Coral and Fish Biocoenosis: Ecological Cells Gradually Maturing in Complexity, Species Composition and Energy Turnover

Long-term observations on coral units and their coral fishes, as well as observations on the growth of colonies of marked corals in the Gulf of Aqaba (Red Sea), have revealed the relationship between coral dimension and complexity and the ecomorphology and sociobiology of the inhabiting fishes. In this study, coral fishes are identified as species that, following the planktonic interval, settle and remain in a selected coral or its immediate vicinity. Parallel with growth of the refuge, the number of fishes in such a coral–fish association (=ecological cell) increases to a certain asymptote, forming a stable symbiotic entity of fish species, specific in composition and biomass. The initial cell begins with 4–5-year-old branching corals that harbors 3–4 cryptic species of gobies (Paragobiodon and Gobiodon species). This is followed by several stages of coral growth and increase in fish species and biomass, especially of pomacentrids, anthiases, pseudochromids and blennies. The water column or functional space from which the fishes harvest plankton, their main food source, starts at around a 0.2m³ column in the young colonies, and ends with a water column of around 500m³ in the mature ecological cells of large coral knolls. At the mature stage, large cells harbor up to 100 diurnal and nocturnal species of fish, permanently using this refuge and forming an integrated collective. The territorial limits of the piscivorous groupers (Cephalopholis species) generally establish the dimensions of these ecological cells. Preliminary data on standard energy metabolism of the fishes for mature ecological cells reveal that 800kg food is required each year to maintain this biomass. The biological composition and stability of fish populations of such ecological cells will depend on the biological and physical stability of the host coral population.     

Metabolic variants of intestinal alkaline phosphatase in relation to fat absorption: in situ demonstration with the organ-specific inhibitors l-phenylalanine and l-homoarginine

Summary The localization of rat intestinal alkaline phosphatase has been studied in relation to fat absorption. The observations support a theory of conversion, within the intestinal mucosa, of intestinal type to liver type alkaline phosphatase when the criterion of differential sensitivity to two amino acid inhibitors, l-phenylalanine, and l-homoarginine, is applied.Following a three hour in vivo exposure to mixtures of oleic acid, sodium taurocholate, and lauric acid, the epithelium becomes depleted of its l-phenylalanine-sensitive, intestinal type alkaline phosphatase. At the same time, enriched activity is seen in the lamina propria; this activity is both particulate and diffuse, and is present both in the connective tissue matrix and in cells, including macrophages, eosinophils and lymphocytes. Most of this enzyme is inhibited by l-homoarginine, a property characteristic of liver type alkaline phosphatase.The localization of enzyme-positive particles 0.5 to 1.0 in diameter in both epithelium and lamina propria appears identical to that of particulate fat. A physical association between transport of absorbed fat and metabolic conversion of intestinal type alkaline phosphatase is postulated.This work was aided in part by grants-in-aid [CA-3332-01, K6-CA-18,453] from the National Cancer Institute, National Institutes of Health, U.S.P.H.S. and the John A. Hartford Foundation, Inc.Pre-doctoral trainee, U.S.P.H.S. grant GM01451.Holder of a Juan Marsh Foundation Fellowship, Lemuel Shattuck Hospital.     

Analysis of T cell receptor Vβ gene usage during the course of disease in patients with chronic hepatitis B

The T cell receptor (TCR) is a heterodimeric molecule expressed on the surface of T cells and recognizes foreign peptides presented by the major histocompatibility complex on the surface of antigen-presenting cells or virusinfected cells. Analysis of TCR usage by T cells which recognize hepatitis B virus (HBV) provides further insight into the participation of T cell populations during the course of disease. In this study, we examined the T-cell-proliferative response and the TCR V gene usage of peripheral blood mononuclear cells in 3 patients with clinical evidence typical of chronic hepatitis B. All 3 patients had significant T-cell proliferative responses against HBV core antigen (HBcAg) during the remission stage, while no responses were detected during the acute exacerbation stage. In addition, the TCR V₇ gene was utilized more frequently in T cells recognizing HBcAg during remission, while TCR V₁ and V₂ were utilized at a higher percentage during acute exacerbation. On the contrary, the T cell proliferative response against HBV surface antigen was undetectable and no specific V gene was utilized more frequently by all 3 patients, regardless of disease state. Our longitudinal studies, although based on a small sample of patients, demonstrate that the population of HBcAg-activated T cells alters during the course of disease in chronic hepatitis B patients.     

Isolation of protoplasts by means of a “species-specific” autolysine inChlamydomonas

Summary Prior to fusion, gametes ofChlamydomonas reinhardii discard their cell walls. This naturally occuring phenomenon has provided the basis for a method of protoplast isolation from both gametes and vegetative cells within the genusChlamydomonas. When synchronized cultures of compatibleChlamydomonas gametes are mixed it is possible, after removal of the cells, to obtain a solution having a high cell wall lytic activity. That vegetative and gamete cells after treatment with this gamete-autolysine are indeed protoplasts has been proved by various light and electron microscopical methods.The species specifity of this autolysine, its difference to the previously described sporangialautolysine (Schlösser 1966) and furthermore its use in the large scale production of protoplasts is also described. Since this wall autolysine is a factor produced by the cells themselves at a particular stage in their life cycle it represents a non-foreign agent in contrast to all other enzymic methods previously employed for protoplast isolation.     

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.     

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.     

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     

Ultrastructure of the new neuromuscular junctions formed during reinnervation of rat soleus muscle by a “foreign” nerve

Summary In rats the fast fibular nerve was transposed to the slow soleus muscle outside the original innervation band. Formation of new neuromuscular junctions was induced by cutting the soleus nerve after different periods of time. The morphological maturation of these junctions was studied by electron microscopy.New neuromuscular junctions do not form when the original innervation is left intact.Three to five days after denervation, vesicle-laden terminal boutons contact muscle fibers with only the basal lamina of the latter intervening. Three weeks after denervation, most boutons are larger and postsynaptic folds are present, although younger stages are also seen. Sixteen weeks after denervation, the neuromuscular junctions appear mature. This corresponds well with electrophysiological findings in the same material.The fully developed neuromuscular junctions sixteen weeks after denervation possess postsynaptic folds similar to those of normal fast muscle fibers. This suggests that the fast fibular nerve rather than the slow soleus muscle fibers determines the morphology of the postsynaptic folds.Possible trophic neuromuscular interactions are discussed.The authors are indebted to Mrs. Jorunn Line Vaaland, Miss Bjørg Riber, and Miss Berit Branil for technical assistance. Dr. T. Lømo and Dr. C. Slater have contributed constructive criticism and advice     

Lateral bud culture of papaya (Carica papaya L.) for clonal propagation

Lateral buds may be preferred to shoot tips for in vitro propagation of papaya because of its unbranched nature. Proliferating shoot cultures from lateral buds appeared extremely compact with shortened internodes and leaf lamina of the cytokinin level (BAP 2 M) reported for multiple shoot production from shoot tips. ZEA (4 M) and 2iP (8 M) although reduced the proliferation rate, resulted in better growth of the shoot from lateral bud. Rooting was observed with IBA 20 M but plantlets so produced remained stunted.     

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).     

Micropropagation of the silk tassel bush,Garrya elliptica Dougl.

Proliferating cultures ofGarrya elliptica were established from shoot tip and nodal explants grown on woody plant medium containing benzyladenine. Optimum numbers of healthy shoots were induced on media containing 1.25 M benzyladenine; higher concentrations caused more hyperhydricity and less extension. Following growth for 28 days on quarter-strength woody plant medium containing 0.2% activated charcoal, 90% of shoots rooted on the same medium lacking charcoal, but with 2.5 M indolebutyric acid. Survival during weaning was poor, and was strongly correlated with the time of year at which plants were removed from culture.Abbreviations WPM McCown & Lloyd's (1980) Woody Plant medium - BA Benzyladenine - IAA Indoleacetic acid - IBA Indolebutyric acid - NAA Naphthaleneacetic acid     

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.     

The effect of different growing conditions on water relations parameters of leaf epidermal cells of Tradescantia virginiana L.

Summary Tradescantia virginiana L. plants were cultivated under contrasting conditions of temperature, humidity, light quality and intensity, and nutrient status in order to investigate the effect of growth conditions on the water relations parameters of the leaf epidermal cells. Turgor pressure (P), volumetric elastic modulus (), half-time of water potential equilibration (T _1/2), hydraulic conductivity (L _p ) were measured with the miniaturized pressure probe in single cells of the upper and lower epidermis of leaves. Turgor differed (range: 0.1 bar to 7.2 bar) between treatments with lowest values under warm and humid conditions and additional supply of fertilizer, and highest values under conditions of low air humidity and low nutrient supply. The volumetric elastic modulus changed by 2 orders of magnitude (range: 3.0 bar to 350 bar, 158 cells), but was only affected by the treatments, in as much as it was dependent on turgor. The turgor dependence of , measured on intact leaves of T. virginiana, was similar to that for cells of the isolated (peeled) lower epidermis, where as a function of turgor was linear over the whole range of turgors. This result has implications for the discussion of pressure/volume curves as measured by the pressure bomb where changes in bulk leaf are frequently discussed as adaptations to certain treatments. The measurements of the hydraulic conductivity indicate that this parameter varies between treatments (range of means: 2.4×10^-6 cm s^-1 bar^-1 to 13.4×10^-6 cm s^-1 bar^-1). There was a negative correlation for L _p in cells of intact leaves as a function of turgor which was altered by the growing conditions. However, a correlation with turgor could not be found for cells from isolated epidermis or cells from a uniform population of plants. The large variation in L _p from cell to cell observed in the present and in previous studies was accounted for in a study of 100 cells from a uniform population of plants by the propagation of measurement errors in calculating L _p . The results suggest that in T. virginiana cellular water relations are changed mainly by the turgor dependence of .     

Immunocytological detection and localization of a peptide reacting with an α-endorphin antiserum in the corticotropic and melanotropic cells of the trout pituitary (Salmo irideus Gibb)

Summary In the pituitary of the trout, the corticotropic and melanotropic cells display a strong immunocytological reaction with -endorphin antiserum. This reaction persists even when a-endorphin antisera treated with -1-24 ACTH or -MSH are used. In the absence of pharmacological tests on the endorphic potencies of the compounds involved in the immunoreaction, it is not yet clear whether this reaction is due to the presence of an -endorphin-like peptide or simply an immunologically related peptide without the properties of endorphin. However, the presence of such peptides in the fish pituitary is interesting from the comparative point of view.     

Fluorescence studies of normal and sickle beta apohemoglobin self-association

The acrylamide quenching of the intrinsic tryptophanyl fluorescence of normal and sickle apohemoglobins has been studied in 0.05 M potassium phosphate buffer,pH 7.5, at 5°C over a protein concentration range from 1 to 50M. Analysis of quenching dynamics revealed a strong dependence on acrylamide concentration for the intrinsic fluorescence of both normal and sickle apohemoglobins, suggesting that one tryptophanyl residue [presumably that at position 37(C3)], was more accessible to collisional quencher than the other tryptophanyl residue [15(A12)]. Additional studies, which altered viscosity and subunit assembly experimental parameters, supported the assignment of residue 37 as the more dynamically accessible residue. Finally, the quenching data were also found to be dependent on protein concentration, implying that this difference in the mobility between the two residues is a sensitive probe of self-aggregation. Extrapolated dynamic quenching constants at low concentration of acrylamide were used to estimate the dimer-monomer equilibrium dissociation constants of normal and sickle apohemoglobins, and were found to be 5.6 and 2.4M, respectively, thus demonstrating distinct self-association properties of _A and _S apohemoglobins.     

Untersuchungen über die Ultrastruktur der Gonaden von Chironomus (Dipt.)

Zusammenfassung Im Ovar von Chironomus sind in Phase 1 des 4. Larvenstadiums polygonal abgeflachte Innenzellen von kleineren Außenzellen umgeben, die Bakteroide und Phagosomen enthalten; zwischen den Innenzellen liegen unregelmäßige Zelltrümmer (keimbahnbegleitende Substanzen). Zu Beginn der Ovariolenbildung werden in Phase 3 durch Spalträume zwei Schichten der Außenzellen voneinander getrennt, von denen die innere (Follikel- und Eikanalepithel) regelmäßige Buchten bildet. In diese Buchten wandern von innen Zellpaare ein, die an synaptischen Komplexen bzw. multiplen Chromatinstrukturen als Ei- und Nährzellen kenntlich sind. Zwischen beiden Zellen sind Fusome häufig, die später in eigentümlicher Weise geschlossen werden. Zwischen den Eikanalzellen entsteht in Phase 5 durch Spaltbildung der Eikanal; in Phase 7 sind die Eikanalzellen auffallend glykogenreich. Kurz vor der Vitellogenese treten im Bereich der Oocyte Membransysteme und annulated lamellae auf; akzessorische Kerne werden als Ausstülpungen des Oocytenkernes gebildet und später abgeschnürt. In Phase 9 sind an der Peripherie der Eizelle Mikrovillisäume und Pinocytosebläschen sichtbar. Die distalen Zellen der Ovariole haben Eioder Nährzellcharakter, sind aber bei Ch. melanotus nicht von Follikelzellen umgeben und werden beim weiteren Ovariolenwachstum reduziert. Trotz extrem geringer Nährzellzahl der Follikel scheint das Chironomus-Ovar funktionell nicht von anderen polytroph meroistischen Insektenovarien unterschieden.

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Ultrastructure of Chironomus (Dipt.) gonads1. Normal development of ovaries during the fourth larval instar

Summary In the ovary of Chironomus during phase 1 of the fourth larval instar, polygonally flattened inner cells are surrounded by smaller outer cells which contain bacteroids and phagosomes. Irregular cell remnants (germ line accompanying substances) lie among the inner cells. At the beginning of ovariole formation in phase 3, two layers of outer cells are separated by the formation of fissures. The inner layer of these cells (follicle- and egg-passage epithelium) forms regular invaginations. Cell pairs, identified as oocytes and nurse cells by synaptic complexes or multiple chromatin structures, wander from inside into the invaginations. Frequently between the two cells are fusomes, which later close in a characteristic manner. During phase 5, an egg passage is formed as a fissure among the egg-passage cells. During phase 7, the egg passage cells are conspicuously full of glycogen. Shortly before vitellogenesis membrane systems and annulated lamellae appear in the region of the oocyte. Accessory nuclei are formed by a tieing-off of projections of the the oocyte nucleus. During phase 9, microvilli and pinocytotic vesicles can be seen at the periphery of the oocyte. The distal cells of the ovariole are of oocyte or nurse cell nature, but in Ch. melanotus they are not surrounded by follicle cells and are reduced during further ovariole growth. In spite of the extremely small number of nurse cells in the follicle, the Chironomus ovary apparently does not differ functionally from other polytrophic meroistic insect ovaries.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Mechanisms of attack and defence at the cell surface: The use of phospholipid bilayers as models for cell membrane

Electrical conductivity across phospholipid bilayers induced by various cytotoxic proteins has been used to analyse the damaging action of such proteins on cells; the protective effect of divalent cations and protons against such attack has also been investigated. The predominant effect of divalent cations and protons is to promote the closed state of membrane pores, i.e. to gate protein-induced lesions.     

Notes on the swim-bladder physiology of cod(Gadus morhua) investigated from the underwater laboratory “Helgoland”

In situ sampling of gas from cod swim-bladders took place during a fortnight's saturation mission with the underwater laboratory Helgoland in May–June 1975. These samples were compared to those done by the conventional method of transporting the fish to the surface for sampling. Based upon these in-situ measurements, the mean O₂-concentration was 55.7% in buoyant cod at 15 m depth. Repeated sampling of the same fish showed a change in gas composition. Compared to the conventional method of transporting fish for sampling to the surface, in-situ sampling gave results with less variation, and indicated that surface-sampling does not give the correct gas composition of buoyant fish at depth of catch.     

Influence of plant growth regulators,basal media and carbohydrate levels on the in vitro development of Pinus ponderosa (Dougl. ex Law.) cotyledon explants

Applications of in vitro screening techniques for Pinus ponderosa resistance to Peridermium harknessii could be beneficial in a tree breeding program. Plant growth regulators, basal media formula and carbohydrate levels were examined to determine the various effects each would have on excised cotyledon growth and development. Proliferating green callus was initiated from cotyledon explants on SH basal medium containing 4.4 M BA:5.4 M NAA and 1% sucrose. Subsequent subculturing onto LS medium supplemented with 44.0 M BA: 5.4 M NAA and 2% sucrose improved callus maintenance. The highest frequency of caulogenesis from cotyledon explants occurred on a modified GD medium containing 44.0 M BA: 0.054 M NAA and 4% glucose. The influence of nitrogen source, osmoticum and medium salt concentrations are discussed relative to callus initiation and caulogenesis.Abbreviations BA 6-benzyladenine - NAA -naphthaleneacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - 2,4-D 2,4-dichlorophenoxyacetic acid - CD Campbell & Durzan - GD Gresshoff & Doy - LP Le Poivre - LS Linsmaier & Skoog - MC McCown - SH Schenk & Hildebrandt     

Increased radioresistance of cells in cultured multicell spheroids. I. Dependence on cellular interaction

Summary Monolayers of six different cell lines were investigated with respect to ionic coupling using micro-electrode techniques. In parallel, survival after Co--irradiation of monolayer- and spheroid cultures of these lines was compared. It was found that spheroids of coupled cell lines were more radioresistant than monolayers (contact effect). However, cell coupling did not enhance the survival of monolayers over single cells. This suggests that the contact effect is a tissue phenomenon requiring cellular interaction but is expressed only under conditions of three-dimensional growth.