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The telescoping plugged catheter in suspected anaerobic infections. A controlled series.

The results of bacterial cultures from transtracheal aspiration and from aspirations obtained by a new telescoping plugged catheter were compared in specimens taken from 13 patients with suspected anaerobic infections of the lower respiratory tract. When all bacterial isolates (potential aerobic pathogens, aerobic nonpathogens, and anaerobes) were counted with transtracheal aspiration as the standard method for comparison, cultures of material obtained by the telescoping plugged catheter technique yielded 22 false negatives and 14 false positives. The results of the 2 methods were in complete agreement in only 3 paired specimens cultured aerobically and in only 6 paired specimens cultured anaerobically. When only potential aerobic pathogens and anaerobes were considered with transtracheal aspiration as the standard method for comparison, cultures of material obtained by the telescoping plugged catheter technique yielded 11 false negatives and 7 false positives, and the results of the 2 methods were in complete agreement in 9 paired specimens cultured aerobically and 6 paired specimens cultured anaerobically. Our results suggest that fiberoptic bronchoscopy using the telescoping plugged catheter technique may be an acceptable method for collecting specimens from the lower respiratory tract, but further studies comparing this method with transtracheal aspiration are needed.

Adult

[In vitro determination of the biological activity of choleragen by the anaerobic dehydrogenase inhibition test in a surviving blastoma cell culture].

A rapid method for the detection and titration of choleragen in vitro in the test allowing one to evaluate dehydrogenase activity in the surviving blastoma culture has been developed. In comparison with the commonly accepted method of using the ligated loop in adult rabbits, the above method is simple, highly sensitive and economical.

Anaerobiosis

Comparison of three methods for detection of group A streptococci in throat swabs.

Group A streptococci are generally detected in throat swabs by (i) rapid antigen tests, (ii) conventional culture, or (iii) combinations of both. Direct fluorescent-antibody testing of a 2-h enrichment broth (FA/EN) was an accepted method for same-day results before the advent of rapid antigen tests. We compared FA/EN in Todd-Hewitt Broth (THB) with conventional culture and a rapid antigen test, TestPack Strep A (TPS). Nine hundred seventy specimens were evaluated in this study. Cultures were performed for 48 h on sheep blood agar (SBA) incubated aerobically and on a selective agar for group A streptococci (SSA) incubated in 5 to 10% CO2. Following a 2-h incubation, the fluorescent-antibody test was performed. A subculture of the centrifuged sediment from the THB enrichment was also done. In comparison with a positive culture on SBA or SSA or subculture of the THB pellet, the sensitivities and specificities of the different methods were as follows: SBA, 92 and 100%; SSA, 92 and 100%; TPS, 68 and 99%; FA/EN, 88 and 98%. The FA/EN method offers the potential for definitive finalized reports on the same day as specimen collection with greater sensitivity than TPS. This study included sequential plating and rapid antigen testing of a single swab. In a separate set of experiments to validate this study design, it was shown that recovery of streptococci from swabs plated sequentially on five plates did not vary with the order of plating and the actual proportion of organisms recovered from a swab on a single plate was only 1%.

Antigens, Bacterial

[Motivation and education].

It was thought for many years that successful motivation in oral health should be based on the knowledge of epidemiology of caries and periodontal diseases and the methods of prevention. One should learn from mistakes: 95 per cent of the Swiss population "suffers" from caries and periodontal disease but apparently this had little impact on prevention. Simple, world-wide accepted methods of prevention are not used by the majority although they provide a pronounced reduction of caries and gingivitis. The number of Swiss communities which offer adequate preventive programs to schoolchildren ranges between 5 to 10 per cent. Although the budgets of school dental services could be lowered by 75%. Reasons for such failures may be inadequate information of dentists, authorities and the population. Not enough information is spread regarding sugarless sweets, or prevention by fluoride. Unsuccessful motivation probably also lies in the inadequate assignment of the roles ofthe patient, his dentist and the social security. So far only the dentist has formulated oral health duties to the patient, e.g. a better oral hygiene, a reasonable healthy nutrition. In the future the patient should have the right to ask the dentist some questions. Is his dentition at the end of a treatment, healthy and ready to render hygienic efforts effective? Patient, doctor and insurances should know that disease still exists if gingival papillae bleed upon using a toothpick. Absence or presence of gingival bleeding are the criteria for health or disease. A check list of the state of oral health and a signed questionnaire concerning the patient's knowledge of oral health are used to improve motivation of both patient and dentist.

Dental Hygienists

Pseudoephedrine absorption from controlled release formulations: absorption rate constant estimation methods.

Five methods of absorption rate (Ka) estimation were compared using data from a previously reported bioavailability study: Wagner-Nelson (WN), asymptotic WN (AWN), the Hendeles-Weinberger modification of the WN (HW), nonlinear regression performed on plasma concentration vs time data with fixed elimination rates (NL), and nonlinear regression performed on the cumulative sum of AUCs obtained during WN analysis (AUCNL). The maximum plasma concentration (Cmax) and the time at which Cmax occurred (Tmax) were predicted with each method's respective Ka's, and these were compared to observed values. The WN and HW were consistently biased in their predictions of Cmax and Tmax, providing slower Ka's than any of the other methods. AWN could only be used for the reference treatment. Both NL and AUCNL provided unbiased estimations, but AUCNL had a high level of imprecision. With this set of data, only NL was an acceptable method for accurately estimating Ka.

Adult

Determination of vitamin D in foods: a review.

Determining vitamin D content in foods is difficult because in natural foods of highest vitamin D activity, and even in vitamin D-fortified foods, only small quantities are present, and many other compounds are extracted along with vitamin D that cause difficulties in purifying the extract or in the spectrophotometry or colorimetry that follows. Several physicochemical methods--such as spectrophotometric, colorimetric, thin-layer chromatographic, adsorption, partition, gas-liquid, and high-performance column chromatographic--have been tried for assay foods for vitamin D, but none of them have been accepted for official or routine use; they are time consuming and expensive, or lack the required sensitivity, precision, or accuracy. Curative biological assays, based on degree of healing of a leg bone of rats previously made rachitic, is the generally accepted method to determine vitamin D content of foods. However, that method also requires too much time and is expensive. The recently developed high-performance liquid chromatographic method may offer the most for establishing a satisfactory physicochemical method for determining vitamin D in foods. Many of the difficulties and problems in assaying foods for vitamin D are discussed.

Animals

An investigation of factors influencing the measurement of creatine phosphokinase activity in serum using coupled enzymatic methods.

1. The factors influencing the measurement of creatine phosphokinase (CPK) activity in serum by coupled enzymatic methods were investigated to establish optimum conditions for this type of assay. Such a study was indicated following observations by the authors of poor performance of commerically produced reagent kits together with the failure of most of the established an well accepted methods to operate under true optimum zero order kinetics in the reaction phase state. 2. The factors invested were the effects of pH, substrate concentrations (creatine phosphate, glucose and NADP+), added auxiliary (hexokinase) and indicator (glucose-6-phosphate dehydrogenase) enzymes, dithiothreitol (DTT) as an activator and conditions of storage of substrate stability. DTT was found to be a suitable activator but not a reactivator of the reaction. The optimum concentrations of creatine phosphate, glucose and NADP+ were found to be 20.0, 20.0 and 2.0 mmol/litre, respectively. Optimum activieies of the enzymes, glucose-6-phosphate dehydrosenase and hexokinase were 1000 and 2000 units/litre, respectively. 3. The between-day precision of the method for measuring serum at pH 6.8 and 30 degrees C at three activity levels under the optimum conditions developed was excellent yielding coefficients of variation ranging from 2.0 to 2.7%.

Creatine Kinase

Diabetic home blood glucose monitoring using a direct reading glucose sensitive test strip.

The aim of this study was to assess the performance of the BM Test Glycemie 20-800. Twelve diabetic patients were studied using the BM Test Glycemie 20-800, compared with one of the commonly available reflectance meters. The correlation coefficient for the BM 20-800 strip versus reference blood glucose result was significantly better than the reflectance meter (p = 0.01). In a separate study, laboratory staff tested both methods and achieved results which were similar to those of the patients using the BM Test Glycemie 20-800, but better than those of the patients using the reflectance meters. The BM Test Glycemie 20-800 is a cheap, reliable and acceptable method for the measurement of blood glucose levels.

Adolescent

Tests alternative to the rabbit bioassay for pyrogens.

The rabbit bioassay is currently the only legally acceptable method to test for pyrogenic contamination of parenteral preparations and medical devices designed to enter the parenterum. However, research efforts have recently made available alternative procedures to test for pyrogens which may have significant value in quality control. Part of the reluctance to accept tests other than the rabbit test may be due to the low visibility of emerging understanding of a significant theory of testing in which it is contended that the parameters of sensitivity and specificity are inversely related and that accuracy is an ideal. The rabbit pyrogen assay will detect 1 to 10 ng of enterobacteriaceal endotoxin (ET). The limulus test will detect 0.01 to 0.1 ng/ml of ET; some of the other tests approach the rabbit assay in sensitivity. Since it is current dogma that pyrogen is equivalent to ET, the basis for the use of ET to standardize pyrogen tests is rationalized. The source of ET in practice is bacterial contamination; therefore, numbers of bacteria that contaminate parenteral preparations can be directly related to potential pyrogenicity. Further, viable counts of bacteria in parenteral preparations, prior to sterilization, is a reliable test for pyrogens. Other tests such as nitroblue tetrazolium reduction and actinomycin-D enhancement of lethality of pyrogen for mice deserve consideration in quality control procedures. The limulus test, the most practical of currently available alternative tests for detection of ET and therefore, of pyrogen has application where the rabbit test cannot be used. Therefore, control personnel must learn of the availability, performance and interpretation of the limulus test. Practical considerations must govern the choice of an alternative test when the use of a test other than the rabbit bioassay is indicated.

Animals

Comparison of radiometric and microbiological assays for the determination of folate in fortified gruel and porridge.

The results obtained by a radiometric assay of folate in fortified gruel and porridge, using a commercially available radioassay kit, were compared with those obtained by a standard microbiological assay with Lactobacillus casei. A lower level of folate was found with the radiometric assay. Comparison of the results obtained with the two methods gave a correlation coefficient of 0.991 when the analysis was performed on samples extracted at the same time. A lower correlation coefficient was obtained between the result if the extractions were carried out on different occasions. The radiometric assay is a acceptable method for the routine determination of folate in fortified gruel and porridge.

Edible Grain

A fluorometric determination of urinary 17-hydroxycorticosteroids using benzamidine.

A fluorometric determination of urinary 17-hydroxycorticosteroids using a reaction of benzamidine with compounds carrying the dihydroxyacetone side chain is described. The fluorescent compounds have excitation and emission maxima at 370 and 480 nm, respectively. The method includes enzymatic hydrolysis with beta-glucuronidase (EC 3.2.1.31, from Escherichia coli) and extraction with methylene chloride and generation of fluorescence in alkaline solution (pH 13.4). The specificity of the reaction was examined and the results were compared with those of an accepted method based on the Porter-Silber reaction (C. C. Porter and R. H. Silber, 1950, J. Biol. Chem. 185, 201-207). The coefficient of correlation was 0.945 with regression line of y = 0.91x + 0.7 mg/day (y, present method; x, Porter-Silber reaction method). Sensitivity of the reaction was 0.5 microgram/ml of standard or sample, mean recovery of cortisol added to five urine samples (5-micrograms addition) was 95%, and the coefficient of variation of the method (five repeated assays of sample with a value of 5.2 mg/liter) was 6.2%.

17-Hydroxycorticosteroids

Further studies with the fluorescent treponemal antibody-absorption double-staining procedure.

A double-staining procedure for the fluorescent treponemal antibody-absorption test, using fluorescein isothiocyanate as a label for the class-specific anti-human globulin and tetramethylrhodamine isothiocyanate as a label for a counterstain reagent, has been described. This method requires the addition of a KP560 barrier filter, with a microscope equipped with vertical illumination, to exclude the rhodamine emission in reading the fluorescein fluorescence. The present study evaluated reversing the dye label for each conjugate in the double-staining procedure, thus eliminating the need for the KP560 filter. It also considered the possibility of shortening the counterstaining time and compared various methods for preparing antigen slides in an attempt to establish a method that increases the number of treponemes per microscopic field. The results indicate that a rhodamine-labeled class-specific anti-human globulin as a primary stain, and a fluorescein-labeled anti-treponemal globulin as a counterstain, provide an acceptable method for performing the fluorescent treponemal antibody-absorption double-staining procedure. Nonfixed antigen slides were held for 16 days in a desiccator or stored in plastic bags with silica gel for 3 weeks; then, with methanol fixation, they were used satisfactorily in the double-staining procedure. A shortened incubation time for the counterstain allowed more rapid slide processing.

Antigens, Bacterial

The management of arthrogryposis.

Arthrogryposis has an evil reputation among orthopedic surgeons, for the deformities are known to be resistant to correction and relapse during growth is common. It is suggested that many of these difficulties have been due to a delay in initiating treatment, to the use of unsuitable methods such as manipulation, and to a lack of appreciation of the pathologic changes. The author's experience with a large number of personally treated cases has convinced him that aggressive surgical treatment in infancy and early childhood is more successful than previously accepted methods of care (Figs. 22, 23).

Arm

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

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

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

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

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