Electron microscopic investigation of the surface membrane structures of the fat-cell and of their changes during depletion of the cell

Summary The main purpose of this paper was to investigate with the electron microscope the structural relationship between the fat cell and the surrounding connective tissue under various functional conditions and thereby to solve questions not decided in the past which concern the membrane of the fat-cell. Outside the well defined plasma membrane two layers were observed, one of lesser electron density next to the plasma membrane and another denser line which separates the former from the connective tissue ground substance. Fundamentally this three-layered surface membrane complex (Robertson) is the same as described by numerous authors in other cells bordering on connective tissue. However, the changes occurring in the surface membrane complex during depletion of fat cells are of special interest. The numerous long processes formed by the cell during the loss of fat in starvation are retracted in extreme depletion. At this time a pericellular space opens between the outer lamella and the plasma membrane. While the less dense material apparently becomes liquified the outer lamella of the surface membrane complex remains in contact with the connective tissue ground substance. These observations made it possible to interpret the surface membrane structures of the fat-cell as consisting, beside the plasma membrane, of a material derived from the ground substance, which is analogous to Robertson's gap substance at the surface of the Schwann cell, and of a limiting membrane toward the ground substance. The nature and possible derivation of the extracellular layers are discussed and the general functional significance of the surface membrane complex is emphasized. These considerations support the repeatedly raised objection against the use of the histological term basement membrane for the submicroscopic structures. During the depletion of the fat cell intensive micropinocytosis occurs regularly in the plasma membrane. It is suggested that the pinching off of numerous pockets may effect the elimination of membrane material in conjunction with the decrease in the surface area which has been found to take place in extreme depletion of the fat cell.This work was performed under the auspices of the U.S. Atomic Energy Commission.     

Anti-diabetic activity of <Emphasis Type="Italic">β</Emphasis>-glucans and their enzymatically hydrolyzed oligosaccharides from <Emphasis Type="Italic">Agaricus blazei</Emphasis>

-Glucans were prepared from Agaricus blazei Murill by repeated extraction with hot water. The average molecular weights of -glucans were 30–50 kDa by gel filtration chromatography. Oligosaccharides (AO), derived from hydrolyzing -glucans with an endo--(16)-glucanase from Bacillus megaterium, were mainly di- and tri-saccharides. Though -glucans and AO both showed anti-hyperglycemic, anti-hypertriglyceridemic, anti-hypercholesterolemic, and anti-arteriosclerotic activity indicating overall anti-diabetic activity in diabetic rats, AO had about twice the activity of -glucans with respect to anti-diabetic activity.     

A female infant with monosomy 21

Summary A female infant with total monosomy 21 identified by Q banding is described. The main clinical features were hypertonia, prominent occiput, hypertelorism, antimongoloid slant of the eyes, broad nose, antimongoloid character of dermatoglyphics. Both parents are phenotypically as well as karyotypically normal.     

Hexahydroindanone derivatives of steroids formed by Rhodococcus equi

Summary A mutant strain of Rhodococcus equi accumulates three metabolites from the androst-4-ene-3,17-dione or from its degradation intermediate, 3a-H-4(3'-propionic acid)-7a-methylhexahydro-1,5-indanedione (MEPHIP). These three metabolites are: 3a-H-4a(3'-propionic acid)-5-hydroxy-7a-methylhexahydro-1-indanone--lactone (HIL); 3a-H-4(3'-trans acrylic acid)-5-hydroxy-7a-methylhexahydro-1-indanone (^2'-5-hydroxy-MEPHIP); and 3a-H-4(3'-hydroxy-3'-propionic acid)-5-hydroxy-7a-methylhexahydro-1-indanone (3'-hydroxy-HIL). The behaviour of this mutant allows us to propose a pathway for degradation of the intermediates, methylperhydroindanone propionates. However, during this degradation, the side-chain propionate was eliminated by a-oxidation mechanism. Offprint requests to: A. Miclo     

Glucoside synthesis by glucoamylase or \-glucosidase in organic solvents

Summary Glucoamylase and -glucosidase have been shown to catalyse respectively - or -glucoside synthesis in water-organic mixtures. The effects of water content, water activity, organic solvent nature and substrate concentration on the reaction yield were studied. Maximum yields were obtained with 10% (v/v) of water in primary alcohol systems and 15% in diol systems. -glucosidase has given the best yields. This enzyme has presented for alcohols a higher affinity than glucoamylase. A difference of stereospecificity was observed with the two enzymes and diols.     

Chloroplast volume: cell water potential relationships and acclimation of photosynthesis to leaf water deficits

Studies were undertaken to determine if there is an association between nonstomatally-mediated acclimation of photosynthesis to low water potential (w) and the maintenance of chloroplast volume during water stress. Spinach plants either kept well watered throughout their growth (non-acclimated), or subjected to water stress such that leaf w dropped to -1.5 megapascals (MPa) and then were rewatered (acclimated) were subjected to drought episodes. During these stress periods, photosynthesis was maintained to a greater extent in acclimated plants as compared to non-acclimated plants at w below -1 MPa.Estimates of internal leaf [CO₂] suggested that photosynthetic acclimation to low w was not primarily due to altered stomatal response. As w dropped from initial values, a decline in steady state levels of ribulose 1,5-bisphosphate (RuBP) occurred in both non-acclimated and acclimated plants. RuBP decline was less severe in acclimated plants.Low w effects on chloroplast volume in non-acclimated and acclimated plants were estimated by measuring the volume of intact chloroplasts isolated from plants in solutions which were made isotonic to declining leaf osmotic potential during the drought episodes. Chloroplast volume was maintained to a greater extent at low w in acclimated, as compared with non-acclimated plants. Although substantial osmotic adjustment occurred in both non-acclimated and acclimated plants, the extent of osmotic adjustment was the same. These data were interpreted as supporting the hypothesis that cellular-level acclimation to low w is associated with chloroplast volume maintenance, and this physiological acclimation is correlated with enhanced photosynthetic capacity of the leaf at low w.Abbreviations [CO₂]_i internal leaf CO₂ concentration - s osmotic potential - RWC relative water content - RuBP ribulose 1,5-bisphosphate - w water potential     

Development of high frequency multiple shoot formation in Persian clover (<Emphasis Type="Italic">Trifolium resupinatum</Emphasis> L.)

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1M BA and 1M IBA or 14.1M BA and 1M IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and -naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8M IBA.     

Water accessibility to plant roots in different soil structures occurring in the same soil type

The capacity of a soil to supply roots with water and nutrients for crop growth is important when defining sustainable land management which implies maintenance of production and reduction of production risks. Not only the amount of available water is important but also its accessibility, which differs among different soil structures. Different structures within one soil series were associated with three types of management: (i) conventional, temporary grassland (Conv), (ii) biodynamic, temporary grassland (Bio) and (iii) conventional permanent grassland (Perm). Transpiration of barley plants, under identical circumstances, and the associated rooting patterns, were measured in five large undisturbed cores from each of the three soil structures. Management had significantly changed bulk density, organic matter content and porosity. Measured transpiration showed significant differences with highest amounts for Perm followed by Conv and lowest amounts for Bio. Rooting pattern characteristics, defined as the relation between a series of hypothetical extraction zones around each root and the volumes of excluded soil were determined for the three structures. These rooting pattern characteristics were most favourable for Perm, followed by Bio and Conv, respectively. The water supply characteristics, defined as the number of days the soil can satisfy a transpiration demand of 5 mm d^-1 as a function of a hypothetical extraction zone, reflects the capacity of the soil to supply roots with water. These water supply characteristics combined with the rooting pattern characteristics were used to quantify the accessibility of soil water. Accessibility was highest for Perm and Conv with 95% and 94% respectively, followed by Bio with 68%. When used in a simulation model and compared with simulations implicitly assuming total accessibility, measured transpirations were better simulated by introducing the expression for water accessibility.     

Clearance rates of bacteria-sized particles by freshwater ciliates,measured with monodisperse fluorescent latex beads

Summary Monodisperse fluorescent latex particles with diameters of 0.57 and 1.04 m have been used to measure the clearance rates of bacteria-sized particles by two ciliates from a Norwegian lake. The clearance rates by Epistylis rotans on these particles were in the range 0.23 to 1.26 l ind^-1h^-1, and by Strombidium sp. 0.26 to 0.90 l ind^-1 h^-1. The clearance rates varied with food level and temperature. Possibilities and limitations of the suggested method are discussed.     

Significance of terminalαDgal(1-4)βDgal residues in urinary tract epithelium

A monoclonal antibody to a terminal residue ofDgal(1-4)Dgal, known as part of the receptor for p-fimbriatedEscherichia coli, was used to study the occurrence of this carbohydrate residue in urinary tract tissue from different animal species and man. Sections of bladder, ureter, and kidney tissue from man, monkey, guinea pig, rat, and mouse were investigated by an avidin-biotin-peroxidase staining method that proved useful for demonstrating carbohydrate structures in tissues. TerminalDgal(1–4)Dgal residues were widely distributed in epithelial cells of the urinary tract in most of the species studied. Cells expressing this carbohydrate were also demonstrated in urinary sediment from healthy women. Experimental infection could not prove that this terminal structure alone was essential for establishment of pyelonephritis.     

Somaclonal variation in insect‐resistant transgenic sugarcane (Saccharum hybrid) plants produced by cell electroporation

A population of 42 transgenic sugarcane ( hybrid, cv. Ja605) clones expressing a truncated cryIA(b) gene from Bacillus thuringiensis was evaluated in field trials under artificial borer (Diatraea saccharalis Fab.) infection. Five clones displaying the highest borer tolerance were selected and analysed with molecular tools (RAPD, AFLP and RAMP) to verify genomic changes. Results of field trials provided evidence both for the expression of the resistance trait and for the occurrence of limited but consistent morphological, physiological and phytopathological variation, as compared with control plants regenerated from dedifferentiated culture without transformation (C1control) or with plants that were clonally propagated in the field (C2control). The five elite transgenic clones, selected for consistent borerresistance and good agronomic traits, were further evaluated in a large scale field trial. It was found that the majority of agronomic and industrial traits were those of the original cv. Ja605, but that a small number of qualitative traits was different. DNA changes were verified in the five selected clones. A total of 51 polymorphic DNA bands (out of the 1237 analysed bands) was identified by extensive AFLP and RAMP analysis, thus showing rare but consistent genomic changes in the transgenic plants, as compared with C1 and C2control plants. It is proposed that the increased variability verified in transgenic plants by field trials and DNA analysis is essentially correlated with cell growth in the dedifferentiated state during the transformation procedure. The results, which are consistent with those published in the case of other transgenic plant populations, are discussed in the context of selecting approaches to gene transfer that minimize somaclonal variation. This is important especially in cases, such as that of sugarcane, where success of backcrosses to restore the original genotype is made difficult by the complex ploidy state of the plant.     

Studies on cellulase production by Clostridium thermocellum

Summary Clostridium thermocellum ATCC 27405 (and its improved cellulase-producing mutant, AS-39) is an anaerobic thermophile that produces endo--glucanase and exo--glucanase when grown on cellobiose or cellulose as major carbon source (Shinmyo et al. 1979). The site of cellulase accumulation was at least 95% extracellular. Optimum conditions for endo--glucanase production in flasks included 1% (w/v) cellobiose, 0.2% (w/v) urea as a nitrogen source, 0.1 M morpholinopropane-sulfonic acid buffer, an initial pH of 7.4, and a yeast extract concentration of 0.6% (w/v). An improved medium (GS medium) was devised for future studies. Xylan was degraded by an extracellular enzyme (s) produced during cultivation on cellobiose, although C. thermocellum does not grow on xylan.     

Comparisons of the Genomes of New Giant Phages Isolated from Environmental Pseudomonas aeruginosa Strains of Different Regions

To study the genome diversity of bacteriophages from geographically distant natural populations, new giant KZ-like Pseudomonas aeruginosa phages isolated in two different regions were compared with earlier known phages of three species (KZ, Lin68, EL). A broad spectrum of lytic activity was demonstrated for all KZ-like phages. Phages of the KZ species proved to be common in natural populations of various regions, while EL- and Lin68-related phages were extremely rare. Most KZ-related phages had unique DNA restriction patterns, but the differences between these were only minor, and the genomes did not contain nonhomologous fragments. The spectrum of capsid polypeptides proved to be conserved in each species, and was proposed as a character necessary and sufficient for express classification of phages with an accuracy of species. Phages isolated in different geographical regions showed no substantial difference. Some phages only slightly differing in DNA restriction pattern from KZ may be used to study the origin of KZ genes coding for orthologs of proteins of unrelated species (other phages, pathogenic bacteria, eukaryotes).     

The RF-cinematogram

We present a spike-triggered averaging method capable of mapping the visual receptive fields of several neurons simultaneously. The stimulation is general and the mapping proceeds automatically without the need to match the stimulation to the cells' preference for position, orientation, direction, etc. The maps are spatiotemporal; receptive field (RF) structures are quantitatively determined in three dimensions: the two dimensions of visuotopic space, and time. The method presented is one of a family of reverse correlation or spike-triggered averaging techniques (DeBoer and Kuyper 1968) capable of revealing linear aspects of stimulus-response coupling. The formal relationship of these methods to stimulus-response crosscorrelation is shown. The analysis is extended to provide some second-order axis-of-motion information (direction marks). The stimulus is a constantly illuminated, randomly jumping bright or dark spot, not an elongated bar. Spot diameters between one-third to 1 × RF width are effective. The method ascertains for each recorded action potential or spike the prior visual field position of the spot. The average or most probable spot positions define the receptive field spatially. Repeating the process for a succession of times prior to observed spikes defines the field temporally, presented here as a succession of spatial maps. We term this portrayal a receptive field cinematogram, RFc or ciné. The RFc reveals and economically portrays the spread of excitability and suppression across the receptive field, culminating in the generation of a spike. RFcs for LGN neurons and for simple cells recorded in cat cortical areas 17 and 18 are presented and interpreted in terms of classic ON/OFF regions. The availability of temporal information permits the separation of an excitatory exit response, generated when a moving bright spot leaves an OFF region, from an excitatory entrance response occurring when a bright spot enters an ON region, because these responses occur at different times (exit responses earlier). Spike emission remains coupled to (cross-correlated with) stimulus events over time periods as long as 96 ms, implying that some stimulus drive or afferent visual input is delayed by as much as 96 ms more than other input. This is a striking instance of temporal dispersion in the visual system. In some cells, said to be spatiotemporally inseparable, the delay (latency) varies systematically across the visual field; i.e., the place for optimal stimulation varies with the time prior to spike emission. In these cells, the RFc shows receptive field structures which move across the visual field over trajectories equal to approximately twice the total conventional RF width. Exit and entrance responses, on the other hand, arise in a simple way from separated ON and OFF RF subregions. ON/ OFF mechanisms thus appear unrelated to spatiotemporal inseparability. The RFc method is easily automated, efficient, and characterizes multiple RFs simultaneously, as required in work with multiple electrode arrays.     

New expanded bed adsorbents for the recovery of DNA

A 20–40 m pellicular high density (3.7 g cm^–3) expanded bed material has been designed for the capture of DNA and other large macromolecules. Anion exchangers fashioned out of these supports exhibited dramatically enhanced DNA binding capacities over commercial anion exchange adsorbents (6 mg ml^–1, c.f. 50 g ml^–1 at 10% breakthrough), due to a combination of small particle and fuzzy surface architecture created through the coupling of polyethylene imine chains.     

Regenerations- und Heilungsversuche an der Proactinula vonEctopleura dumortieri (Athecatae-Anthomedusae) unter Anwendung der Zeittransformation

Zusammenfassung 1. Entwicklungsstadien vonEctopleura dumortieri van Beneden (Tubularidae, Athecatae-Anthomedusae) vom 8. Teilungsschritt (256 Blastomere) an bis zur Proactinula (Sternchen-Stadium) mit bereits beweglichen Tentakeln wurden mikrochirurgischen Eingriffen ausgesetzt.2. Einschnitte in den Körper der Proactinula zwischen zwei Tentakeln heilen in zwei Stunden zu, ohne Spuren der Verletzung zu hinterlassen.3. Trotz Amputation sämtlicher Tentakel eines Sternchens entsteht eine sich normal verhaltende Proactinula, jedoch mit reduzierter Tentakelzahl.4. Völlige Durchtrennung eines Sternchens wird gut vertragen. In jedem Fall bilden die Teilstücke Tentakel aus, die sich meist normal strecken und bewegen.5. Ob Regeneration oder Neubildung von Fangarmen nach Verletzungen vorliegt, läßt sich nur durch Versuche an frühen Entwicklungsstadien entscheiden, und zwar nicht für den einzelnen Tentakel, sondern nur an Hand der Summe aller Tentakel sämtlicher Teilstücke; diese beträgt oft mehr als das Doppelte der Maximalzahl, welche eine normale Proactinula zu bilden vermag.6. Aus allen 10 Teilstücken eines Sternchens entstanden kleine Proactinulae mit 3 bis 4 normalen Tentakeln, zum Teil deutlich erkennbarer aboraler Stielanlage und vorwachsender Basis der Oraltentakel.7. Aus nicht zu kleinen Teilstücken können ganze Polypen entstehen, die sich mit langem Stiel festsetzen; sie sind zum Beutefang befähigt.8. Die Vitalität der Regenerate und Neubildungen sowie ihr Bewegungsverhalten werden durch Zeitraffer-Filmaufnahmen kontrolliert.

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Regeneration and healing experiments on the proactinula ofEctopleura dumortieri (Athecatae-Anthomedusae) by employing time transformation

Micro-surgical operations were performed on different developmental stages ofEctopleura dumortieri van Beneden (Tubularidae, Athecatae-Anthomedusae). The tested stages, i.e., 8th cell-division until proactinula (Sternchenstadium), revealed a considerable capacity for healing, regeneration and formation of new body parts. After cutting the embryo up — into a maximum of 10 parts — all fragments assumed the shape of a sphere, formed tentacles and, in part, aboral stem-anlagen as well as the bases of the oral tentacles. The fragments usually form twice as many tentacles as do the normal actinula-stages. If the fragments are not too small, polyps are formed which may settle down and take up food. These polyps differ from normal ones only in that they have fewer tentacles. Vitality and locomotory behaviour of regenerates and newly formed body parts were assessed by means of time lapse movie pictures.

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Herrn Professor Dr.Friedrich Krüger zum 65. Geburtstag am 18. August 1967 gewidmet.     

Ecophysiological analysis of woody species in contrasting temperate communities during wet and dry years

This study employed an intensive sampling regime in which leaf gas exchange and tissue-water relations were measured simultaneously on the same leaf at midday on 19 tree species from three distinct forest communities during wet (1990) and dry (1991) growing seasons. The study sites were located on a xeric barrens, a misic valley floor, and a wet-mesic floodplain in central Pennsylvania, United States. The xeric, mesic, and wetmesic sties had drought-related decreases in gravimetric soil moisture of 53, 34 and 27%, respectively. During the wet year, xeric and mesic communities had high seasonal mean photosynthetic rates (A) and stomatal conductance of water vapor (g _wv) and low midday leaf water potential (), whereas the wet-mesic community had low A and g _wv and high midday . The mesic and wet-mesic communities had dry year decreases in predawn , g _wv and A with the greatest drought effect occurring in the mesic community. Regression analysis indicated that species from each site that exhibited high wet-year A and g _wv tended to have low midday . This trend was reversed only in the mesic community in the drought year. Despite differences in midday , all three communities had similar midday leaf turgor pressure (_p) in the wet year attributable to lower osmotic potential at zero turgor ( ⁰ ) with increasing site droughtiness. Lower wet year ⁰ in the xeric community was due to low symplast volume rather than high solute content. Species with the lowest ⁰ in the wet year often did not have the lowest ¹⁰⁰ possibly related to differences in tissue elasticity. Moreover, increased elasticity during drought may have masked osmotic adjustment in ¹⁰⁰ but not in ⁰ , via dilution of solutes at full hydration in some species. Despite the sampling regime used, there were no relationships between gas exchange and osmotic and elastic parameters that were consistently significant among communities or years. This result questions the universal, direct effect of osmotic and elastic adjustments in the maintenance of photosynthesis during drought. By including a large number of species, this study provided new insight to the ecophysiology of contrasting forest communities, and the community-wide impact of drought on contrasting sites.     

Die Biologische Anstalt Helgoland Gegenwart und Zukunft

Ohne ZusammenfassungIn der Septemberausgabe des Der Helgoländer (offizielles Veröffentlichungsorgan des Vereins Helgoland e. V. — Geschichte und Kultur der Deutschen Bucht und zugleich Sonderausgabe der Cuxhavener Allgemeinen für die Insel Helgoland) erschienen anläßlich des 75 jährigen Bestehens der Biologischen Anstalt Helgoland insgesamt 19 Beiträge über unsere Anstalt. In etwas veränderter Form wird hier der Einleitungsaufsatz — mit freundlicher Genehmigung der Cuxhavener Allgemeine — abgedruckt.     

Molecular analysis of the inheritance of the S-5 locus,conferring wide compatibility in Indica/Japonica hybrids of rice (O. sativa L.)

RFLP analysis was conducted on a population derived from a three-way cross to determine the location of the hybrid sterility locus, S-5, in relation to mapped molecular markers and to identify markers that would be useful for selection in breeding. S-5 is of interest to rice breeders because it is associated with spikelet sterility of F₁ hybrids in Indica/Japonica crosses. Identification of an S-5 allele which confers fertility in Indica/Japonica hybrids when introgressed into either the Indica or the Japonica parent has been reported. Varieties carrying this S-5 ⁿ allele are known as wide compatibility varieties (WCV). Our data suggests that RFLP marker RG213 on chromosome 6 is closely linked to the S-5 locus and can be efficiently used to identify wide compatibility (WC) lines. RG213 is a single-copy genomic clone that detects three bands of different molecular weights in DNA from Japonica (Akihikari) and Indica (IR36) varieties and WC line (Nekken 2). We demonstrate that the three alleles detected by this marker could be used to trace the inheritance of the wide compatible phenotype in breeders' material.     

Forskolin Modulates Acetylcholine Receptor Gating by Interacting with the Small Extracellular Loop Between the M2 and M3 Transmembrane Domains

1. Forskolin acts as an allosteric modulator of muscle-type nicotinic acetylcholine receptors. Receptors from mouse muscle and Torpedo electroplax demonstrate differential sensitivity to inhibition by forskolin. Previous work from this laboratory suggested that the subunit is responsible for this differential sensitivity.2. We have used a series of mouse/Torpedo species-chimeric subunits to further define the site of forskolin interaction with the subunit. Analysis of the patterns of forskolin inhibition of receptors containing mouse/Torpedo chimeric subunits along with the mouse , , and subunits suggests that forskolin interacts with the small extracellular domain that links the M2 and M3 transmembrane domains (the M2–M3 linker).3. We suggest that the M2–M3 linker domain plays an important role in the transduction of ligand binding to the conformational changes that result in channel opening.