„Der epidurale Blood Patch: ein Auftrag für den versierten Anästhesisten?“

Die Anaesthesiologie -     

Fibroblast growth factor-2 but not Mel-CAM and/or beta3 integrin promotes progression of melanocytes to melanoma

A variety of melanoma-associated antigens have been identified that mediate adhesion, growth, proteolysis, and modulation of immune response. However, the mechanisms by which human normal melanocytes become malignant are not clearly understood. Among the most consistent observations is the up-regulation of fibroblast growth factor-2 (FGF-2) and of the adhesion molecules beta3 integrin and Mel-CAM during melanoma progression. To evaluate the potential role of FGF-2, beta3 integrin and Mel-CAM in melanoma development we overexpressed FGF-2, beta3 integrin and Mel-CAM in normal human melanocytes using replication-deficient adenoviruses as a gene delivery vehicle. Fibroblast growth factor-2 overexpressing melanocytes in monolayer culture displayed cytological atypia. Furthermore, in human skin reconstructs where the physiological milieu is recreated in vitro, FGF-2-overexpressing melanocytes exhibited marked proliferation, upwards migration, cluster formation and type IV collagen expression within the epidermal compartment, simulating early radial growth phase melanoma. In contrast, overexpression of beta3 integrin and/or Mel-CAM in melanocytes did not affect their biological behaviour in human skin reconstructs. The described results of the current and previous studies emphasise the key role of FGF-2 in melanoma development and progression, underscoring the promise of FGF-2 as a target for therapy.     

The effect of staphylococcal alpha toxin on the superprecipitation and Ca2+-uptake of isolated arterial smooth muscle preparations

The effects of Staphylococcal alpha toxin from culture filtrates of Staphylococcus aureus, strain Wood 46, on superprecipitation of actomyosin and Ca²⁺-uptake of microsomes and fragments of plasma membrane from bovine carotid arteries were followed. It was found that alpha toxin inhibits the active Ca²⁺-uptake by smooth muscle microsomes and fragments of plasma membrane and liberates bound calcium. The toxin causes in the absence of free Ca²⁺ superprecipitation of actomyosin and activation of ATP-ase. The liberation of calcium from cellular structures and the inhibition of Ca²⁺ transport into these structures by alpha toxin could be responsible for its spasmogenic action.     

Comportement des acides aminés plasmatiques libres chez des chiens normaux et des chiens pancréatectomisés après administration intraveineuse rapide de l-leucine

Resume L'injection intraveineuse rapide de l-leucine entraîne chez les chiens normaux une hypoglycémie, une chute du taux plasmatique du potassium et du phosphore inorganique, une hyperaminoacidémie initiale fort transitorie, et une l'hypoaminoacidémie survenante après. Chez les chiens traités au préalable par chlorpropamide l'hypoglycémie, l'hypokaliémie et l'hypoaminoacidémie entraînées par l'injection de leucine sont plus marquées - à différent degré; par contre la fugace hyperaminoacidémie et l'hypophosphatémie sont du même rang que dans le groupe précédent. Chez les chiens pancréatectomisés, l'injection rapide de leucine provoque un abaissement minime de la glycémie et n'influence pas le taux plasmatique du potassium; chez ces animaux la chute du taux des acides aminés dans le plasma est inférieure par rapport aux animaux sains, tandis que l'hyperaminoacidémie initiale et l'hypophoshatémie sont les mêmes.L'injection intraveineuse rapide de la l-valine et de l-alanine aux chiens normaux entraîne un abaissement de la glycémie et de la kaliémie inférieur en comparaison avec les mêmes effects de la leucine, lors que l'hypophosphatémie est du même rang. Ces acidesaminés provoquent également un accorissement initial de la concentration d'autres acidesaminés dans le plasma, montrant des différences en comparaison avec les modifications faisantes suite à l'injection de la leucine. La l-valine produit également un abaissement secondaire du taux de certains acides-aminés, tandis que l'on n'observe pas ce phénomène après l'injection de l-alanine. Dans la discussion on a évoqué la possibilité d'une influence de la leucine alimentaire sur l'utilisation des hydrates de carbone et des protéines digérés.

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Summary The quick intravenous administration of l-leucine produces in normal dogs a hypoglycemia, a decrease of potassium and of inorganic phosphorus, an initial and very transient increase of blood aminoacid plasma values and afterwards a decrease of blood aminoacids. In dogs previously treated with chlorpropamide, the decrease of glycemia, kaliemia and aminoacidemia are more evident, and in different degrees: on the contrary the transient decrease of blood aminoacids and the decrease of blood phosphorus are of he same degree as the other group's. In pancreatectomized dogs the quick administration of leucine produces a very small decrease of glycemia and it does not affect the potassium plasma values: in these animals the fall of the aminoacids in plasma is inferior to the one that happens in healthy animals, while the initial hypoaminoacidemia and hypophosphatemia are of the same degree.The quick intravenous administration of l-valine and l-alanine in normal dogs produces a decrease of glycemia and kaliemia inferior compared with the same effects of leucine, while the hypophosphatemia is of the same degree. Valine and alanine produce an initial increase of the aminoacid blood levels too, showing afterwards some differences from the changes that take place after the leucine administration. L-valine produces a secondary decrease of some aminoacid levels too, while such phenomenon cannot be observed after l-alanine injection. During the discussion, the possibility that food leucine may affect the utilization of carbohydrates and digested proteins is taken into consideration.

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Zusammenfassung Die rasche i.v. Injektion von l-Leuzin verursacht bei normalen Hunden eine Hypoglykaemie, Abfall des Plasmaniveaus von K und inorganischem P, anfaengliche, rasch voruebergehende Hyperaminoazidaemie und darauffolgende Hypoaminoazidaemie. Bei den zuvor mit Chlorpropamid behandelten Hunden sind die Hypoglykaemie, Hypokaliaemie und die Hypoaminoazidaemie — mit unterschiedlichem Grad — ausgepraegter; hingegen sind die voruebergehende Hyperaminoazidaemie sowie die Hypophosphataemie gleichen Grades, wie bei der vorhergehenden Gruppe. Bei den pankreatektomierten Hunden bewirkt die rasche Leuzin-Injektion einen minimalen Abfall des Blutzuckers und beeinflusst nicht das Plasmaniveau von K. Bei diesen Tieren ist der Abfall des Plasmaniveaus der Aminosaeuren geringer als bei gesunden Tieren, waehrend die anfaegliche Hyperaminoazidaemie und Hypophosphataemie die gleichen sind.Die rasche i.v. Injektion von l-Valin und l-Alanin bewirkt bei den normalen Hunden einen Abfall des Blutzuckers und der Kaliaemie, der geringer ist als der von Leuzin hervorgerufene, waehrend die Hypophosphataemie gleichen Grades ist. Diese Aminosaeuren rufen auch eine anfaengliche Erhoehung der Konzentration der anderen Aminosaeuren im Plasma hervor und offenbaren somit Unterschiede in Bezug auf die nach Leuzin-Injektion auftretenden Veraenderungen. Das l-Valin produziert ferner einen sekundaeren Abfall des Niveaus einiger anderer Aminosaeuren, waehrend man dieses Phaenomen nach l-Alanin-Injektion nicht beobachtet. In der Diskussion wird auf die Moeglichkeit eines Einflusses des alimentaeren Leuzins auf die Verwertung der verdauten Kohlehydrate und der Proteine hingewiesen.

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Resumen La inyección endovenosa rápida de l-leucina en perros normales determina una hipoglicemia, una caída del nivel plasmático del potasio y del fósforo inorgánico, una hiperaminoacidemia inicial my transitoria y una sucesiva hipoaminoacidemia. En los perros previamente tratados con clorpropamida, la hipoglicemia, la hipocaliemia y la hipoaminoacidemia son más marcadas, en grados diversos; en cambio, la fugaz hiperaminoacidemia y la hipofosfatemia son del mismo grado que en el grupo precedente. En los perros pancreatectomizados, la inyección rápida de leucina provoca una mínima disminución de la glicemia y no influye sobre el nivel plasmático del potasio; en estos animales, la caída del nivel plasmático de los aminoácidos es inferior a la de los animales sanos, mientras la hiperaminoacidemia inicial y la hipofosfatemia son las mismas.La inyección endovenosa rápida de l-valina y de l-alanina en perros normales determina una menor disminución de la glicemia y de la caliemia en comparación con los mismos efectos de la leucina, mientras la hipofosfatemia es del mismo grado. Estos aminoácidos provocan igualmente en el plasma un aumento inicial en la concentración de los otros aminoácidos, mostrando algunas diferencias con respecto a las modificaciones que se verifican después de la inyección de leucina. La l-valina también produce una reducción secundaria del nivel de algunos aminoácidos, mientras dicho fenómeno no se observa después de inyectar la l-alanina. En la discusión se illustra la posibilidad de influencia de la leucina alimenticia en la utilización de los hidratos de carbono y de las proteínas digeridas.

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Riassunto L'iniezione endovenosa rapida di l-leucina determina in cani normali una ipoglicemia, una caduta del livello plasmatico del potassio e del fosforo inorganico, una iperaminoacidemia iniziale, molto transitoria, ed una successiva ipoaminoacidemia. Nei cani trattati preventivamente con cloropropamide, l'ipoglicemia, l'ipokaliemia e l'ipoaminoacidemia sono più marcate, in gradi diversi; in cambio la fugace iperaminoacidemia e l'ipofosfatemia sono dello stesso grado del gruppo precedente. Nei cani pancreatectomizzati, l'iniezione rapida della leucina provoca un abbassamento minimo della glicemia e non influenza il livello plasmatico del potassio; in questi animali la caduta del livello plasmatico degli aminoacidi è inferiore a quanto avviene negli animali sani, mentre la iperaminoacidemia iniziale e l'ipofosfatemia sono le stesse.L'iniezione endovenosa rapida della l-valina e della l-alanina nei cani normali determina un abbassamento della glicemia e della kaliemia inferiore in confronto agli stessi effetti della leucina, mentre l'ipofosfatemia è dello stesso grado. Questi aminoacidi provocano ugualmente un aumento iniziale nella concentrazione degli altri aminoacidi nel plasma, mostrando quindi delle differenze rispetto alle modifiche che si verificano dopo l'iniezione di leucina. La l-valina produce ugualmente un abbassamento secondario del livello di alcuni aminoacidi, mentre tale fenomeno non si osserva dopo l'iniezione della l-alanina. Nella discussione viene ricordata la possibilità d'influenza della leucina alimentare sull'utilizzazione degli idrati di carbonio e delle proteine digerite.

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Rapport présenté pendant le 1° Congresso Nazionale della Società Italiana di Diabetologia, le 16 févriér 1966 à Catania.     

Compound 48/80 and the healing of wounds in rats. The effect of timing of drug injections

Wound breaking strength was measured in Long-Evans rats in two settings: in one experiment animals received Compound 48/80 i.p. for 8 consecutive days, starting 1 day before infliction of wound and in the second experiment 48/80 was given for 6 days before wounding. Wound breaking strength was always measured on the seventh day after wounding. There was a significant increase in wound breaking strength in animals treated before incision was made, as compared to controls, saline-treated animals (p less than 0.01). When 48/80 was given one day before incision was made, the breaking strength was significantly decreased (p less than 0.0002). Possible explanation for these opposite results is discussed.     

The adhesion molecule L1 (CD171) promotes melanoma progression

The adhesion molecule L1 is expressed in primary melanomas and cutaneous metastases in contrast to melanocytic nevi and melanocytes, and is significantly associated with metastatic spread. Recent studies have demonstrated that in carcinomas L1 expression is associated with sustained activation of the extracellular signal-regulated kinase (ERK) pathway and upregulation of ERK-dependent, motility- and invasion-associated gene products including alphavbeta3 integrin. The objective of this study was to further investigate the role of the adhesion molecule L1 in melanoma progression, and to evaluate whether targeting the L1 adhesion molecule would have therapeutic effects against invasive melanoma growth. Using human melanoma cells from different stages of progression in monolayer and organotypic human skin culture mimicking the pathophysiological environment of cutaneous melanoma, we found that (1) L1 expression mostly correlates with melanoma progression and alphavbeta3 integrin expression, (2) overexpression of L1 in early radial growth phase melanoma cells promotes conversion from radial to vertical growth phase melanoma without upregulation of alphavbeta3 integrin expression, and (3) suppression of L1 function significantly reduces migration and invasion of melanoma cells, but does not completely block invasive melanoma growth. Altogether, L1 plays a critical role in melanoma invasion and progression and offers therapeutic potential in combination with conventional anticancer agents.