The membrane systems of the cardiac muscle cell of Meganychtiphanes norvegica (M. Sars), euphauciacea,crustacea

Summary The membrane systems of cardiac muscle cells of the euphausiacean Meganychtiphanes norvegica are described. Transverse tubules are found both at the Z-band level (T_z-tubules) and at the H-band level (T_h-tubules). Within the sarcomere narrow longitudinal tubules branch off from the T_z-tubules. At the H-band level these tubules expand forming flattened cisternae in dyadic and triadic couplings with the sarcoplasmic reticulum (SR). Adjacent myofibrils are separated by a well developed SR. Modifications of the SR are seen at the H-band level where junctional cisternae are formed.     

Muscle remodeling in adult snapping shrimps via fast-fiber degeneration and slow-fiber genesis and transformation

Bilateral asymmetry of the paired snapper/pincer claws may be reversed in adult snapping shrimps (Alpheus heterochelis). Removal of the snapper claw triggers transformation of the contralateral pincer claw into a snapper and the regeneration of a new pincer claw at the old snapper site. During this process the pincer closer muscle is remodeled to a snapper-type, and these alterations have been examined with the electron microscope. There is selective death of the central band of fast fibers, accompanied by an accumulation of electron-dense crysttaline bodies in the degenerating fibers. Two principal types of hemocytes (amebocytes and coagulocytes) invade the area and the degenerating muscle fibers. New myotubes also appear in this central site. The myotubes are characterized by a prolific network of presumptive sarcoplasmic reticulum and transverse tubules, nascent myofibrils, and crystalline bodies. The myotubes are innervated by many motor nerve terminals, and they subsequently differentiate into long-sarcomere (8–12 m), slow muscle fibers. Remodeling of the central band, therefore, occurs by degeneration of the fast fibers and their replacement by new slow fibers. Remnants of the degenerating fast fibers act as scaffolding for the myotubes which originate from adjacent satellite cells. The crystalline bodies may represent protein stores from the degeneration of the fast fibers, recycled for use in the genesis of new fibers. The invading hemocytes appear to play several roles, initially phagocytosing the fast muscle fibers, transporting the crystalline bodies into the new myotubes, and acting as stem cells for the new muscle fibers. Apart from the central band of fibers, the remaining pincer-type slow fibers with sarcomere lengths of 5–7 m are transformed via sarcomere lengthening into snapper-type slow fibers with sarcomere lengths of 7–12 m. Thus, during claw transformation in adult snapping shrimps, the pincer closer muscle is remodeled into a snapper closer muscle by selective death of the fast-fiber band, replacement of the fast-fiber band by new slow fibers, and transformation of the existing slow fibers to an even-slower variety. Note. This paper is dedicated to the fond memory of Professor M.S. Laverack whose enjoyment of biological research and gentle encouragement of such endeavours touched all those who knew him.     

Ultrastructure of the neurogenic heart of Limulus polyphemus

Summary The ultrastructure of Limulus cardiac muscle was examined. The hearts were fixed in situ by perfusion with isotonic glutaraldehyde solution while in relaxed, contracted, or stretched states. The sarcomeres are relatively long, varying in length from about 2.5 to 6.6 . The average A-band length is 2.46 . M lines are absent, and H zones are poorly distinguished. Thick and thin filament diameters average about 200 Å and 50 Å, respectively; each thick filament is surrounded by 8–12 thin ones. Superficial invaginations of the sarcolemma occur, making contact with the Z lines of the outermost myofibrils. There is an extensive sarcoplasmic reticulum and transverse (T) tubules. Some T tubules run longitudinally and some open into deep sarcolemmal invaginations which extend into the fiber interior. The T tubules swell markedly in hypertonic solution. Single neurons and small bundles of neurons are observed in close apposition with myocardial cells. Intercalated disks are found in Limulus heart at regions of contact between contiguous myocardial cells lying end to end; semitight or gap junctions are essentially absent. Prominent differences in sarcomere lengths sometimes occur across the disk, thus indicating that the disks demarcate cells functionally. Hence, in addition to direct motoneuron activation, there may be some transfer of excitation across the intercalated disks in accord with our previous finding that propagating, overshooting action potentials can be induced in this heart.Supported by grants from the American Heart Association and from the Public Health Service (HE-11155 and HE-05815). I thank Mrs. Jan Redick for expert technical assistance.     

The membrane systems of the cardiac muscle cell of Tmetonyx cicada O. Fabricius (Crustacea,Amphipoda)

Summary The membrane systems of the cardiac muscle cell of the amphipod Tmetonyx cicada (O. Fabricius) are described. The sarcolemma invaginates and forms a transverse network of tubules at the level of the Z band. Narrow longitudinal tubules branch from the network and connect to another transverse network of tubules at the H band level, where dyadic and triadic junctions are formed with the sarcoplasmic reticulum. Adjacent myofibrils are normally separated by a well developed double layer of the sarcoplasmic reticulum. In areas where the myofibrils closely approach the outer sarcolemma, peripheral couplings have been found at the level of the H band.     

Ultrastructure of the heart muscle cells of the cuttlefish Rossia macrosoma (Delle Chiaje) (Mollusca: Cephalopoda)

Summary The muscle cells of the ventricle, the branchial heart and the branchial heart appendages of Rossia macrosoma (Delle Chiaje) are studied. The ventricle myocardium has three muscle layers, while the other two organs exhibit a loose arrangement of muscle cells. The muscle cells of the ventricle, the branchial heart and the branchial heart appendages are similar in structure. The nuclei are surrounded by myofibrils. In the myofibrils A-, I- and discontinuous Z-bands are seen. The diameters of the thick filaments are 300–400Å, their length varies from 1.7 to 3.9 . Thin filaments have a diameter of approximately 85Å. The ratio between thick and thin filaments is roughly 1 to 11.The SR runs mostly as a longitudinal network within the myofibrils. A few short T-tubules are observed in the Z-regions. Peripheral and internal couplings exist. The latter are few in number.Intercalated discs are small and rarely observed. They have been found in all three organs. A difference in the function of these organs is not reflected in the ultrastructure of the intercalated discs. These discs are often of the interdigitating type with interfibrillar junctions and unspecialized regions. Peripheral couplings are seen at the unspecialized regions. The intercalar surfaces of the muscle cells shoulder off into the lateral surface, and the transition between the two surfaces is not a sharp one. Attachment plaques are found scattered over the whole sarcolemma.     

Ultrastructure of the myocardial cell and its membrane systems in the adult fly Calliphora erythrocephala (Insecta: Diptera)

Summary Calliphora erythrocephala has cross-striated cardiac muscle cells with A, I and Z-bands. The diameters of the myosin and actin filaments are 200–250 Å and 85 Å respectively and the length of the myosin filaments (A-band) is approximately 1.5 . Usually 8–10 actin filaments surround each myosin filament.The myocardial cells show a well-developed membrane system and interior couplings. A perforated sheet of SR envelopes the myofibrils at the A-band, dilates into flattened cisternae at both A-I band levels before it merges into a three-dimensional net-work between the actin filaments of the I-bands and between the dense bodies of the discontinuous Z-discs. The T-system consists of broad flattened tubules running between the myofibrils at the A-I band levels forming dyads with the SR-cisternae. Longitudinal connections between the transverse (T-) tubules often occur.It is suggested that this well-developed SR may be an adaptation to facilitate a rapid contraction/relaxation frequency by an effective Ca²⁺ uptake.     

Novel maltotriose esters enhance biodegradation of Aroclor 1242 by Burkholderia cepacia LB400

The objective of this research was to evaluate the effect of enzymatically synthesized maltotriose fatty acid monoesters (Ferrer, M., et al. 2000 Tetrahedron 56, 4053–4061) on Aroclor 1242 solubilization and biodegradation. Three forms of the surfactant, laurate, palmitate and stearate monoester, were tested. Potential enhancement of solubilization of hydrophobic substances mediated by these non-ionic surfactants was exploited in this study. A polychlorinated biphenyl (PCB) degrading organism, Burkholderia cepacia LB400, was also selected. It was found that all surfactants were effective in solubilizing Aroclor 1242 but the rate of Aroclor 1242 biodegradation proceeded rapidly only in the presence of 6-O-palmitoylmaltotriose. For example, the addition of 48 mg 6-O-palmitoylmaltotriose/l increased the apparent solubility from 140 to 305 g/l. As a result, only 8% of the Aroclor remained at the end of 24 h incubation. In contrast, 49.2% of the Aroclor 1242 remained in the absence of surfactant. It appears that maltotriose fatty acid monoesters can significantly increase the bioavailability, and thereby accelerate the biodegradation of highly chlorinated PCBs, particularly Aroclor 1242, by Burkholderia cepacia LB400. The possibility of obtaining these biodegradable surfactants with high yield, easy recovery and high purity by using a new enzymatic methodology, makes maltotriose esters available for bioremediation purposes.     

Relations between hydrological variables and year-class strength of sportfish in eight Florida waterbodies

Hydrological variables have influenced fish recruitment in lakes, reservoirs and rivers. We evaluated how annual and seasonal hydrological variables were related to year-class strength (i.e., residuals from catch curves) of sportfish across eight Florida waterbodies (four rivers and four lakes). Multiple regression equations computed for black bass Micropterus spp. were combined across rivers and year-class strength was negatively related to spring median flow rates and in some cases positively related to winter median flow rates (all p0.10). Conversely, Lepomis spp. residuals combined from the rivers indicated that year-class strength was positively related to median flow rates in the fall prior to spawning and negatively related to post-spawn fall median flow rates (all p0.10). Fish recruitment combined across lakes were not related to water levels in this study, although within lake relationships did occur in some instances. Ecological implications of this work include regulations such as minimum flows and levels (MFLs) regarding sportfish species. Impacts of hydrology on year-class strength of sportfish were stronger in rivers than in lakes for these Florida systems. High flows at least once every 3years in the fall may allow inundation of floodplain habitat, providing favorable environmental conditions for Lepomis spp. reproduction. Setting MFLs during periods of drought (i.e., 3years or more) should consider impacts to short-lived species such as Lepomis spp.     

Polarized distribution of γ interferon-stimulated MHC antigens and transferrin receptors in a clonal cell line isolated from Fisher rat thyroid (FRT cells)

Summary The fine structure of single identified muscle fibers and their nerve terminals in the limb closer muscle of the shore crab Eriphia spinifrons was examined, using a previous classification based on histochemical evidence which recognizes a slow (Type-I) fiber and three fast (Type-II, Type-III, Type-IV) fibers. All four fiber types have a fine structure characteristic of crustacean slow muscle, with 10–12 thin filaments surrounding each thick filament and sarcomere lengths of 6–13 m. Type-IV fibers have sarcomere lengths of 6 m while the other three types have substantially longer sarcomeres (10–13 m). Structural features of nerve terminals revealed excitatory innervation in all four fiber types but inhibitory innervation in Type-I, Type-II, and Type-III fibers only. Thus fibers with longer sarcomeres receive the inhibitor axon but those with shorter sarcomeres do not. Amongst the former, synaptic contact from an inhibitory nerve terminal onto an excitatory one, denoting presynaptic inhibition, was seen in Type-I and Type-II fibers but not in Type-III and Type-IV fibers. Inhibitory innervation of the walking leg closer muscle is therefore highly differentiated: some fibers lack inhibitory nerve terminals, some possess postsynaptic inhibition, and some possess both postsynaptic and presynaptic inhibition.     

Different susceptibilities of complex-, hybrid- and high-mannose-type α1- inhibitor and α1-acid glycoprotein to endo-β-N-acetylglucosaminidase F and peptide:N-glycosidase F

Endo--N-acetylglucosaminidase F (endo F, EC 3.2.1.96) and peptide:N-glycosidase F (PNGase F, EC 3.2.2.18) fromFlavobacterium meningosepticum were used for the deglycosylation of ₁-proteinase inhibitor and ₁-acid glycoprotein carrying oligosaccharide side chains of the complex-, high-mannose- and hybrid-type. High-mannose-and hybrid-type glycoproteins were obtained by the incubation of rat hepatocyte primary cultures with 1-deoxymannojirimycin or swainsonine, respectively. It was found that endo F cleaves hybrid- and high-mannose-type ₁-proteinase inhibitor and ₁-acid glycoprotein at pH 4.5 as well as at pH 8.5 in the presence or absence of 1% octyl--d-glucopyranoside. Complex-type ₁-proteinase inhibitor or ₁-acid glycoprotein were not cleaved by endo F even in the presence of octyl--d-glucopyranoside.PNGase F was found to cleave complex-, hybrid- and high-mannose-type oligosaccharide side chains of ₁-proteinase inhibitor and ₁-acid glycoprotein at pH 4.5 and pH 8.5 in the presence of 0.75% octyl--d-glucopyranoside. The deglycosylation of both protein substrates was very poor without detergents.Abbreviations Endo F endo--N-acetylglucosaminidase F (EC 3.2.1.96) - PNGase F peptide:N-glycosidase F (EC 3.2.2.18) Dedicated to Prof. Dr. Wolfgang Gerok on the occasion of his 60th birthday     

Discovery,localization, and sequence characterization of molecular markers for the crown rust resistance genes <Emphasis Type="Italic">Pc38, Pc39</Emphasis>, and <Emphasis Type="Italic">Pc48</Emphasis> in cultivated oat (<Emphasis Type="Italic">Avena sativa</Emphasis> L.)

Molecular markers for the crown rust resistance genes Pc38, Pc39, and Pc48 in cultivated oat (Avena sativa L.) were identified using near-isogenic lines and bulked segregant analysis. Six markers for Pc48, the closest being 6 cM away, were found in a Pendek-39 × Pendek-48 (Pendek3948) population, but none was found in a Pendek-48 × Pendek-38 (Pendek4838) population. Three markers for Pc39 were found in the Pendek3948 population, one of which cosegregated with the gene. This same marker was found to be 6 cM away from the gene in an OT328 × Dumont (OT328Du) population. Nine markers for Pc38 were found in the Pendek4838 population, eight of which are within 2 cM of the gene. One other marker for Pc38 was found in the OT328Du population; however, comparative mapping suggests that the Pc38 region in OT328Du is in a different location than that in Pendek4838. A number of markers unlinked to the genes under study formed linkage groups in both the Pendek3948 and Pendek4838 populations. Four of these show homology or homoeology to each other and to the Pc39 region in Pendek3948. Two RFLP clones closely linked to Pc38 code for a putative leucine-rich repeat transmembrane protein kinase and a cre3 resistance gene analogue. This study provides information to support molecular breeding in oat, and contributes to ongoing research into genomic regions associated with fungal pathogen resistance.     

Lack of effect of toxic aluminium concentrations on nitrogen fixation by nodulatedStylosanthes species

Summary Effects of three solution aluminium concentrations (0, 25, and 100M) on nitrogen fixation by well-nodulated plants ofStylosanthes hamata, Stylosanthes humilis andStylosanthes scabra are reported. Plants were inoculated with Rhizobium CB756 and grown for 21 days in an aluminium-free nutrient solution at pH 5.3 before imposition of the aluminium treatments.Nitrogen fixation was measured both by the increase in total nitrogen content of the plants and acetylene reduction in roots of plants harvested at 10 and 20 days after imposition of the aluminium treatments. Solution aluminium concentrations as high as 100M, had no detrimental effect on nitrogen fixation in any species.     

Contribution to the knowledge of the enzymatic activity of yeasts of clinical interest

The determination of the enzymatic activity of the yeasts has been applied to the identification of species, specially that ofCandida albicans. In order to know its usefulness in species of clinical interest, we have tested the commercial system API ZYM (Bio Mérieux) on 500 isolated strains of different organic samples, belonging to eight genera and twenty species. All the strains showed positivity to Phosphatase alcaline, Esterase (C4), Esterase lipase (C8), Leucine arylamidase and Phosphatase acid, and negativity to Lipase (C14), Trypsin, Chymotrypsin, -galactosidase, -glucoronidase, -manosidase and -fucosidase. Fourteen enzymatic activity patterns were obtained considering the substrates with variable results for the whole of the strains: Valine arylamidase, Cystine arylamidase, Naphthol-AS-BI-phosphohydrolase, -galactosidase, -glucosidase, -glucosidase and N-acetyl--glucosaminidase. In the majority of the species, the enzymatic profile did not have very specific results since it is usually shared by more than one species.C. albicans is that which presents the greatest number of enzymatic variations, some of these are similar to those of other common clinical species, such asCandida krusei, Candida parapsilosis andCandida tropicalis. This system is proposed as a rapid method for identification and as an epidemiological marker of medically important yeasts.Abbreviations AGL -glucosidase - BGA -galactosidase - BGL -glucosidase - CAA Cystine arylamidase - NAG N.Acetyl--glucosaminidase - PHO Naphthol-AS-BI-phosphohydrolase - VAA Valine arylamidase     

Interactions of membrane excitability mutations affecting potassium and sodium currents in the flight and giant fiber escape systems of Drosophila

Summary We have studied the influence of the K⁺-current mutations eag and Sh and the Na⁺-current mutation nap ^ts upon two well-defined neural circuits that underlie flight and an escape response in Drosophila, recording from dorsal longitudinal and tergotrochanteral muscles. Mutations of Sh and eag affected refractory period and following frequency, but not latency, of the jump-and-flight escape response. The nap ^ts mutation altered these 3 physiological parameters of the jump (TTM), but not the flight (DLM), branch, suggesting differences in the vulnerability of different circuit components to the mutation. In contrast to their interaction in some other systems, nap ^ts did not counteract the effects of eag and Sh upon these physiological parameters in eag Sh; nap triple mutants.In eag Sh double mutants, in which multiple K⁺ currents may be diminished, flight muscles showed abnormal rhythmic activity not associated with flight, and some flies also had an abnormal wings-down posture. The low-frequency spikes probably originated in the flight muscle motoneurons, but the coordination between muscle fibers during this non-flight activity was distinct from flight. Nevertheless, in spite of the presence of this non-flight activity in resting eag Sh flies, those animals with normal wing posture were also able to fly, with a normal pattern of muscle activity. This suggests that in these mutants, the DLM motoneuron circuit is able to switch between two patterns of output, non-flight activity and flight. In eag Sh; nap triple mutants, the non-flight activity and abnormal wing posture were absent, indicating that a reduction of Na⁺ current counteracts the hyperexcitable influence of the K⁺-current mutations in this circuit.Abbreviations CGF cervical giant fiber - DLM dorsal longitudinal muscle - eag ether à go-go - FF ₅₀ following frequency with 50% response - nap ^ts no action potential — temperature sensitive para paralytic - PSI peripherally synapsing interneuron - Sh Shaker TTM tergotrochanteral muscle     

Agrobacterium-mediated transformation of Solanum phureja

A population of transgenic plants was produced by the transformation of internodal explants of Solanum phureja, DB337/37 (the cultivar Mayan Gold) using an Agrobacterium tumefaciens LBA4404-based vector containing a phytoene synthase gene (crtB). The regeneration strategy utilised a two-step protocol, with a 12-day callus induction stage mediated by 1.07 M -napthaleneacetic acid (NAA), 7.10 M zeatin riboside and 0.06 M gibberellic acid (GA3), followed by a prolonged (up to 90 day) shoot induction stage on medium containing 0.11 M NAA, 7.10 M zeatin riboside and 0.06 M GA3 supplemented with kanamycin at 50 mg l^–1 as the selection agent. Southern analysis of the transgenic population revealed that the transgene copy number varied between one and five in the lines tested. Northern blot analysis showed significant expression of the introduced crtB gene in some lines during tuber development. Cytological analysis of the material showed a high incidence of chromosome doubling in the transgenic population with over 80% of all lines tested having doubled their chromosome complement during the transformation process.     

Morphological,physiological and enzymatic characteristics of cephalosporin acylase-producing Arthrobacter strain 45-8A

A bacterial strain producing cephalosporin acylases was isolated from soil. The morphological and physiological properties of this strain suggest that it belongs to the genus Arthrobacter, and the isolate was therefore designated Arthrobacter strain 45-8A. Substrate specificity of the enzyme was examined. The enzyme can convert both cephalosporin C and 7-(4-carboxylbutan-amino)cephalosporanic acid to 7-aminocephalosporanic acid. An interesting feature of the acylases is their temperature-dependent regulation. Activity of acylases was detected in strain 45-8A grown at temperature below 30 °C, but was not observed at higher temperature. Arthrobacter strain 45-8A did not exhibit -lactamase activity, even though its resistance to cephalosporin C was very strong (>2000 g/ml). This is quite beneficial for its application in the manufacture of 7-aminocephalosporanic acd.Abbreviations used NBHAB 2-Nitro-5-(6-bromohexanoylamino)-benzoic acid - NIPAB 2-Nitro-5-phenylacetaminobenzoic acid - CPC cephalosporin C - GL-7ACA 7-(4-carboxybutanamino)cephalosporanic acid - 6-APA aminopenicillanic acid - 7-ACA 7-aminocephalosporanic acid - PDAB p-Dimethylaminobenzaldehyde     

Plant regeneration via somatic embryogenesis in Styrian pumpkin: cytological and biochemical investigations

Somatic embryo formation was induced from cotyledon explants of Styrian pumpkin (Cucurbita pepo L. subsp. pepo var. styriaca Greb.) by using a solid MS medium supplemented with 16.11M NAA and 4.44M BA or 26.85M NAA and 13.32M BA. The callus proliferation was more efficient on medium supplemented with 26.85M NAA and 13.32M BA. In contrast, the embryogenic response was higher on medium with lower concentrations of growth regulators (16.11M NAA and 4.44M BA). The time needed for embryo induction did not depend on medium composition. Embryos in globular stage were transferred to three different maturation media, containing 2.89M GA₃ in combination with 0.54M NAA, 11.42M IAA and growth regulator-free medium. The germination rate was the highest when embryos were cultured on medium with 11.42M IAA. Plantlets grown on this medium achieved maturity suitable for transplantation into soil within 9 to 10weeks. The regenerated plants were successfully transferred into field and developed fertile flowers and set fruits. Biochemical analysis showed significant lower total glutathione levels among in vitro grown plantlets compared to seedlings grown in soil. When the plantlets were transferred into soil, they reached a normal size within a month and the glutathione concentration was comparable to seed-derived plants at the same developmental stage. Transmission electron microscopy was used to investigate possible differences in the ultrastructure of cells from callus cultures, and leaf cells of regenerated and seed-derived plants. Differences in the ultrastructure were found within chloroplasts which contained only single thylakoids, large starch grains and small plastoglobuli in callus cells in comparison to leaf cells, which possessed a well developed thylakoid system, small starch grains and large plastoglobuli.     

Cucurbitacins and predation of the spotted cucumber beetle, Diabrotica undecimpunctata howardi

Two hypotheses about the ralationship of diabroticina beetles and plants in the family Cucurbitaceae are tested: (1) evolution of sensory receptors for cucurbitacins by diabroticina beetles was in part due to the predator protection offered by ingestion of these compounds, and (2) commercial varieties of cucurbitacin-producing cucumber offer Diabrotica undecimpunctata howardi Barber chemical protection from some potential sources of natural controlSpotted cucumber beetles fed either Marketmore 70 cucumber which contains cucurbitacin-C or Marketmore 72 which totally lacks cucurbitacin were presented to four species of vertebrate predators that commonly occur in the summer and/or winter habitats of D. u. howardi: Bufo americanus, B. fowleri, Peromyscus maniculatus, Colinus virginianus. None of the four species were significantly deterred from preying on beetles that had eaten Marketmore 70 cucumber. These results do not support either of the two hypotheses. The limitations of these negative results as evidence for refutation of the first hypothesis are discussed.

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Résumé Deux hypothèses concernant les relations entre les Diabroticina et les Cucurbitaceae ont été examinées: (1) l'évolution chez les Diabroticina des récepteurs pour les Cucurbitacines est due en partie à la protection contre les prédateurs apportée par la consommation de ces produits (2) les variétés modernes de concombres produisant de la cucurbitacine fournissent à D. undecimpunctata howardi Barber une protection chimique contre quelques sources potentielles de lutte biologique.Des D. undecimpunctata howardi, ayant consommé soit des concombres Marketmore 70 contenant de la cucurbitacine C, soit des concombres Marketmore 72 qui ont totalement perdu leur cucurbitacine, ont été présentés à quatre espèces de vertébrés prédateurs, qui s'observent fréquemment dans les habitats d'été ou d'hiver de D. undecimpunctata howardi soient Bufo americanus, B. fowleri, Peromyscus maniculatus, Colinus virginianus. Aucune de ces quatre espèces n'est significativement dissuadée d'attaquer les coléoptères qui on consommé les concombres Marketmore 70. Ces résultats n'étayent aucune des deux hypothèses. La discussion porte sur les limites de ces résultats négatifs comme preuve de la réfutation de l'hypothèse 1.

     

Utilization of aromatic amino acids as nitrogen sources in Brevibacterium linens: an inducible aromatic amino acid aminotransferase

The first step of the utilization of the aromatic amino acids as sole nitrogen sources by Brevibacterium linens strain 47 was found to be a transamination. The deaminated metabolites of the amino acids were detected in culture supernatants, and the enzyme activity was identified in cell free extracts. The cells contained increased aromatic amino acid aminotransferase activities on growth on the aromatic amino acids as sole nitrogen sources. Two aromatic aminotransferases (AT-I and AT-II) were separated upon diethylaminoethyl-Trisacryl M column chromatography of cell free extracts. Only AT-I was responsible for the increased level of aromatic amino acid aminotransferase activity of induced cells. The results suggested a catabolic role of AT-I in vivo.Abbreviations DNP dinitrophenyl - HPLC high performance liquid chromatography - PLP pyridoxal-5-phosphate     

Continuous culture and synchronization ofHyphomicrobium sp. B-522

A technique was developed for synchronization ofHyphomicrobium sp. strain B-522. Bacteria were grown in continuous culture with methanol (0.1%; v/v) growth limiting. Vitamin B₁₂ (2.5 g/l) was necessary to obtain steady state growth. The critical dilution rate wasD _c =0.112; maximum cell output occurred atD=0.105 (Dx=30 mg l^-1 h^-1). Continuous cultures ofHyphomicrobium B-522 atD=0.110 were used to obtain cells for synchronization experiments. Synchronization was achieved by trapping young hyphal and budding cells in a glass wool column, while the initial swarmer cells were allowed to pass through. By semicontinuously rinsing the system, newly produced swarmers could be sampled in the effluent. The mean length of these synchronous swarmer cells was 1.25 m (s=±0.13 m; range 0,6 m) as compared to 1.40 m (s=±0.21 m; range 1.2 m) for swarmer cells of the continuous culture. Division of synchronous swarmer populations was completed after 7 h; the synchronization index was 0.76.