Rat lens gamma-crystallins. Characterization of the six gene products and their spatial and temporal distribution resulting from differential synthesis

We have isolated, purified and characterized six individual gamma-crystallin polypeptides present in the rat lens. Comparison of their amino acid compositions with the known structure of the six gamma-crystallin genes permits a one-to-one correspondence to be made between each protein synthesized and the encoding gene. This demonstrates that each of the six genes is actually expressed in vivo. Two classes of three gamma-crystallins each, which we have designated classes gamma ABC and gamma DEF, are known to exist, on the basis of internal sequence homology. We have measured the temperature-dependent phase-separation characteristics of solutions of the six purified gamma-crystallins, and find that the three members of the gamma DEF class (gamma 2-2, gamma 3-1 and gamma 4-1) are all cryo-proteins with relatively high phase-separation temperatures, whereas the three gamma ABC crystallins (gamma 1-1, gamma 1-2 and gamma 2-1) do not show phase separation above -7 degrees C. We have measured the spatial distribution in rat lens of each of the alpha-, beta- and gamma-crystallins as a function of age from 1 to 420 days, using size-exclusion and ion-exchange high-pressure liquid chromatography (HPLC). Our findings in the cortical layer permit us to establish the differential synthesis of each of the crystallins during lens development. Particular attention has been devoted to the spatial and temporal distribution of the six individual gamma-crystallins. Up to birth, synthesis of the three components of the gamma DEF class predominates, and in particular that of gamma 2-2. In subsequent development the three components of the gamma ABC class assume a greater proportion of monomeric crystallins synthesized, while beta s-crystallin synthesis predominates in late development. Our analysis of different layers within single lenses provides novel information on spatial gradients of the water-soluble and water-insoluble protein fractions as a function of age. We consider the consequences of these findings for lens transparency and opacity in both rat and mouse lens. We show that the high concentrations of gamma DEF-crystallins appear to be responsible for the opacity known to occur in young rat lenses. We conclude from these observations that close control of the differential synthesis of gamma-crystallins plays an important role in maintaining lens transparency during development.     

Permanent suppression of phase separation cataract in calf lens using amine modification agents

Low temperature induced opacification (cold cataract) of the nucleus of young mammalian lenses is associated with a phase separation of proteins in the lens cell cytoplasm. Calf lenses were treated with a variety of imido-esters and N-hydroxysuccinimide-esters, which react specifically with amino groups. Many potent inhibitors of phase separation cataract were identified which lower the opacification temperature by 6 degrees C or more. Lenses generally remain clear, colorless and soft. Furthermore, suppression of the cold cataract temperature is permanent upon removal of excess reagent.     

On the ancestral form of true organ of fructification in the saprophytic stage of the dermatophytes of the faviform group: Favomicrosporon pinettii,N.SP. and its perfect form: Anixiopsis stercoraria (hansen) hansen, 1897

Summary The discovery of the hidden, built-in macroconidia in the four members of the Faviform Group of the dermatophytes, i.e.,Achorion schoenleinii, Trichophyton violaceum, Trichophyton verrucosum andMicrosporon ferrugineum, is described.To bring the hidden, built-in macroconidia to full fructification, i.e., to force the production of imperfect and perfect organs of fructification (macroconidia, cleistothecia), two entirely different techniques have been used: 1) the hair-soil method, 2) the yeast extract method.The two techniques, entirely independent from each other, yielded the same result: the ancestral form of the four members of the Faviform Group of dermatophytes. The imperfect form is described asFavomicrosporon pinettii,Benedek, 1965, sp. nov. The perfect form isAnixiopsis stercoraria (Hansen)Hansen, 1897.The ancestral form was found not only in and cultured from the strains of those dermatophytes derived from pathological material, but it was also recovered from its saprophytic habitat, from the soil (potting soil).

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Zusammenfassung Die Entdeckung der verborgenen, eingebauten Makrokonidien in den vier Representanten der Faviformen Gruppe der Dermatophyten, i.e.Achorion schoenleinii, T. violaceum, T. verrucosum, Microsporon ferrugineum, wird beschrieben.Um die verborgenen, eingebauten Makrokonidien zur vollen Fruchtbildung zu bringen, i.e. um die Produktion der imperfekten und perfekten Organe der Fruktifikation (Makrokonidien, Kleistothecien) zu erzwingen, sind zwei völlig verschiedene Methoden benutzt worden: 1) die Haar-Erde-Methode, und 2) die Hefeextrakt-Methode.Beide Methoden, völlig unabhängig von einander, haben zu demselben Ergebnis geführt, i.e. zur Entdeckung der Urform von den vier Representanten der Faviformen Gruppe der Dermatophyten. Die imperfekte Form wird alsFavomicrosporon pinettii,Benedek, 1965, sp. nov. beschrieben. Die perfekte Form istAnixiopsis stercoraria (Hansen)Hansen, 1897.Die ancestrale Form wurde nicht nur aus den Stämmen jener Dermatophyten gezüchtet, die aus pathologischen Produkten gewonnen worden sind, sondern auch aus dem natürlichen Habitat: von der Erde (potting soil).

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Résumé La découverte des macroconidies occultes et encastrées dans les quatre membres du Groupe Faviforme des dermatophytes:Achorion schoenleinii, Trichophyton violaceum, Trichophyton verrucosum etMicrosporon ferrugineum, est décrite.Pour forcer les macroconidies occultes et encastrées à la fructification complète, i.e. de produire des organs de reproduction imparfaits et parfaits, macroconidies et cleistothecia, on a fait l'usage de deux techniques complètement différentes: 1) des cheveux sur sol et 2) de l'extraction de levure.Toutes les deux méthodes, complètement indépendantes l'une de l'autre, ont produit le même résultat: la forme ancestrale des quatre membres de la Groupe Faviforme des dermatophytes. La forme imparfaite est décrite commeFavomicrosporon pinettii,Benedek, 1965, sp. nov. et la forme parfaite commeAnixiopsis stercoraria (Hansen)Hansen, 1897.La forme ancestrale a été trouvée non seulement dans les souches des dermatophytes indiquées et en cultivées, provenantes des produits pathologiques, mais aussi du sol, du terrain jardinier.

     

Application of Fourier transform infrared spectroscopy to studies of aqueous protein solutions.

Modern protein Fourier transform infrared (FT-IR) spectroscopy has proven to be a versatile and sensitive technique, applicable to many aspects of protein characterization. The major practical drawback for the FT-IR spectroscopy of proteins is the large absorbance band of water, which overlaps the amide I resonances. D2O is often substituted for H2O in infrared experiments. Removal of water from protein samples can be complicated and tedious and potentially lead to denaturation, aggregation, or sample loss. Solvent removal by dialysis is difficult for suspensions and sols. A new method called the D2O dilution technique (Ddt) is described which simplifies the sample preparation step and improves the solvent subtraction. The effect of the D2O concentration on the IR spectrum of aqueous solutions of several model proteins was studied. Dilution of aqueous samples with D2O yields good quality spectra. The Ddt has been evaluated for quantitative analysis using standard proteins and its applicability to solutions and suspensions of a genetically engineered malaria antigen is demonstrated. Use of resolution-enhancement techniques with spectra in mixed solvents has also been investigated.     

Fragmenta mycologica

Summary The historical developmental stages of the hairbaiting technique are traced through the past decades.Since three investigators have substantially contributed to the development of this technique, it is proposed that this technique should in the future be known as theToma-Karling-Vanbreuseghem hairbaiting method or abbreviated as in the anagram: ToKaVa-hairbaiting method.

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Zusammenfassung Die historischen Entwicklungsstufen der Haarköder-Methode werden durch die Jahrzehnte verfolgt.Da drei Forscher zum Aufbau dieser Methode am wesentlichsten beigetragen haben, wird es vorgeschlagen, dass diese Methode in Zukunft als dieToma-Karling-Vanbreuseghem Haarköder-Methode benannt werde oder kurz, mit einem Anagramm, als die ToKaVa-Haarködermethode.

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Résumé L'auteur discute les étapes historiques du développement de la méthode en usant la kératinophilie des dermatophytes pour les cultiver.Puisqu'il y a trois investigateurs qui ont contribué, par excellence, au développement de cette méthode, on propose que cette téchnique soit dénommée au future comme d'appât de cheveux deToma-Karling-Vanbreuseghem ou, bref, la méthode d'appât de cheveux de ToKaVa.

     

A comparative dynamic study of the effectiveness of gastric cytoprotection by vitamin A, De-Nol, sucralfate and ulcer healing by pirenzepine in patients with chronic gastric ulcer (a multiclinical and randomized study)

The gastric cytoprotective effects of vitamin A, De-Nol and sucralfate were compared with the effectiveness of pirenzepine in healing ulcer in patients with chronic gastric ulcer. A total of 100 patients was randomized into different groups: the patients were treated with antacids, vitamin A (3 X 50.000 IU), De-Nol liquid (4 X 5 ml), sucralfate (4 X 1 g) or pirenzepine (3 X 50 mg). The treatment was continued for 4 weeks. At the beginning, 2 and 4 weeks after starting treatment the patients were subjected to endoscopy and the size of the ulcer was measured planimetrically. The ulcer-healing effect of De-Nol liquid was significantly better than that of the antacids (p less than 0.01). Ulcer size was reduced significantly in all groups (p less than 0.01), however, at the end of the study the gastric ulcers were smallest in the De-Nol treated group (p less than 0.001). The dynamics of ulcer healing in the second week was most favourable in the patients receiving vitamin A (p less than 0.01). The present data point to the cytoprotective effects of De-Nol liquid, vitamin A and sucralfate and to their ability of healing chronic gastric ulcers.     

A new dermatophyte,Thallomicrosporon kuehnii Benedek, 1963

Summary A new species of a dermatophyte,Thallomicrosporon kuehnii Benedek, 1963, is described. In contradistinction to all other known dermatophytes, it reproduces exclusively by thallospores: 1) large, fusiform, macroconidia-like structures, 2) arthrospores and, 3) chlamydospores. Microconidia are present, rodlike or pyriform. The elicitation of a perfect form of fructification in hair-on-soil cultures was unsuccessful.

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Zusammenfassung Eine neue Art der Dermatophyten,Thallomicrosporon kuehnii Benedek, 1963, wird beschrieben. Sie unterscheidet sich von allen anderen Hauptilzen durch den Umstand, dass sie sich ausschliesslich durch Thallosporen reproduziert: 1) Formen, ähnlich fusiformen Makrokonidien; 2) Arthrosporen und 3) Chlamydosporen. Mikrokonidien werden gebildet. Das Hervorbringen der perfekten Phase der Fruktifikation in Kulturen von Haar an Humus ist nicht gelungen.

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Résumé Une espèce nouvelle des dermatophytes,Thallomicrosporon kuehnii Benedek, 1963, est présentée. Elle est distincte de tous les autres dermatophytes par le fait de sa reproduction exclusive par des thallospores: 1) des structures imitant des macroconidies fusiformes; 2) des arthrospores et 3) des chlamydospores. Des microconidies sont présentes. L'élicitation de la phase perfecte de fructification en culture des poils en sol n'a pas réussi.