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PubMed2005

Analysis of the human saliva proteome.

Interest in the characterization of the salivary proteome has increased in the last few years. This review discusses the different techniques and methodologies applied to the separation and identification of salivary proteins. Nowadays, proteomic techniques are the state of the art for the analysis of biologic materials and saliva is no exception. 2D electrophoresis and tryptic digest analysis by mass spectrometry are the typical methodology, but new approaches using 2D liquid chromatography/mass spectrometry methods have already been introduced for saliva analysis. Due to their important physiologic role in the oral cavity, low-molecular-weight proteins and peptides are also included in this article and the methodologies discussed.

Animals↗
PubMed1996

Olfactory control of mosquito behaviour.

The following papers will tell us much about our current knowledge of certain aspects of the olfactory control of mosquito behaviour-specifically host-locating behaviour. More importantly, as we read the papers, we should pay attention to the insights they will provide about what is not known and to the questions that our lack of knowledge should raise. The authors will also provide us with some new approaches and methodologies that we can apply to the study of mosquito physiology and behaviour.

Animals↗
PubMed2001

Simultaneous detection of multiple cytokines from conditioned media and patient's sera by an antibody-based protein array system.

We have developed a novel technique for high-throughput simultaneous screening of multiple cytokine expression based on a protein array system. Our method has the advantage of showing the specificity of enzyme-linked immunosorbent assays, sensitivity of enhanced chemiluminescence (ECL), and high-throughput of microspot. In this system, the cytokine array membranes were created by spotting capture antibodies onto the membranes. The membranes were then incubated with biological samples such as conditioned media and patient's sera. The bound proteins were then recognized by biotin-conjugated antibodies and detected by horseradish peroxidase-conjugated streptavidin coupled with ECL. Experiments demonstrated that 24 cytokines from conditioned media and patient's sera could be simultaneously detected using this new approach. This methodology should allow us to develop many high-density protein array systems to detect a variety of proteins. To validate and quantitate the expression of key molecules in a wide range of samples, we have developed conditioned medium arrays to evaluate hundreds and even thousands of samples from individual cells and patients in a single microarray. The combinations of protein arrays and conditioned medium arrays or serum arrays will provide a powerful tool to identify the protein expression profiles and rapidly validate their expression in many types and numbers of samples.

Antibodies↗
PubMed2002

Assessment of the sustainability of technology by means of a thermodynamically based life cycle analysis.

Life cycle analysis is one of the tools in the assessment of the sustainability of technological options. It takes into account all effects on the ecosystem and the population which may endanger the possibilities of current and future generations. However, the main bottleneck in current LCA methodologies is the balancing of different effects, being all quantified on different scales. In this work, a methodology is proposed, which allows one to quantify different effects of the production, consumption and disposal of goods, and services on a single scale. The basis of the methodology is the second law of thermodynamics. All production, consumption and disposal processes affecting the ecosystem and the population, are quantified in terms of loss of exergy. The exergy content of a material is the maximum amount of energy which can be transformed into work at given environmental conditions. Next to the elaboration of the methodology, the new approach is illustrated by examples of the production of synthetic organic polymers, inorganic building insulation materials and different waste gas treatment options.

Conservation of Natural Resources↗
PubMed1979

Depression and monoamine oxidase.

1. The first part of the paper is devoted to a critical review on the possible relationships between depression and monoamine oxidase. 2. This study describes the results of an investigation of MAO-activity in depression, using new approaches and methodology. This methodology was developed because the literature data indicated that a) previous results are difficult to compare because of varying methods and diverse target populations used; b) previous methodological deficiencies do not allow to draw definite conclusions about the relationship between MAO-activity and depression. 3. The present investigation selected 35 psychiatric patients according to clearly defined diagnostic criteria (20 endogenous depressive, 10 neurotic depressive, and 5 manic patients) matched to 25 healthy control-subjects. 4. The Michaelis-constant (Km), the maximum reaction speed (Vmax), and the 50% enzyme inhibition by tranylcypromine (IC50) of platelet-MAO were determined during and after recovery from the depressive or manic episode using 3 substrates (tyramine, tryptamine, and phenylethylamine). 5. The present investigation, in contrast to conventional methodology, utilized three different substrates at different concentrations. Significant correlations were demonstrated for the Vmax-values of each of the three substrates, whereas the Km and the IC50 (tranylcypromine)-values varied for each substrate. 6. The results show that there were no differences between the characteristics of the platelet-MAO in depressive or manic patients and those of normal subjects. Furthermore, treatment with tricyclic antidepressants had no effect on MAO-activity. A previous investigation indicated that the MAO-properties in human brain tissue were similar to that in human platelets.

Adult↗
PubMed2006

Assessing the impact of leather industries on the quality of water discharged into the East China Sea from Wenzhou Watersheds.

Ammonium nitrogen and total germanium are among the main pollutants in the wastewater discharged from the leather industry. The intake of high concentrations of ammonium nitrogen and/or total germanium harms human health and biological species, as is well documented in literature. This paper focuses on assessing the trends of ammonium nitrogen and total germanium concentrations through time in two watersheds (Aojiang and Oujiang) in the Wenzhou metropolitan area of Zhejiang Province and their relationships with the released wastewater using regression and correlation statistics. The paper also utilizes the integrated pollution index to evaluate water quality in the two watersheds. Preliminary results show that, from 1992 to 1998 in the Aojing watershed, the concentrations of ammonium nitrogen and total germanium increased 13 and 14 times, respectively, decreasing somewhat after 1998, while between 1992 and 1997 in the Oujiang watershed, the concentrations increased, then decreased after 1997. The concentrations of ammonium nitrogen and total germanium are positively related to the amount of released wastewater. The concentrations of ammonium nitrogen and total germanium exceeded water standards 12 and 3 times, respectively, in Pingyang county of the Aojiang watershed, 14 and 3.3 times in Lucheng District of the Oujiang watershed, and 14 and 3.8 times in the Ouhai Oujiang watershed, respectively. In Pingyang county of the Aojiang watershed, the water quality degraded from Type III in 1992 to over Type V in 2003, and in the Oujiang watershed, the water quality degraded from Type II to over Type IV in 1999, when they were compared with the water quality standards. The water quality slightly improved in 2003 for the Oujiang watershed. It appears that pollution did have a direct positive correlation with leather industry production in the Pingyang Aojing watershed, while there was a negative correlation between the two in the Oujiang watershed. In these two watersheds, the integrated pollution index did not appear to relate to population dynamics and agricultural production. This paper also discusses the current new methodologies and approaches adopted nationally and internationally to reduce the contaminants and purify the environment for maintaining a sustainable and healthier environment in Wenzhou.

China↗
PubMed2026

Functional perturbation reveals context-dependent contributions of nuclear receptors to drug-induced hepatic steatosis.

Drug-induced hepatic steatosis is mediated by diverse molecular mechanisms, yet several nuclear receptors have been proposed as molecular initiating events or early key events within adverse outcome pathways for hepatic steatosis. However, direct functional evidence supporting these mechanistic roles in human-relevant experimental systems remains limited. The present study evaluated the contribution of selected nuclear receptors to drug-induced hepatic steatosis using complementary human hepatic in vitro models. Stable short hairpin RNA-mediated knockdown of individual nuclear receptors was established in HepG2 and differentiated HepaRG cells, followed by exposure to representative steatogenic drugs, including valproic acid, amiodarone, tamoxifen, and rifampicin. In parallel, primary human hepatocyte spheroids were used to compare drug-induced lipid accumulation with direct pharmacological activation of individual nuclear receptor pathways. While depletion of multiple nuclear receptors markedly affected oleic acid-induced lipid accumulation, drug-induced steatogenic responses exhibited predominantly selective and compound-specific receptor dependencies. In differentiated HepaRG cells, nuclear receptor depletion influenced basal lipid homeostasis more strongly than valproic acid-induced lipid accumulation. Conversely, direct activation of liver X receptor and peroxisome proliferator-activated receptors α and γ in primary human hepatocyte spheroids induced robust lipid accumulation, whereas most steatogenic drugs produced comparatively modest responses. These findings demonstrate that the contribution of individual nuclear receptors to drug-induced hepatic steatosis is highly compound- and context-dependent and cannot be explained by a single conserved receptor pathway. This study provides functional evidence from complementary human-relevant hepatic models that supports refinement of hepatic steatosis adverse outcome pathways and highlights the value of targeted perturbation strategies for mechanistic toxicology.

Adverse outcome pathway, HepaRG↗
PubMed2002

Methodological approach towards the definition of new storage conditions for inert wastes.

In 1997, the French Ministry of Environment launched studies aiming to define a specific regulation concerning inert waste disposal in order to limit potential impact of such facilities on the environment by fixing minimum requirements. A model (chemical model/hydrodynamic model) was developed to determine dumping conditions. This model was then applied on two defined scenarios (landfill surface, effective rainfalls...) in order to study the sulphate concentrations in aquifer system immediately downstream from the storage facility. Results allow us to determine in which conditions the sulphates concentrations are compatibles with the potentially drinkable character of the groundwater. They more specifically concern the nature of the waste disposed of, the efficient rainfalls and the landfill area.

Environmental Monitoring↗
PubMed2001

Disability methodology redesign: considerations for a new approach to disability determination.

Disability determination meets important societal needs, involving billions of dollars and millions of people every year. However, disability determination decisions often are incorrect, and the high proportion of decision appeals and reversals creates additional administrative expense and difficulty for the people that the disability determination system is intended to support. Projects funded by the United States Social Security Administration explored these issues and developed new conceptual models and tools to improve the accuracy and fairness of disability determination. This paper provides an introduction to the projects and the papers in this special issue of the Journal of Occupational Rehabilitation.

Disability Evaluation↗
PubMed2000

A new procedure for evaluation of renal function without urine collection in rat.

BACKGROUND: A new procedure to improve the accuracy of inulin assessment and renal glomerular filtration rate (GFR) avoiding urine sampling was compared and validated versus the reference procedure (with urine sampling and Anthrone reaction) in conscious unrestrained male Wistar rats. METHODS: The hemodynamic study consisted of a priming dose of inulin (16 mg/kg) and para-aminohippurate (PAH; 8 mg/kg) followed by an infusion of inulin (36 mg/mL) and PAH (5.8 mg/mL) at a rate of 0.055 mL/min until steady-state conditions were reached (105 min). Inulin concentrations from samples were determined by a new enzymatic assay and Anthrone reaction. PAH concentrations were determined according to the standard method described by Smith et al. RESULTS: A high correlation was found between GFR and renal blood flow (RBF) values calculated using the alternative (without urine sampling) and the reference (with urine sampling) clearance techniques (r = 0.98, P < 0.001, and r = 0.97, P < 0.001, respectively). Moreover, a significant and positive correlation between the values obtained from enzymatic and Anthrone inulin assessments was found (r = 0.99, P < 0.001). Likewise, the values of the 95% confidence interval (mean +/- 2 SD) for the enzymatic inulin assay showed a good agreement with those achieved with Anthrone (1.14 +/- 0.21 and 1.14 +/- 0.19 mL. min-1. 100 g-1 rat body weight, respectively). CONCLUSIONS: This new approach has methodological and experimental advantages with respect to traditional procedures, making it a useful tool, not only for research purposes but also in the clinical setting.

Animals↗
PubMed2006

Children's environment and health in Latin America: the Ecuadorian case.

Environmental health problems of children in Latin America and Ecuador are complex due to the close relationship that exists between social and environmental factors. Extended poverty and basic problems, such as the lack of drinking water and sanitation, are common. Infectious diseases are the greatest cause of morbidity and mortality among children. Development in industry and the introduction of chemical substances in agriculture add new risks including pesticide use, heavy metal exposure, and air pollution. Major problems can be divided into (a) lack of basic infrastructure, (b) poor living conditions, (c) specific environmental problems, and (d) child labor. Reproductive health disorders are frequent in developing countries like Ecuador. Issues related to children's health should consider new approaches, creative methodologies, and the search for independent predictors to separate environmental from social problems. Only with knowledge of the specific contribution of each factor, can it be possible to develop a strategy for prevention.

Child↗
PubMed2001

Current status of three-dimensional echocardiography.

Volume rendered three-dimensional echocardiography (3DE), a rapidly emerging imaging method, offers clinicians and surgeons a new perspective for visualizing the heart. Using both transthoracic and transesophageal approach this new methodology enables qualitative and quantitative information of cardiac disorders to be obtained. 3DE image projections have allowed visualization of intracardiac structures and great vessels and delineation of their pathology in a comprehensive manner. Experimental and clinical studies suggest that 3DE is likely to play a valuable role in the evaluation of various cardiac and flow disorders and in planning interventions and surgery. In addition, 3DE has been applied to derive quantitative measurements of volume, mass and dimensions of the left and right ventricles and also other cardiac lesions, such as atrial and ventricular septal defects. Every aspect of 3DE is in continuous evolution. Faster and more highly automated image processing could make the technique more easily applicable in the clinical scenario.

Echocardiography, Three-Dimensional↗
PubMed2003

The role of social worker in a team of rehabilitation: methodological approach.

This article introduces a new concept of a social worker role in rehabilitation team in Lithuania and presents a methodological approach for investigation of this role as well. THE GOAL of this study is to establish a standardized method for evaluation of social worker roles', significance for disabled people in a home-based rehabilitation setting, and to evaluate the most useful roles of social worker according to the needs of disabled people. The research method of this study was designed, based on a review of Lithuanian and foreign literature. Fifty (25 male, 25 female) disabled persons, who received home-based rehabilitation services, were observed and questioned according to this method. The results of the study show that the most important roles of a social worker in home-based rehabilitation are those of a broker, an enabler and a teacher. CONCLUSION. It was discovered that there is a need for follow-up qualification program of social workers in a rehabilitation team in order to develop additional skills that correspond to the roles of a teacher, an enabler and a broker, with application of this method for home-based rehabilitation of disabled people.

Persons with Disabilities↗
PubMed2000

Flow-cytometric characterization of normal versus psoriatic epidermis using improved cell separation methodology.

Recently a new approach for epidermal cell characterization was developed: three-parameter flow cytometrical analysis of pure and complete epidermal cell suspensions prepared from punch biopsies followed by dermoepidermal separation by thermolysin. The aim of the present communication is the comparison between psoriatic lesional skin and normal skin using this new approach with respect to the percentage of suprabasal keratinocytes (keratin 10+ cells), mesenchymal cells, including the infiltrate cells (vimentin+ cells) and the percentage of basal cells in SG2 M phase, in order to validate this methodology in studies on psoriatic skin. Punch biopsies were taken from 7 healthy volunteers and in 7 psoriatic patients 4 biopsies were taken in each of them from comparable lesions. The present study reconfirmed that the percentage of basal keratinocytes in psoriasis was increased and the percentage of keratin 10+ cells was substantially decreased as compared to normal skin. The new methodology revealed data with a narrow range. In psoriatic lesional skin the intra individual variation was less compared to the inter individual variation.

Case-Control Studies↗
PubMed1996

Methodology for a randomised controlled trial of preschool vision screening. A new approach with the 'ALSPAC' project.

We present the methodology of a population-based Randomised Controlled Trial, comparing an intensive programme of primary preschool vision screening by orthoptists with the usual non-specialist screening. The aims of the trial are to compare the effectiveness and costs of intensive orthoptic screening with non-specialist measures. The orthoptic screening programme will be evaluated both as a composite package and in terms of the screening value of the individual tests at specific ages. This trial is nested within a large population-based longitudinal study. Additional demographic and developmental data on the children in the trial are therefore available. The results of the trial will be used to help clarify which methods of preschool ophthalmic population screening are best in terms of disease detection and cost efficiency.

Amblyopia↗
PubMedDate not supplied

A new approach to phenotyping disseminated tumor cells: methodological advances and clinical implications.

At the time of primary therapy (surgery, systemic chemotherapy and/or radiation), disseminated tumor cells in the bone marrow can be found in almost one-third of patients with cancer of the breast, ovary, esophagus, stomach, colon, and other solid tumors. Whereas the prognostic impact of the mere presence of these cells is still a matter of debate, it has been shown that expression of tumor-associated antigens in disseminated tumor cells is linked to more aggressive disease. Therefore, further characterization of disseminated tumor cells at the protein and gene level has become increasingly important. To date, the most common detection method for disseminated tumor cells in the bone marrow is an immunocytochemical approach using cytokeratin-directed antibodies for detection of epithelial cells and the APAAP system for their visualization. We have established a new double immunofluorescence technique enabling simultaneous detection, phenotyping, and antigen quantification of disseminated tumor cells. Mononuclear cells from bone marrow are enriched by Ficoll gradient centrifugation and cytospins are prepared. Double immunofluorescence is performed using antibodies against cytokeratins 8/18/19 (mAb A45B/B3) and the uPA receptor CD87 (pAb HU277). CD87 expression is recorded by confocal laser scanning microscopy (CLSM) using fluorescence labeled latex beads as the reference; staining intensities of all the scans are then summed and quantified (extended focus). This protocol, originally designed for disseminated tumor cells in bone marrow, can also be applied to disseminated tumor cells in blood, to leukapheresis cells or to cells present in malignant ascites or other malignant effusions. The tumor cells detected may be used for gene and mRNA analyses. Furthermore, disseminated tumor cells also represent interesting targets for clinical studies on patient prognosis or prediction of therapy response as well as for specific tumor-biological therapies.

Biomarkers, Tumor↗
PubMed1987

The therapeutic efficacy of critical care units from two perspectives: a traditional cohort approach vs a new case-control methodology.

The therapeutic efficacy of critical care units-whether they do more good than harm and for whom--has not been established, except for patients who are admitted for life-sustaining interventions, such as mechanical support of ventilation. However, most patients are admitted for observation, and to facilitate intervention if deterioration occurs or complications develop. The objective of this study was to determine whether direct admission to critical care units reduced mortality rates. The population under study consisted of all 604 patients admitted to the medical service during a one month period. At the time of admission, the responsible residents rated patients as to how sick and stable they were. These ratings of illness severity and stability have been shown to be the most significant predictors of in-hospital mortality and morbidity, respectively; they were employed to stratify the patients prognostically. The first analysis utilized the entire cohort of 604 patients. After patients who would have been ineligible for entry into a trial were removed, direct admission to the unit was associated with a reduced mortality in only one group of patients: the unstable, moderately ill (p less than 0.05). "Unstable, severely ill" patients had high mortality rates when admitted to the floor or units, and stable patients (mildly or moderately ill) did equally well when admitted to either location. A further analysis revealed a possible explanation for these findings. Among the unstable, moderately ill patients, the rate of deterioration of pre-existing problems was significantly lower among patients directly admitted to the unit (p less than 0.05), whereas the rate of new complications did not differ.(ABSTRACT TRUNCATED AT 250 WORDS)

Critical Care↗