The isolation of barley-aleurone protoplasts

Summary A procedure is described for the isolation of large numbers of viable aleurone protoplasts. After treatment with Onozuka cellulase protoplasts were obtained which were surrounded by a thin, Onozuka-resistant wall. These cells were termed spheroplasts. Treatment of spheroplasts with Glusulase digested away the residual wall, yielding naked protoplasts. Measurements of respiration using a Clarke-type oxygen electrode indicated that aleurone cells isolated by this procedure were viable.Work supported by National Science Foundation grant GB-27468.     

Plant regeneration via somatic embryogenesis in Styrian pumpkin: cytological and biochemical investigations

Somatic embryo formation was induced from cotyledon explants of Styrian pumpkin (Cucurbita pepo L. subsp. pepo var. styriaca Greb.) by using a solid MS medium supplemented with 16.11M NAA and 4.44M BA or 26.85M NAA and 13.32M BA. The callus proliferation was more efficient on medium supplemented with 26.85M NAA and 13.32M BA. In contrast, the embryogenic response was higher on medium with lower concentrations of growth regulators (16.11M NAA and 4.44M BA). The time needed for embryo induction did not depend on medium composition. Embryos in globular stage were transferred to three different maturation media, containing 2.89M GA₃ in combination with 0.54M NAA, 11.42M IAA and growth regulator-free medium. The germination rate was the highest when embryos were cultured on medium with 11.42M IAA. Plantlets grown on this medium achieved maturity suitable for transplantation into soil within 9 to 10weeks. The regenerated plants were successfully transferred into field and developed fertile flowers and set fruits. Biochemical analysis showed significant lower total glutathione levels among in vitro grown plantlets compared to seedlings grown in soil. When the plantlets were transferred into soil, they reached a normal size within a month and the glutathione concentration was comparable to seed-derived plants at the same developmental stage. Transmission electron microscopy was used to investigate possible differences in the ultrastructure of cells from callus cultures, and leaf cells of regenerated and seed-derived plants. Differences in the ultrastructure were found within chloroplasts which contained only single thylakoids, large starch grains and small plastoglobuli in callus cells in comparison to leaf cells, which possessed a well developed thylakoid system, small starch grains and large plastoglobuli.     

Screening of white-rot fungi for biological pretreatment of wheat straw for biogas production

Summary Twenty two basidiomycetes, mostly white rot fungi, were grown on wheat straw. Lignin-, cellulose-, and hemicellulose-degradation was recorded in order to find a species growing on lignin preferably. The oyster-mushroomPleurotus sp. florida showed fastest delignification of all tested fungi.Straw pretreated by this fungus was fermented anaerobically to biogas. The gas yield produced from myco-straw is twice the amount from untreated straw.     

Over expression of integrin α 5 β 1 in human hepatocellular carcinoma cell line suppresses cell proliferation in vitro and tumorigenicity in nude mice

Integrin 5 1 and 2 1 are the major integrin receptors in human hepatocytes. However, in human hepatocellular carcinoma cells it was found that the expression of integrin 5 1 was decreased and another integrin 6 1 increased. In this study, the SMMC7721 human hepatocellular carcinoma cells cotransfected or singlely transfected with integrin 5 and/or 1 cDNAs were established, and designated 5 1.6-7721, 5.3-7721, and 1.6-7721 cell lines, respectively. Transfection with cDNAs of integrin 5 and 1 subunits resulted in the overexpression of each integrin and modified biological properties, including a slowed growth rate, changes in the cell cycle from 15.5% of control cells in the G2/M phase to 12.1%, 9.6% and 9.4% in 5.3-7721, 1.6-7721, 5 1.6-7721, respectively, and a decrease in the Cell Mitosis Index from 1.6 in controls to 0.96, 0.95, and 0.72, and 34%, 28% and 52% derived from colony forming ability, respectively. Tumorigenicity was also tested in nude mice with inoculation of cells subcutaneously. Tumor masses growing in nude mice following inoculation with 1.6-7721,and 5 1.6-7721 cells weighed only 52% or 31% those of control cells. These results indicated that deletion or low expression of integrin 5 1 may play an important role in the development of hepatocellular carcinoma. Therefore, induction of expression of the integrin 5 1 in malignant cells could be a potential means of treating hepatocellular carcinoma.     

Genetic variation detected by use of the M13 “DNA fingerprint” probe in Malus,Prunus, and Rubus (Rosaceae)

Summary Recently, DNA fingerprints have been reported in a wide array of organisms. We used the M13 repeat probe on several genera and species in the angiosperm family Rosaceae. Four apple cultivars could be differentiated when any one of five restriction enzymes was used to analyze minisatellite DNA. Similarly, four individual trees of Prunus serotina (black cherry) exhibited different fingerprints with each of four enyzmes. A total of 14 Rubus (blackberries and raspberries) plants representing four species were investigated with two enzymes. Extensive inter-and intraspecific variation was found. However, some closely growing plants had identical fingerprints, probably due to their being derived through vegetative propagation.     

A model for light adaptation: Producing Weber's law with bleaching-type kinetics

An adaptation model having two stages is introduced and its mathematical properties are examined. The two stages are the adaptive process (parameter K _b), which has bleaching-type kinetics, and the response function (parameters K _r and n), which incorporates response saturation. In order to study the increment threshold functions generated by the adaptation model the concept of a detector is required. It is demonstrated that without an adaptive process the compression hypothesis, in the form of the difference equation, produces increment threshold functions which saturate and do not obey Weber's law. It is then shown that an adaptive process with bleaching-type kinetics can prevent saturation and produce Weber's law behavior provided that the adaptive strength of the system exceeds the detector sensitivity.     

C(alpha) chemical shift tensors in helical peptides by dipolar-modulated chemical shift recoupling NMR

The C chemical shift tensors of proteins contain information on the backbone conformation. We have determined the magnitude and orientation of the C chemical shift tensors of two peptides with -helical torsion angles: the Ala residue in G*AL (=–65.7°, =–40°), and the Val residue in GG*V (=–81.5°, =–50.7°). The magnitude of the tensors was determined from quasi-static powder patterns recoupled under magic-angle spinning, while the orientation of the tensors was extracted from C–H and C–N dipolar modulated powder patterns. The helical Ala C chemical shift tensor has a span of 36 ppm and an asymmetry parameter of 0.89. Its ₁₁ axis is 116° ± 5° from the C–H bond while the ₂₂ axis is 40° ± 5° from the C–N bond. The Val tensor has an anisotropic span of 25 ppm and an asymmetry parameter of 0.33, both much smaller than the values for -sheet Val found recently (Yao and Hong, 2002). The Val ₃₃ axis is tilted by 115° ± 5° from the C–H bond and 98° ± 5° from the C–N bond. These represent the first completely experimentally determined C chemical shift tensors of helical peptides. Using an icosahedral representation, we compared the experimental chemical shift tensors with quantum chemical calculations and found overall good agreement. These solid-state chemical shift tensors confirm the observation from cross-correlated relaxation experiments that the projection of the C chemical shift tensor onto the C–H bond is much smaller in -helices than in -sheets.     

Harzkonservierte fossile Vogelfedern aus der untersten Kreide

Zusammenfassung Harzkonservierte Fossilien ermöglichen bei Anwendung adäquater Methoden die morphologische Analyse der Feinmerkmale bis zur Auflösungsgrenze des Lichtmikroskops, Beobachtung in verschiedenen Ebenen und Richtungen, und somit konkrete Rückschlüsse auf die Wirkung und Bedeutung der Einzelelemente und des Gesamtgefüges.Eine so eingehende funktionsmorphologische Analyse mit Berücksichtigung der Positionsvariation (graduell verschiedene Gestaltung in gesetzmäßiger Abhängigkeit von der Lage innerhalb der Gesamtfeder) der Einzelelemente wie Abzweigungs-, Knick-, Neigungswinkel, Krümmung, Länge, Dicke, Querschnitt, Dichte, Differenzierungsgrad der verschiedenen Abschnitte von Rhachis, Rami, Radii inklusive Häkchen und Cirren wird erstmals für fossile Vogelfedern geliefert (hier als Abriß zu einer dokumentarisch und thematisch ausführlicheren Darstellung in Stuttgarter Beiträge zur Naturkunde).Diese Federn entstammen der untersten Unterkreide und sind damit nur relativ wenig jünger alsArchaeopteryx. Sie weisen extrem differenzierten Aufbau auf, der auf hohe flugtechnische und wärmeisolierende Leistungsfähigkeit schließen läßt.Die hier vorgelegten funktionsmorphologischen Ermittlungen an fossilen Körperkonturfedern mögen auch zu einer intensiveren Analyse der bis jetzt stark vernachlässigten Untersuchung ganz normaler Körperfedern rezenter Vögel anregen. Erst dann, nach umfassender Kenntnis ihrer Ausgestaltung innerhalb der verschiedensten rezenten Vogelgruppen, läßt sich überzeugend begründen, ob und wieweit die hier vorgelegten Federn dieses Unterkreide-Vogels noch ursprüngliche Elemente (Plesiomorphien) oder ihnen eigene Sonderbildungen (Autapomorphien) aufweisen; das gilt sowohl für morphologische wie für funktionelle Elemente der Gesamtstruktur.

---

Resin-preserved fossil bird's feathers from the Lowermost Cretaceous

Summary Parts of some feathers, originating from a single bird, were discovered in our collections of Lower Cretaceous amber from the Lebanon mountains — which, in general, contains the oldest terrestrial microfossils preserved with all morphological details.These contour feathers of the trunk, which are nearly as old as Archaeopteryx (Lowermost Cretaceous: Neocomian/Uppermost Jurassic: Kimmeridigian) were studied with magnifications of 500–900 in several levels by a special technique. (In normal fossils, i.e., impressions, the granulation of the sediment and the fossil's bulky carbon remainders cause a blurred image even at a magnification of merely 100).Special emphasis was laid on the study of the individual elements' gradual variation, depending on the respective position within the total feather (position variation). Where appropriate, an analysis of lengths, quantity, degree of differentiation, angle of inclination, break, and branching, cross-sectional view, curvature, etc. of the rhachis, rami, distal and proximal radii, barbicles, hooklets, etc. were undertaken. [Through measurements of the depth of details the effects caused by a sloping position (apparent variation) may be precisely separated from the real variation.]On the basis of such a detailed knowledge of structure and relative position a thorough functional analysis of the single elements as well as the total system is given.Principal features: The production of plain stability in the feather's center, and of flexibility in its apical and lateral rims; dispersion of forces in case of pressure or a pulling load; function of the hooklets (which donot serve as an interlocking mechanism while the feather is in the normal resting position, but function with increasing braking action only when a neighboring ramus diverges to a precisely defined extent from its resting position) including the mechanism of their unhooking; devices for the avoidance of harmful hooking into contacted parts of other feathers; production of maximal stability by minimal air resistance, and of minute chambers (<0,00001 mm³) with still air for optimal heat isolation.Apart from this abstract, further information, accompanied by numerous figures, will be given in a later paper in Stuttgarter Beiträge zur Naturkunde.

---

---

Veränderte Fassung eines am 11. 10. 1971 gehaltenen Vortrages auf der 83. Jahresversammlung der Deutschen Ornithologengesellschaft in Bonn.     

Microaerophily

Objections to previous definitions of anaerobiosis and micro-aerophily are discussed, and a new definition of micro-aerophily is proposed. A survey of physiological characteristics of the micro-aerophilic bacterium Campylobacter sputorum spp. bubulus is given, and the concept of micro-aerophily is illustrated by the results of a study on the oxygen metabolism and oxygen sensitivity of this bacterium.     

Intracellular sites of the synthesis of sweet potato mitochondrial F1ATPase subunits

Summary Five subunits (-, -, -, - and -subunits) of the six -and -subunits) in the F₁ portion (F₁ATPase) of sweet potato (Ipomoea batatas) mitochondrial adenosine triphosphatase were isolated by an electrophoretic method. The - and -subunits were not distinguishable immunologically but showed completely different tryptic peptide maps, indicating that they were different molecular species. In vitro protein synthesis with isolated sweet potato root mitochondria produced only the -subunit when analyzed with anti-sweet potato F₁ATPase antibody reacting with all the subunits except the -subunit. Sweet potato root poly(A)⁺RNA directed the synthesis of six polypeptides which were immunoprecipitated by the antibody: two of them immunologically related to the -subunit and the others to the - and -subunits. We conclude that the -subunit of the F₁ATPase is synthesized only in the mitochondria and the -, - and -subunits are in the cytoplasm.     

Das Orientierungssystem der V?gel II. Heimfinden und Navigation

Zusammenfassung Die Analyse des Navigationssystems der Vögel bezieht sich meist auf Versuche mit Brieftauben. Kramers Karte-Kompaß-Prinzip geht davon aus, daß die Heimrichtung zunächst als Kompaßkurs bestimmt wird. Dies wurde durch zahlreiche Zeitumstimmungsversuche bestätigt. Damit erscheinen nur zwei Navigationsstrategien möglich: (1) Benutzen von Weginformation, die auf einem Richtungsreferenzsystem beruht, und (2) Benutzen von Ortsinformation, deren Richtungsbeziehung zum Ziel bekannt ist.Junge Tauben, denen während der Verfrachtung der Zugang zu magnetischer Information verwehrt worden war, flogen ungerichtet ab. Dies spricht für eine Navigationsstrategie, bei der die Richtung des Hinwegs mit dem Magnetkompaß gemessen wird. Diese Art von Wegintegration stellt aber nur eine erste Strategie sehr junger, unerfahrener Tauben dar; sobald wie möglich benutzen die Tauben Ortsinformation, denn diese erlaubt Nachbestimmung des Heimkurses und damit das Korrigieren von Fehlern.Navigation anhand von Ortsinformation bedeutet, daß Vögel den Heimkurs aufgrund von Faktoren am Auflaßort bestimmen. Die modernen Vorstellungen berücksichtigen die zentrale Rolle eines äußeren Referenzsystems und gehen von einer Navigationskarte aus, die einer kompaßmäßig ausgerichteten mentalen Repräsentation des Verlaufs der Navigationsfaktoren entspricht. Der Heimkurs wird aus den Unterschieden zwischen den Werten am Auflaßort und denen am Heimatort abgeleitet. Da es sich bei den Navigationsfaktoren um Gradienten handelt, können die Vögel deren Verlauf extrapolieren und ihren Heimkurs auch in unbekanntem Gelände bestimmen. Unregelmäßigkeiten im Gradientenverlauf führen zu Abweichungen von der Heimrichtung, sogenannten Ortsmißweisungen. In der Nähe des Heimatorts wird die Navigationskarte durch eine entsprechende Mosaik-Karte von Landmarken ergänzt.Beide Karten werden durch Lernvorgänge erstellt. Junge Tauben verschaffen sich die notwendige Information auf spontanen Flügen, indem sie Information über den zurückgelegten Weg mit Ortsinformation verknüpfen und gemeinsam in den Karten abspeichern. Bei Tauben wird die Navigationskarte etwa im dritten Lebensmonat funktionstüchtig, aber sie wird auch später noch ständig erweitert und verbessert. Ihre Größe hängt von der Erfahrung der Vögel ab. Die Karte enthält offenbar Entfernungsangaben und erlaubt freie Flüge zwischen beliebigen Zielen. In einigen Punkten entspricht sie der in der psychologischen Literatur diskutierten kognitiven Karte.Das Navigationssystem der Vögel ist dadurch gekennzeichnet, daß sie sich mit Hilfe eines einfachen angeborenen Mechanismus, des Magnetkompaß, viele andere Faktoren durch Lernvorgänge nutzbar machen und diese in komplexe Mechanismen wie die Karten integrieren.

---

The orientation system of birds — II. Homing and navigation

Summary The analysis of the navigational system of birds is largely based on experiments with displaced homing pigeons. Kramer's map-and-compass model assumes that the home direction is first established as a compass course. This is confirmed by numerous clock-shift experiments. Consequently, only two types of navigational strategy appear possible: (1) the use of route-specific information based on an external reference or (2) the use of site-specific information whose directional relationship is familiar to the birds.The use of route-specific information is indicated by the finding that young birds deprived of magnetic information during displacement were disoriented, whereas birds receiving the same treatment at the release site were not. This suggests that birds navigate by recording the direction of the outward journey with their magnetic compass, determining the home course by reversing it. This strategy of path integration with the help of an external reference, however, is used only by very young, inexperienced pigeons during an early phase in the development of the navigational system. As soon as birds become more experienced and are able to use site-specific information, they give up route-specific information in favor of the former. The reasons for this change in strategy lie in the fact that using site-specific information enables birds to redetermine their home course as often as necessary, thus allowing the correction of initial mistakes.Site-specific information means that birds can derive their home course from factors perceived at the release site. The present models on navigation acknowledge the crucial role of an external reference system by proposing the navigational map to be a directionally oriented mental representation of the spatial distribution of at least two navigational factors, which are assumed to be environmental gradients. The birds determine their home course by comparing the local values of these factors with the remembered home values. Gradients can be extrapolated beyond the range of direct experience, which explains the birds' ability to home from distant, unfamiliar sites. Deviations from the true home direction, so-called release site biases, as frequently observed in pigeons and other bird species, may be attributed to unforeseen irregularities in the distribution of the navigational factors, which cause birds to misinterpret their position. Near home, the navigational or grid map is supplemented by the mosaic map, which is supposed to be a directionally oriented mental representation of the distribution of familiar landmarks.Both maps are based on experience. Young birds obtain the relevant information during spontaneous flights by combining information on the route travelled with information on the location of prominent landmarks and the direction of environmental gradients. In pigeons, the maps become functional during the third month of life. They are continuously enlarged and updated also in later years. The total size of the navigational map appears to depend on the spatial range of the birds' experience. The maps seem to include information on distance; they are not restricted to homing, but allow free movements between arbitrary goals. In several respects, the model of the avian navigational map is similar to the concept of the cognitive map discussed in psychological literature, the main differences being the larger distances involved, the representation of continuous environmental gradients instead of separate entities, and the essential role of an external reference.The navigational system of birds is thus characterized as utilizing a wide variety of environmental cues. Learning processes, which are based on a simple, innate mechanism, the magnetic compass, integrate these cues and form complex, experience-based mechanisms, such as the maps.

     

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.     

Detection and partial characterization of two antigenically distinct β-amylases in developing kernels of wheat

A -amylase (EC 3.2.1.2) was identified in the outer pericarp (P) of developing seeds of wheat (Triticum aestivum L.) and compared with the well known -amylase which is synthesized during seed development in the starchy endosperm (E). The enzyme P already exists in the tissues before anthesis and vanishes at the time when E starts to accumulate. The isoelectric-focusing patterns of P and E are very similar. The relative molecular weight (M_r) of P is slightly higher than that of E (66 and 64.5 kDa, respectively). Both P and E exhibit common epitopes in addition to epitopes specific for each of them. The two enzymes were identified in small amounts in the green tissues of the developing seeds (inner pericarp and testa). No antigenic difference was detected between P and the -amylases of roots and leaves.Abbreviations P pericarp -amylase - E endosperm -amylase - IS1 anti--amylase immune serum - IS2 anti- and anti- amylase immune serum - IS3 anti- amylase immune serum - IEF isoelectric focusing - IgG immunoglobulin G The authors thank Dr. P. Ziegler (Universität Bayreuth, FRG) for stimulating discussion and for useful suggestions during the writing of the text. The authors thank Miss C. Mayer for her skillful technical assistance.     

α-Thalassemia and the production of different α chain variants in heterozygotes

The production of five chain variants (Hb G-Georgia, Hb St. Luke's, Hb Lloyd, Hb Montgomery, and Hb G-Philadelphia) in heterozygotes was evaluated through hematological observations, hemoglobin quantification, and biosynthetic studies. All heterozygotes for Hb St. Luke's and Hb Lloyd and most heterozygotes with Hb G-Georgia and Hb Montgomery had normal hematology and average / values of about 1.1. They were assigned a normal genotype (^G/), although the proportions of Hb St. Luke's and Hb G-Georgia were low (10 to 13%) and those of Hb Lloyd and Hb Montgomery twice as high (20%). Data from short-term incubations confirmed this genotype for some of these heterozygotes. Isolated Hb St. Luke's and Hb G-Georgia gave low ^G/ values (0.2 and 0.3) indicating that these Hb variants were defective at the level of Hb assembly. Isolated Hb Montgomery and Hb G-Philadelphia, however, gave higher ^G/ values of 0.6 and 0.8, respectively. A second type of variability existed among Hb G-Georgia (20 vs. 13%), Hb Montgomery (28 vs. 20%), and Hb G-Philadelphia (47 vs. 34%) heterozygotes, in whom the levels of Hb G differed. The occurrence of higher levels of these three chain heterozygosities was associated with hematological or biosynthetic evidence of a mild or moderate chain deficiency due to an -thalassemia-2 heterozygosity (^G/⁰ or ^0G/) or a homozygosity (^0G/⁰), respectively.This study was supported in part by USPHS Research Grants HLB-05168 and HLB-15158.     

Induction of the Synthesis of Endo-1,4-β-xylanase and β-Galactosidase in Original and Recombinant Strains of the Fungus Penicillium canescens

The induction of synthesis of the secreted enzymes endo-1,4--xylanase (EC 3.2.1.8) and -galactosidase (EC 3.2.1.23) in original and recombinant Penicillium canescens strains has been studied. In all producer strains, the synthesis of these enzymes was induced by arabinose and its metabolite arabitol. The two enzymes differed in the concentration of arabinose required for induction: the synthesis of -galactosidase was most pronounced at 1 mM, whereas maximum synthesis of endo-1,4--xylanase was observed at 5–10 mM. An increase in the number of endo-1,4--xylanase copies in the high-copy-number strain of the fungus suppressed the synthesis of -galactosidase; the synthesis of endo-1,4--xylanase in the high-copy-number recombinant producing -galactosidase was affected to a lesser extent. The amount of enzymes synthesized did not depend on the saccharide used as the sole source of carbon for growing the mycelium prior to its transfer to the inducer-containing medium.     

Histoenzymatic characterization of sub-types of Type I fibres in fast muscles of rats

Summary The histochemical activities of succinic dehydrogenase (SDH), myofibrillar Adenosine triphosphatase (ATPase) and alpha glycerophosphate dehydrogenase were studied in serial sections of rat vastus lateralis (red) (RVL), gastrocnemius and diaphragm muscles. Three main fibre-types were distinguished. The Type I fibres of RVL and gastrocnemius muscles fell into two distinct groups: one category-Type IA showed very low ATPase activity. The second category of Type IB fibres displayed moderate ATPase reaction. The Type IA fibres were divisible into two sub-groups when tested for SDH reaction. Type IA₁ fibres possessed a homogenous distribution of diformazan·granules throughout the fibre: Type IA₂ fibres displayed characteristic moth-eaten pattern of diformazan localization. The diaphragm muscle did not show either Type IB or Type IA₂ varieties. The great majority of Type I fibres were sub-type IA₁ in the three fast muscles studied. It is also demonstrated here that an inherent heterogeneity exists between Type I fibres of diaphragm and leg muscles in regard to -GPD localization. This histochemical data emphasizes the fact that subdivision of Type I striated muscle fibres of mammalian animals into two sub-types is only approximate and that a further subcategorization is possible.     

Zur Feinstruktur von Cuticula und Epidermis beim Flußkrebs Orconectes limosus während eines Häutungszyklus

Zusammenfassung Die Cuticula an der Innen- und Außenseite der Branchiostegite des Flußkrebses besteht wie für Arthropoden üblich aus Epi- und Procuticula. Sowohl die Epicuticula als auch die Procuticula von Innen- und Außenseite unterscheiden sich im Feinbau wesentlich voneinander. An der Innenseite ist die Epicuticula einfach gebaut; Die Procuticula ist lamelliert und zeigt meist die bogenförmigen Muster von Mikrofibrillen. Die Epicuticula an der Außenseite weist in den in dieser Arbeit untersuchten Entwicklungsstadien einen sehr viel komplizierteren Feinbau auf, der in der Entwicklung gewissen Änderungen unterliegt. In der wiederum lamellierten Procuticula an der Außenseite sind die Mikrofibrillen zu Balken gebündelt. Die Ausrichtung der Mikrofibrillen dreht sich innerhalb einer Lamelle um 180°. Durch die Procuticula ziehen Fortsätze der Epidermiszellen, außerdem Stäbe der sog. Verbindungsstrukturen.Die Bildung der Cuticula an der Innenseite konnte weitgehend vollständig verfolgt werden; sie ist gut mit der Bildung der Cuticula bei verschiedenen Insekten vergleichbar.Die Bildung der Cuticula an der Außenseite konnte dagegen nur von Beginn der Abscheidung der Proouticula bis zur Häutung verfolgt werden. Kurz vor Beginn der Cuticulaabscheidung kommt es in den Epidermiszellen zu einer stärkeren Entwicklung des rauhen ER. Während der gesamten von uns verfolgten Bildungsstadien sieht man Vesikel mit dichtem Inhalt besonders in der Nähe des Zellapex. Sie geben anscheinend hier ihren Inhalt, Cuticulamaterial, nach außen ab. Sie stammen wohl aus Golgibereichen. Auch Stachelsaumbläschen (coated vesicles) kommen regelmäßig vor, deren genetischer Zusammenhang mit multivesikulären Körpern diskutiert wird. Bei der Abscheidung der fibrillären Cuticulasubstanzen spielen besondere Differenzierungen der Zell oberfläche, — kappenartige Verdichtungen der Zelloberfläche, meist an der Spitze kleiner Mikrovilli — eine wesentliche Rolle.

---

The ultrastructure of cuticle and epidermis in the crayfish Crconectes limosus during a moulting cycle

Summary The cuticle of the inside and outside of the branchiostegites of the crayfish consists of an epicuticle and a procuticle — as common in arthropods. Concerning their ultrastructure epicuticle and procuticle differ essentially from each other on both the inside as well as the outside. On the inside the epicuticle is built plainly; the procuticle is laminated, and, mostly it shows the arched patterns of microfibrils. In those developmental stages investigated in this project the epicuticle of the outside shows a much more intricated ultrastructure, since during formation it is subject to certain changes. On the outside the procuticle is also laminated; the microfibrils are bundled up to bars. The alignment of those microfibrils within one lamella is twisted for 180°. The procuticle is penetrated by processes of epidermal cells and by rods of the so-called connecting structures.The formation of the cuticle on the inside was observed completely; it is comparable to the forming of the cuticle in several insects. However, the formation of the cuticle on the outside was only observed from the beginning of the procuticular development up to the moulting.Shortly before formation of the cuticle the development of rough ER in the epidermal cells seems to be intensified. In all of developmental stages observed there appear vesicles with dense contents mainly situated nearby the cell apex. At this site they evidently deliver their contents — cuticular materials — to the outside of the cell; they probably originate in the Golgi areas. There occur coated vesicles regularity, too; their genetic relation to multivesicular bodies is discussed. Special differentiation on the cell surface i.e. dome-like consolidations of the cell surface mainly placed at the tip of small microvilli are of great importance for the secretion of the cuticle substances.

     

Activation of peritoneal macrophages in patients with gynecological malignancies by sizofiran and recombinant interferon-γ

We investigated the influence of the combined use of sizofiran, a-1,3-glucan and a recombinant interferon- (rIFN-) upon biological activities of peritoneal macrophages (M). The number of peritoneal M and the production of cytokines (interleukin-1, interferon- and tumor necrosis factor) was increased by the combined treatment. Fully activated peritoneal M based on the increased number of elongated pseudopods were observed by electromicroscope. Sizofiran seems to assure a sufficient supply of M to kill tumor cells in the peritoneal cavity and co-administered rIFN- seems to directly stimulate the accumulated M in addition to its direct cytotoxicity against tumor cells. This combination therapy may be a step to the prevention of the recurrence of gynecological malignancies including ovarian cancer, after a negative second-look laparotomy.Abbreviations rIFN- recombinant interferon- - IL-1 interleukin-1 - TNF tumor necrosis factor - SLL second look laparotomy     

Curdlan and other bacterial (1→3)-β-d-glucans

Three structural classes of (13)--d-glucans are encountered in some important soil-dwelling, plant-associated or human pathogenic bacteria. Linear (13)--glucans and side-chain-branched (13,12)--glucans are major constituents of capsular materials, with roles in bacterial aggregation, virulence and carbohydrate storage. Cyclic (13,16)--glucans are predominantly periplasmic, serving in osmotic adaptation. Curdlan, the linear (13)--glucan from Agrobacterium, has unique rheological and thermal gelling properties, with applications in the food industry and other sectors. This review includes information on the structure, properties and molecular genetics of the bacterial (13)--glucans, together with an overview of the physiology and biotechnology of curdlan production and applications of this biopolymer and its derivatives.