation by Doppler echocardiography. Validation with simultaneous cardiac catheterization.

BACKGROUND: The instantaneous pressure gradient between the left ventricle and left atrium during systole can be calculated from the mitral regurgitation Doppler velocity curve. The purpose of our study was to determine the accuracy of measuring the time constant of relaxation (TAU) derived from the Doppler mitral regurgitation signal by comparing it with simultaneous high-fidelity left ventricular pressure measurements in humans. METHODS AND RESULTS: Twenty-five patients had continuous-wave Doppler mitral regurgitation recordings performed with simultaneous high-fidelity left ventricular pressure measurements. Fifteen of these patients had measurements of six to eight beats at various RR intervals. Doppler velocity curves were converted to left ventricular pressure curves by different methods through application of the modified Bernoulli equation at 3-msec intervals. The correlation between catheter-derived and Doppler-derived TAU was best when a zero asymptote and knowledge of the left ventricular end-diastolic pressure were used. A less optimal but acceptable method used the addition of 20 mm Hg to the Doppler-derived ventriculoatrial gradient. Use of a nonzero asymptote for calculation of TAU yielded poor correlation between catheter and Doppler measurements. The correlation of percentage change in Doppler-derived TAU plotted against percentage change in catheter-derived TAU was poor. CONCLUSIONS: The descending limb of the Doppler-derived mitral regurgitation velocity signal can be used as a semiquantitative estimate of the rate of ventricular relaxation. This method requires knowledge of left atrial pressure and may not be sufficiently accurate for detecting small changes in the rate of relaxation on a beat-to-beat basis.

Atrial Function, Left↗

Random-effects Cox proportional hazards model: general variance components methods for time-to-event data.

Proportional hazards regression models are commonly used to study factors associated with time-to-event data. Because many complex genetic diseases exhibit variation in age at onset, it is important to have the capability to perform survival analyses on data collected from individuals whose observations are correlated due to shared genes or environment. While there are widely accepted methods for variance components analysis for simple quantitative traits, a parallel methodology for survival data has not been available. This manuscript outlines a method to perform variance component analyses under general random effects proportional hazards models. This method is based on a Laplace approximation, and makes computation for correlated time-to-event data feasible. The correlated frailty models described here can be used to perform genetic analyses, and other analyses with structured random effects, on age-at-onset data in a manner analogous to standard variance components methods for quantitative traits. We illustrate the use of the method by examining the heritability of breast cancer in a large familial cohort study. We also perform variance components linkage analyses on data simulated for the Twelfth Genetic Analysis Workshop (GAW12), and further examine the performance of this method for linkage analysis in a simulation study. The breast cancer analyses support significant heritability of disease age-at-onset that is of moderate size. The variance component linkage analyses successfully identify the location of the disease genes that were simulated to have a direct impact on age-at-onset. The methods outlined here make it possible to perform general variance components analyses on time-to-event endpoints, even on large data sets, in a computationally efficient manner.

Age of Onset↗

Quadruplet pregnancy in IVF.

The frequency of multiple pregnancy following IVF with the presently accepted methods is ten times higher than in natural cycles, and is identical to that following induction of ovulation with HMG. In spite of the fact that multifetal pregnancy exposes both fetus and mother to certain dangers, most couples joining IVF programs expose themselves to, and generally accept the risk of, multiple pregnancy, since this is their last resort for having a baby of their own. We report such a case of quadruplet pregnancy after the IVF procedure and review the literature.

Abortion, Induced↗

Use of WuScope for exchange of endotracheal tube in a patient with difficult airway.

OBJECTIVE: To describe a method for the exchange of a defective endotracheal tube using the WuScope in patients with difficult airways who cannot tolerate interrupted ventilation. STUDY DESIGN: Case report. METHODS: Detailed description of proposed modality for the endotracheal tube exchange. RESULTS: Exchanging a defective endotracheal tube in patients with a "difficult airway" and compromised oxygenation can be a challenging task. Performing fiberoptic visualization with the WuScope and using a "double intubation" technique may be an acceptable method for endotracheal tube exchange in some clinical situations. CONCLUSION: By using our technique the exchange of an endotracheal tube in a patient with a difficult airway may be achieved without interrupting the ventilation.

Anesthesia, General↗

[Oropharyngeal flora. Epidemiologic survey of prevalence].

OBJECTIVES: Epidemiological surveys which are not frequently carried out in medical practice should provide useful information for the choice of antibiotics to be prescribed in community-acquired infections particularly with the recent development of therapeutic difficulties due to resistant strains. We therefore analyzed the prevalent pharyngeal flora in a general patient population. METHODS: The study was conducted during a single 24-hour period in 1991 by 43 general practitioners and included 645 subjects consulting for benign affections. No patient selection was made. Two pharyngeal swabs were obtained from each subject and cultured in aerobic and anaerobic conditions. Internationally accepted methods for identifying bacteria in pharyngeal samples all performed by one well-equipped laboratory. Beta-lactamase activity was determined with the nitrocephine technique, both directly and after culture. RESULTS: Patient age varied from 16 to 45 years; most (68.5%) consulted for reasons other than ear-nose-throat affections. Only 41 patients (4.3%) consulted for sore throat and 65.4% had not received antibiotics for at least 6 months. Haemophilus influenzae was found in 59.6% of the patients, 20% of the strains were beta-lactamase producers as were 83.7% of the Moraxella catarrhalis strains identified. CONCLUSION: These factors are indicators of potential risk of therapeutic failure when using beta-lactams unstable to beta-lactamases for the treatment of pharyngeal infections.

Adolescent↗

Ocular manifestations of Sotos syndrome.

BACKGROUND: Sotos syndrome (cerebral gigantism) is a congenital syndrome characterized by large body size for patient age, advanced bone age, and unusual facial characteristics with varying levels of cognitive, developmental and perceptual deficits. While more than 150 cases have been reported, only a single case report has been published in the ophthalmic literature. This study briefly reviews the pertinent aspects of this syndrome and reports on the medical, physical, developmental and ocular manifestations of 32 subjects. METHODS: Our patient population was obtained from several schools and colleges of optometry and private offices and clinics of optometrists and ophthalmologists. The Sotos Syndrome Support Association assisted in obtaining patients for this study as well. All children had been previously diagnosed as having SS. Commonly accepted methods for evaluating young, non-verbal, and/or handicapped children were used. RESULTS: Our findings indicate that moderate to high refractive error (hyperopia), nystagmus, and strabismus (esotropia) are commonly associated conditions of this syndrome. CONCLUSIONS: Since many of our subjects exhibited these amblyogenic precursors, a routine optometric vision evaluation should be an essential part of the individual educational and habilitation plan for all children with Sotos syndrome.

Abnormalities, Multiple↗

[A case report of Konno's operation for discrete type subaortic stenosis associated with previous VSD closure and repair of coarctation].

A patient, 1 year and 3 month old boy, underwent Konno's operation for severe discrete type subaortic stenosis, which had developed after the coarctation repair (Subclavian Flap Method) at 15 day old and VSD closure at 21 day old. Konno's operation was performed successfully with SJM 21A and postoperative course was uneventful. The discrete type subaortic stenosis may also develop after VSD closure complicating with coarctation complex. Konno's operation is an acceptable method and stenotic left ventricular outflow tract can be enlarged sufficiently in even young infants. This is the youngest patient treated successfully with Konno's operation in Japan.

Aorta, Thoracic↗

Contrast sensitivity determined with the spatial bandwidth equalization technique: threshold, suprathreshold, and spatiotemporal measurements.

Contrast sensitivity functions were obtained from normal subjects using a spatial bandwidth equalization (SBE) technique and the more conventional display method. Static sensitivity measurements obtained with the two methods were in good agreement. However, when the patterns were counterphase flickered , sensitivity to the lower spatial frequencies was enhanced using the conventional method while sensitivity measured with the SBE technique was slightly depressed. The SBE method also was used to investigate suprathreshold contrast perception with static and flickering gratings. In general, the perception of contrast was independent of spatial frequency content of the stimulus, especially with increasing contrast levels and flicker frequencies. These studies have shown that the SBE technique is an acceptable method to assess static contrast sensitivity and suprathreshold contrast perception.

Humans↗

Evaluation of the E test for antimicrobial susceptibility testing of Porphyromonas gingivalis.

The antimicrobial susceptibilities of 81 recent clinical Porphyromonas gingivalis isolates and two reference strains were determined by the E test, a new method, and were compared with the minimal inhibitory concentrations for these strains obtained by the reference agar dilution method on supplemented Brucella blood agar. The following agreements were obtained: benzylpenicillin 100%, ampicillin 96%, cefaclor 82%, cefuroxime 91%, erythromycin 93%, clindamycin 99%, tetracycline 66%, doxycycline 89%, metronidazole 77% and ciprofloxacin 77%. Very major discrepancies were observed with ciprofloxacin. This study indicates that the E test is an acceptable method to determine the susceptibility of P. gingivalis for most antimicrobials.

Ampicillin↗

The 'induction' dose of thiopentone. A method of study and preliminary illustrative results.

Study of the minimal dose requirements for induction of anaesthesia poses great problems which are solved by the use of a standard administration technique and the abolition of the eyelash reflex as an endpoint. This has been used in 2206 consecutive unselected inductions, in which variables considered to be likely to influence the dosage were recorded. Milligram per kilogram is the most acceptable method of expressing the average dosage of thiopentone. Doses follow a right skew distribution. Women required a significantly lower average dose of thiopentone than men, while obese patients required less than others. Moderate or heavy drinking increased the induction dose but the use of tobacco did not have any influence. The most important factors governing dosage are the physical fitness of the patient and the premedication used. Patients in ASA grades 1 and 2 required significantly more thiopentone than those in grades 3 and 4. This effect is as great as that of premedication in which an opiate with a phenothiazine or hyoscine markedly reduced the induction dose. It was more important than the patient's pre-operative condition with respect to sedation or apprehension. Small doses of opiates or benzodiazepines do not have as much effect on dosage.

Adolescent↗

Achieving and maintaining pregnancy after age 40.

Demographic data from the United States show the need for testing ovarian reserve in a population that is increasingly delaying reproduction. The childless rate for U.S. women over the age of 35 has nearly doubled to 25% since 1925. Yet, in 1994 approximately one-third of all first births were by women over the age of 35. The first-line treatment for the aging effect on the ovarian reservoir of oocytes is ovulation induction. Study results reveal that a delivery rate of only 2.4% was achieved at age 42, and in 119 cycles of therapy in women above that age there were no live births. The clomiphene citrate challenge test is one method of dynamic testing of ovarian reserve. It has the advantages of simplicity and reduced cost when compared with the gonadotropic stimulatory test. In one test group utilizing the clomiphene citrate challenge test only 4.8% conceived at any age following an abnormal test. As a prognostic test with an approximate 95% accuracy rate for negative predictive value it has much to offer. With oocyte donation becoming more widespread as an acceptable method of achieving pregnancy, clomiphene citrate challenge testing should be performed more frequently, in order to spare couples the emotional and financial sequelae of multiple attempts at ovulation induction and IVF when expected pregnancy rates fall below a level acceptable to both patient and physician.

Adult↗

Magnetic resonance imaging in detection and functional assessment of coronary artery disease.

The past few years have brought significant improvements in the field of cardiovascular magnetic resonance imaging (MRI), which evolved from an experimental technique to a clinically accepted method of coronary artery disease detection (stress MRI) and viability assessment. In this article, we describe current MRI technology for detection and functional assessment of ischemia, such as dobutamine/atropine MRI, perfusion techniques, viability, and flow reserve in native coronary arteries and grafts. With further refinement in the technology, wide acceptance of cardiovascular MRI is anticipated in clinical practice.

Atropine↗

Review of methodologies for the determination of nonvolatile N-nitroso compounds in foods.

An attempt has been made to briefly review methods available for the determination of total N-nitroso compounds, N-nitrosamides, N-nitrosamino acids, and miscellaneous other nonvolatile N-nitroso compounds in foods and beverages, giving special emphasis to the progress made during the past five years. It appears that a wide variety of acceptable methods are available for N-nitrosamino acids and simple hydroxylated N-nitrosamines, but none of them has yet been adequately validated. Only limited progress has been made for the analysis of N-nitrosamides, N-nitroso sugar amino acids, and other N-nitroso compounds. Various mass spectrometric methods available for the determination and confirmation of these compounds are also discussed.

Food Analysis↗

Comparison of computer-assisted learning with tutorial teaching in a group of first-year dental students.

Two identical groups of first-year dental students were instructed in the pathology of dental caries and periodontitis using computer-assisted learning (CAL) or tutorial teaching (TT). A cross-over arrangement, in which the first group was taught about dental caries by CAL and periodontitis by TT and the second group in the reverse order, allowed comparison of the two methods. The comparison included a knowledge test and completion of a questionnaire. CAL and TT were equally effective as far as acquisition of knowledge was concerned. Students felt pressurized with CAL and had problems with note-taking. They liked a teacher to be present. Nevertheless, CAL was an acceptable method of instruction and was more economical, in terms of staff involvement, than TT.

Computer-Assisted Instruction↗