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A rapid sensitive monoclonal assay for lipid A in solution.

Lipid A is the toxic component of bacterial endotoxins (LPS) and LPS has been thought to be clinically important in Gram-negative bacterial sepsis, as well as in non-bacteremic states where endotoxemia of enteric origin may be deleterious. The presently accepted method of detecting both LPS and its common lipid A moiety is the Limulus lysate amebocyte assay (LAL), but this test cross-reacts with non-bacterial antigens and serum contains natural inhibitors to the reaction. As an extension of previous work using a polyclonal antibody, we have developed a rapid and sensitive monoclonal antibody assay for lipid A. This 3-step inhibition ELISA is reproducible in aqueous solutions and capable of detecting less than 10 pg/ml of this shared toxic endotoxin component. The assay does not detect intact LPS but acid hydrolysis releases the active lipid A for reaction. While already valuable in detecting lipid A in biological solutions, the presence of naturally occurring anti-lipid A immunoglobulins in serum interfere with the reaction and cause false positives in this inhibition assay. Clinical usefulness of the assay will depend on removal of these antibodies from serum prior to testing.

Antibodies, Monoclonal↗

Fluctuation test for two-stage mutations: application to gene amplification.

The determination of mutation rates is an important experimental procedure for characterizing mutation processes. The accepted method of determining mutation rates, the fluctuation test, was introduced by Luria and Delbrück in 1943. Since then it has been applied to various microorganisms and cells. The Luria-Delbrück test is based on a restrictive hypothesis of mutations being due to single irreversible events. However, some inherited changes in phenotype, like gene amplification, may be due to two or more genetic changes, some of which may be reversible. The Luria-Delbrück model of mutation was compared to other models which included reversibility and more than one mutation stage. The Luria-Delbrück model has been confirmed to be consistent with the original bacteriophage resistance data. However, for gene amplification this model gives incompatible estimates of mutation rates by the P0 and r methods. Relaxing the hypotheses of the single-stage models did not improve the fit. In contrast, a two-stage reversible model provided a fit. Analysis of gene amplification data by the two-stage reversible model provides new information, including estimates of rates for each of the two forward stages and of the reverse step.

Animals↗

Postpartum bacteriuria. A multicenter evaluation of different screening procedures and a controlled short-course treatment trial with amoxycillin.

A total of 10,909 puerperal women from 6 different hospitals were screened for bacteriuria by culture of voided midstream urine (MSU), and a significant growth was found in 881 patients (8.1%). In 731 cases the urine was reexamined by using suprapubic aspiration (SPA), and in only 354 (48%) of the samples the diagnosis of bacteriuria was confirmed. The contamination rate of the MSU samples varied from 46 to 69% between the different hospitals, indicating that in the postpartum period positive MSU findings would necessitate more thorough examination in order to confirm the diagnosis of urinary tract infection. In our study, suprapubic aspiration was found to be a simple and acceptable method without any side effects. Confirmed bacteriuria occurred in 3.2% of the women. Operative delivery (Cesarean section, forceps and vacuum extractor delivery), epidural anesthesia and bladder catheterization increased the risk of bacteriuria in the postpartum period. Only 27% of the women with positive bladder urine complained of dysuria and this symptom was significantly more common in women who had been catheterized. 230 patients with confirmed bacteriuria with amoxycillin-sensitive bacterias participated in a randomized short-course treatment trial: 114 women received 3 days treatment with amoxycillin (1.5 g/day), 116 received the traditional 10 days therapy (750 mg amoxycillin/day). Both antibiotic regimens were observed to be effective with a cure rate of 96 and 98%, respectively. Short-course antibiotic treatment should thus be recommended to puerperal women with urinary tract infections since this avoids prolonged drug exposure to the lactating mother.

Amoxicillin↗

Reductions in cost and cesarean rate by routine use of external cephalic version: a decision analysis.

OBJECTIVE: To describe currently accepted methods for managing the term breech pregnancy, and to predict the delivery outcomes and costs of each method. METHODS: Literature review was used to derive four options for the peripartum management of the term breech pregnancy. Using decision-analysis techniques, we calculated the predicted delivery outcomes and costs associated with each option. RESULTS: Applying external cephalic version to all term breech pregnancies without contraindications to labor or version, and allowing a trial of labor for eligible women who fail version, results in a 25% cesarean rate, at a cost of $8071 per case. Delivering unsuccessful versions by scheduled cesarean results in a 32% cesarean rate, at a cost of $8276 per case. In contrast, a strategy not using external version but allowing a trial of labor for those mothers who meet eligibility criteria results in a 63% cesarean rate, at a cost of $8755 per case. Routinely scheduling a cesarean when a breech is identified at term results in an 89% cesarean rate, at a cost of $9544 per case. CONCLUSIONS: Although the liberal use of vaginal delivery for term breech pregnancies has been suggested as one way of lowering the cesarean rate, the addition of routine external cephalic version to the management strategy will result in more vaginal deliveries and lower costs than strategies that allow vaginal delivery but do not include an attempted cephalic version. Routine cesarean without attempted external cephalic version results in excessive operative deliveries.

Breech Presentation↗

Oral assessment of the "risk" patient.

Certain patients may be considered as being at "risk" with respect to the possibility of development of significant diseases of either a local or a systemic nature as a direct result of the presence of oral infection. The problems relating to the development of bacteremia subsequent to surgery in the patient with cardiovascular disease are well appreciated. Thought should be given to the possibilities of such developments in relation to the presence of periapical pathosis. It is demonstrated that a definitive evaluation of the status of the nonvital tooth cannot be made by the presently accepted methods of clinical and radiographic examination.

Disease↗

Management of mandibular trauma with rigid internal fixation.

The use of rigid internal fixation has become an acceptable method for the treatment of fractures and continuity defects of the mandible. This technique has a wide variety of applications, including stabilization of mandibular segments for the application of bone grafts. The primary advantage of rigid fixation is that it obviates the need for maxillomandibular fixation associated with the treatment of mandibular trauma by traditional methods. This article reviews twenty cases in which rigid internal fixation was used for the treatment of fractures and continuity defects of the mandible. In only one case was early removal of the implant necessary as a result of infection.

Adult↗

Adjustment of DLCO for varying COHb, and alveolar PO2 using a theoretical adjustment equation.

The diffusing capacity of the lung for carbon monoxide (DL) is affected by changes in alveolar partial pressure of oxygen (PAO2), hemoglobin concentration (Hb), and carboxyhemoglobin concentration (COHb). A number of investigators have derived empiric adjustment equations to account for changes in these variables. We evaluated an adjustment of DL for changes in COHb and PAO2 using a single equation derived from Roughton and Forster's original definitions (J. Appl. Physiol., 1957). Unadjusted DL values declined significantly with rising COHb (-0.938%/percent COHb increase, P less than 0.0001) and rising PAO2 (-0.343%/mm Hg PAO2 increase, P less than 0.0001). Adjusted measured DL using the derived equation showed no significant change with changing COHb and PAO2 levels and provides an acceptable method for adjustment of DL for the effects of varying COHb and PAO2 levels from the standard conditions of COHb% = 0, and PAO2 = 110 mm Hg. Since a similar equation has previously been used to adjust for changes in DL due to anemia, we propose to use a single equation which is theoretically derived and empirically verified to adjust DL measurements for changes in COHb, PAO2 and hemoglobin.

Carbon Monoxide↗

Testing the indeterminacy of linear color mechanisms from color discrimination data.

It have previously been reported that, for some choices of the fixed spatial and temporal characteristics of test stimuli, it was possible to estimate the spectral sensitivities of chromatic mechanisms from chromatic discrimination data alone. If mechanism sensitivities could be reliably estimated for any choice of test stimuli characteristics, the influence of spatial and temporal factors on chromatic discrimination performance could be directly measured. Previous studies, using test stimuli with other spatio-temporal characteristics, have found equi-discrimination contours whose ellipsoidal shapes seem to preclude estimation of mechanisms. Since there is no commonly-accepted method for testing the adequacy of ellipsoidal fits of chromatic equi-discrimination contours, it is possible that alternative psychophysical procedures combined with more powerful statistical tests could detect the pattern of deviations from ellipticality reported previously. In this paper, we described psychophysical tests and statistical analyses that, taken together, provide a more powerful test of the indeterminacy of mechanisms than previous methods. We develop a method based on analysis of residuals for detecting the pattern of deviations from ellipticality. We apply these tests under fixed experimental conditions similar to those in which other researchers have found ellipsoidal equi-discrimination contours. For these conditions, for any of the tests performed, we do not reject the hypothesis that equi-discrimination surfaces are ellipsoidal.

Color Perception↗

The use of a 51Cr technique to detect gastrointestinal microbleeding associated with nonsteroidal antiinflammatory drugs.

Of techniques used to evaluate gastrointestinal (GI) bleeding, use of radiochromium (51Cr)-tagged erythrocytes is the most quantitative and scientifically acceptable method. The value of this technique as well as systematic errors possible with its use are discussed. The medical literature concerning 51Cr evaluation of GI microbleeding with naproxen therapy is critically reviewed. We suggest that future studies using this technique be parallel, randomized, double-blind, and include a 1-week placebo baseline phase for all subjects. Treatment with nonsteroidal antiinflammatory drugs (NSAIDs) should last 3 to 4 weeks. A parallel group of subjects should receive placebo throughout the study. For valid statistical analyses, randomization must achieve baseline comparability of weight, height, age, and sex in the treatment groups. Data transformations may be necessary to satisfy the assumptions of the statistical model. Following these guidelines will enable investigators to better evaluate GI microbleeding during treatment with naproxen or other NSAIDs, and, hopefully, to establish the safety profiles of these drugs.

Anti-Inflammatory Agents, Non-Steroidal↗

The influence of sample age on collagen-induced platelet aggregation in whole blood.

Whole blood electrical aggregometry (WBEA) has become an accepted method to gain quick information on platelet disorders. Compared to the optical method WBEA is closer to physiology and less complicated, but on the other hand more difficult to standardize. Different approaches have been attempted in the past to improve the reliability and practicability of this technique. The influence of sample age has not been defined so far. In a first step a mathematical modelling program was established, which is able to characterize the aggregation curves obtained after collagen stimulation. In a mathematical analysis various characteristics of the curve function were calculated and their sensitivity for aging investigated. Regression was performed for each characteristic, and correction factors defined. Our results indicate, that whole blood specimen for collagen induced aggregation can be used without correction factor up to 30 minutes. Data obtained with an age exceeding half an hour have to be corrected following a quadratic regression.

Blood Preservation↗

Control of renal arterial bleeding after percutaneous biopsy.

Transcatheter arterial embolization is becoming an accepted method of controlling massive acute hemorrhage, including that secondary to blunt and penetrating renal trauma. Two patients with retroperitoneal bleeding postrenal biopsy and angiographically proved pseudoaneurysms at the biopsy sites had successful embolization without sequelae. Experimental and clinical experience with transcatheter embolization is reviewed. It appears that this modality is a safe, effective way of controlling renal hemorrhage.

Adult↗

Pulse actinomycin D scheduling in nonmetastatic gestational trophoblastic neoplasia: cost-effective chemotherapy.

The currently accepted method of treatment of patients with nonmetastatic gestational trophoblastic neoplasia (NMGTN) is single-agent chemotherapy. While methotrexate and actinomycin D have shown superiority over other cytotoxic agents, doses of drug and length of treatment are subject to controversy. Recently, the demonstration of the long half-life of actinomycin D in both animal models and humans has necessitated a reevaluation of the method of administration of this drug for NMGTN. Following the demonstration of minimal severe toxicity of pulse actinomyin-D scheduling by E. S. Petrilli and C. P. Morrow (Actinomycin D toxicity in the treatment of trophoblastic disease, Gynecol. Oncol. 9, 18-22 (1980), 12 consecutive patients with nonmetastatic gestational trophoblastic neoplasia were treated. The remission rates, toxicity, and cost effectiveness of pulse actinomycin-D scheduling in these 12 patients are reported.

Adolescent↗

Gynecological and childbearing needs of lesbians.

This descriptive study explored how self-identification as lesbian ("coming out") affected gynecological and childbearing needs and experiences. Twenty interviews were conducted using a researcher-designed interview schedule. The majority of lesbians disclosed their sexual/affectional identity to providers ostensibly to negate heterosexual assumptions. Fear and the unpleasantness of coming out influenced the majority to postpone gynecological care or to seek lesbian-sensitive providers. Half of the participants had gone to ob/gyn appointments accompanied by the partner. Traditional health history questions about marital status, sexual activity, and birth control elicited inaccurate information from participants. Participants believed important qualities in providers to be: sensitivity, knowledge about lesbian sexuality, and female gender. Providers need to learn clues of lesbian identification or coming out; they should also strive to use inclusive gender words and remove heterosexual assumptions. Seventy percent of participants desired children; alternative insemination was an accepted method of conception to most. Midwives can play a special role with lesbians desiring pregnancy and children.

Attitude of Health Personnel↗

Estimation of nasal epithelial lining fluid using urea as a marker.

BACKGROUND: The luminal surface of respiratory mucous membranes is lined with an epithelial lining fluid (ELF) layer. Previous attempts to determine ELF volumes in airways have used dyes or freely diffusible molecules such as urea, yet have not led to a universally accepted method. The nasal mucous membrane provides an accessible area to examine whether urea is an appropriate marker of respiratory ELF volume. METHODS AND RESULTS: Collection of undiluted nasal secretions after either glandular stimulation or induction of vascular permeability confirmed that plasma urea and nasal urea concentrations are equivalent. Baseline ELF volume was calculated as 800 microliters/nostril. The calculated molar concentrations of urea in ELF did not vary with either methacholine or histamine challenge. CONCLUSIONS: These data indicate the plasma, interstitial, glandular, and ELF urea concentrations are equivalent and, therefore, that urea is a useful marker of ELF volume in the nasal mucosa.

Adult↗

Fetal survival in the cow after pregnancy diagnosis by palpation per rectum.

It is desirable to determine whether a cow has failed to become pregnant as early as possible, preferably prior to 50 d after insemination. Although palpation per rectum has been the generally accepted method of pregnancy diagnosis in cattle, the procedure may be a significant iatrogenic cause of fetal attrition. In a study conducted at a North Florida dairy from January through June 1982, pregnancy was determined in 192 Holstein-Friesian cows by measuring low milk progesterone (P4) content on day of insemination (Day 0) and elevated P4 on Days 21 and 24. Pregnant cows were randomly assigned to a treatment and a control group. Cows in the treatment group (n=85) were palpated per rectum twice between Days 42 and 46 after insemination. Cows in the control group (n=107) were not palpated until both groups were palpated at Day 90. Palpation, done by two experienced clinicians, consisted of palpation of fetal fluid fluctuation, identification of the amniotic vesicle, and slipping of the chorioallantoic membranes. In both groups of cows fetal viability was monitored by milk P4 content. Last milk (5 to 15 ml) was collected from one front quarter on Days 0, 21, and 24 and twice weekly thereafter through Day 63. Milk was defatted by centrifugation and the fat-free milk progesterone content measured by a radioimmunoassay without extraction. The milk P4 test was 80.0% accurate in determining pregnancy in the palpated and nonpalpated cows. In the cows palpated on Days 42 to 46, pregnancy rates declined by 7.5% as determined by palpation at Day 90, or by 11.4% as determined by milk P4 content through Day 63 (both values P<0.05). Cows that were not palpated on Days 42 to 46 showed a 1.9% increase or a 4.3% decline in pregnancy rates as determined by the same criteria. Before palpation, at Days 42 to 46, pregnancy rates were better in cows that were inseminated in winter (January to March) than in spring (April to June) (82.3% vs 61.6%; P<0.05); P4 content was higher (winter>spring=2.13 ng/ml vs 1.38 ng/ml; P<0.05). First-lactation cows had higher P4 values on Days 21 and 24 than older cows (P<0.01).

Journal Article↗

The effect of insecticide-treated bed nets on mortality of Gambian children.

Insecticide treatment of bed nets ("mosquito nets") may be a cheap and acceptable method of reducing the morbidity and mortality caused by malaria. In a rural area of The Gambia, bed nets in villages participating in a primary health-care (PHC) scheme were treated with permethrin at the beginning of the malaria transmission season. Additionally, children aged 6 months to 5 years were randomised to receive weekly either chemoprophylaxis with maloprim or a placebo throughout the malaria transmission season. We measured mortality in children in PHC villages before and after the interventions described, and compared this with mortality in villages where no interventions occurred (non-PHC villages). About 92% of children in PHC villages slept under insecticide-treated bed nets. In the year before intervention, mortality in children aged 1-4 years was lower in non-PHC villages. After intervention, the overall mortality and mortality attributable to malaria of children aged 1-4 in the intervention villages was 37% and 30%, respectively, of that in the non-PHC villages. Among children who slept under treated nets, we found no evidence of an additional benefit of chemoprophylaxis in preventing deaths. Insecticide-treated bed nets are simple to introduce and can reduce mortality from malaria.

Antimalarials↗