The use of probit-method for an estimation of toxic effects of combined toxicants at low concentration levels

In the present report the result of investigation of combined operating of propyl clone of Arminum is esteemed at effect on different intensity levels (LD, ED, Relatively Safe Level of Compound), with three-orthocresylphosphate and chlorophose in experience on white mice. The tendency of more expressed potentiation of toxic effect is detected at low levels of toxicant effect. Based on the dates of literature and the mathirials of the inner investigation, the expediency of the count toxicometric and statistical parameters received by a probit-method at a 5% level responses of the bioanswer is routined at a quantitative assessment of effects of combined operating of xenobiotics on low levels of effect.     

The evolutionary significance of phase-separated microsystems

The source, preparation, and properties of phase-separated systems such as lipid layers, coacervate droplets, sulphobes, and proteinoid microspheres are reviewed. These microsystems are of interest as partial models for the cell and as partial or total models for the protocell. Conceptual benefits from study of such models are: clues to experiments on origins, insights into principles of action and, in some instances, presumable models of the origin of the protocell. The benefits to evolution of organized chemical units are many, and can in part be analyzed. Ease of formation suggests that such units would have arisen early in primordial organic evolution. Integration of these various concepts and the results of consequent experiments have contributed to the developing theory of the origins of primordial and of contemporary life.Invited paper. Presented at the International Seminar Origin of Life, 2–7 August 1974, Moscow, U.S.S.R.     

Capacitive microsystems for biological sensing

The growing interest in personalized medicine leads to the need for fast, cheap and portable devices that reveal the genetic profile easily and accurately. To this direction, several ideas to avoid the classical methods of diagnosis and treatment through miniaturized and label-free systems have emerged. Capacitive biosensors address these requirements and thus have the perspective to be used in advanced diagnostic devices that promise early detection of potential fatal conditions. The operation principles, as well as the design and fabrication of several capacitive microsystems for the detection of biomolecular interactions are presented in this review. These systems are micro-membranes based on surface stress changes, interdigitated micro-electrodes and electrode-solution interfaces. Their applications extend to DNA hybridization, protein-ligand binding, antigen-antibody binding, etc. Finally, the limitations and prospects of capacitive microsystems in biological applications are discussed.     

The use of cyclic voltammetry for the evaluation of antioxidant capacity

Low-molecular weight antioxidants (LMWAs) play a major role in protecting biological systems against reactive oxygen-derived species and reflect the antioxidant capacity of the system. Cyclic voltammetry (CV), shown to be convenient methodology, has been validated for quantitation of the LMWA capacity of blood plasma, tissue homogenates, and plant extracts. Analysis of the CV tracing yields the values of (i) the biological oxidation potential, E and E(1/2), which relate to the nature of the specific molecule(s); (ii) the intensity (Ia) of the anodic current; and (iii) the area of the anodic wave (S). Both Ia and S relate to the concentration of the molecule(s). LMWA components of human plasma and animal tissues were identified and further validated by reconstruction of the CV tracing and by high-performance liquid chromatography-electrochemical detection. To reflect the oxidative stress status, the use of an additional parameter, R, has been proposed. R represents the level (%) of oxidized ascorbate (compared with total ascorbate) and is measured by high-performance liquid chromatography-electrochemical detection. All these parameters were monitored in healthy human subjects as well as in chronic (diabetes mellitus) and acute care patients (subjected to total body irradiation before bone marrow transplantation). The electroanalytical methodologies presented here could be widely employed for rapid evaluation of the status of subjects (in health and disease) for monitoring of their response to treatment and/or nutritional supplementation as well as for screening of specific populations.     

The use of jackknifing for the evaluation of geographic profiling reliability

The use of geographic profiling (GP), based on “Rossmo's formula”, a technique derived from criminology, has been previously proven to be effective in assessing the origin of invading species. The application on Caulerpa taxifolia showed the most probable center of spread of the invasion. This article discusses a method of assessing the degree of robustness of the results obtained with Rossmo's method.To provide an evaluation of the reliability of geographic profiling results we used the jackknife technique, randomly eliminating part of the data set for a given number of replicates (500) in order to analyze the obtained result for each replicate. In GP the results are a series of images with geoprofiling prioritization, each produced with one of the replicates. These images can be summarized in three different ways: (1) OR, depicting all the high probability pixels from the series of replicates; (2) AND, depicting only those high probability pixels present in every replicate; and (3) MEAN, depicting the mean color value for each pixel calculated from all the replicates. We show that jackknifing can be a useful method to increase robustness of GP analysis in criminology, epidemiology and biological invasions. Summarizing jackknifing results with the OR logical operator yields the highest sensitivity and worst specificity, while the use of the AND operator increases specificity but reduces sensitivity. Using the mean of the pixel values maintains the visualization of the areas of highest priority (specificity), while also showing the surrounding area with varying colors, analogous to confidence limits.     

The use of scientometric parameters for the evaluation of scientific contributions

This paper deals with the application of scientometric parameters in the evaluation of scientists, either as individuals or in small formal groups. The parameters are divided into two groups: parameters of scientific productivity and citation parameters. The scientific productivity was further subdivided into three types of parameters: (i) total productivity, (ii) partial productivity, and (iii) productivity in scientific fields and subfields. These citation parameters were considered: (i) impact factors of journals, (ii) impact factors of scientific fields and subfields, (iii) citations of individual papers, (iv) citations of individual authors, (v) expected citation rates and relative citation rates, and (vi) self-citations, independent citations and negative citations. Particular attention was payed to the time-dependence of the scientometric parameters. If available, numeric values of the world parameters were given and compared with the data about the scientific output of Croatian scientists.     

Chromatographic methods for the determination of toxicants in faeces

Modern chromatographic techniques and their application in the determination of toxic compounds in faeces are reviewed. Faecal analysis may be of importance in toxicokinetic studies of xenobiotics in order to determine factors such as metabolism, body burden and major routes of elimination. Compounds of interest include various food constituents, drugs and occupational or environmental factors. Further, various mutagenic or carcinogenic compounds which are excreted by faeces have been indicated to represent risk factors for colorectal cancer. In this context, the chromatographic determination of the endogenously generated fecapentaenes and bile acids, both postulated etiological factors in colorectal carcinogenesis, is reviewed. For fecapentaene determination, several high-performance liquid chromatographic (HPLC) methods are available; however, the applicability of some of these methods is limited owing to insufficient separation of various isomeric forms or discrimination between fecapentaenes and their precursors. For the determination of bile acids in faeces, many chromatographic procedures have been reported, and the characteristics of the most relevant methods are compared and discussed. It is concluded that separation by gas chromatography (GC) in combination with mass spectrometry provides the highest selectivity and sensitivity. A relatively rapid alternative analysis for the determination of total and aqueous faecal bile acids is proposed. Further, methods for the determination of polycyclic aromatic hydrocarbons (PAHs) are reviewed. Although the use of radiolabelled PAHs in animal studies has many advantages, it cannot be applied for human biological monitoring and HPLC and GC provide sensitive alternatives. An HPLC method for the determination of non-metabolized PAHs in faeces is described.     

Thin-film IrOx pH microelectrode for microfluidic-based microsystems

Microsensors are valuable tools to monitor cell metabolism in cell culture volumes. The present research describes the fabrication and characterization of on-chip thin-film iridium oxide pH microsensors with dimensions of 20 microm x 20 microm and 20 microm x 40 microm suitable to be incorporated into nl volumes. IrOx thin films were formed on platinum microelectrodes by electrochemical deposition in galvanostatic mode. Anodically grown iridium oxide films showed a near super-Nernstian response with a slope of -77.6+/-2 mV/pH at 22 degrees C, and linear responses within the pH range of 4-11. Freshly deposited electrodes showed response times as low as 6s. Long-term studies showed a baseline drift of 2-3 mV/month, which could easily be compensated by calibration. This work demonstrated for the first time the use of planar IrOx pH microelectrodes to measure the acidification rate of CHO and fibroblast cells in an on chip cell culture volume of 25 nl with microfluidic control.     

The use of primary hepatocyte cultures for the evaluation of chemoprotective agents

Primary cultures of human and rat hepatocytes are widely used in pharmacotoxicological research. This model presents the advantages of retaining liver function for at least a few days, expressing both phase 1 and phase 2 enzymes, and responding to inducers. Recently we made use of primary hepatocytes to investigate the effects of chemoprotective agents on drug-metabolizing enzyme expression and activities. The treatment of rat and human hepatocytes with two chemoprotective agents, oltipraz (a synthetic derivative of 1,2-dithiole-3-thione) and sulforaphane (an isothiocyanate found in broccoli), clearly demonstrated that both of these compounds are inducers of glutathione transferases and transient inhibitors of cytochrome P450, suggesting that these two compounds could exert their chemoprotective effects by both reducing the formation of reactive metabolites of chemicals and enhancing their inactivation.     

The use of genetic tools for the evaluation of a potential migration barrier for the bullhead

Microsatellite analysis and computer simulations strongly suggested that a culvert, i.e . a connection between two river stretches by a narrow tube underneath an artificial channel, was not a migration barrier for the endangered bullhead Cottus gobio .     

The effect of organic toxicants on sensitivity of intestinal glycosidases to Cu and Zn in juvenile roach

The chronic effect (duration of exposure 218 days) of polychlorinated biphenyls (PCBs) and the prolonged effect of the short-term action of chlorophos or of N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) during embryogenesis upon the sensitivity of intestinal glycosidases to Cu and Zn was studied in roach (Rutilus rutilus (L.) underyearlings. The Cu⁺² and Zn⁺² ions at concentrations ranging from 0.1 to 25 mg/L in vitro cause a 10–77% decrease in amylolytic activity in the intestinal mucosa of control roach. An elevated level of PCBs (50.8 ng/g wet weight of food and 426 ng/g dry weight of ground) increased the sensitivity of glycosidases to Cu and Zn. The embryotoxic action of chlorophos at concentrations of 1 × 10⁻⁶−1 × 10⁻² mg/L in most cases increased the inhibitory effect of Cu but decreased that of Zn. As a rule, MNNG (3 × 10⁻⁷−3 × 10⁻¹ mg/L) reduced the glycosidase sensitivity to the effect of metal ions. The magnitude and direction of the effect depend on the nature and concentration of toxicants.     

Evaluation of various toxicants in rabbit whole-embryo culture using a new morphologically-based evaluation system

In an effort to advance the use of whole-embryo culture (WEC) techniques in the rabbit, we recently developed a gestational-age-based quantitative morphologic evaluation system for rabbit embryos. In the current study, we applied this new morphological scoring system to assess the development of rabbit gestational day (gd) 9 embryos exposed for 48 hr in WEC to the teratogens ethanol (EtOH, 154 mM), 6-aminonicotinamide (6AN, 0.15 mM), and methoxyacetic acid (MAA, 5.0 mM), and the nonteratogen penicillin G (PG, 2.0 mM). Each teratogen at the concentration tested markedly inhibited morphological development, as indicated by significantly lower morphologic scores (10.1+/-0.05, EtOH; 10.2+/-0.05, 6AN; and 9.8, MAA) relative to controls (10.6+/-0.04), and resulted in an increased percentage of malformed embryos (53%, EtOH; 57%, 6AN; 90%, MAA; and 3%, control). Embryonic growth, as measured by head length, somite number, and total embryonic protein, was significantly decreased by each teratogen. The abnormalities produced by teratogen exposure, which included brain, somite, and facial defects, were often similar to those produced following in vivo exposure in rabbits and rodents, and/or in vitro exposure in rodents. In contrast to the teratogen exposure groups, PG had no effect on embryo growth parameters, or on malformation rate (6%), although a slight but statistically significant decrease in morphology score (10.5+/-0.03) was noted. Our preliminary studies demonstrate the usefulness of the morphology evaluation system by quantifying graded differences in development, and indicate that rabbit WEC may be a useful adjunct to rodent WEC in gaining insights regarding differential interspecies sensitivity.     

The use of environmental media evaluation guides for screening environmental contamination

In conducting health assessments at hazardous‐waste sites, ATSDR staff must identify environmental chemical contaminants that pose a potential public health hazard. To assist health assessors in this effort, ATSDR has developed environmental media evaluation guides (EMEGs). EMEGs have been developed for water, soil, and air. The EMEG values are expressed as a range to account for the range in exposures in different segments of the population. The use of EMEGs provides health assessors with a consistent strategy for selecting environmental contaminants to be further evaluated in the health assessment process.     

The evaluation of the temperature effect and hydrolysis conditions for amino acid analysis

A systematic investigation of the optimal temperature and hydrolysis time for amino acid analysis has been carried out under various conditions. It is found that some simplification and increase in speed relative to the conventional protocol of employing vacuum-sealed tubes and 110 C/24-72 hour hydrolysis can be achieved without loss of accuracy and performance in amino acid analyses of proteins and peptides. The effects of hydrolysis temperature and heating time on the recoveries of various labile and hydrophobic amino acids are exemplified in the hydrolysis of oxidized ribonuclease A, lysozyme and lens crystallin. The method provides a rapid processing of multiple samples within hours instead of days with the potential for the total automation of amino acid analysis starting from the preparation of protein hydrolysates.