Highly specialized Cretaceous beetle parasitoids (Ripiphoridae) identified with optimized visualization of microstructures

Extremely miniaturized longipedes insects (body length c. 0.3 mm) embedded in two pieces of Cretaceous amber from Myanmar are described and interpreted. Using inverted fluorescence and light microscopy for detailed analysis of microstructures, the inclusions were identified as primary larvae of the beetle family Ripiphoridae, subfamily Ripidiinae. While the structure of thoracic and abdominal segments including appendages corresponds well with the groundplan known in recent members of Ripidiinae, a curved prosternal ridge with prominent spines (each c. 5 μm), the reduced condition of stemmata and antennae and the lack of sharp mandibles are unique features within the entire family, apparently apomorphies of the longipedes larvae. A sinuate prosternal edge with a dense row of spines (prosternoctenidium) might be homologous with ‘head ctenidia’ in some previously described miniaturized conicocephalate larvae, but further investigation is needed. The morphological differences between the head of longipedes larvae and extant Ripidiinae are interpreted as adaptations to different groups of hosts and life strategies. Palaeoethology of the longipedes larvae is briefly discussed. In addition, the systematic placement of conicocephalate larvae from Canadian, Myanmar and Russian Cretaceous ambers, already interpreted by various authors as primary instars within Coleopterida (assigned to either Strepsiptera or to the coleopteran Tenebrionoidea: Ripiphoridae), is discussed.     

Comparative mineral and hormonal analyses of wild type and TLS somaclonal variant derived from oil palm (Elaeis guineensis Jacq. var. tenera) tissue culture

Comparative mineral and hormonal analyses were made on tissue culture derived truncated leaf syndrome and wild type oil palm seedlings. Mineral analysis confirmed that Boron, Zinc and chlorophyll levels were significantly lower in truncated leaf syndrome leaves than those of wild type. Hormonal analysis also revealed various cytokinin derivatives such as trans-zeatin, trans-zeatin riboside, trans-zeatin O-glucoside and trans-zeatin riboside 5??mono phosphate were significantly higher in truncated leaf syndrome leaves compared to wild type leaves. Brassinolide level was also significantly higher in truncated leaf syndrome leaves than those of the wild type. These observations suggest that the truncated leaf syndrome abnormality could be associated to high cytokinin and brassinosteroid production which affects the uptake of Boron and Zinc.     

Untersuchungen über die Inaktivierung und Radikalbildung an suspendiertem Trypsin nach UV-Bestrahlung

Zusammenfassung Es wird eine Methode zur UV-Bestrahlung von Enzymsuspensionen in Alkohol angegeben. Durch Eintauchen der UV-Lampe in die homogene Suspension läßt sich auf einfache Weise die vom Enzym absorbierte Energie bestimmen. Die Inaktivierung wird in Abhängigkeit von der Bestrahlungsdauer gemessen und mittels der ESR-Spektroskopie die Radikalbildung untersucht. Die ESR-Spektren zeigen, daß eine sauerstoff- und wasserfreie Äthanolsuspension vakuumähnliche Bestrahlungsbedingungen liefert. Man erhält ein Radikal am-Kohlenstoff und ein Schwefelradikal vom Typ RS ·. Es wurden die Quantenausbeuten für die Inaktivierung _i und für die Radikalbildung _r ermittelt. Für Trypsin finden wir nach Bestrahlung mit UV-Licht der Wellenlänge 254 nm _i=2,4·10_–2 und _r =1,7·10_–3. Daraus ergibt sich für die Anzahl der Radikale pro inaktiviertem Molekül ein Wert von 0,07, der nahelegt, daß zwischen der gemessenen Inaktivierung und den noch vorhandenen Radikalen kein direkter Zusammenhang besteht. Untersuchungen mit einem kontinuierlichen UV-Spektrum ergaben dieselbe Radikalzahl pro inaktiviertem Molekül, wobei jedoch die Schwefelradikalausbeute geringer ist als nach Bestrahlung mit der Linie 254 nm. Bestrahlung mit Wellenlängen > 300 nm bewirkte eine teilweise Löschung der durch die Linie 254 nm erzeugten Radikale.

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Studies on inactivation and radical formation after UV-irradiation of trypsin in suspension

Summary A method for UV irradiation of suspensions of enzymes in alcohol has been described. The UV lamp was dipped into the suspension which was stirred during irradiation in order to provide a homogeneous exposure of the enzymes and to facilitate the determination of the energy absorbed in the enzymes. The inactivation has been studied as a function of exposure time, radical formations being analysed by way of EPR-spectroscopy. The EPR spectra indicated that oxygen- and waterfree suspensions in ethanol provide vacuumlike conditions for UV irradiation. A radical at the C position and a sulfur radical of the RS · type were observed. Quantum yields for the inactivation ( _i) and for radical formation ( _r) were obtained. UV irradiation of trypsin at 254 nm yielded _i=2,4·10^–2 and _r=1,7·10^–3. As a result the value of 0.07 free radicals per inactivated molecule leads to the suggestion that there is no direct relation between the inactivation and the observed radical formation. The same number of free radicals per inactivated molecule was obtained after irradiation with a continuous UV spectrum, the yield of sulfur radicals however was lower than in the 254 nm investigation. Irradiation with > 300 nm partially quenches the radicals produced at 254 nm.

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Abschließend möchte ich des im Februar 1969 verstorbenen Prof. Dr. Kurt Sommermeyer gedenken, dem ich für die Anregung zu diesem Thema und für wertvolle Diskussionen zu danken habe. Dem Deutschen Akademischen Austauschdienst danke ich für das Stipendium, das mir die Durchführung der Arbeit im Radiologischen Institut ermöglicht hat.     

Reproductive anatomy, manipulation of ovarian activity and non-surgical embryo recovery in suni (Neotragus moschatus zuluensis)

Marked disparity in the uterine horn dimensions and relative degrees of caruncle development in suni suggested that exclusive or predominant dextral implantation occurs in association with bilateral ovulatory activity. Daily urinary measurements of pregnanediol-3 alpha-glucuronide revealed an oestrous cycle of approximately 21 days in length. Ovarian activity was controlled for synchronization of oestrus by using progestagen-impregnated intravaginal sponges and multiple ovulations were induced by using exogenous gonadotrophin therapy. An effective transcervical uterine catheterization technique was developed for the non-surgical collection of embryos. The efficiency of embryo recovery performed 5 days after sponge removal was 50.0%.     

Empirical testing of cryoconite granulation: Role of cyanobacteria in the formation of key biogenic structure darkening glaciers in polar regions

Cryoconite, the dark sediment on the surface of glaciers, often aggregates into oval or irregular granules serving as biogeochemical factories. They reduce a glacier's albedo, act as biodiversity hotspots by supporting aerobic and anaerobic microbial communities, constitute one of the organic matter (OM) sources on glaciers, and are a feeder for micrometazoans. Although cryoconite granules have multiple roles on glaciers, their formation is poorly understood. Cyanobacteria are ubiquitous and abundant engineers of cryoconite hole ecosystems. This study tested whether cyanobacteria may be responsible for cryoconite granulation as a sole biotic element. Incubation of Greenlandic, Svalbard, and Scandinavian cyanobacteria in different nutrient availabilities and substrata for growth (distilled water alone and water with quartz powder, furnaced cryoconite without OM, or powdered rocks from glacial catchment) revealed that cyanobacteria bind mineral particles into granules. The structures formed in the experiment resembled those commonly observed in natural cryoconite holes: they contained numerous cyanobacterial filaments protruding from aggregated mineral particles. Moreover, all examined strains were confirmed to produce extracellular polymeric substances (EPS), which suggests that cryoconite granulation is most likely due to EPS secretion by gliding cyanobacteria. In the presence of water as the only substrate for growth, cyanobacteria formed mostly carpet-like mats. Our data empirically prove that EPS-producing oscillatorialean cyanobacteria isolated from the diverse community of cryoconite microorganisms can form granules from mineral substrate and that the presence of the mineral substrate increases the probability of the formation of these important and complex biogeochemical microstructures on glaciers.     

Transport and dynamics of molecules dissolved in maize root cortex membranes

Translational diffusion of a fluorescent sterol probe was measured in the plasma membranes of protoplasts isolated from cortical cells of the primary root of maize seedlings. The apparent lateral diffusion coefficient was typically observed to be nearly insensitive to temperature, while the mobile fraction increased with increasing temperature. These fluorescence photobleaching recovery (FPR) measurements were compared with the electron paramagnetic resonance (EPR) spectra of the methyl ester of 13-doxyl palmitic acid in membranes of corn root tissue in situ. The complex spectra observed with this probe were analyzed as weighted sums of simpler spectra of various order parameters and rotational correlation times. The reconstituted spectra calculated from the model show that EPR also detects a mobile (less ordered, fluid) fraction, distinguished by the order parameter S=0.1 to 0.2, which becomes more abundant as temperature increases and is qualitatively comparable to the mobile fraction determined by the FPR method. The observed results on the mobile fractions and the diffusion rates for translational (FPR) as well as rotational (EPR) motions are interpreted in terms of membrane organization, thus providing information on the population and structural patterns of the coexisting domains with a special emphasis on the response of the membrane to temperature changes.This work was supported in part by grants from the Ministry of Science and Technology of the Republic of Slovenia and the International Research Program of the U.S. Department of Agriculture (USDA-JF 814-51) to M.S., and by grants from the Competitive Grants Program of the U.S. Department of Agriculture (88-37264-3807 and 90-37264-5471) to E.A.N.     

Potentiation of antitumor effects of tumor necrosis factor α and interferon γ by macrophage-colony-stimulating factor in a MmB16 melanoma model in mice

The efficacy of systemic infusion of recombinant human macrophage-colony-stimulating factor (M-CSF) in combination with local treatment with human recombinant tumor necrosis factor (TNF) and mouse recombinant interferon (IFN) was studied in vivo on a subclone of B16 melanoma (MmB16) in mice. Short-term intravenous administration of M-CSF at a dose of 10⁶ units daily had no antitumor effect in vivo. Similarly, local treatment of tumor with TNF (5 g daily) did not produce any therapeutic effect. However, simultaneous administration of the same dose of TNF with IFN (1000 units daily) resulted in a synergistic effects manifested by the retardation of tumor growth. Addition of systemic infusion of M-CSF to the local therapy with TNF and IFN induced further augmentation of antitumor efficacy and delayed progression of MmB16 melanoma. The strengthened antitumor effect of combination therapy including M-CSF, TNF and IFN was most probably due to the increased release of monocytes from the bone marrow, their recruitment into the site of tumor growth and subsequent local stimulation of their antitumor activity.     

Eurotium (Aspergillus) repens metabolites and their biological activity

Eurotium repens mycelium cultivated under static conditions was used to isolate and identify metabolities—echinulin, physcion, erythroglaucin, flavoglaucin and asperentin; the filtrate of the culture yielded asperentin 8-methylether. The broadest biological activity spectrum was displayed by asperentin which had antibacterial and antifungal effects and, at a concentration of 86 ώg/ml, caused 50 % mor7 tality inArtemia saline larvae. The highest cytotoxicity towards HeLa cells was found in physcion which caused 50 % growth inhibition at a concentration of 0.1 ώg/ml.