Substrate specifity of the hexose carrier in the plasmalemma of Chenopodium suspension cells probed by transmembrane exchange diffusion

Substrate specifity of the proton-driven hexose cotransport carrier in the plasmalemma of photoautotrophic suspension cells of Chenopodium rubrum L. has been studies through the short-term perturbation of ¹⁴C-labelled efflux of 3-O-methyl-d-glucose. Efflux, occurring exclusively via carrier-mediated exchange diffusion, is trans-stimulated by the substrate and trans-inhibited by the glucose-transport inhibitors phlorizin (K _1/2=7.9 mM) and its aglucon phloretin (K _1/2=84 μM); with both inhibitors, 3-O-methyl-d-glucose efflux may be blocked completely. Trans-stimulation of efflux (up to fourfold) by a variety of the d-enantiomers of neutral hexoses, including glucose (K _1/2=48 μM), 3-O-methyl-d-glucose (K _1/2=139 μM), and fructose (K _1/2=730 μM), but not by, for instance, d-allose, and l-sorbose, shows that carrier-substrate interaction critically involves the axial position at C-1 and C-3, respectively. We suggest that substrate binding by the Chenopodium hexose carrier involves both hydrophobic interaction with the pyran-ring and hydrogen-ion bonding at C-1 and C-3 of the d-glucose conformation.     

The in vitro metabolism of benzo[a]pyrene by polychlorinated and polybrominated biphenyl induced rat hepatic microsomal monooxygenases

The metabolism of benzo[a]pyrene by halogenated biphenyl-induced rat hepatic microsomal monooxygenases was determined using a high pressure liquid chromatographic assay system. Incubation of benzo[a]pyrene with microsomes from rats pretreated with phenobarbitone or phenobarbitone-type inducers (2,2',4,4',5,5'-hexachlorobiphenyl, 2,2',4,4',6,6'-hexachlorobiphenyl, 2,2',5,5'-tetrachlorobiphenyl, 2,2',4,4',5,5'-hexabromobiphenyl, and 2,2',5,5'-tetrabromobiphenyl) resulted in increased overall metabolism of the hydrocarbon (less than fourfold) into phenolic, quinone, and diol metabolites, with the most striking increase observed in the formation of 4,5-dihydro-4,5-dihydroxybenzo[a]pyrene. In contrast, the metabolism of benzo[a]pyrene by microsomes from rats induced with 3-methylcholanthrene or 3,3',4,4'-tetrachlorobiphenyl resulted in a greater than 10-fold increase in overall benzo[a]pyrene metabolism, with the largest increases observed in the formation of the trans-7,8- and -9,10-dihydrodiol metabolites of benzo[a]pyrene. However, in comparison to control and phenobarbitone-induced microsomes, the oxidative conversion of benzo[a]pyrene by microsomes induced with 3-methylcholanthrene and 3,3',4,4'-tetrachlorobiphenyl into the 6,12-quinone was substantially inhibited. Previous reports have shown that the commercial halogenated biphenyl mixtures, fireMaster BP-6, and Aroclor 1254 are mixed-type inducers and that microsomes from rats pretreated with these mixtures markedly enhance the overall metabolism of benzo[a]pyrene. Not surprisingly, the metabolism of benzo[a]pyrene by microsomes from rats pretreated with the mixed-type inducers, 2,3,3',4,4'-penta-,2,3,3',4,4',5-hexa-, and 2',3,3',4,4',5-hexa- chlorobiphenyl was also increased and the metabolic profile was similar to that observed with fireMaster BP-6 and Aroclor 1254 induced microsomes.     

Verhalten der Glasaale(Anguilla anguilla) bei ihrer Wanderung in den Ästuarien deutscher Nordseeflüsse

Zusammenfassung 1. Das Verhalten von GlasaalenAnguilla anguilla L., die bei Tage im oberen Tidegebiet der Elbe an der Oberfläche und im Schwarm wanderten, wurde beobachtet.2. Aus diesen und früheren Untersuchungen in der Elbe geht hervor, daß bei Ebbe die Aale ihre Tageswanderung am Ufer etwas unterhalb der Flutstromgrenze in den frühen Morgenstunden beginnen und täglich in den Vormittagsstunden wieder abbrechen. Sie verbergen sich dann im Sand oder unter den Steinen.3. Die täglich weiter flußaufwärts begonnenen Oberflächenwanderungen enden vermutlich an der Tidegrenze oder etwas oberhalb davon und umfassen damit einen Bereich von etwa 50 km.4. Glasaalschwärme wurden am häufigsten an den Tagen, die dem Mitternachtstidehochwasser an der Flutstromgrenze folgten, festgestellt. Diese Erscheinung läßt sich aus dem Verhalten der Glasaale erklären, negativ phototaktisch und passiv mit dem Flutstrom flußaufwärts zu treiben.5. Die Aale halten während der Tageswanderung engen Uferkontakt und lassen sich in ihrer Wanderrichtung, außer bei starker Turbulenz, durch gleichlaufende Strömungen nicht irritieren.6. Das Elbestauwehr wird, außer gelegentlich an der Schleuse, während des beschriebenen Entwicklungsabschnittes nur bei Hochwasser überwunden; dann können die Aale im Bereich überfluteten Geländes in das Gebiet oberhalb des Stauwehrs gelangen.7. Die in der Elbe nachgewiesenen Verhaltensweisen bestätigen sich nach früheren Beobachtungen anderer Autoren und eigenen Ermittlungen in den Flüssen Weser und Ems.8. Das periodische Auftauchen der Glasaale an der Wasseroberfläche im oberen Tidebereich wird dadurch erklärt, daß sich aus der vorangegangenen Zeit im wechselnden Flut- und Ebbstrom ein innerer Rhythmus erhalten hat, der abwechselnd eine positive und negative Rheotaxis auch ohne äußeren Zeitgeber noch einige Tage in Erscheinung treten läßt.9. Die Frage, wie die Aale im äußeren und inneren Küstenabschnitt Ebbe-oder Flutströmungen perzipieren und welche Bedeutung möglicherweise lunaren Einflüssen zukommt, wird diskutiert.

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Behaviour of elvers(Anguilla anguilla) during their migration in estuaries of German North Sea rivers

Upon observing the tidal section of the Elbe River, it was found that the elvers (Anguilla anguilla L.), gathering in shoals immediately below the surface of the water, moved only in the early part of the morning. The area of such below-the-surface movements probably ranges from approximately the uppermost point reached by the flood stream to the uppermost point of measurable vertical tidal fluctuations. In the Elbe this area spreads over a distance of about 50 kilometers. Subsequent to such early day movements, the elvers bury themselves in the river bottom. They are passively carried upstream and, following a midnight high tide, appear at the upper boundary of the flood stream. Above this boundary the elvers swim against the current and close to the river bank. If the direction of the downstream current occasionally changes, for example, due to the special morphology of the river banks, the elvers swim with it, but, in principle, they do not alter their upstream direction of swimming. The elvers reach the section above the weir near Geesthacht only when the water is extremely high and the surrounding land flooded, or, though more seldom, through the sluice. Behavioural patterns and hydrographical relationships similar to those described for the Elbe were found by other authors to exist in the Weser and Ems Rivers. The periodic emergence of the elvers to the surface in the upper tidal region can be explained by previous experiences and resulting internal rhythms, which do not need reinforcements by external factors over shorter periods of time. The question of how ebb and flood streams are perceived in outer and inner coastal waters by the elvers, as well as possible direct and indirect influences through the moon, are discussed.

     

Apoptosis: the skin from a new perspective

In this review we present skin biology from the perspective of apoptosis. We stress that apoptosis acts as an important homeostatic and defence mechanism in the developing and mature epidermis. Programmed cell death functions in establishing the architecture of the human epidermis and its appendages during development by deletion of stage-specific cells and in the adult epidermis by elimination of excess and abnormal cells. Arguments are presented to support the hypothesis that known regulators of keratinocyte growth may act as survival factors which suppress the cell death pathway. Surviving cells continue to divide until they encounter anti-proliferative factors. Then, unless cells are severely injured and die of necrosis, they will terminally differentiate to death or will die by apoptosis. The mechanisms controlling keratinocyte maturation are co-ordinated with cell position within the epidermal strata. Inappropriate regulatory signals or response of a cell inappropriate to its state will activate apoptosis. Parallels between terminally differentiating keratinocytes and apoptotic cells imply that terminal differentiation and apoptosis proceed along the same death pathway. For terminally differentiating cells, however, this pathway is more elaborate because it allows expression of tissue- and differentiation-specific genes. A model is presented that integrates apoptosis and keratinocyte growth and differentiation.     

Avian feather development: relationships between morphogenesis and keratinization

Morphogenesis and expression of the alpha and beta keratin polypeptides are controlled by epidermal-dermal interactions during development of avian skin derivatives. We have examined the relationship between morphogenesis of the embryonic feather and expression of the feather alpha and beta keratins by routine histology, indirect-immunofluorescence, and SDS-PAGE. Initially beta keratins are expressed only in the feather sheath. Following barb ridge morphogenesis beta keratins can be detected in the barb ridge, coincident with the differentiation of barb ridge cells into eight distinct morphological types. Beta keratinization occurs in gradients; from feather apex to base, and from periphery of the barb ridge to the interior. The onset of beta keratinization in the barb ridges is paralleled by an increase in the major feather beta keratin polypeptides, as detected by SDS-PAGE. The alpha keratins are present in both the periderm and feather sheath at early stages of feather development, but become greatly reduced after hatching, when the down feather emerges from the sheath.     

The lysolipid sphingosine modulates pyrophosphatase activity in tonoplast vesicles and isolated vacuoles from a heterotrophic cell suspension culture of Chenopodium rubrum

The proton pumping activity of the tonoplast (vacuolar membrane) H⁺-ATPase and H⁺-pyrophosphatase (H⁺-PPase) has been studied on a tonoplast-enriched microsomal fraction and on intact vacuoles isolated from a heterotrophic cell suspension culture of Chenopodium rubrum L. in the presence of the lysosphingolipids D-sphingosine, psychosine (galactosylsphingosine) and lysosulfatide (sulfogalactosyl-sphingosine). Sphingosine strongly stimulates (K_a= 0.16 μ M ) the PPase activity, assayed both as ΔpH formation across the tonoplast vesicle membrane, and as reversible clamp current measured by the whole-vacuolar mode of the patch-clamp technique. Psychosine showed a minor, and lysosulfatide no stimulatory effect. No effect upon the ATPase activity has been observed. No sphingosine-induced change could be observed in the affinity of the PPase for its substrate (apparent K_m= 10 μ M MgPPi). We tentatively conclude that sphingosine, which is known as a potent inhibitor of the protein kinase C in animal cells, may be a regulator of the plant vacuolar PPase.     

Native plasmids of Fusobacterium nucleatum: characterization and use in development of genetic systems

Three native plasmids of Fusobacterium nucleatum were characterized, including DNA sequence analysis of one plasmid, pFN1. A shuttle plasmid, pHS17, capable of transforming Escherichia coli and F. nucleatum ATCC 10953 was constructed with pFN1. pHS17 was stably maintained in the F. nucleatum transformants, and differences in the transformation efficiencies suggested the presence of a restriction-modification system in F. nucleatum.