La chromatographie sur couche mince: Principes et possibilités

Résumé Après un bref rappel des avantages de la chromatographie de surface sur la chromatographie sur colonne, on compare les avantages et les inconvénients respectifs de la chromatographie sur couche mince et de la chromatographie sur papier. Les méthodes générales de la chromatographie sur couche mince (CCM) sont présentées (préparation des couches, types de couches, dépot des échantillons à analyser, CCM analytique, CCM préparative, moyens de développement des chromatogrammes). Les principes généraux des divers types de chromatographie (adsorption, partage direct, partage inversé, relargage, exclusion-diffusion, échange d'ions, spécifique) qui peuvent être mis en oeuvre sont rappelés et des exemples concrets sont donnés dans chaque cas. Les problèmes de l'électrophorése et de la chromato-electrophorése sur couche mince sont également évoqués. Bien que cette méthode analytique soit le plus simple et le puissant moyen de séparation des corps en mélange, il est nécessaire de rappeler que bien des erreurs de conception sont à la base de son manque de développement dans certains domaines (chromatographie des substances hydrosolubles par exemple). A titre d'exemple on rappelle que la première chromatographie de relargage et la première chromato-electrophorése basse tension n'ont été réalisées que tout récemment.

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Summary After a brief summary of the advantages of surface chromatography compared to column chromatography, the respective advantages and inconveniences of thin-layer and paper chromatography are compared. The general methods of thin-layer chromatography are presented (preparation of the layers, types of layers, deposition of the samples to be analyzed, analytical TLC, preparative TLC, methods for developing the chromatograms). The general principles of the various types of chromatography (adsorption, direct partition, reverse partition, salting out, exclusion-diffusion, ion-exchange, specific chromatography) that may be used are reminded of, and concrete examples given in each case. The problems of electrophoresis and thin layer chromato-electrophoresis are also treated. Though this analytical method be the simplest and most powerful means for the isolation of components in mixtures, it is necessary to remind that quite a number of errors in the conception are responsible for its lack of development in certain fields (water soluble substances chromatography, for instance). As an example one might remember that the first salting-out chromatography, and the first low-tension chromatoelectrophoresis have been developed only very recently.

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Zusammenfassung Die Vorzüge der Oberflächen-Chromatographie gegenüber der Säulen-Chromatographie wurden kurz aufgezählt und die Besonderheiten der Dünnschicht- und der Papier-Chromatographie verglichen. Die Grundlagen der DC werden erklärt (Vorbereitung der Schicht, Arten der Schicht, Auftragen der Proben, analytische DC, vorbereitende DC, Entwicklungsverfahren der Chromatogramme). Anschließend werden die grundsätzlichen Verfahren der verschiedenen Typen von Chromatographie erörtert (Adsorption, direkte Trennung, inversierte Trennung, Aussalzen, Diffusion, Ionenaustausch, spezifische Verfahren), die praktisch infrage kommen. Für jeden Typ werden Beispiele gegeben. Die Probleme der Elektrophorese und der Elektrophorese-DC werden ebenfalls behandelt. Obwohl diese Analysenmethode das einfachste und stärkste Trennverfahren für Stoffgemische ist, so muß daran erinnert werden, daß ihre Entwicklung in gewissen Gebieten durch Fehler in der Auffassung gehemmt worden ist, zum Beispiel ihre Anwendung für wasserlösliche Stoffe. Verfasser weist darauf hin, daß die erste Aussalz-DC und die erste Niederspannungs-Chromato-Elektrophorese erst ganz kürzlich realisiert wurden.

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Avec la collaboration technique de MademoiselleCh. Thommegay et de MademoiselleA.-M. Drapier.     

Response of Bluebunch Wheatgrass and Medusahead to Defoliation

Our objective was to determine the short-term response of bluebunch wheatgrass and medusahead to defoliation of wheatgrass designed to stimulate regrowth through tillering. We hypothesized that defoliating bluebunch wheatgrass by 20% at the 3 to 3.5 leaf stage followed by a 50% defoliation at peak standing crop would increase its tillering and biomass production. Consequently, we expected a reduction of the density and biomass of medusahead over that of bluebunch wheatgrass defoliated 50% at peak standing crop. Treatments included four initial medusahead densities (200, 333, 444, 600 plants · m^-2) created by hand-pulling and three defoliation regimes factorially arranged (12 treatment combinations) in a randomized complete-block design and replicated four times at two sites. In 2006 and 2007, defoliation was accomplished by hand-clipping bluebunch wheatgrass 1) by 50% once at peak standing crop (late June); 2) by 20% at the 3 to 3.5 leaf stage, then again to 50% at peak standing crop (mid May, late June); or 3) plants were not clipped. Density was sampled in 2006 and 2007, and biomass was harvested only at Star Mountain (near Riverside, Oregon) in 2007 because Warm Springs (near Drewsey, Oregon) was burned by a wildfire before final 2007 data could be collected. In 2006, no treatments applied at either site detectably altered the number of tillers produced by bluebunch wheatgrass nor did they affect bluebunch wheatgrass density or biomass in 2007 at Star Mountain. Changes in medusahead density were not detected in 2006, but this annual invasive grass increased in density and biomass in 2007 at Star Mountain in plots receiving two defoliations. The relatively short growing period caused by summer drought and the relative intolerance of bluebunch wheatgrass to grazing make the twice-over grazing an unlikely practice for arid rangelands in the western United States. In fact, it could possibly increase the risk of annual grass invasion.     

Kinetics of substrate oxidation and hydrogen peroxide production by Mycoplasma mycoides subsp. mycoides Large Colony (LC) type and Mycoplasma mycoides subsp. capri

Mycoplasma mycoides subsp. mycoides Large Colony (LC) type is a pathogen of goats causing contagious agalactia and respiratory disease, found on all continents where small ruminants are kept. It shares close genetic characteristics with M. mycoides subsp. capri. Substrate oxidation by 22 strains of M. mycoides subsp. mycoides LC from nine countries was compared with that of eight strains of M. mycoides subsp. capri from five countries. There was considerable similarity in the substrates used, but substrate saturation coefficients (K_s) varied for different substrates. Substrate utilization patterns and K_s values did not (1) significantly differentiate the LC strains from each other, (2) show any correlation with geographical origin, or (3) distinguish the LC strains from the capri strains. These results support previous studies justifying the reclassification of these subspecies as a single species.     

Evaluation of trypanosomiasis control alternatives using an epidemiologic simulation model

The effect of various disease-vector control alternatives on the prevalence of trypanosomiasis in southwest Ethiopia was examined with the aid of an epidemiologic model. The alternatives considered included vegetation clearing by manual labor, game elimination, insecticidal spraying from airplanes coupled with settlement, use of sterile male Glossina, avoidance of tsetse infested areas, increasing the resistance of the cattle population, therapy, and combinations of the above methods. First, the currently estimated endemic prevalence of trypanosomiasis (27.3%) in the simulated area of Ethiopia was established and maintained over a 10 year period. Then, various control alternatives were introduced and the simulation was run for an additional 10 years to observe the effect of these control alternatives on the prevalence. The combined use of vegetation clearing, insecticides, therapy, and settlement (or resettlement) was the most effective and feasible method of trypanosomiasis control for the simulated situation.     

Papillomavirus-associated basosquamous carcinoma in an Egyptian fruit bat (Rousettus aegyptiacus).

A 5-yr-old female Egyptian fruit bat (Rousettus aegyptiacus) had a small raised pigmented mass removed from the lateral canthus of the left eye. Six additional variably sized, raised, smooth to cauliflower-like skin masses were observed randomly distributed throughout the left wing membranes. Four masses were removed and diagnosed microscopically as basosquamous carcinomas and papillomas. Additional masses, removed 6 mo and 1 yr later, showed bony invasion and squamous differentiation. Immunohistochemistry detected positive intranuclear staining for bovine papillomavirus antibody in all samples. Polymerase chain reaction done on DNA extracts from formalin-fixed, paraffin-embedded tumor tissue amplified a 450 base-pair segment analogous to the L1 region of human papillomavirus types 96 and 5. Basic Local Alignment Search Tool analysis of sequenced amplicons suggests a novel chiropteran papillomavirus. To our knowledge, this is the first report of papillomavirus-associated carcinoma in a chiropteran species.     

Correlation between loss of efficacy of macrocyclic lactone heartworm anthelmintics and P-glycoprotein genotype

Macrocyclic lactone (ML) molecules have been used for heartworm control for more than 25 years. However, in recent years, there have been reports of loss of efficacy of ML heartworm preventatives against Dirofilaria immitis in some locations in the United States. Macrocyclic lactone resistance is a common problem in nematode parasites of livestock, and more recently, evidence of ivermectin resistance has been reported in the human filarial nematode Onchocerca volvulus. In this study, four D. immitis sample groups from the United States with different treatment histories were investigated for evidence of ML-driven genetic selection. DNA from individual adult worms and microfilariae was amplified by polymerase chain reaction to investigate a gene encoding a P-glycoprotein, a protein class known to be involved in ML pharmacology. A significant correlation of a GG-GG genotype with ivermectin response phenotype was found. Moreover, a significant loss of heterozygosity was found in a low responder group; loss of heterozygosity is commonly seen in loci when a population has been under selection. Further studies are required to confirm ML resistance in heartworm populations. However, the genetic changes observed in this study may be useful as a marker to monitor for ML resistance in D. immitis.