Treatment of the yeast<Emphasis Type="Italic"> Rhodotorula glutinis</Emphasis> with AlCl<Subscript>3</Subscript> leads to adaptive acquirement of heritable aluminum resistance

When aluminum (Al) was added to a culture, growth of Rhodotorula glutinis IFO1125 was temporarily arrested, showing longer lag phases, depending on the Al concentrations (50–300 M) added, but the growth rates were not affected at all. Resistant strains obtained by one round of plate treatment containing Al reverted the resistance level to the wild-type level when cultivated without Al. Repeated Al treatments, however, induced heritable and stable Al resistance, the level of which was increased up to 4,000 M by stepwise increments in Al concentrations. Thus, the heritable Al resistance adaptively acquired was due neither to adaptation nor to mutation, but to a mechanism which has yet to be studied. Heritable Al resistance seemed to release the Al inhibition of magnesium uptake.     

Evidence of β-carotene 7,8(7′,8′) oxygenase (β-cyclocitral,crocetindial generating) in Microcystis

A -carotene oxygenase is described which occurs in the Cyanobacterium Microcystis. It cleaves -carotene and zeaxanthin specifically at the positions 7,8 and 7,8, while echinenone and myxoxanthophyll are not affected. The oxidative cleavage of -carotene leads to the formation of -cyclocitral and crocetindial and that of zeaxanthin to hydroxy--cyclocitral and crocetindial in nearly stoichiometric amounts. Oxidant is dioxygen as has been demonstrated by high incroporation (86%) of ¹⁸O₂ into -cyclocitral. -Carotene oxygenase is membrane bound, sensitive to sulfhydryl reagents, antioxidants and chelating agents. Iron seems to be an essential part of the enzyme activity. Cofactors necessary for the reaction could not be detected.Abbreviations TLC thin layer-chromatography - PIPES piperazine-N,N-bis-(2-ethanesulfonate) Na - TES 2{[tris-(hydroxymethyl)-methyl]-amino} ethanesulfonic acid Dedicated to Professor G. Drews on occasion of his 60th birthday     

Hybridoma growth,metabolism, and product formation in HEPES-buffered medium: I. Effect of passage number

Shake flask studies with a murine hybridoma cell line show drastically increased cell yields per mL of culture medium as function of passage number (cell age), whereas the maximum antibody concentrations at the end of batch growth decreased from 180 g/mL to 75 g/mL after 10 months of serial propagation in flasks. The results are presented in the light of differences in cell metabolism with glucose, lactate, ammonia, glutamine, and selected amino acids being monitored.     

Erythrocytenenzyme der Primaten

Zusammenfassung Die Adenosindesaminasen der Primaten zeigen eine genetisch determinierte Variabilität. Bei der Untersuchung von 327 subhumanen Primaten (289 Simiae der Alten Welt, 38 Prosimiae) konnten wir neun Adenosindesaminase-Varianten nachweisen, die auf Grund ihrer unterschiedlichen elektrophoretischen Wandergeschwindigkeit als ADA 6, ADA 4, ADA 2, ADA 2, ADA 1, ADA 3, ADA 5, ADA 5 und ADA 7 bezeichnet werden. Die Verteilung der verschiedenen Phänotypen wurde ermittelt.

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Red cell enzymes of primatesAdenosine deamisnase (EC: 3.5.4.4.)

Summary The polymorphism of adenosine deaminase has been investigated in 327 subhuman Primates (289 Old World Monkeys and 38 Prosimians). Nine adenosine deaminase variants were found to be present, which on the basis of their different electrophoretic mobilities were designated ADA 6, ADA 4, ADA 2, ADA 2, ADA 1, ADA 3, ADA 5, ADA 5 and ADA 7. The distribution of these various phenotypes has been estimated.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Art- und Mischgesang von Nachtigall und Sprosser (Luscinia megarhynchos, L. luscinia)

Zusammenfassung Die Gesänge beider Arten unterscheiden sich in der Strophenlänge [Nachtigall (Ng) im Mittel 3,1 s, Sprosser (Sp) etwa 6 s], im Strophenaufbau (Gesamtzahl der Phrasen, Anzahl und Lage von echten Phrasen) und in einer beim Sp hörbar langsameren Vortragsweise (Silben·s^–1), sowie im Auftreten von jeweils artcharakteristischen Phrasen. Die hier untersuchten Sp weichen in den Silbentypen ihrer Kastagnetten- und Schnatter-Phrasen stark von einer finnischen Population ab (Dialektbildung beim Sp). Mischsänger trugen in unterschiedlichen Anteilen arteigene Strophen, solche der Zwillingsart und Mischstrophen vor. Während ihre Artstrophen weitgehend denen normal singender Artgenossen entsprachen, waren die Fremdstrophen in allen untersuchten Parametern sehr ähnlich denen des Vorbildes. Eine geringe gegenseitige Beeinflussung beider Gesangsformen kann jedoch nicht ausgeschlossen werden. Von 239 untersuchten Sp- waren 67 (28%) Mischsänger. Ihr Anteil blieb über neun Jahre gleich. Wahrscheinlich gab es weitere Individuen mit einem unerkannten geringen Anteil fremder Lautmuster. Unter mehr als 200 Ng fand sich ein Mischsänger; dies ist der erste Nachweis bei dieser Art. Der Mischgesang ist wahrscheinlich auf Fehler beim Gesangslernen zurückzuführen. Die Kopierfehler werden innerhalb der Sp-Population tradiert. Sp-Mischsänger waren in 12 Fällen erfolgreich konspezifisch verpaart. Möglicherweise wird der Mischgesang beim Sp durch einen Selektionsdruck auf die Ausbildung interspezifischer Reviere begünstigt (character convergence). Sechsmal kam es zur Bastardierung mit der Zwillingsart, wobei der Mischgesang der beteiligten als hauptsächliche Ursache angesehen wird. Dennoch ist gegen Sp-Mischsänger wahrscheinlich keine Selektion wirksam. Mischgesang verschafft ihnen möglicherweise einen von der Populationsdichte abhängigen Selektionsvorteil.

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Species-specific song and mixed singing of Nightingale and Thrush Nightingale (Luscinia megarhynchos, L. luscinia)

Summary In Schleswig-Holstein (northern FRG), the sibling species Nightingale (Ng) and Thrush Nightingale (Sprosser, Sp) are sympatric. Here their respective ranges form the most northwestern part of the zone of overlap. In 1976–1984, the song of both species was investigated. Species-specific songs (Strophen) differ in length (Ng mean 3,1 s, Sp about 6 s), structure (total number of phrases, number and position of real phrases built up by repeated syllables), and by a slower speed (syllables·s^–1) in the Thrush Nightingale's songs. Both species dispose of species-characteristic phrases. In the castanet phrases and in the rattling phrases the Thrush Nightingales showed a quite different syllable type repertoire compared with a Finnish population (song dialect in this species). Mixed singers performed species-specific songs as well as songs which were heterospecific or mixed up of vocal patterns of both species. Their species-specific songs were highly equivalent to those of normal singing conspecifics, whereas all investigated parameters of the heterospecific songs were very similar to those of the imitated model. A possible slight mutual influence of both forms on each other can, however, not be excluded. Out of 239 Thrush Nightingales examined 67 (28 %) were mixed singers. The annual proportion in the population remained more or less constant during nine years. Presumably there were further individuals having only a small unrecognized amount of alien song patterns in their repertoire. In the Nightingale, only one out of more than 200 studied was a mixed singer. This is the first proof for mixed song in this species.Mixed singing is most probably due to misdirected song learning. The copying errors are transmitted within the Thrush Nightingale population by cultural tradition. In 12 cases mixed singing paired successfully with conspecific . Mixed song is here possibly favoured by a selective pressure on the development of interspecific territories (character convergence). In 6 cases, however, where hybridisation with the sibling species was observed, these mixed breedings could mainly be attributed to mixed singing of the involved. Nevertheless, there is probably no selection against mixed singing Thrush Nightingales. Dependent on population density it might even be advantageous for the birds to exhibit mixed song.

     

Cytoplasmic gel and water relations of axon

Summary A previous method of measuring the swelling pressure ( _g ) of the cytoplasmic gel of the giant axon ofLoligo vulgaris was refined. The estimates of _g made with the improved method were consistent with those made with the earlier method. In these methods the activity of the solvent in the gel is measured by increasing the activity of the solvent in the internal phase of the gel by application of hydrostatic pressure to the gel directly. Comparable values for the activity of the solvent in the gel were obtained also by an alternate method, in which the deswelling of the gel is measured upon decreasing the activity of the solvent in the external phase by addition of a nonpenetrating high mol wt polymer (i.e., Ficoll).Additional support was obtained for the earlier suggestion that _g contributes to the swelling and shrinkage pattern of the whole axon. In part, the new evidence involved two consecutivedirect measurements of intraxonal pressure. The first measurement was that of a mixed pressure composed of _g and _m ( _m being the effective osmotic pressure due to the intra-extraxonal gradient in the activity of mobile solutes). The subsequent measurement was that of _g alone. The latter measurement was made feasible by destroying the axolemma, thereby eliminating the contribution of _m . An estimate of _m was obtained by subtracting _g from the total pressure measured initially. The _m determined by the above method was two orders of magnitude smaller than the theoretical osmotic pressure. This is consistent with the _m determined previously, where osmotic intra-extraxonal filtration coefficients were compared to the hydrostatic. The mixed pressure experiments lend credence to the idea that the substantial contribution of _g to the water relations of the whole axon is due to _g being of the same order of magnitude as _m .The degree of free swelling of axoplasmic gels was studied as a function of pH, salt concentration, and hydration radius of the anion of the salt used. The swelling increased with an increase in the reciprocal of the hydration radius, a decrease in salt concentration, and at pH below or above 4.5.The nature of the constraints to the free swelling of axoplasm in axons immersed in seawater was studied. With the seawater employed, these constraints appeared to be due more to the retractive forces of the sheath than to _m .     

Ascorbic acid spares α-tocopherol and prevents lipid peroxidation in cultured H4IIE liver cells

Ascorbic acid, or vitamin C, can recycle -tocopherol in lipid bilayers, but even sparing of -tocopherol has not been a consistent finding in intact cells. Therefore, we tested the ability of ascorbate loading to spare -tocopherol and to prevent lipid peroxidation of cultured H4IIE rat liver cells. Although -tocopherol was undetectable in H4IIE cells, its cell content was increased by overnight incubation with -tocopherol in culture. Cells incubated with ascorbate 2-phosphate accumulated ascorbate to concentrations as high as 0.6 mM after overnight loading, but also released ascorbate into the medium. Ascorbate loading of -tocopherol-treated cells spared -tocopherol in a concentration-dependent manner during overnight incubation. Lipid peroxidative damage, measured as a decrease in fluorescence of cell-bound cis-parinaric acid, was decreased in cells loaded with either -tocopherol or ascorbate 2-phosphate, and showed an additive effect. These results suggest that ascorbate loading of H4IIE cells spares cellular -tocopherol and either directly or through recycling of -tocopherol prevents lipid peroxidative damage due to oxidant stress in culture.     

Cytokinesis in the green alga Eremosphaera viridis: Plasmalemma formation from “open membranes”

Summary Cytokinesis in the unicellular chlorococcalean alga Eremosphaera viridis de Bary has been investigated by electron microscopy of thin sections. The new plasmalemmata of the daughter cells in this organism form centrifugally within a phycoplast. Unlike other cell division systems each new plasmalemma is formed, not by the fusion of vesicles, but rather by the fusion of open membranes which are characteristically heavily stained. Measurements of these open membranes reveal that they are 11 nm thick with a central 4,5 nm unstained portion. The possible origin of these open membranes as burst-open vesicles has been suggested from the presence of intensely straining vesicles in the vicinity of the cell equator. Calculations of vesicle and open membrane surface areas support this contention.     

Saturated C21–C33Hydrocarbons Are Involved in the Self-Regulation of Pseudomonas fluorescensAdhesion to a Glass Surface

One of the two putative groups of antiadhesins was identified in Pseudomonas fluorescensby the method of gas chromatography–mass spectrometry. A mixture of high-molecular unbranched hydrocarbons (HC) with a chain length from 21 to 33 carbon atoms reduced cell adhesion to a glass surface. These HC accumulated in the culture liquid to a total concentration of 10–15 g/l; the concentrations of individual HC ranged from 0.1 to 3.0 g/l. After the addition of individual HC to the bacterial culture, the number of cells attached to the glass surface decreased. This decrease in cell adhesion was due to the enhanced aggregation of the bacterial cells, which promoted mechanical (hydrodynamic) cell detachment from the surface.     

The effects of testosterone, 5α-dihydrotestosterone, 3α-androstanediol,and 3β-androstanediol on epithelial fine structure of the rabbit epididymis in organ culture

Summary The fine structure of the corpus epididymidis of the rabbit has been studied following organ culture. Various modifications of tissue preparation and culture conditions were examined to obtain good maintenance of cellular integrity as well as to preserve sperm fertilizing ability.After 5 to 7 days in culture in the absence of hormonal support, the epididymal epithelium showed signs indicative of cellular regression. Such changes included shrinkage of the cells, loss of the border of stereocilia, decrease in smooth endoplasmic reticulum, and an increase in autophagic vacuoles. The presence of androgens in culture media prevented cellular regression to varying degrees, depending on the hormone utilized. With regard to maintenance of cellular integrity, potency of the androgens tested was as follows: 5-dihydrotestosterone >= 3-androstanediol > testosterone > 3-androstanediol. Addition of insulin to dihydrotestosterone-containing cultures resulted in no improvement in maintenance.Phagocytosis of spermatozoa by epithelial cells was observed in cultured tubules and the degree of spermiophagy was inversely proportional to successful maintenance of fine structural characteristics of epithelial cells.The morphological findings reported here correlate well with the fertilizing ability of spermatozoa from cultured epididymis as reported in an accompanying communication.     

Lipid production by Aspergillus oryzae from starch substrates

Summary Reports that starch is a poor substrate for lipid production are attributed to the low available C:N ratio which occurs because starch is not directly available to the microbial cell. If cultural conditions were established which gave rapid and extensive amylase activity, sufficient starch hydrolysis would occur to give a high ratio of available C:N, conditions favourable to lipid accumulation. This hypothesis was tested experimentally with Aspergillus oryzae and mycelium with 37% lipid content was obtained with starch as sole carbon source in defined culture media.     

Streptomycin resistance is inherited as a recessive Mendelian trait in a Nicotiana sylvestris line

Summary The SR180 cell line has been isolated in a callus culture derived from a haploid Nicotiana sylvestris (n = X = 12) plant by its ability to proliferate on a selective medium containing 2,000 g/ml streptomycin sulphate. From the cell line diploid plants have been regenerated. The SR180 selfs are resistant to streptomycin. Streptomycin sensitivity in F1, and a 31 (sensitive to resistant) segregation in F2 indicate that resistance in the SR180 mutant is the result of a recessive Mendelian mutation.     

Interspecific differences in aluminium tolerance in relation to root cation-exchange capacity

Genotypic differences in aluminium (Al) tolerance hold considerable promise in overcoming an important limitation to plant growth in acid soils. Little is known, however, about the biochemical basis of such differences. Extracellular properties, particularly low root cation-exchange capacity (CEC), have been associated with Al tolerance, since roots of low CEC adsorb less Al than do those of high CEC. A solution culture study was conducted in which 12 plant species (monocots and dicots) were grown in solution culture of low ionic strength (ca 2 mM) for 8 d at four Al concentrations (0, 16, 28 and 55 M). The species differed significantly in Al tolerance as shown by differences in root length. Root length relative to that of the same species grown in the absence of Al varied from 6 to 117% at 16 M Al, and from 6 to 75% at 28 M Al. Species tolerance of Al was not closely associated with differences in root CEC. Although in some species Al sensitivity was associated with high adsorption of Al during a 10- or 40-min exposure to Al (expressed on a fresh mass or root length basis), this was not a good predictor of Al tolerance across all species studied.     

Simultaneous overproduction of extracellular endoglucanase and intracellular β-glucosidase by genetically engineered Bacillus subtilis

Summary Simultaneous overproduction of intracellular -glucosidase and extracellular endoglucanase was attempted by constructing two artificial operon systems comprising the -glucosidase-endoglucanase gene(E) or the endoglucanase--glucosidase gene(E) under the control of a strong engineered promoter, BJ27U88 and expressing them in Bacillus subtilis DB104. Two artificial operon systems contained 30 bp or 5 bp gap between the termination codon of the upstream gene and the SD sequence of the downstream gene, respectively. These operon systems were expressed well in B. subtilis and overproduced the -glucosidase cell extract as well as the endoglucanase supernatant. The level of expression in the operon system was almost the same as that in a single expression system.     

The conventional short-circuiting technique under-short-circuits most epithelia

Summary The relationships among ion current, membrane potential difference, and resistance of an epithelium are studied. The short-circuit technique introduced by Ussing and Zerahn does not completely short circuit the epithelium if the series resistance parallel to the cell layer between the voltage electrodes is not properly compensated. The residual potential difference across the epithelial cell layer in the short-circuit state is proportional to both the measured short-circuit current and the resistance of the diffusion barriers not compensated. In the conventionally short-circuited small intestinal mucosa the villus and crypt areas are hypo-polarized to different degrees rather than simultaneously hyper- and hypo-polarized. Short-circuiting the whole tissue reduces but does not abolish the passive net ion movement across the tissue. Measurements of the electrical properties of the whole and denuded rat distal small intestine in HCO₃-Ringer solution containing 10mm glucose reveal that the measured short-circuit current has under-estimated approximately 33% of the true short-circuit current and that the passive net Na flux from serosa to mucosa and Cl flux from mucosa to serosa are not negligible in the short-circuit state.     

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.     

Benthic algal populations respond to aluminum,acid, and aluminum-acid mixtures in artificial streams

Elevated aluminum (Al) concentrations are often associated with acid-stressed aquatic ecosystems, so it has been unclear whether acidic water or elevated Al is more responsible in changing community composition. Experiments were done to investigate effects of acidification and increased Al on the abundance of benthic algae in artificial streams supplied with natural water and nominal treatments of (a) pH 4.8, (b) 500 µg l^-1 Al, or (c) the mixture of pH 4.8 and 500 µg l^-1 Al compared to a control without added Al or acid. These treatments are referred to as Acid, Al-only, Acid + Al, and the control, respectively. In the Acid treatment the abundance of two diatoms, two green algae, dry weight biomass, and chlorophyll a decreased; one diatom and one filamentous blue-green alga increased. In the Al-only treatment, densities of two diatoms, one green alga, one blue-green alga, dry weight biomass, and chlorophyll a increased. In the Acid + Al treatment, abundances of one green alga, two blue-green algae, and concentrations of chlorophyll a decreased below the levels observed in the Acid treatment. Acid and Al concentrations were altered by each other and by chemical and biological processes in the stream system. Species of diatoms, green algae, and blue-green algae responded individually to treatments and mixtures of acid and Al. Shifts in the abundance of species may change food web relationships for higher-level consumers, and algae may be useful biomonitors of ecological stress.     

Requirement of chondroitin sulfate/dermatan sulfate recognition in midkine-dependent migration of macrophages

Midkine (MK) is a heparin-binding growth factor that promotes cell migration, cell growth and cell survival. The promotion of migration of inflammatory cells, especially macrophages, by MK is involved in formation of a vascular abnormality, i.e. neointima formation. MK-induced migration of peritoneal exudate macrophages was inhibited by heparin, chondroitin sulfate E and dermatan sulfate, but not by chondroitin sulfate D or chondroitin 6-sulfate. Digestion of macrophages with chondroitinase ABC as well as chondroitinase B decreased the migratory activity. However, heparitinase digestion showed only slight effects. These results indicated that a chondroitin sulfate, i.e. an E-type oversulfated structure with dermatan sulfate domain, is involved in MK-induced migration of macrophages. Although a chondroitin sulfate proteoglycan, receptor-type protein tyrosine phosphatase (PTP ), participates in MK-induced migration of neurons and osteoblasts, PTP was not detected in macrophages. The MK-induced migration was inhibited by PP1, wortomanin, PD 98059 and vanadate, indicating that the downstream signaling system, which includes Src, PI3 kinase and ERK as important components, is shared with other MK signaling systems in which PTP is involved.