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PubMed1993

Application of flow cytometry to rapid microbial analysis in food and drinks industries.

In food and drinks industries, the time required for conventional tests can lead to substantial delays in product release to the market. Flow cytometry (FCM) has been used in conjunction with viability markers for rapid counting of yeast, mould and bacterial cells in food products. A single-parameter flow cytometer has proved applicable to the rapid detection of low numbers of microbial contaminants in finished products. The excellent correlation between FCM results and product quality shelf-life expiry date has allowed the establishment of realistic quality control criteria for rapid positive release of product. Used for the monitoring of microbial biomass during manufacturing processes, flow cytometry allowed a direct assessment of bacterial growth. The reproducibility of the results and the proven correlation with standard plate count method obtained in industrial conditions make FCM a good predictive method for product and process quality control.

Bacteria↗
PubMed1999

An exact test for all-way interaction in a 2M contingency table: application to interval capture-recapture estimation of population size.

An exact conditional test for an M-way log-linear interaction in a fully observed 2M contingency table is formulated. From this is derived a procedure for interval estimation of the total count N in a 2M contingency table, one of whose entries is unobserved. This procedure has an immediate application to interval estimation of the size of a closed population from incomplete, overlapping lists of records, as in capture-recapture analysis of epidemiological data. Data on the prevalence of spina bifida in live births in upstate New York in 1969-1974 illustrate this application.

Biometry↗
PubMed1997

Treatment of prolonged chemotherapy induced severe thrombocytopenia with recombinant human interleukin-3--a report on four cases.

Interleukin-3 (IL-3) is a multipotent hematopoietic growth factor, which exhibits stimulatory effects on leucocytes, reticulocytes and platelets. Due to its pronounced induction of megakaryopoiesis, IL-3 is thought to be a cytokine with the potential to prevent and to overcome chemotherapy-induced thrombocytopenia. We report on four cases (two of metastatic breast cancer, one of metastatic ovarian cancer and one of Hodgkin's disease) with prolonged chemotherapy-induced thrombocytopenia in whom rhIL-3 in combination with either recombinant human (rh) granulocyte macrophage colony stimulating factor (GM-CSF) or rh granulocyte colony stimulating factor (G-CSF) was administered. In all cases, a steady and clinically significant increase in platelet counts could be observed. No major side effects, neither due to the application of rhIL-3 nor due to rhGM-CSF or rhG-CSF, occurred; only flu-like symptoms were seen, which could effectively be treated with paracetamol. This report highlights the efficacy of combined treatment with rhIL-3 plus rhGM-CSF or rhG-CSF in chemotherapy-induced thrombocytopenia, where megakaryopoiesis could be stimulated efficiently by rhIL-3. Based on this experience, the authors conclude that established thrombocytopenia as a major side effect of myelosuppressive chemotherapy should be considered as an indication for the use of rhIL-3 in interventional treatment. Further investigations in this area are encouraged.

Adult↗
PubMedDate not supplied

Exchange blood transfusion and on-line plasma exchange for sepsis in infants.

During the last 6 years, 19 infants with sepsis have been treated with exchange blood transfusions 47 times, and 13 patients survived. However, this procedure is compromised by antibody formation against WBCs and platelets. Accordingly, a miniature system for on-line plasma exchange (PE) between septic infant and healthy parent has been developed and evaluated using a canine model. Escherichia coli, at a dose of 5 X 10(9) CFU/ml/kg, was injected intravenously into 17 puppies that were divided into 3 groups; untreated; sham treated; and PE in which 80 ml/kg of plasma in each septic puppy was replaced during 2 hr with fresh plasma simultaneously obtained from healthy adult dogs. Four of 5 puppies survived in the PE group, while all other puppies died within 24 hr. In the PE group, viable cell counts of E. coli and endotoxin decreased significantly, and opsonic activity improved. This system is effective and applicable for treatment of sepsis in infants.

Animals↗
PubMed2005

A prognostic index for systemic AIDS-related non-Hodgkin lymphoma treated in the era of highly active antiretroviral therapy.

BACKGROUND: The established International Prognostic Index for lymphomas has not included patients with systemic AIDS-related non-Hodgkin lymphoma. OBJECTIVE: To establish the most appropriate prognostic index for use in patients with systemic AIDS-related non-Hodgkin lymphoma. DESIGN: A prospective study involving univariate and multivariable analyses of patients with AIDS-related non-Hodgkin lymphoma whose data were used to examine standard and new criteria for survival after diagnosis. SETTING: The Chelsea and Westminster cohort of HIV-1-infected persons. PATIENTS: 9621 HIV-positive patients, 111 in whom AIDS-related non-Hodgkin lymphoma was treated after 1996, in the era of highly active antiretroviral therapy (HAART). INTERVENTION: Cox proportional hazards regression analysis to determine the prognostic significance of multiple clinicopathologic variables. RESULTS: Survival of patients with AIDS-related non-Hodgkin lymphoma has increased in the HAART era (log-rank chi-square, 9.23; P = 0.002). Univariate analyses using the established International Prognostic Index factors of age, tumor stage, lactate dehydrogenase level, Eastern Cooperative Oncology Group performance status, and number of extranodal sites were confirmed to be significant variables. Regression modeling for patients in whom disease was diagnosed after 1996 revealed only 2 independent predictors of death: International Prognostic Index risk group and CD4 cell count. These predictors yielded 4 internally validated risk strata with predicted 1-year survival rates of 82%, 47%, 20%, and 15% (P < 0.001). Prognostic risk scores in the highest quartile yielded a likelihood ratio for death of 7.90 (hazard ratio, 1.0), whereas a prognostic score less than 1.0 yielded a likelihood ratio of 0.23 (hazard ratio, 0.15 [95% CI, 0.06 to 0.33]). LIMITATIONS: The sample was small, and different HAART regimens were used. CONCLUSIONS: For patients with AIDS-related non-Hodgkin lymphoma that was diagnosed in the era of HAART, application of the International Prognostic Index remains useful. The addition of CD4 cell count provides further independent prognostic information. Patients who present with AIDS-related non-Hodgkin lymphoma and a low CD4 cell count have a poor prognosis; this information can be used to guide therapeutic options.

Antiretroviral Therapy, Highly Active↗
PubMed2000

Comparison of methods to investigate microbial populations in soils under different agricultural management.

The microbial community in sludge-treated and nearby untreated soils was investigated using direct microscopic counting, plate culture, carbon substrate utilisation (Biolog), and fatty acid methyl esters. Long-term sewage sludge applications had resulted in higher concentrations of organic carbon and had altered other factors in the soil, including the concentration of metals. Bioluminescence was inhibited in assays of the sludge-treated soil, although microbial counts were similar in all soils. A detailed analysis of carbon substrate utilisation patterns and fatty acid methyl esters showed qualitative differences in the microbial populations. This work shows that a variety of approaches are required to assess microbial communities in soil where, despite large differences in land management, the populations are similar in size and overall composition.

Journal Article↗
PubMed1995

Heterogeneity and reproducibility of microvessel counts in breast cancer.

BACKGROUND: Microvessel counting has proven to be of prognostic value in breast cancer, as shown in different retrospective studies. However, methodology has not been studied widely, and this must be done before the method can become clinically applicable. The aim of this study was to determine the degree of heterogeneity and reproducibility of microvessel counts (MC) in breast cancer. EXPERIMENTAL DESIGN: In 10 cases of breast cancer, the available blocks (2-4 blocks) containing invasive tumor parts were selected, and four sections (4 microns) were cut with interdistances of 100 microns. In each section, two or three invasive areas (0.5 x 0.5 cm) were demarcated. Microvessels, visualized by immunohistochemistry (CD31 Ab), were counted by one observer in 10 systematically selected fields of vision (400 x magnification). Furthermore, microvessels were counted in four fields with the highest microvessel density ("hot spots"). Coefficients of variation (CV) were calculated for the different sampling levels. RESULTS: Repeated MC yielded high intra- and interobserver reproducibility (correlation coefficients > 0.92). For the systematic counting method, CV between MC from different areas within one section was on average (17.1% (0.7-52.1). When comparing MC from corresponding areas in different sections from the same black, CV was on average 14.7% (0.5-41.9), and for MC from different blocks of the same tumor, CV was on average 25.8% (9.9-44.6). Nested ANOVA showed an approximately equal contribution to the total variance of the different sampling levels, except for the variation between sections (not significant). For the hot spot MC, CV for different sections from the same block was on average 11.1% (0.7-29.5) and for different blocks from the same tumor, 24.2% (5.7-54.9). Nested ANOVA showed that variation between different blocks from the same tumor contributed most to the total variance. CONCLUSIONS: There is a noteworthy heterogeneity in MC between different areas from the same section, between corresponding areas in different sections from the same block, and between different blocks from the same tumor. Consequently, one must carefully scan all the available tumor material in each case for the best hot spot. The hot spot approach is efficient and reproducible, but only a comparative prognostic evaluation can show whether it is clinically more useful than systematic counts.

Breast Neoplasms↗
PubMed2005

Non-parametric estimation of the odds ratios for continuous exposures using generalized additive models with an unknown link function.

The generalized additive, model (GAM) is a powerful and widely used tool that allows researchers to fit, non-parametrically, the effect of continuous predictors on a transformation of the mean response variable. Such a transformation is given by a so-called link function, and in GAMs this link function is assumed to be known. Nevertheless, if an incorrect choice is made for the link, the resulting GAM is misspecified and the results obtained may be misleading. In this paper, we propose a modified version of the local scoring algorithm that allows for the non-parametric estimation of the link function, by using local linear kernel smoothers. To better understand the effect that each covariate produces on the outcome, results are expressed in terms of the non-parametric odds ratio (OR) curves. Bootstrap techniques were used to correct the bias in the OR estimation and to construct point-wise confidence intervals. A simulation study was carried out to assess the behaviour of the resulting estimates. The proposed methodology was illustrated using data from the AIDS Register of Galicia (NW Spain), with a view to assessing the effect of the CD4 lymphocyte count on the probability of being AIDS-diagnosed via Tuberculosis (TB). This application shows how the link's flexibility makes it possible to obtain OR curve estimates that are less sensitive to the presence of outliers and unusual values that are often present in the extremes of the covariate distributions.

AIDS-Related Opportunistic Infections↗
PubMed2000

Rapid detection and quantification of Cryptosporidium baileyi oocysts in feces and organs of chickens using a microscopic slide flotation method.

A simple semiquantitative microscopic slide flotation (MSF) method using modified Sheather's sugar solution (MSSS) is presented for the rapid detection and quantification of Cryptosporidium baileyi oocysts in the feces and mucosal and/or organ scrapings of chickens. Oocyst shedding was evaluated by examination of the surface of coverslips, and the average quantitative score (0-5) recorded for 10 microscopic fields (magnification x250) is reported. The equivalence between these scores and the actual number of oocysts counted per gram of feces was assessed (rs = 0.89; P < 0.001). The applicability of this method was tested by comparison of the kinetics of oocyst shedding in feces of inoculated chickens with those reported by other authors working under similar conditions. In organs the MSF method was compared to histology. Fewer false-negative results were obtained using MSF versus the histology method. The MSF method was particularly more efficient in tracheae with low levels of infection and in the lungs, regardless of the level of infection. The MSF method was also very efficient in detecting oocysts in air sacs from chickens with aerosacculitis. It provides a specific and sufficiently sensitive, simple, rapid, reliable, and low-cost means of diagnosing C. baileyi in the feces and organs of chickens. This method can be used in the routine diagnosis of cryptosporidia in chickens, and it could be extended to other avian species and used in epidemiology studies to evaluate the prevalence of cryptosporidiosis in fowl.

Animals↗
PubMedDate not supplied

Effects of a 1 per cent hydrocortisone conditioner on the prevention of immediate and late-phase reactions in canine skin.

Ten laboratory beagles were used to determine whether a 1 per cent hydrocortisone conditioner applied topically for three consecutive days would inhibit IgE-mediated immediate and late-phase reactions induced in their skin. The trial was blinded, controlled with the product's vehicle and designed with a crossover. It consisted of three phases: one period without treatment (control phase) and two periods of treatment with either the active ingredient or the vehicle. Immediate and late-phase reactions were induced by the intradermal injection of rabbit anti-canine IgE polyclonal antibodies. Twenty minutes after the intradermal challenge, the diameter of the wheal, but not the erythematous flares, were significantly reduced after the application of the active product. In contrast, IgE-mediated cutaneous late-phase reactions, evaluated by measurements of dermal thickness and eosinophil counts six hours after challenge and the numbers of dermal CD3-positive T lymphocytes after 24 hours, were not reduced by its application.

Administration, Cutaneous↗
PubMed2006

Platelet function after single [153Sm]EDTMP therapy in prostate cancer.

AIM: Aim of the study was to assess whether [153Sm] EDTMP therapy at a low-dose is associated with platelet activation. METHODS: In 29 patients suffering from metastatic prostate cancer platelet count and various platelet function parameters have been monitored for 2 months after a single (the first) application of 1.1 GBq mCi [153Sm]EDTMP. RESULTS: After 3 days insignificant signs of platelet activation (increase in malondialdehyde, adenosine diphosphate-induced platelet aggregation, decreased platelet sensitivity) occur, normalizing rapidly. At the nadir of platelet count (3-4 weeks) platelet aggregation response in non-count adjusted samples is somewhat lower, while activity per cell (count adjusted samples) is unchanged. Platelet proteins do not change at all. Insignificant activation of platelet function at day 3 is interpreted as an indicator of mild oxidation injury, late aggregation response changes in non-adjusted samples seem only to reflect temporarily decreased number of circulating platelets. Samarium-153-EDTMP therapy at doses (1.1 GBq) used in the Vienna-protocol is not associated with a significantly altered functional behavior of platelets. CONCLUSIONS: We conclude that a single dose of 1.1 GBq 153Sm-EDTMP does not significantly affect in vivo and ex vivo platelet function.

Aged↗
PubMed1980

Quantitation of proteins in the femtomole range by hemagglutination using an electronic particle counter.

By using an electronic particle counter equipped with two counting channels, it is possible to simultaneously count total particles and agglutinated particles in an erythrocyte population. Using these two parameters, an empirically useful measure of the degree of agglutination in a sample (the agglutination index) can be generated. This method has been used to accurately and precisely determine human serum IgG concentrations in an inhibition of hemagglutination assay. As little as 1 microgram/ml of IgG could be measured in this way. Agglutination and inhibition of agglutination assays with other systems indicate that this method will be widely applicable.

Animals↗
PubMed2001

Application of semiconductor light sources for investigations of photochemical reactions.

Semiconductor light sources, like laser diodes or ultrabright light emitting diodes, are widely used in optical spectroscopy. In this presentation an overview of applications in photochemistry is given. Since the beginning of the 1990s an increasing number of publications with the application of semiconductor light sources appeared. Three different techniques were used: single photon counting with short pulses, phase-modulation fluorometry using a conventional modulation spectrometer, or a lock-in amplifier. Using continuous wave laser diodes in the visible region, which are available from 690 to 630 nm (and, recently, down to 400 nm), a new compact fluorescence spectrometer was developed in our laboratory. Using the phase fluorometric method, measurements down to 100 ps are now possible. Values can be measured in steps of 10 ps with good reproducibility using a high-frequency signal generator and a GHz digital storage oscilloscope. Several investigations have been carried out applying this technique including time-resolved detection of crude oil as an example for possible practical applications.

Biochemistry↗
PubMed1996

Predictors of survival in human immunodeficiency virus type 1-seropositive intravenous drug users.

In an ongoing prospective study of street-recruited intravenous drug users (IDUs) in Miami, Fla., 116 human immunodeficiency virus type 1 (HIV-1)-infected IDUs were monitored for up to 7 years. This provided an opportunity to evaluate baseline immunological parameters as potential predictors of survival among HIV-1-infected IDUs. As expected, HIV-1-infected IDUs who had an advanced stage of the disease (Centers for Disease Control and Prevention classification III or IV); p24 antigenemia; human T-cell leukemia virus type 1/2 seropositivity; low CD4 counts (< or = 200); low hemoglobin (< or = 14), high serum immunoglobulin A (IgA) (> 500 mg/dl), or high serum IgG (> or = 3,500 mg/dl) levels; or low proliferative responses to pokeweed mitogen (< or = 1,500 cpm) and to phytohemagglutinin (< or = 80,000 cpm) at baseline had worse survival rates. Results from multivariate Cox's models of survival showed that the baseline serum IgG level, serum IgA level, and CD4 count independently predict survival in HIV-1-infected IDUs. Cross-validation procedures verified the above-mentioned findings. These findings support the routine consideration of serum immunoglobulin levels in addition to CD4 count, especially in early evaluation of disease stage, as these evaluations may modify application of prophylaxis and treatment for HIV-1-infected IDUs. We recommend consideration of use of serum IgG and IgA as immunological markers for long-range prediction of survival in HIV-1-infected IDUs. These determinations are less onerous and more appropriate for use in field studies and financially less favored settings.

Adult↗
PubMed1980

[Effect of the antirheumatic drug Sulindac on thrombocyte function (author's transl)].

The effect of a treatment with the antirheumatic drug Sulindac (from 200 to 400 mg/day over 4 weeks) on thrombocyte function (bleeding time, platelet retention, platelet aggregation induced with suitable concentrations of ADP, epinephrine, collagen and Ristocetin) on thrombocyte count and thromboplastin time was investigated in 20 patients. In 10 healthy volunteers collagen-induced platelet aggregation was measured after oral application of 200 mg Sulindac. Analysis of the data obtained by the tests revealed no unwanted or harmful effects of Sulindac on platelet functions, platelet count and prothrombin time. No significant difference between the data before and during treatment was found. No medical drop out was registrated.

Collagen↗
PubMed1984

Histological changes in rabbits after application of medicaments and cosmetic bases (III).

An experimental study is reported of the irritant potential of oil of purcellin , isopropyl palmitate, nonyl phenol, Modulan , Amerchol and Acetulan when massaged in fixed amounts into the rabbit's skin daily for 30 days. Skin biopsies provided data on the number of epithelial cell layers, cell counts of the superficial dermis, and the fibre and basement membrane changes. All the test compounds induced changes in the form of increased number of epithelial cell layers and of the papillary dermis cell count, more marked for Acetulan or Amerchol . The difficulties in the evaluation of the results and their applications to human pathology are discussed.

Animals↗
PubMed1995

Prognostic factors for survival in advanced non-small-cell lung cancer: univariate and multivariate analyses including recursive partitioning and amalgamation algorithms in 1,052 patients. The European Lung Cancer Working Party.

PURPOSE: This study attempted to determine the prognostic value for survival of various pretreatment characteristics in patients with nonresectable non-small-cell lung cancer in the context of more than 10 years of experience of a European Cooperative Group. PATIENTS AND METHODS: We included in the analysis all eligible patients (N = 1,052) with advanced non-small-cell lung cancer registered onto one of seven trials conducted by the European Lung Cancer Working Party (ELCWP) during one decade. The patients were treated by chemotherapy regimens based on platinum derivatives. We prospectively collected 23 variables and analyzed them by univariate and multivariate methods. RESULTS: The global estimated median survival time was 29 weeks, with a 95% confidence interval of 27 to 30 weeks. After univariate analysis, we applied two multivariate statistical techniques. In a Cox regression model, the selected explanatory variables were disease extent, Karnofsky performance status, WBC and neutrophil counts, metastatic involvement of skin, serum calcium level, age, and sex. These results were confirmed by application of recursive partitioning and amalgamation algorithms (RECPAM), which led to classification of the patients into four homogeneous subgroups. CONCLUSION: We confirmed by our analysis the role of well-known independent prognostic factors for survival, but also identified the effect of the neutrophil count, rarely studied, with the use of two methods: a classical Cox regression model and a RECPAM analysis. The classification of patients into the four subgroups we obtained needs to be validated in other series.

Algorithms↗
PubMed2005

The safety of thiamethoxam to pollinating bumble bees (Bombus terrestris L.) when applied to tomato plants through drip irrigation.

Thiamethoxam, mainly sold under the trademark of Actara, is a neonicotinoid widely used in covered vegetables for the control of aphids and whiteflies. In these crops, and particularly in covered tomatoes, bumble-bees are used for cross-pollination as an alternative to labour intensive manual techniques. In this study, made on tomatoes grown in separated greenhouse plots in Murcia, Southern Spain, thiamethoxam was applied through drip irrigation at a rate of 200 g ai/ha, and as a split application of the same rate, to evaluate the effects on pollinating bumble bees compared to a foliar application of a toxic standard. The results showed that the toxic foliar standard had a clear effect on the pollination of tomato flowers, declining to zero pollination two weeks after application, whereas both the single and split drip irrigation applications of Actara had no effect on pollination when compared to the control plots. The count of dead adults and larvae did not show any differences between the treatments, whereas the measurement of sugar water consumption was shown to correlate well with pollination. The consumption of sugar water declined in the toxic standard plots by 69% with respect to the control, whilst the decline in lower dose drip irrigation application was only 3%. In regard to hive weight, and number of adults and brood after destructive sampling; there were no statistical differences between the treatments but a negative effect of the foliar treatment was observed. Based on these results we can conclude that a split application of Actara applied in drip irrigation to the soil/substrate has no effect on the bumble-bees used in tomatoes for pollination.

Agriculture↗