Sample pretreatment and determination of non steroidal anti-inflammatory drugs (NSAIDs) in pharmaceutical formulations and biological samples (blood, plasma, erythrocytes) by HPLC-UV-MS and micro-HPLC.

The article discusses the qualitative and quantitative determination of non-steroidal anti-inflammatory drugs like salicin, salicylic acid, tenoxicam, ketorolac, piroxicam, tolmetin, naproxen, flurbiprofen, diclofenac and ibuprofen by reversed phase high performance liquid chromatography (RP-HPLC) and micro-HPLC (micro-HPLC) hyphenated with UV-absorbance and mass spectrometric detection. Both detection methods delivered calibration plots with good linearity (r(2) > 0.9800), limits of detection in the low nanogram range and recovery rates between 94 and 104 %. For the analysis of biological samples such as blood, plasma and erythrocytes liquid-liquid extraction (LLE) and solid phase extraction (SPE) on the basis of new synthesized glycidylmethacrylate/divinylbenzene copolymer (GMA/DVB) particles and commercially available material on the basis of poly(divinylbenzene-co-N-vinylpyrrolidone) copolymer were investigated. Finally the use of a micro-HPLC system with separation columns in the range of 8 cm x 200 microm I.D. for the determination of non-steroidal anti-inflammatory drugs (NSAIDs) is presented, emphasizing on the type of column and sample amount needed.

Electrodynamics of microtubular motors: the building blocks of a new model.

Microtubules are ubiquitous components of the cytoskeleton. They participate in many motility processes ranging from intracellular transport or chromosome movement during mitosis to ciliary and flagellar beating. The biophysical mechanism inherent in the generation and control of movement in all these motility phenomena has not yet been entirely elucidated. The authors propose a new model based on a charge transfer mechanism capable of shedding a new light on the molecular foundations of all motility processes. Electron transfer along the microtubular lattice is responsible for activation and control of all microtubule-associated ATPases (i.e. force generating enzymes). Microtubules are thus shown to be the basic motors of cell dynamics. The model is first applied to intracellular transport and ciliary and flagellar beating. Through two additional examples, the authors show the heuristic capabilities of the suggested hypothesis. The application of charge transfer control to the Protozoan Euglena gracilis leads to a plausible model capable of accounting for its phototactic response mechanism. Furthermore, the model allows a new interpretation of the electrophysiological response in vertebrate photoreceptors.

[Invariability of the H-Y antigen expression in the heterogametic sex of some amphbians and evidence for sexual dimophrism of the antigen expression in Pelodytes punctatus D (Amphibia, Anura)]. / Sur la constance de l'expression de l'antigène H-Y chez le sexe hétérogamétique de quelques Amphibiens et sur la mise en évidence d'un dimorphisme sexuel de l'expression de cet antigène chez l'Amphibien Anoure Pelodytes punctatus D.

The H-Y antigen is studied in some Amphibians whose sexual genetic constitution is known. Thus, in Pelurodeles waltlii, Ambystoma mexicanum, Xenopus laevis and Rana ridibunda, the invariability of the H-Y antigen expression in the heterogametic, sex is confirmed. In Pelodytes punctatus the genetic sex is unknown but the existence of the H-Y antigen in phenotypic males leads to the conclusion of a male heterogamety of the XY type.

H-Y antigen expression in temperature sex-reversed turtles (Emys orbicularis).

H-Y antigen has been used as a marker for the heterogametic sex and is assumed to be an organizing factor for the heterogametic gonad. In the turtle Emys orbicularis, H-Y antigen is restricted to the female cells, indicating a female heterogamety (ZZ/ZW) sex-determining mechanism. Moreover, the sexual differentiation of the gonads is temperature sensitive, and complete sex reversal can be obtained at will. In this framework the relationships between H-Y antigen, temperature, and gonadal phenotype were studied. Mouse H-Y antiserum was absorbed with blood and gonadal cells of control wild male and female adults, and with blood and gonadal cells from three lots of young turtles from eggs incubated at 25-26 degrees C (100% phenotypic males), at 30-30.5 degrees C (100% phenotypic females), or at 28.5-29 degrees C (majority of females with some males and intersexes). The residual activity of H-Y antiserum was then estimated using an immunobacterial rosette technique. In adults, both blood cells and gonadal cells were typed as H-Y negative in males and as H-Y positive in females. In each of the three lots of young, blood cells were H-Y negative in some individuals and H-Y positive in others. The proposed interpretation is that the H-Y negative individuals were genotypic males (ZZ) and the H-Y positive were genotypic females (ZW). The gonads of these animals were then pooled in different sets according to their sexual phenotype and to the presumed genotypic sex (i.e., blood H-Y phenotype). Testicular cells were typed as H-Y negative in genotypic males as well as in the presumed sex-reversed genotypic females; likewise, ovarian cells were typed as H-Y positive in genotypic females as well as in the presumed sex-reversed genotypic males. These results provide additional evidence that H-Y antigen expression is closely associated with ovarian structure in vertebrates displaying a ZZ/ZW sex-determining mechanism.

Temperature-dependent gonadal differentiation in the turtle Emys orbicularis: concordance between sexual phenotype and serological H-Y antigen expression at threshold temperature.

As in many other turtles, the sexual differentiation of gonads in embryos of Emys orbicularis is temperature-sensitive, 100% phenotypic males being obtained below 27.5 degrees C and 100% phenotypic females above 29.5 degrees C. The expression of the serologically defined H-Y (SD-H-Y) antigen at both low and high temperatures has been shown to be different in gonads and in blood : in gonads, it is closely associated with ovarian structure, whereas in blood it is independent of sexual phenotype and appears to indicate sexual genotype. Both sexes differentiate at 28.5 degrees C, suggesting that at this intermediate (threshold) temperature, sexual differentiation of gonads conforms with sexual genotype. To test this hypothesis, the expression of SD-H-Y antigen has been carried out in blood cells of Emys individuals raised from eggs incubated at the threshold temperature (28.5 degrees C). All phenotypic males typed SD-H-Y negative, whereas most phenotypic females typed SD-H-Y positive. From this concordance between sexual phenotype of gonads and SD-H-Y phenotype of blood, we postulate that a ZZ male/ZW female mechanism of genotypic sex determination is revealed at the threshold temperature for gonad differentiation in Emys.

H-Y antiserum recognizes male-specific and non-specific components in human lymphocytes.

Serological assays for H-Y antigen, which is male-specific in mammals, all give slightly positive but non-negligible results when females are tested. All assays used to this day share this characteristic; furthermore they do not analyse the pattern of antigens recognized by the antisera. We used the Western blot technique in an attempt to do this, and observed that three components (relative molecular mass: 15-20 kilodaltons (kD)) were recognized both in females and in males tested, and a single specific band of 32-34 kD relative molecular mass in males. This confirms the existence of a serologically detectable male antigen on human lymphocytes, and explains the existence of a certain level of antigen expression in females tested by different techniques, including the indirect immunofluorescence and ELISA used here for comparison.