Ultrastruktur der Thyreozyten der Ratte im alimentären Tocopherol- und Ubichinonmangel und nach Substitution

Zusammenfassung Die Follikelepithelzellen der Rattenschilddrüse zeigen drei Funktionsphasen, die normalerweise nicht nebeneinander auftreten: 1. Hormonsynthese und Sekretion, 2. relative Inaktivität und 3. Kolloidresorption. Während die Resorption von Thyreotropin induziert wird, scheint die Proteinsynthese vom hypophysären Stimulus unabhängig zu sein. Die Ultrastrukturveränderungen, die an einigen Zellorganellen (Mitochondrien, rauhes endoplasmatisches Retikulum) der Rattenschilddrüse im Tocopherol-Chinon (T-Q)-Mangel zu beobachten sind, stören offenbar den Ablauf ihrer Funktionsphasen. Der alimentäre T-Q-Mangel beeinflußt die Schilddrüse möglicherweise auf zwei verschiedenen Wegen: 1. in Abhängigkeit von der Antioxidanseigenschaft der Tocopherole über den peripheren Stoffwechsel der Schilddrüsenhormone; 2. über eine Beteiligung der Metaboliten mit Chinonstruktur am mitochondrialen Enzymsystem. — Die im T-Q-Mangelzustand auftretenden strukturellen Veränderungen der Thyreozyten können durch Substitution der Mangeldiät mit d,1--Tocopherol oder -Tocopherolchinon wieder normalisiert werden.

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Ultrastructure of the rat thyroid epithelial cells during alimentary tocopherol and ubiquinone deficiencies and after substitution

Summary Normal thyroid follicular epithelial cells in the rat show three functional phases: (1) synthesis of protein and secretion into the follicular lumen; (2) relative inactivity; (3) resorption of the colloid. The resorption of the colloid is stimulated by the thyrotropic hormone, whereas colloid protein synthesis appears to be independent of pituitary stimulus. Ultrastructural alterations, present in some cellular components during alimentary tocopherolubiquinone deficiency, reflect disturbances in the normal functional phases. The deficiency possibly influences the function of the thyroid gland in two ways: in affecting the metabolism of thyroid hormones which seems to depend on the antioxidant properties of tocopherols, and/or, in affecting the participation of the quinoid forms in the mitochondrial enzyme system. Substitution with either d,1--tocopherol or -tocopherolquinone reverses the alterations and disturbances produced by the dietary deficiency.

Herrn Prof. Dr. Dr. h.c. W. Bargmann, Kiel, gewidmet.     

Untersuchungen zur Bildung der Vomerspange beiSalamandra salamandra (L.)

Zusammenfassung BeiSalamandra salamandra wurde die Entwicklung der Vomerspangen anhand von Exstirpations- und Transplantationsversuchen vor und während der Metamorphose untersucht. Die Vomerspange bildet sich aus Bindegewebe, das am Caudalrand des Larvenvomer liegt. Exstirpierte Vomerteile werden nicht regeneriert. Ein kurzer caudaler Abschnitt der larvalen Vomerzahnleiste genügt für die vollständige Ausbildung der Zahnleiste der Vomerspange. Wird die Spitze der auswachsenden Vomerspange reseziert, bleibt die Spange unvollständig. Die Bildung des Definitivvomer bleibt davon unberührt. In die Subcutis der Rumpfhaut transplantiertes Gewebe der sich bildenden Vomerspange bleibt zwar erhalten, wächst aber nicht weiter. In den Spangenbildungsbereich eingefügte Rumpfhaut verhindert das Auswachsen der Vomerspange.

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Investigations on the development of the vomerine bar inSalamandra salamandra (L.)

Summary The development of the vomerine bar inSalamandra salamandra was studied by amputation and transplantation procedures before and during metamorphosis. The bar of the vomer arises from the connective tissue near the caudal margin of the larval vomer. The larval vomer does not regenerate after amputation. A short caudal part of its dental lamina is sufficient for the outgrowing of the complete dental lamina of the vomerine bar. If the tip of the outgrowing vomerine bar is removed, the bar will remain incomplete, which procedure has no influence on the development of the definitive vomer. The material of the developing vomerine bar transplanted to the subepithelial connective tissue of the skin remains intact, but does not continue to grow. Fragments of the skin transplanted to the region from which the bar of the vomer arises prevent the development of the bar.

     

Ultrastruktur des Neurohaemalorgans im Nervus corporis allati II vonAcheta domesticus

Summary InAcheta domesticus the proximal part of the nervus corporis allati II (Nca II) is differentiated as a neurohemal organ und consists of several hundred fiber profiles. The neurosecretory region is confined to the peripheral layer and contains axons with different vesicular inclusions. The liberation of neurohormones is accomplished by exocytosis and the formation of synaptoids. Structures resembling synaptic ribbons were observed in contact with axons, glial profiles and interstitial stroma. The central area of the nerve contains only non-neurosecretory axons of various sizes. Connection with the subesophageal ganglion is attained by only 9 large axons of the central region.Zusammenfassung BeiAcheta domesticus ist der proximale Abschnitt des Nervus corporis allati II (Nca II) als Neurohaemalorgan mit mehreren hundert Faserprofilen ausgebildet. Die neurosekretorische Region ist auf die Peripherie des Nerven begrenzt und enthält Fasern mit unterschiedlichen vesikulären Einschlüssen. Die Freisetzung der Neurohormone erfolgt exocytotisch und durch Bildung von Synaptoiden. Es werden stiftchenförmige synapsenähnliche Kontaktstellen mit Axonen, Gliaprofilen und dem interzellulären Stroma beschrieben. Der zentrale Teil des Nerven führt nicht-neurosekretorische Axone unterschiedlichen Durchmessers, von denen lediglich 9 große Fasern die Verbindung mit dem Unterschlundganglion herstellen.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Über die Ultrastruktur des „Nervus opticus“ und des Ganglion opticum I von Daphnia pulex

Zusammenfassung Die Opticusfasern (Neuriten der Rezeptorzellen) und das Ganglion opticum I von Daphnia pulex wurden elektronenmikroskopisch untersucht. Die 8 Neuriten jeweils eines Ommatidiums werden in einem Bündel zusammengefaßt, in dem die Neuriten nur unvollständig von einem Gliafortsatz umhüllt sind, das Bündel jedoch vollständig von einer Basalmembran bedeckt ist. Die Neuriten weisen quervernetzte Mikrotubuli auf. In der Peripherie des Ganglion opticum liegen große und kleine unipolare Nervenzellen, deren Fortsätze ins zentrale Neuropil ziehen, wo sie u.a. synaptische Kontakte mit den Neuriten bilden. Es werden 3 Formen von Synapsen beschrieben: 1. eine bisher nicht beschriebene Synapsenform, 2. Synapsen von gewöhnlichem Typ und 3. Ribbon Synapsen. Die peripher gelegenen Gliazellen umhüllen die Nervenzellen und senden lamelläre Fortsätze in das Neuropil, wo sie sich den benachbarten Zellelementen derart anpassen, daß der extrazelluläre Raum zu einem System von Interzellularfugen eingeengt wird. Außer den beschriebenen Zellformen kommen weniger häufig neurosekretorische Nervenzellen vor, deren Fortsätze an der Ganglionoberfläche nur von Basalmembranen bedeckt sind. Ferner sind selten multipolare Ganglienzellen zu finden.

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On the ultrastructure of the optic nerve and the ganglion opticum I of Daphnia pulex

Summary Optical fibres (neurites of receptor cells) and ganglion opticum I of Daphnia pulex were studied electron microscopically. The 8 neurites of each ommatidium are bundled by a complete wrapping of basement membrane, while each neurite is incompletely enveloped by a glial process. The neurites contain transversally interconnected microtubules. Processes of large and small unipolar nerve cells situated at the periphery of the ganglion reach the central neuropile, where they establish synaptic contacts, f.i., with optical fibres. 3 types of synapses occur: 1. one type of synapse which has not yet been described, 2. synapses of the usual type, 3. ribbon synapses. Glial cells situated peripherally in the ganglion envelope the nerve cells. Their lamellar processes projecting into the neuropile adapt their surfaces to all neighboring elements so that the extracellular space is reduced to a labyrinth of narrow intercellular clefts. The number of multipolar ganglion cells and neurosecretory elements is relatively small. The processes of neurosecretory cells contact the surface of the ganglion where they are covered by a basement membrane.

     

Über die Struktur der Pilzkörper des Insektenhirns

Zusammenfassung Die Organisation der -Loben der Pilzkörper im Gehirn von Acheta domesticus L. wird nach licht- und elektronenmikroskopischen Befunden beschrieben. Der säulenartige Faserkomplex des -Lobus besteht aus Fortsätzen von Pilzkörperzellfasern (intrinsischen Fasern, IF) und pilzkörperfremden Fasern (extrinsischen Fasern, EF), die in den Lobus eindringen. Die feinen IF durchziehen den Lobus hauptsächlich parallel zu seiner Längsachse, während die EF zumeist senkrecht zur Längsachse angeordnet sind. Der Lobus erscheint von seiner Peripherie bis zu seinem Zentrum durch IF-Zonen gegliedert. Die Verteilung der EF weist auf eine zusätzliche Ordnung von der Basis zur Spitze des Lobus hin. Zahlreiche polarisierte Synapsen verbinden IF mit EF. Die IF zeigen Vesikelanhäufungen und präsynaptische Apparate besonders in Erweiterungen, die auch in Golgi-Präparaten lichtmikroskopisch zu sehen sind. Es werden zwei EF-Typen unterschieden: 1. Postsynaptische EF (zahlreich) und 2. EF mit prä- und postsynaptischen Kontakten, die nur in einigen Regionen des -Lobus gefunden wurden. Präsynaptische IF konvergieren auf dendritische EF, die Verbindungen mit anderen Teilen des Hirns und des Nervensystems herstellen. Funktionelle Gesichtspunkte werden diskutiert.

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On the structure of the mushroom-bodies in the brain of insectsII. Synaptic connexions in the alpha-lobe of the cricket Acheta domesticus L.

Summary The organization of the -lobes of the corpora pedunculata in the brain of the cricket Acheta domesticus L. has been investigated in the light and electron microscopes. The cylindrical fibre complex is composed of branches of mushroom-body fibres (intrinsic fibres) and extrinsic fibres, which penetrate the -lobe. Intrinsic fibres (IF) run through the -lobe in the same direction, but not strictly parallel to each other or to the axis of the -lobe. Extrinsic fibres (EF) and their fine branches are often arranged perpendicular to the axis of the -lobe. There is some evidence that different IF zones occur in the -lobe when passing from its periphery to its centre. The distribution of EF may reflect a structural order when passing from the base of the lobe to its top. Numerous polarized synapses connect the IF with the EF. The IF show clusters of vesicles and presynaptic figures especially in their blebs, which can be seen in Golgi preparations for light microscopy. Two types of EF are distinguished on the basis of their synaptic junctions: (1) postsynaptic EF (abundant) and (2) EF with pre- and postsynaptic sites (perhaps restricted to some regions of the -lobe). Presynaptic IF converge on EF, which may transfer excitation from the -lobe to different parts of the brain and nervous system.

     

Zur Feinstruktur der Spezialzellen von normalernährten und hungernden Aeolidiern (Gastr. Nudibranchia)

Zusammenfassung Die Spezialzellen im Bindegewebe der Kolben von Eubranchus farrani und Trinchesia granosa (Aeolidoidea) bilden Gruppen, die stets von einer Basallamina umschlossen werden. Die Zellen werden nicht durch desmosomale Bildungen untereinander verbunden. In den oft geweiteten Interzellularlakunen findet sich feinstflockiges Material. Spezialzellen normal ernährter Schnecken besitzen ein außerordentlich reiches, geordnetes, rauhes endoplasmatisches Retikulum und einen großen Kern mit großem Nucleolus. Golgi- und Vakuolenapparat der Zellen bleiben klein. Mit zunehmender Dauer des Fastens wird das rauhe ER durch ungeordnetes, glattes endoplasmatisches Retikulum ersetzt, wächst der Golgiapparat und nimmt die Zahl der Autolysosomen zu. Die feinstrukturellen Unterschiede zwischen Spezialzellen, Blasenzellen und Nervenzellen werden beschrieben, die Funktion der Spezialzellen abschließend diskutiert.

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An electron-microscope study of cellules spéciales in normally nourished and starved aeolids (Gastr. nudibranchia)

Summary Cellules spéciales of Aelidoidea have been investigated in the cerata of Eubranchus farrani and Trinchesia granosa. All groups of cellules spéciales are limited by a basal lamina. Opposing membranes are not connected by specialized structures as desmosomes etc. The intercellular cleft often is widened and contains a fine floccular substance. In normally nourished animals the cellules spéciales show a rich, highly organized, rough surfaced reticulum, while there are only a few vacuoles and a small Golgi Complex. In starved animals mainly smooth reticulum is found, but the Golgi Apparatus has enlarged and autolysosomes are abundant. The fine structural differences of cellules spéciales, globular cells and nerve cells are described, the function of the cellules spéciales is discussed.

Mit dankenswerter Unterstützung durch den Schweizerischen Nationalfond zur Förderung der Wissenschaften und die Deutsche Forschungsgemeinschaft. Ich danke ferner für die guten Arbeitsmöglichkeiten in der Zoologischen Abteilung der Stazione Zoologica Napoli und Frau A. Merkelbach (Tübingen) für technische Assistenz.     

Stabilität des Phagen PS8 in Crown-Gall-Gewebekulturen von Vinca rosea L.

Zusammenfassung Aus einem A. tumefaciens, B6, infizierten Sproß von Vinca rosea wurde eine Tumorgewebekultur hergestellt, die in 2 verschiedenen Tests bakterienfrei war. Beim Plattieren von Homogenaten dieser Kultur gegen den phagensensitiven Stamm B6-806 von A. tumefaciens wurden keine Phagen gefunden. Auch Hitzeschock, Bestrahlung mit UV oder Behandlung mit Mitomycin induzierten keinen Phagen. Das Homogenat selbst inaktiviert zugefügten Phagen PS8 nicht.Bei 26° C verliert PS8 seine Aktivität in 13 Wochen (von 10⁷PS8/ml auf 0). Wenn PS8 einer wachsenden Tumor-Gewebekultur zugefügt wird, verliert er rascher an Aktivität. Dies ist z. T. Folge der Verdünnung des Phagen durch die wachsende Gewebekultur, z. T. Folge einer Wechselwirkung zwischen Phage und Gewebe. In der Tumorkultur wurde keine Vermehrung des Phagen PS8 beobachtet.Die Ergebnisse werden im Hinblick auf tumorigene Eigenschaften von PS8 diskutiert.

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Stability of phage PS8 in crown-gall tissue cultures of Vinca rosea L.

From Vinca rosea shoots a tumor tissue culture (Agrobacterium tumefaciens B6) was prepared which was bacteria-free in 2 experimental tests. This culture did not contain any phages when homogenates of the tissue were plated against the phage-sensitive strain B6-806 of Agrobacterium tumefaciens. Also heat shock, UV irradiation or treatment with mitomycin C did not induce any phage.The homogenate itself did not interfere with the activity of added phage PS8. At room temperature the bacteriophage PS8 looses activity within 13 weeks (10⁷ PS8/ml to 0).If PS8 is added to a growing tumor tissue culture the loss of activity is more rapid, partly because the phage is diluted by the growing culture, partly because some interaction between tissue culture and phage activity. No propagation of PS8 in the tumor tissue culture was observed.These results are discussed in relation to the hypothesis of PS8 as the tumor inducing principle.

     

Die fluchtreaktionen der ErbsenlausAcyrthosiphon pisum ausgel?st durch Alarmpheromon und zus?tzliche reize

Zusammenfassung Der Duft von (E)--Farnesen veranlaßte in hohen Konzentrationen 84% ungeflügelter Erbsenläuse (Acyrthosiphon pisum (Harris)) innerhalb von 10 Min., die Stechborsten aus dem Blattgewebe zu ziehen. Jedoch verließen nur 12% in 25 Min die Pflanze, die übrigen liefen auf der Pflanze umher und siedelten wieder auf ihr.Das Alarmpheromon konnte in seiner Wirkung gefördert oder gehemmt werden durch zusätzliche, nichtchemische Reize. Bestimmte taktile Reize erhöhten die Wirkung des Siphonensekrets auf 100%. Andererseits verringerten Erschütterungen oder optische Reize die Reaktion. Larven vonCoccinella septempunctata L. beeinflußten die Reaktion der Läuse auf das Farnesen nicht, die Wirkungen durch Feinde und Farnesen erwiesen sich als additiv.(E)--Farnesen scheint nicht die einzige Komponente im Alarmpheromon der Erbsenlaus zu sein, denn der Anteil der Fallreaktionen an der Gesamtreaktion war beim (E)--Farnesen, unabhängig von seiner Konzentration, kleiner als bei Siphonensekret.

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Summary High levels of (E)--farnesene caused 84% of apterous pea aphids to withdraw their stylets from the plant tissue within 10 min. But only 12% left the plant within 25 min, the others were running on the plant and settled down again.The effect of alarm pheromone could be decreased and increased with additional, non-chemical stimuli. Special tactile stimuli increased the effect of siphunculi droplets secretion. On the other hand, vibration or optical stimuli decreased the reaction of the aphids. Larvae ofCoccinella septempunctata L. did not influence the reaction to farnesene, the escape responses to enemies and to farnesene seem to be additive.(E)--farnesene is probably not the only component in the alarm pheromone of the pea aphid as the percentage of reacting aphids showing the falling response was lower with (E)--farnesene —independent of the concentration — than with the siphunculi fluid.

     

Attraction of the bark beetle Tomicus piniperda and some other bark- and wood-living beetles to the host volatiles α-pinene and ethanol

The attraction of some bark- and ambrosia beetles as well as associated beetles to the host volatiles -pinene and ethanol was studied in field tests with flight barrier traps. Tomicus piniperda (L.) (Scolytidae), Thanasimus formicarius (L.) (Cleridae), and Rhizophagus ferrugineus (Payk.) (Rhizophagidae) were attracted by -pinene, while Hylurgops palliatus (Gyll.) and Trypodendron lineatum (Oliv.) (Scolytidae) were attracted by ethanol and Epuraea spp. (Nitidulidae) by both -pinene and ethanol. Combinations of -pinene and ethanol attracted high numbers of H. palliatus, T. lineatum, R. ferrugineus, Epuraea spp., and Glischrochilus spp. (Nitidulidae) and the catches increased with increasing release rates of ethanol. By contrast, lower numbers of T. piniperda were caught in traps baited with combinations of -pinene and ethanol than in traps baited with -pinene alone, and the catches of this species decreased with increasing release rates of ethanol. Traps baited with a combination of -pinene and ethanol or with -pinene alone caught similar numbers of T. formicarius. The results are discussed on the basis of species differences in preference for breeding substrate.

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Zusammenfassung Die Anlockung mehrerer Borkenkäfer und assoziierter Käferarten zu den flüchtigen Wirtsstoffen -Pinen und Äthanol wurde in Feldversuchen mit Flugbarrierenfallen studiert. Tomicus piniperda (L.) (Scolytidae), Thanasimus formicarius (L.) (Cleridae) und Rhizophagus ferrugineus (Payk.) (Rhizophagidae) wurden durch -Pinen angelockt, die Borkenkäfer Hylurgops palliatus (Gyll.) und Trypodendron lineatum (Oliv.) durch Äthanol und die Epuraea-Arten (Nitidulidae) durch sowohl -Pinen als auch Äthanol.Kombinationen von -Pinen und Äthanol lockten viele Individen von H. palliatus, T. lineatum, R. ferrugineus, Epuraea spp. und Glischrochilus spp. (Nitidulidae) an, und die Fänge nahmen mit zunehmender Äthanol-Abgabe zu. Umgekehrt wurden weniger T. piniperda in Fallen mit Kombinationen von -Pinen und Äthanol gefangen als in Fallen mit -Pinen allein, und die Waldgärtner-Fänge nahmen mit zunehmender Äthanol-Abgabe ab. Die Fänge von T. formicarius in Fallen mit einer Kombination von -Pinen und Äthanol unterschieden sich nicht von denen in Fallen mit nur -Pinen.Die Ergebnisse werden auf der Grundlage der Unterschiede zwischen den Arten in der Wahl des Brut-substrats besprochen.

     

Purification and partial characterisation of and α-tocopherol-binding protein from rabbit heart cytosol

An -tocopherol-binding protein has been isolated and purified from rabbit heart cytosol. The purified protein had an apparent molecular mass of 14,200, as derived from SDS-PAGE. The content of the protein in rabbit heart was around 11.8 g per g of tissue. The binding of -tocopherol to the purified protein was rapid, reversible, and saturable. Neither nor tocopherol could displace the bound -tocopherol from the protein, suggesting a high specificity for -tocopherol. -Tocopherol-binding protein did not bind oleate. Transfer of -tocopherol from liposomes to mitochodria was stimulated 8-fold in the presence of the binding protein, suggesting that this protein may be involved in the intracellular transport of -tocopherol in the heart.     

Homocysteine stimulates the expression of monocyte chemoattractant protein-1 in endothelial cells leading to enhanced monocyte chemotaxis

The effect of intraperitoneal administration of tocopherol (100 mg/kg wt/24 h) on ascorbate (0.4 mM) induced lipid peroxidation of mitochondria and microsomes isolated from rat liver and testis was studied. Special attention was paid to the changes produced on the highly polyunsaturated fatty acids C20:4 n6 and C22:6 n3 in liver and C20:4 n6 and C22:5 n6 in testis. The lipid peroxidation of liver mitochondria or microsomes produced a significant decrease of C20:4 n6 and C22:6 n3 in the control group, whereas changes in the fatty acid composition of the tocopherol treated group were not observed. The light emission was significantly higher in the control than in the tocopherol treated group. The lipid peroxidation of testis microsomes isolated from the tocopherol group produced a significant decrease of C20:4 n6 , C22:5 n6 and C22:6 n3, these changes were not observed in testis mitochondria. The light emission of both groups was similar. The treatment with tocopherol at the dose and times indicated showed a protector effect on the polyunsaturated fatty acids of liver mitochondria, microsomes and testis mitochondria, whereas those fatty acids situated in testis microsomes were not protected during non enzymatic ascorbateFe²⁺ lipid peroxidation. The protector effect observed by tocopherol treatment in the fatty acid composition of rat testis mitochondria but not in microsomes could be explained if we consider that the sum of C20:4 n6 + C22:5 n6 in testis microsomes is 2-fold than that present in mitochondria.     

Relationship between soluble tumor necrosis factor (TNF) receptors and TNFα during immunotherapy with interleukin-2 and/or interferon α

Eleven metastatic cancer patients were studied during three different regimens of immunotherapy with interleukin-2 (IL-2) and/or interferon (IFN): group A received 4 days of IL-2 i.a. infusion (n=3), group B IFN s.c. during 5 days (n=4), followed on day 3 by 5 days of a continuous IL-2 i.v. infusion, and group C had 4 days of IL-2 i.v. infusion together with s.c. IFN on days 1 and 4 (n=4). Soluble tumor necrosis factor receptors (sTNFR) p55 and p75 and TNF concentrations in serum were analyzed before therapy and daily during 8 days of the first therapy cycle. sTNFR was measured by radioimmunoassay. sTNFR p55 increased in all patient groups from a baseline value of 5.2±0.9 ng/ml to a maximum of 13.6±1.2 ng/ml by days 3–4 (P=0.003). sTNFR p75 increased from 7.6±1.1 ng/ml to peak values of 30.1±2.6 ng/ml in groups A and B (P=0.02). In group C the sTNFR p75 response was weak (NS). In group B, the increase of both p55 and p75 occurred only after addition of IL-2 to IFN. TNF increased weakly during treatment with IFN alone (group B); it rose strongly during IL-2 and the combined treatment (groups A-C) from 8±2 pg/ml to 115±13 pg/ml (P=0.003). In group B, it reached the maximum 24 h after addition of IL-2 to IFN and decreased thereafter. there was a significant relationship between TNF and sTNFR p55 or sTNFR p75 in groups A and C, (P=0.001), but not in group B. Group C was also investigated during the third therapy cycle. The increase of sTNFR p75 was stronger (P=0.01) and that of TNF weaker than in the first cycle; the sTNFR p55 response was similar in both cycles. In conclusion sTNFR p55 and p75 are rapidly induced during IL-2 and IL-2+IFN treatment, the increase of sTNF receptors parallels or exceeds that of TNF and may influence the immunomodulatory effects of TNF during cytokine therapy.     

Peptide modification by incorporation ofα-trifluoromethyl substituted amino acids

Summary Metabolic stabilization of pharmacologically active peptides can be achieved by incorporation of sterically hindered non-natural amino acids, e.g. C ^, -disubstituted amino acids.-Trifluoromethyl substituted amino acids, a subclass of C ^, -disubstituted amino acids, also fulfil this requirement while featuring additional properties based on the electronic influence of the fluorine substituents.This review summarizes the results concerning the stability of peptides containing-TFM amino acids towards proteolysis by-chymotrypsin. Furthermore, configurational effects of-TFMAla on the proteolytic stability of peptides are explained using empirical force field calculations. The influence of-TFMAla incorporation on the secondary structure of selected tripeptide amides is compared to the effects exerted by its fluorine-free analogue, aminoisobutyric acid.Finally, results on metabolic stabilization and biological activity of modified thyrotropin releasing hormone are interpreted.     

A robust and cost-effective method for the production of Val, Leu, Ile (δ1) methyl-protonated 15N-, 13C-, 2H-labeled proteins

A selective protonation strategy is described that uses [3-2H] 13C -ketoisovalerate to introduce (1H- methyl)-leucine and (1H- methyl)-valine into 15N-, 13C-, 2H-labeled proteins. A minimum level of 90% incorporation of label into both leucine and valine methyl groups is obtained by inclusion of 100 mg/L -ketoisovalerate in the bacterial growth medium. Addition of [3,3-2H2] -ketobutyrate to the expression media (D2O solvent) results in the production of proteins with (1H-1 methyl)-isoleucine (>90% incorporation). 1H-13C HSQC correlation spectroscopy establishes that CH2D and CHD2 isotopomers are not produced with this method. This approach offers enhanced labeling of Leu methyl groups over previous methods that utilize Val as the labeling agent and is more cost effective.     

The effect of reserpine on the pars intermedia of the rat pituitary

Summary Reserpine has a stimulatory effect on the pars intermedia of the rat pituitary, probably mediated by its action on regulatory catecholaminergic nerves. The effect of single intraperitoneal injections of 0.1–20 mg/kg b.w. of reserpine was studied in adult male rats. Reserpine at a dose of 2 mg/kg b.w. induced degranulation, orientation of the secretory granules along the cell membrane and loss of formaldehyde-chloral-induced fluorescence, accompanied by an activation of the granular endoplasmic reticulum and the Golgi apparatus. With higher doses progressive degranulation and loss of fluorescence were observed. The effect was, however, heterogeneous, and with all doses cells displaying normal ultrastructure and normal fluorescence were regularly present.To study the release of granular products (containing a different components of the pro-opiomelanocortin chain) from individual cells, formaldehyde-chloral induced fluorescence and -MSH- and -endorphin immunoreactivies were demonstrated in consecutive sections from pituitaries of rats given 8 mg/kg body weight of reserpine 24 h before sacrifice. The results indicate coordinated release of these granular products at the cellular level after reserpine treatment.This work was supported by Finska Läkaresällskapet     

Structure of α-α-Hairpins with Short Connections in Globular Proteins

The analysis of conformations of more than 100 --hairpins with closely packed helical segments and connections up to four amino acid residues in length was carried out. Five types of the connections were revealed, and their and values on the Ramachandran map were found. Each type of --hairpins was shown to have a unique sequence pattern for hydrophobic and hydrophilic residues.     

Heteronuclear correlation experiments for the determination of one-bond coupling constants

Spin-state selective experiments, HSQC-/ and CT-HMQC-/, are proposed for the simple and rapid measurement of scalar one-bond coupling constants in two-dimensional,¹ H-detected ¹⁵N-¹H or¹³ C-¹H correlation experiments based on HSQC and HMQC schemes. Pairs of subspectra are obtained, containing either the high-field or the low-field component of the doublet representing the one-bond coupling constant. The subspectral editing procedure retains the full sensitivity of HSQC and HMQC spectra recorded without heteronuclear decoupling during data acquisition, with a spectral resolution similar to that of decoupled spectra.     

Evolution of parallel β/α-barrel enzyme family lightened by structural data on starch-processing enzymes

The parallel /-barrel domain consisting of eight parallel -sheets surrounded by eight -helices has been currently identified in crystal structures of more than 20 enzymes. This type of protein folding motif makes it possible to catalyze various biochemical reactions on a variety of substrates (i.e., it seems to be robust enough so that different enzymatic functionalities could be designed on it). In spite of many efforts aimed at elucidation of evolutionary history of the present-day /-barrels, a challenging question remains unanswered: How has the parallel /-barrel fold arisen? Although the complete sequence comparison of all /-barrel amino acid sequences is not yet available, several sequence similarities have been revealed by using the highly conserved regions of -amylase as structural templates. Since many starch-processing enzymes adopt the parallel /-barrel structure these enzymes might be useful in the search for evolutionary relationships of the whole parallel eight-folded /-barrel enzyme family.     

Beziehungen zwischen Carnitinstoffwechsel und Fettsäureassimilation bei Pseudomonas putida

Zusammenfassung Der Carnitinstoffwechsel und einige Beziehungen zum Fettsäurestoffwechsel wurden mittels der Wachstumskontrolle, der Bestimmung von Metaboliten und des Nachweises von Enzymaktivitäten in Pseudomonas putida untersucht. Der Stamm wuchs auf -Butyrobetain, D,L-und L-Carnitin, Glycinbetain, Cholin, D,L-Norcarnitin, D,L--Amino--hydroxybutyrat und D,L--Hydroxybutyrat. Obwohl der Stamm unverzweigte Fettsäuren von 2–16 C-Atomen zu untzen vermag, konnte er nur auf O-Acyl-L-carnitinen von 10 oder mehr C-Atomen in der Acylgruppe wachsen. Zugabe von Carnitin stimulierte das Wachstum auf langkettigen Fettsäuren.Die Bildung von Trimethylamin stieg, wenn Carnitin oder -Butyrobetain nur C-Quellen waren, und sank, wenn diese Trimethylammoniumverbindungen sowohl C-als auch N-Quellen waren. L-Carnitin induzierte sowohl die Carnitindehydrogenase als auch die -Hydroxybutyratdehydrogenase. -Butyrobetain als C-und N-Quelle induzierte ebenfalls die Carnitindehydrogenase. Im Rohextrakt betrug die spezifische Aktivität der -Hydroxybutyratdehydrogenase entsprechend dem Wachstum auf L-Carnitin oder D,L--Hydroxybutyrat 0,7 oder 1,6 Mol · min^-1 · mg^-1. Glycinbetain, Glucose und langkettige Fettsäuren reprimierten die Synthese beider Enzyme. Abhängig von der N-Quelle wird L-Carnitin offensichtlich auf zwei unterschiedlichen Stoffwechselwegen abgebaut.

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Interrelationships between carnitine metabolism and fatty acid assimilation in Pseudomonas putida

The carnitine metabolism and some relations to the fatty acid metabolism were studied in Pseudomonas putida by means of control of growth, analysis of metabolites, and determination of enzyme activities. The strain grew on -butyrobetaine, D,L-and L-carnitine, glycinebetaine, choline, D,L-norcarnitine, D,L--amino--hydroxybutyrate, and D,L--hydroxybutyrate. Although the strain used straight-chain fatty acids of 2–16 C-atoms, it was only able to grow on O-acyl-L-carnitines of 10 or more C-atoms in the acylgroup. Addition of carnitine stimulated the growth on long-chain fatty acids.The formation of trimethylamine increased, if L-carnitine or -butyrobetaine were the only carbon sources, and decreased, if these trimethylammonium compounds were carbon as well as nitrogen sources. L-Carnitine induced the carnitine dehydrogenase as well as the -hydroxybutyrate dehydrogenase. -Butyrobetaine as carbon and nitrogen source induced the carnitine dehydrogenase, too. In the crude extract the specific activities of -hydroxybutyrate dehydrogenase were 0.7 or 1.6 moles·min^-1·mg^-1 after growth on L-carnitine and D,L--hydroxybutyrate, respectively. The synthesis of both enzymes was repressed by glycinebetaine, glucose and long-chain fatty acids. Dependent on the nitrogen source L-carnitine was catabolized via two different pathways.

     

Phase I study of TNFα AutoVaccIne in Patients with metastatic cancer

We evaluated the safety and immunogencity of a novel vaccine directed against autologous TNF in a Phase I fixed dose escalation trial. The vaccine consisted of two recombinant TNF proteins, with specific peptides replaced by foreign immunodominant T cell epitopes from tetanus toxoid. The main objectives were to establish a safe dose and evaluate the vaccines ability to raise neutralising TNF antibodies. Secondary objectives were improvements in body weight and tumour response. Thirty-three patients were vaccinated with three doses (20, 100, or 400 g) of TNF vaccine at 2-weekly intervals adjuvanted with aluminium hydroxide. Anti-TNF antibody titres were measured by both a RIA, using soluble native TNF as the antigen, and by an ELISA using immobilized partly denatured TNF. Eleven patients (33%) had mild grade1/2 injection site reactions at the higher doses. In 10 of 20 patients, serum antibodies recognize denatured TNF in the ELISA, whereas, antibody titres against native TNF in the RIA were undetectable. This suggests that the production process had partly denatured the vaccine preventing the formation of cross-reacting antibodies to native TNF. In conclusion, TNF vaccine was able to elicit vaccine specific antibodies. However, since the antibodies were only able to cross-react with partly denatured TNF, evaluation of safety and tumour responses to the TNF vaccine was compromised.