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Effect of granulocyte colony-stimulating factor treatment on ex vivo neutrophil functions in nonneutropenic surgical intensive care patients.

Granulocyte colony-stimulating factor (G-CSF) preferentially stimulates growth and differentiation of neutrophil precursors and activates neutrophil functions. The aim of the present study was to investigate the functional response of the neutrophil to exogenous recombinant human G-CSF (rHuG-CSF) in nonneutropenic patients. In 30 surgical intensive care unit patients with severely impaired wound healing, leukocyte differential count, plasma G-CSF level, and a broad spectrum of neutrophil functions were monitored before (day 0), throughout (days 1 and 5), and at days 1 and 5 after stopping G-CSF treatment. G-CSF application resulted in a 3.5-fold increase in peripheral blood granulocyte count at day 5 of treatment. The mean plasma G-CSF level rose from 48 to a maximum of 2314 pg/ml at day 1 of G-CSF therapy. Neutrophil chemotaxis and stimulated lysozyme release were decreased throughout G-CSF treatment, whereas respiratory burst activity, phagocytic activity, and intracellular calcium concentration were enhanced by G-CSF. Neutrophil membrane depolarization remained unaffected. The increased count and activation state of neutrophils were associated with clinical improvement in most of these patients. Thus, G-CSF may be a useful adjuvant treatment for nonneutropenic patients with severely impaired wound healing.

Adult↗

Capture-recapture method for estimating misclassification errors: application to the measurement of vaccine efficacy in randomized controlled trials.

BACKGROUND: The measure of efficacy is optimally performed by randomized controlled trials. However, low specificity of the judgement criteria is known to bias toward lower estimation, while low sensitivity increases the required sample size. A common technique for ensuring good specificity without a drop in sensitivity is to use several diagnostic tests in parallel, with each of them being specific. This approach is similar to the more general situation of case-counting from multiple data sources, and this paper explores the application of the capture-recapture method for the analysis of the estimates of efficacy. METHOD: An illustration of this application is derived from a study on the efficacy of pertussis vaccines where the outcome was based on > or =21 days of cough confirmed by at least one of three criteria performed independently for each subject: bacteriology, serology, or epidemiological link. Log-linear methods were applied to these data considered as three sources of information. RESULTS: The best model considered the three simple effects and an interaction term between bacteriology and epidemiological linkage. Among the 801 children experiencing > or =21 days of cough, it was estimated that 93 cases were missed, leading to a corrected total of 413 confirmed cases. The relative vaccine efficacy estimated from the same model was 1.50 (95% confidence interval: 1.24-1.82), similar to the crude estimate of 1.59 and confirming better protection afforded by one of the two vaccines. CONCLUSION: This method allows supporting analysis to interpret primary estimates of vaccine efficacy.

Adolescent↗

Prevention of radioactive indicator and viral particle transmission with an ointment barrier.

OBJECTIVE: To determine the efficacy of a lanolin-based gel in preventing radioactive particle and viral penetration. DESIGN: Paired, stacked filter discs were held in a stainless steel support, and the gel was applied manually to the upper surface of the upper filter. Indicator solution containing either radioactive viral particles (3H-labeled simian virus 40 or 3H-labeled woodchuck hepatitis virus) or 20 microliters or 100 microliters of 32P-labeled radioactive compounds of much lower molecular weight then were applied to the upper filter. The filter discs were separated after 30 minutes, and the lower disc was examined for radioactivity in a liquid scintillation counter. RESULTS: Transmission of radioactive particles was statistically significantly reduced by the application of the ointment on the upper filter (from 6.7 +/- 0.1 x 10(5) counts per minute [cpm] to 88 +/- 38 cpm). Transmission of both labeled viral particles also was reduced to a similar degree. CONCLUSIONS: Application of protective ointment to the filters significantly reduces transmission of radioactive viral particles and smaller radioactive compounds through filter discs. Use of this ointment may offer similar mechanical protection against the transmission of viruses between patient and healthcare provider.

Equipment Contamination↗

Application of the correct information unit analysis to the naturally occurring conversation of a person with aphasia.

The Correct Information Unit (CIU) analysis for measuring the communicative informativeness and efficiency of connected speech (Nicholas & Brookshire, 1993) was applied to the naturally occurring conversation of a person with moderate aphasia. Results indicated that, in this instance, reliable CIU measures could not be obtained. Intrarater reliability for CIU and %CIU was low, reaching only 72%, and interrater reliability was never greater than 63%. However, reliability of word counts was good. Post hoc analysis of rater disagreements in application of the CIU analysis revealed that the majority (72%) resulted from insufficiencies in the scoring rules that were originally designed to measure single speaker connected discourse. Two descriptive categories of disagreements were identified: interpretations of informativeness and absence of rules. The remaining 28% of disagreements were attributable to human error in the application of scoring rules. Comparison of findings with previous research and implications for future research are discussed.

Aphasia↗

Antiproliferative effects and insulin-like growth factor-I expression in Balb-C 3T3 fibroblasts after treatment with somatostatin and gonadotropin-releasing hormone analogs.

The present study was aimed to compare antiproliferative effects of somatostatin (SS) and gonadotropin-releasing hormone analogs (GnRHa) on a fibroblast cell line. Proliferation index, cell count, viability of the cells and insulin-like growth factor-I (IGF-I) immunoreactivity were determined after treatment with either SS (100 microM/ml), GnRHa (35 nM/ml) or SS and GnRHa of Balb-C 3T3 mouse fibroblasts. It was found that the proliferation index, cell count, viability and IGF-I immunoreactivity were not affected by GnRHa treatment as compared with no treatment (p > 0.05). Application of SS to the fibroblasts resulted in a significant reduction in proliferation index, cell count, and IGF-I immunoreactivity as compared with GnRHa treatment and no treatment, but it had no effect on cell viability. The labelling index in SS-treated cells was significantly reduced as compared with combined treatment with SS and GnRHa. In conclusion, a direct effect of GnRHa on fibroblast cells in culture could not be demonstrated. SS had direct inhibitory effects on cell proliferation possibly via inhibition of IGF-I effects without affecting cell viability.

3T3 Cells↗

The use of a surface adhesion immunofluorescent (SAIF) method for the rapid detection of Yersinia enterocolitica serotype O:3 in meat.

This study examined the attachment kinetics of Yersinia enterocolitica serotype O:3 to determine the optimum conditions for its isolation from meat enrichment systems using a novel surface adhesion technique. Minced beef was inoculated with Y. enterocolitica at an initial level of 10 cfu g-1 and incubated at 25 degrees C in an enrichment broth. Yersinia was recovered from enriched samples on polycarbonate membranes by surface adhesion and enumerated using immunofluorescence microscopy. The surface adhesion immunofluorescence technique (SAIF) had a minimum detection limit of approximately 4.0-4.5 log10 cfu ml-1 and provided good correlation between the estimation of the numbers of Yersinia in the enrichment broth derived from plate counts on Yersinia Selective agar (CIN) and those determined by SAIF (r2 = 0.94; rsd = +/- 0.21). A derived regression equation of the SAIF count vs plate counts was used to predict Y. enterocolitica numbers in spiked meat samples stored at 0 degree C for up to 20 d. The numbers as predicted by the SAIF method showed good correlation with counts derived by plating techniques (r2 = 0.78; rsd = +/- 0.42). The application of the SAIF technique for the rapid detection of Y. enterocolitica serotype O:3 from meat is discussed.

Animals↗

Measuring proliferation in breast cancer: practicalities and applications.

Various methods are available for the measurement of proliferation rates in tumours, including mitotic counts, estimation of the fraction of cells in S-phase of the cell cycle and immunohistochemistry of proliferation-associated antigens. The evidence, advantages and disadvantages for each of these methods along with other novel approaches is reviewed in relation to breast cancer. The potential clinical applications of proliferative indices are discussed, including their use as prognostic indicators and predictors of response to systemic therapy.

Antigens, Nuclear↗

Dibromochloropropane: epidemiological findings and current questions.

Dibromochloropropane, DBCP, has had a seminal role in our current understanding of how to prevent chemical risks to health. Early toxicological studies showed its special impact on the testes, and detection of its mutagenic potency was soon followed by demonstration of its carcinogenicity to animals. Its commercial use as a useful nematocide ignored, at first, these warnings from the laboratory. When production workers, first in California and then in Israel, found they were sterile as a result of their exposure, this was convincing evidence that prevention had failed. The evidence, azospermia, oligospermia, and gonadotrophin response to testicular damage, were found first in production workers. In agricultural applicators in California who used the material, decreased sperm count and increased gonadotrophin levels were found. While production in California, Texas, and Israel was halted, studies continued and so, it seems, did use. Our first Israeli study was of workers on banana crops, and we found convincing evidence of increased spontaneous abortion in their family histories. Follow-up studies among production workers in Israel showed that some recovered testicular function, but among their offspring, otherwise in good health, there was a predominance of females. Those who did not recover from azospermia were those with high levels of follicle stimulating hormone. However data for production workers did not show increased spontaneous abortion. Nor have any studies so far shown increased birth defects or increased infant mortality. Unfavorable reproductive outcomes can be due to other agents, as shown by sprayers in Dutch orchards having hypofecundity (increased interpregnancy period) and sex ratio changes; but the agents responsible have not yet been identified. These experiences have lead to the general acceptance of some implications: (1) Paternal exposures can lead to a variety of unfavorable effects on reproductive outcome, including infertility, spontaneous abortion, and altered sex ratio. (2) Patterns of effects of potent agents in production workers and in applicators or users of chemicals may differ. (3) Although human carcinogenesis has not yet been confirmed, unfavorable reproductive outcomes are a reasonable early indicator of such risk. (4) Shifts in sex ratios of populations may be a subtle sign of more serious risks. (5) Continued use of an agent such as this under circumstances in which these UROs cannot be prevented is unconscionable.

Agriculture↗

PCS System I: point counting stereology programs for cell biology.

PCS System I (PCS) is a set of four software modules designed to simplify the application of stereology to problems in cell biology. It is written in BASIC for the Tektronix 4052A microcomputer (Beaverton, OR). A Counting Module collects raw data counts in either a Density Mode (points, intersections, transections, profiles) or a Boundary Mode (intersections with complete nuclear profiles). This information is stored on tape or disk data files and can also be printed. Three analysis modules use data files created with the Counting Module. The Density Module uses Density Mode data files to calculate volume, surface, length, and numerical densities. The B Numerical Density Module uses both Boundary Mode and Density Mode data to calculate the means for the boundary, diameter, and surface area of a nuclear compartment. The mean nuclear surface area is then used with the nuclear surface density to estimate the nuclear numerical density, which, in turn, is used to calculate surface areas of membrane compartments in average cells and in 10(6) cells. The Format Module reformats raw data files for analysis with Tektronix statistical software.

Biometry↗

Effect of an antibacterial varnish and amine-fluoride/stannous fluoride (AmF/SnF2) toothpaste on Streptococcus mutans counts in saliva and dental plaque of children.

The purpose of this study was to evaluate the effect of the simultaneous application of a chlorhexidine and thymol-containing varnish (Cervitec) and an amine fluoride/stannous fluoride containing toothpaste (Meridol) on Streptococcus mutans counts in saliva and dental plaque of school children 12-14 years of age, during a six-week period. The children were separated into group 1 (Cervitec varnish + fluoride-containing toothpaste), group 2 (Cervitec varnish + Meridol toothpaste), and group 3 (Meridol toothpaste alone). Over the six weeks the greatest improvement in salivary Streptococcus mutans count occurred in group 2. Overall, a statistically significant decrease in total microbiological count, and Streptococcus mutans was found in all three groups.

Adolescent↗

Relative number of cells projecting from contralateral and ipsilateral nucleus isthmi to loci in the optic tectum is dependent on visuotopic location: horseradish peroxidase study in the leopard frog.

The leopard frog optic tectum is the principal target of the contralateral retina. The retinal terminals form a topographic map of the visual field. The tectum also receives bilateral topographic input from a midbrain structure called nucleus isthmi. In this study we determined the relative strength of n. isthmi projections to different loci in the tectum. Horseradish peroxidase (HRP) was applied at single superficial tectal locations in a series of leopard frogs. The application sites were distributed across the tectum. Retrogradely filled cells were counted in ipsilateral and contralateral nucleus isthmi. Although all regions of the tectum receive input from both n. isthmi, the relative number of labeled cells in the two n. isthmi is dependent on visuotopic location. Input to the rostromedial tectum representing the visual field ipsilateral to the labeled tectum comes primarily from the contralateral n. isthmi. Input to the caudolateral tectum representing the visual field contralateral to the labeled tectum originates mostly from the ipsilateral n. isthmi. Tectal application sites representing the visual midline had approximately equal numbers of labeled cells in the two n. isthmi. The results are similar at postapplication survival times ranging from 2 to 14 days. Using application of HRP to rostral tectum and application of nuclear yellow to caudal tectum, we show that the anisotropy in isthmi labeling is not due to take up of these labels by isthmotectal fibers passing through the application sites that terminate elsewhere.

Animals↗

Surgical scrub and skin disinfection.

The role of pre-operative disinfection of the surgeon's hands and the skin of the operative site in the prevention of wound infection remains uncertain. The normal resident skin flora, consisting mainly of coagulase-negative staphylococci and aerobic and anaerobic diphtheroids, is an uncommon cause of infection except in prosthetic operations. Staphylococcus aureus is rarely a resident on normal skin other than the perineum, and is mostly present on the hands as a transient acquired from the nose. Nevertheless, it seems rational to kill or remove all transients on the hands of the surgeon and reduce residents to low levels. Surgical skin disinfection is usually assessed by measuring the reduction in organisms on the hands immediately after disinfection, after repeated applications of the disinfectant and after wearing gloves for two to three hours. The hands are commonly sampled in a bowl or plastic bag containing Ringer's or a similar solution and relevant neutralizers, or by the use of glove washings. A standardized technique is necessary to provide a statistical comparison between agents. Antiseptic detergents, chlorhexidine or povidone iodine show immediate reductions in bacterial counts of 70% to 80%, increasing to 99% after repeated application. Hexachloraphene and triclosan detergents show a lower immediate reduction but a good residual effect. Seventy percent ethyl or 60% propyl alcohol, with or without an antiseptic, show an immediate reduction of over 95%, and in excess of 99% on repeated application. Residual levels of organisms tend to be lower after repeated alcohol treatment than following the use of antiseptic detergents.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriological Techniques↗

Virtual microscopy:applications to hematology.

Virtual microscopy is the simulation of microscopy over a computer network. A virtual slide is a giant digital image file of a glass slide that can be displayed, panned, zoomed, and focused in a virtual slide viewer on a computer screen. Virtual slides represent a revolutionary advance over glass slides. They are easy to file, store, retrieve, annotate, and mark and can be preserved indefinitely. Furthermore, they are easy to duplicate and distribute and can be integrated into electronic patient records. Large virtual slides can be readily transmitted to users over a standard broadband connection. With the recent introduction of viewers that can focus virtual slides, virtual microscopy can simulate all the functions of real microscopy. Virtual microscopy has significant advantages over real microscopy in education and in proficiency testing. In education, virtual microscopy enables "anytime, anywhere" learning and has been favorably received by students and teachers. In proficiency surveys, all users view the same image, virtual slides are easy to distribute, and the slides do not deteriorate. Potential applications for hematology proficiency surveys include blood and bone marrow morphology, differential cell counts, cytochemistry and immunocytochemistry, detection of malarial parasites, and other tests. Virtual microscopy enables proficiency surveys of critical clinical parameters, such as the bone marrow blast count, and implementation of "locate and identify" exercises. It is conceivable that with the next generation of technological developments, virtual microscopy can be extended to diagnostic applications. Important goals are to minimize slide file size without loss of relevant detail, to establish diagnostic equivalence, and to automate virtual slide capture with high throughput for integration into laboratory information systems. Key factors that will drive implementation include user-friendliness, cost, data storage requirements, and throughput speed. Implementation may have constructive effects on teaching and learning, the peer-to-peer consultative process, and diagnostic accuracy and performance.

Hematology↗

Length of survival and lymphocyte percentage in women with mammary cancer as a function of psychotherapy.

50 patients suffering from terminal mammary cancer and visceral metastases received chemotherapy, and in addition half of them received psychotherapy, while half did not. A lymphocyte count was undertaken seven times in all, once before initiation of chemotherapy, the others after six applications of the therapy. As hypothesized, the 25 women who received psychotherapy lived longer and had significantly higher lymphocyte counts on Occasions 5, 6 and 7 than did the 25 women who did not receive psychotherapy. Of the types of psychotherapy employed, a specially designed form of behaviour therapy was significantly more helpful than dynamic psychotherapy in prolonging life.

Antineoplastic Combined Chemotherapy Protocols↗

The benefit of stereology for quantitative radiology.

A knowledge of stereology (i.e. proper sampling), the opportunities provided by computers for image analysis (i.e. image segmentation, image registration, data base exploration, 3D reconstruction), and the strengths (i.e. non-invasive) and limitations (i.e. finite resolution, image artefacts) of medical imaging equipment must all be combined for reliable quantitative magnetic resonance imaging (MRI), the goal of which is to obtain a deeper understanding of the structure, function, life cycle and evolution of the human body, especially the brain, and a more objective diagnosis of disease and assessment of its response to treatment. In this article we illustrate the first of these requirements. We describe the application of proper sampling strategies and efficient computer-based counting procedures for obtaining unbiased estimates of volume by the Cavalieri method and of surface area from vertical sections. In particular, we estimate the volume of a brain tumour from Cavalieri sections, the volume of grey matter in the cerebral hemispheres from Cavalieri slices and the surface area of the cerebral cortex from vertical sections. The estimates obtained are mathematically unbiased. In each case, we assess the precision of the estimates empirically. Application of formulae available for predicting the precision of volume estimates obtained using the Cavalieri sections and slices methods is also described.

Algorithms↗

Covariance components models for longitudinal family data.

A longitudinal family study is an epidemiological design that involves repeated measurements over time in a sample that includes families. Such studies, that may also include relative pairs and unrelated individuals, allow closer investigation of not only the factors that cause a disease to arise, but also the genetic and environmental determinants that modulate the subsequent progression of that disease. Knowledge of such determinants may pay high dividends in terms of prognostic assessment and in the development of new treatments that may be tailored to the prognostic profile of individual patients. Unfortunately longitudinal family studies are difficult to analyse. They conflate the complex within-family correlation structure of a cross-sectional family study with the correlation over time that is intrinsic to longitudinal repeated measures. Here we describe an approach to analysis that is relatively straightforward to implement, yet is flexible in its application. It represents a natural extension of a Gibbs-sampling-based approach to the analysis of cross-sectional family studies that we have described previously. The approach can be applied to pedigrees of arbitrary complexity. It is applicable to continuous traits, repeated binary disease states, and repeated counts or rates with a Poisson distribution. It not only supports the analysis of observed determinants, including measured genotypes, but also allows decomposition of the correlation structure, thereby permitting conclusions to be drawn about the effect of unobserved genes and environment on key features of disease progression, and hence to estimate the heritability of these features. We demonstrate the efficacy of our methods using a range of simulated data analyses, and illustrate its practical application to longitudinal blood pressure data measured in families from the Framingham Heart Study.

Blood Pressure↗

Immune modulating effects of low doses of cyclophosphamide and keyhole limpet hemocyanin on peripheral blood immune parameters in patients with metastatic renal cell carcinoma.

Assessing the peripheral blood immune status, we had found evidence of immunosuppression in metastatic renal cell carcinoma. Since immunosuppression might interfere with the outcome of immunotherapy, it was submitted to further investigation. Immunosuppression can be mediated by T suppressor cells, which for their part are inhibited by low-dose cyclophosphamide (Cy). We tested whether the immunomodulating effects of the biological response modifier keyhole limpet hemocyanin (KLH) on cellular and humoral immune parameters would be intensified by low-dose Cy. 10 patients were given 300 mg/m2 Cy i.v. 3 days before application of 1 mg KLH i.m. up to 8 times in 4-weekly intervals. Immune parameters were assayed twice pre-therapeutically, and prior to and 1 day and 1 week after each KLH injection. In contrast to the results obtained with KLH-mono-application, lymphocyte subsets-with the exception of T4 cell counts and the T4/T8 ratio-remained stable during the first 4 months of observation. There were increases in immunoglobulins and in the immunoactivation markers tumor necrosis factor, neopterin and beta 2-microglobulin. The tendencies found here differed from those found in a previous study on patients who were given KLH alone, and were similar to those found in patients with nonmetastatic disease who received KLH alone. However, the increases of the activation markers during KLH+Cy application were at best half as much as in patients with nonmetastatic disease. In conclusion, immunosuppression was influenced, but not counterbalanced by low-dose Cy. Most probably other mediators of immunosuppression than T suppressor cells, for example prostaglandin E2, must be considered as well.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗

Efficacy of directed misting application of a peroxygen disinfectant for environmental decontamination of a veterinary hospital.

OBJECTIVE: To evaluate effectiveness of 4% peroxymonosulfate disinfectant applied as a mist to surfaces in a large animal hospital as measured by recovery of Staphylococcus aureus and Salmonella enterica serovar Typhimurium. DESIGN: Field trial. SAMPLE POPULATION: Polyester transparencies inoculated with bacteria. PROCEDURE: Polyester transparencies were inoculated with S aureus or S Typhimurium and placed in various locations in the hospital. After mist application of the peroxygen disinfectant, viable bacterial numbers were quantified and compared with growth from control transparencies to assess reduction in bacterial count. RESULTS: When applied as a mist directed at environmental surfaces contaminated with a geometric mean of 4.03 x 10(7) CFUs of S aureus (95% confidence interval [CI], 3.95 x 10(7) to 4.11 x 10(7)) or 6.17 x 10(6) CFUs of S Typhimurium (95% Cl, 5.55 x 10(6) to 6.86 x 10(6)), 4% peroxymonosulfate reduced the geometric mean number of viable S aureus by 3.04 x 10(7) CFUs (95% Cl, 8.6 x 10(5) to 1.7 x 10(6)) and S Typhimurium by 3.97 x 10(6) CFUs (95% Cl, 8.6 x 10(5) to 3.5 x 10(6)). CONCLUSIONS AND CLINICAL RELEVANCE: Environmental disinfection with directed mist application of a 4% peroxymonosulfate solution was successful in reducing counts of bacterial CFUs by > 99.9999%. Directed mist application with this peroxygen disinfectant as evaluated in this study appeared to be an effective and efficient means of environmental disinfection in a large animal veterinary hospital and would be less disruptive than more traditional approaches to intensive environmental cleaning and disinfection.

Aerosols↗