Microaerophily

Objections to previous definitions of anaerobiosis and micro-aerophily are discussed, and a new definition of micro-aerophily is proposed. A survey of physiological characteristics of the micro-aerophilic bacterium Campylobacter sputorum spp. bubulus is given, and the concept of micro-aerophily is illustrated by the results of a study on the oxygen metabolism and oxygen sensitivity of this bacterium.     

Computerized scheme for the reaction of hemoglobin with ligands

A scheme for the reaction of hemoglobin with ligands is described, which postulates the functional heterogeneity of the chains, considers all possible combinations of the distribution of the ligand on the four chains of hemoglobin, and does not require simplifying assumptions about the hemoglobin reactivity. Ten tetrameric species are considered, together with 16 reactions between these species, each with an on and an off rate constant. The dissociation of hemoglobin tetramers into dimers is also considered, with four on and four off rate constants for the reactions between dimers, and ten equilibrium constants for the reactions between tetramers and dimers. Moreover, some side reactions, such as the trapping of ligands by a hemoglobin competitor, are included. A FORTRAN program, suitable for microcomputers, is described for handling this scheme, with some examples showing its advantages.     

Characterization of two beta-tubulin genes from Geotrichum candidum.

Summary The -tubulin genes G1 and G2 from the phytopathogenic hemiascomycete Geotrichum candidum were found to be highly diverged in amino acid sequence from those of other filamentous fungi. G1 and G2 were also divergent from each other, with the coding regions sharing only 66% nucleotide sequence homology and 64% amino acid identity. However, the proteins shared 82% similarity and only 25 of the 161 non-identical amino acid substitutions were non-conservative. The organization of G1 is similar to other fungal -tubulin genes, but G2 has several unusual features; it has 2 amino acid additions in the N-terminal 40 residues and must employ an uncommon 5 splice junction sequence in preference to an overlapping perfect consensus. The amino acid change found to confer benomyl resistance in Neurospora crassa was also present in G2. G1 has four introns which are located similarly to those of -tubulin genes in other fungi. G2, however, has a single intron in a unique location. Translational fusions employing the 5 non-coding regions of the two Geotrichum -tubulin genes were made with the hygromycin phosphotransferase gene and shown to function in Schizosaccharomyces pombe and Trichoderma hamatum. However, G. candidum could not be transformed with these or other tested plasmids commonly used for fungal transformation.     

Rapid clonal propagation of guava through in vitro shoot proliferation on nodal explants of mature trees

In vitro clonal propagation of guava Banaras local was achieved by culturing nodal explants of mature trees on Murashige and Skoog (MS) revised medium supplemented with 4.5 M 6-benzyladanine (BA) alone or in combination with either 0.6 M indole-3-acetic acid (IAA), 0.5 M indole-3-butyric acid (IBA) or 0.3 M gibberellic acid (GA₃). Multiple shoots were induced to form by enhancement of axillary branching and BA (4.5 M) without any auxin and gibberellin was found to give best shoot multiplication rate. In this medium 3–6 shoots developed on explants collected from field-grown plants and 5–10 shoots developed on explants taken from in vitro proliferated shoots within 12 wk of culture. A prior transfer of shoot clumps to a medium containing a lower concentration of BA (0.5 M) before harvesting of cuttings for rooting allowed rapid extension growth and increased the number of usable shoots per culture. Adventitious rooting occurred after subculturing excised shoots on a medium containing 1/2 strength MS salts, 1.5% sucrose, 1 M each of IBA and -naphtha-leneacetic acid (NAA), and 1 gl^-1 activated charcoal. Regenerated plantlets were successfully established on soil.     

Identification of the LH and TSH-secreting cells in the pituitary gland of the rhesus monkey

Summary Pituitary glands from juvenile (pre-pubertal) and adult male and female rhesus monkeys were examined following immunocytochemical staining with antisera to the beta subunits of ovine luteinizing hormone (LH) and of human thyroid stimulating hormone (TSH). The LH antiserum reacts with a cell that is PAS-positive, occurs singly and is randomly distributed throughout the pars distalis. The diameter of these cells is approximately 11.5 m. They do not seem to vary in number in either juveniles (pre-pubertals) or adults, or in males or females. There appears to be fewer LH cells in the pituitary glands of pregnant and lactating females. In addition to staining cells in the pars distalis, the antiserum also reacts with a population of cells located in the pars tuberalis.The cells that stain with the anti-TSH serum are confined primarily to the pars distalis. They are approximately 15.8 m in diameter and are generally found in groups or clusters located in the anterior and medial regions of the gland. The TSH cells vary in number from one animal to another; however, this variability is unrelated to the age or the sex of the animals. No demonstrable changes occur in the number of TSH cells during pregnancy or lactation.Supported by NIH General Research Support Grant RR05654The author wishes to express appreciation to the Hormone Distribution Program of NIAMDD for the preparations of ovine FSH, TSH and human TSH, and to Drs. H. Papkoff for the ovine LH and LH, L. Reichert for the human FSH, J. Vaitukaitis for the anti-human TSH, and L.A. Sternberger for the PAP complex     

Transglycosylation reactions by exoglycosidases from the termite Macrotermes subhyalinus

The ability of four exoglycosidases (-galactosidase, -glucosidase, -glucosidase and invertase) from the termite Macrotermes subhyalinus to catalyse tranglycosylation reactions was tested using lactose, cellobiose, maltose and sucrose as glycosyl donors and 2-phenylethanol as glycosyl acceptor. The experimental conditions were optimized in relation to the time course of the reaction, pH and concentrations of glycosyl donor and acceptor. Whereas the hydrolytic activity was largely predominant over the transferase activity with -galactosidase and -glucosidase, the transglycosylation activity represented 68% with -glucosidase. In addition, as demonstrated by the transglycosylation product formed, the hydrolysis of sucrose was catalysed by -glucosidase and not by invertase. On the basis of this work, -glucosidase from M. subhyalinus appears to be a valuable tool for the preparation of neoglycoconjugates.     

Presence of hybridizing DNA sequences homologous to bovine acidic and basicβ-crystallins in all classes of vertebrates

Summary The eye lens-crystallins in cow and chicken are encoded by a family of at least six genes. In order to assess the distribution of the corresponding genes among other vertebrates we hybridized -crystallin sequences (A2, A3/A1, A4, B1, B2, B3), isolated from a bovine lens cDNA library, to Southern blots on whichEcoR1-digested chromosomal DNA was blotted from different vertebrate species. These included human, chimpanzee, calf, rat, pigeon, duck, monitor lizard, toad, trout, and lamprey. Positive hybridization signals were found in the representatives of virtually all classes of vertebrates. The basic B-crystallins gave hybridization signals in more species than the acidic A ones. In monitor lizard and toad the weakest hybridization signals for basic crystallin probes were found. For acidic crystallin probes the distribution pattern was more simple; among cold-blooded vertebrates a signal for A2 was found in trout and lamprey, for A4 in trout, and for A3/A1 only in toad. The results demonstrate that the duplications leading to the -crystallin gene family occurred before or during the earliest stages of vertebrate evolution.     

Effects of trichloroethylene,tetrachloroethylene and dichloromethane on enzymatic activities in soil

Summary Enzyme assays for -glucosidase, -acetylglucosaminidase, phosphatase, phosphodiesterase, and proteinase were made in soil samples incubated for two months after contamination with trichloroethylene, tetrachloroethylene, and dichloromethane. These volatile chlorinated hydrocarbons were added at doses of 10, 100, and 1000 g per 100 g dry soil, respectively. Almost no effect was observed in soil sample contaminated with 10 g of the chemicals when compared with control soil. When 100 g of the volatile chlorinated hydrocarbons was added, the activity of -glucosidase, -acetylglucosaminidase and, in part, also of proteinase, was reduced during the first 28 days of incubation but returned to the same or slightly higher level than in the control soil after 2 months. Trichloroethylene, tetrachloroethylene, and dichloromethane at a concentration of 1000 g per 100 g soil primarily inhibited activity of all enzymes under test. However, after two months, the enzymatic activities especially in soil samples contaminated with tetrachloroethylene and dichloromethane were found to be at the same or higher level than in the control soil.     

<Emphasis Type="Italic">Tuamotuichthys bispinosus</Emphasis>, a new genus and species from off Tuamotu Islands,South Pacific Ocean (Ophiidiformes,Bythitidae)

A new bythitid genus and species, Tuamotuichthys bispinosus, is described from an adult male (88mm SL), caught by dredge in 1990, off Morane Atoll, Tuamotu Archipelago (23°0843 S, 137°0919 W, 536m). It is characterized by having large scales on head and body, two opercular spines, few gill rakers (total 11, 3 of them long), depressed head, vertically expanded posterior maxilla, and a short predorsal (29% SL), each of which resembles several known genera (Calamopteryx, Cataetyx, Diplacanthopoma, Grammonus, and Lucifuga). The combination of character states, however, does not fit any of the known bythitid genera.     

A Comment on La Vaque and Rossiter

The paper by La Vaque and Rossiter recommends that placebo-controlled trials only be conducted when best proven treatments are not available. In this comment, I review evidence suggesting that identifying best proven treatments is a complex task involving many variables and that making such a decision entails many real-world difficulties.     

Validation of an ELISA test kit for the detection of ochratoxin A in several food commodities by comparison with HPLC

An ELISA Microtiter Plate, Ochratoxin Test called AgraQuant^® was validated to measure ochratoxin A in a range from 2 to 40ppb in corn, milo, barley, wheat, soybeans and green coffee. The test is performed as a solid phase direct competitive ELISA using a horseradish peroxidase conjugate as the competing, measurable entity. For the test method, ochratoxin A is extracted from ground samples with 70% methanol and sample extracts plus conjugate are mixed and then added to the antibody-coated microwells. After 10min incubation at room temperature, the plate is washed and enzyme substrate is added and allowed to incubate for an additional 5min. Stop solution is then added and the intensity of the resulting yellow color is measured optically with a microplate reader at 450nm. Results obtained from internal validation studies assessing accelerated stability indicate a 1year shelf life; accuracy and precision are comparable to HPLC from 0 to 80ppb and limit of detection in corn is 1.9 ppb and other food commodities is up to 3.8 ppb. Comparison of the method to HPLC, ability to detect individual ochratoxins, and ruggedness of the test kits determined this test to be rugged from 18 to 30°C, sensitive, accurate, precise and effective comparable to HPLC for measuring ochratoxin A ranging from 2 to 40ppb in several commodities.     

Alterations in certain lysosomal glycohydrolases and cathepsins in rats on dexamethasone administration

Glucocorticoids have been used in the treatment of a number of diseases where immunological intolerance plays a predominant role. Since immunological intolerance points to the involvement of lysosomal enzymes and glucocorticoids are known to affect their activities, we have attempted to study the effect of these steroids on cardiac and renal enzymes. Dexamethasone, a glucocorticoid, is administered subcutaneously to male Wistar rats at a dosage of 2.5 mg/kg/week on alternate days for two weeks. After withdrawing the steroid, the animals are monitored for one week to oversee the recovery process. Total and free activities of glycohydrolases and cathepsins in serum, heart and kidney are assayed on the days 4, 8, 12, 16 of dexamethasone administration and also on days 4 and 8 following discontinuation of the steroid. During dexamethasone administration, a significant decrease in both the free and total activities of -glucuronidase, -N-acetyl glucosaminidase, -galactosidase, -galactosidase, -mannosidase, cathepsin B and cathepsin D are observed in heart and kidney, but the enzyme levels are shown to increase in serum. On withdrawal of the steroid, the activities of -glucuronidase, -N-acetyl glucosaminidase, -galactosidase are found to be increased in heart and kidney, whereas, the activity of -mannosidase remains within normal values. Thus, it could be seen that dexamethasone alters the pattern of glycohydrolases and cathepsins, which are involved in protein degradation.     

β-Glucuronidase activity in human T and B lymphocytes and the Tμ and Tγ subpopulations

Summary The -glucuronidase staining characteristics of isolated T cell populations and the T and T enriched fractions derived of them were studied. T lymphocytes were obtained from purified T lymphocytes by ox-IgG rosette sedimentation. The rosette-forming cells in the pellet were referred to as T lymphocytes, whereas the lymphocytes in the interface were referred to as T depleted or T lymphocytes. B cells were studied on rosetted mononuclear cells with either mouse erythrocytes or with Staphylococcus Aureus (Cowan I) bacteria, after a preceeding polyvalent anti-human Ig treatment of the cells. While B cells showed mostly no reactivity, T and T cells were respectively characterised by a dot-like and granular pattern of reactivity. These findings are in agreement with those observed by others after -naphthyl-acetate esterase or acid phosphatase staining. Within the T lymphocyte fraction, the T non-, non lymphocytes seemed to have a granular pattern of reactivity. The same staining pattern was found in non-B, non-T lymphocytes.     

UDP-GlcNAc: Galβ3GalNAc-Mucin: (GlcNAc → GalNAc) β6-N-Acetylglucosaminyltransferase and UDP-GlcNAc: Galβ3(GlcNAcβ6) GalNAc-Mucin (GlcNAc → Gal)β3-N-Acetylglucosaminyltransferase from Swine Trachea Epithelium

Summary Two specific -N-acetylglucosaminyltransferases involved in the branching and elongation of mucin oligosaccharide chains, namely, a 1,6 N-acetylglucosaminylsaminyltransferase that transfers N-acetylglucosamine from UDP-N-acetylglucosamine to Gal3GalNAc-Mucin to yield Gal3(GlcNAc6)GalNAc-Mucin and a 3-N-acetylglucosaminyl transferase that transfers N-acetylglucosamine from UDP-N-acetylglucosamine to Gal3(GlcNAC6)GalNAc-mucin to yield GlcNAc3Gal3 (GlcNAc6)GalNAc-Mucin were purified from the microsomal fraction of swine trachea epithelium. The 1,6-N-acetylglucosaminyltransferase was purified about 21,800-fold by procedures which included affinity chromatography on DEAE columns containing bound asialo Cowper's gland mucin glycoprotein with Gal1,3GalNAc side chains. The apparent molecular weight estimated by gel filtration was found to be about 60 Kd. The purified enzyme showed a high specificity for Gal1,3GalNAc chains and the most active substrates were mucin glycoproteins containing these chains. The apparent Km of the 6-glucosaminyltrans-ferase for Cowper's gland mucin glycoprotein containing Gal1,3GalNAc chains was 0.53 µM; for UDP-N-acetylglucosamine, 12 µM; and for Gal 1,3GalNAc NO₂ø, 4 mM. The activity of the 6-glucosaminyltransferase was dependent on the extent of glycosylation of the Gal3GalNAc chains in Cowper's gland mucin glycoprotein.The best substrate for the partially purified 3-Glucosaminyltransferase was Cowper's gland mucin glycoprotein containing Gal1,3(GlcNAc6)GalNAc side chains. This enzyme showed little or no activity with intact sialylated Cowper's gland mucin glycoprotein or derivatives of this glycoprotein containing GalNAc or Gal1,3GalNAc side chains.The radioactive oligosaccharides formed by these enzymes in large scale reaction mixtures were released from the mucin glycoproteins by treatment with alkaline borohydride, isolated by gel filtration on Bio-Gel P-6 and characterized by methylation analysis and sequential digestion with exoglycosidases. The oligosaccharide products formed by the 6- and 3-glucosaminyltransferases were shown to be Gal3(GlcNAC6) GalNAc and GlcNAc3 Gal3(GlcNAC6)GalNAc respectively.Taken collectively, these results demonstrate that swine trachea epithelium contains two specific N-acetylglucosaminyltransferases which catalyze the initial branching and elongation reactions involved in the synthesis of O-linked oligosaccharide chains in respiratory mucin glycoproteins. The first enzyme a 6-glucosaminyltransferase converts Gal3GalNAc chains in mucin glycoproteins to Gal3(GlcNAc6)GalNAc chains. This product is the substrate for a second 3-glucosaminyltransferase which converts the Gal3(GlcNAc6)GalNAc chains to GlcNAc3Gal(GlcNAc6)GalNAc chains in the glycoprotein. The 3-glucosaminyltransferase did not utilize Gal3GalNAc chains as a substrate and this results in an ordered sequence of addition of N-acetylglucosamine residues to growing oligosaccharide chains in tracheal mucin glycoproteins.Abbreviations NeuNAc N-acetylneuraminic acid - GalNAcol N-acetylgalactosaminitol - CGMG Cowper's gland mucin glycoprotein - GalNAc-CGMG Cowper's gland mucin glycoprotein containing GalNAc side chains O-glycosidically linked to serine or threonine - Gal3GalNAc-CGMC Cowper's gland mucin glycoprotein containing Gal3GalNAc side chains - MES 2-(N-morpholino) Ethane Sulfonic acid - PBS Phosphate Buffered Saline     

Genetics of Ia-like alloantigens in chickens and linkage withB major histocompatibility complex

Two sets of backcross matings were performed to test for linkage between genes coding for the Ia-like antigens (Ia) and the B erythrocyte antigens (Ea-B) of the chicken. Evidence is presented which indicates that the la antigens are determined by a single codominant locus and that theEa-B and Ia loci are on the same chromosome. Failure to detect a single recombinant between theEa-B and Ia loci out of 208 progeny suggests close linkage of the two genes with a map distance of up to about 2 centimorgans. The Ia genes are thus included in theB major histocompatibility complex of the chicken.     

Beiträge zur funktionellen Morphologie der Neurohypophyse

Zusammenfassung Beidseitige Adrenalektomie und Hypophysektomie führen bei der Ratte zu gleichartigen histologischen Veränderungen in der Zona externa infundibuli. In beiden Fällen treten in der normalerweise weitgehend goniorinegativen Zona externa infundibuli große Mengen gomoripositiver Granula auf. Sie scheinen Fasern anzugehören, die senkrecht zur Längsachse des Infundibulum verlaufen.Die Befunde werden als weiterer Hinweis dafür betrachtet, daß die gomoripositiven Substanzen der Zona externa infundibuli eine Bedeutung für die Steuerung der Nebennierenrindenfunktion haben.

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Studies on the functional morphology of the neurohypophysisII. Comparison of histological changes in the median eminence of the rat after bilateral adrenalectomy and hypophysectomy

Summary Bilateral adrenalectomy and hypophysectomy in the rat produce similar histological changes in the outer layer of the median eminence. In both cases, abundant gomoripositive granules are observed in the outer layer, which normally reacts gomorinegative. The gomoripositive granules seem to belong to fibres running vertically through the infundibulum.These findings are regarded as a further indication, that the gomoripositive substances of the outer layer of the median eminence play a significant role in controlling adrenocortical function.

     

Ultrastructure of four types of striated muscle fibers in the atlantic hagfish (Myxine glutinosa,L.)

Summary Four types of striated muscle fibers with distinctive ultrastructure were defined in the Atlantic hagfish (Myxine glutinosa, L.): white, intermediate, and red fibers of m. parietalis, and red fibers of m. craniovelaris.White fibers are thick, contain very few mitochondria and fat vacuoles, and possess distinct and separate myofibrils with thin Z-disks and distinct M-lines. Intermediate fibers are thinner, possess largely similar myofibrils that often are even better separated due to a higher content of fat vacuoles and especially mitochondria and glycogen granules. Red fibers of m. parietalis contain large amounts of mitochondria, fat vacuoles, and glycogen granules. Their myofibrils possess M-lines, and although branching more, the myofibrils of red fibers conform with a Fibrillenstruktur pattern like those of white and intermediate fibers. Red fibers of m. craniovelaris are very thin, possess many smaller fat vacuoles, and large amounts of mitochondria and glycogen granules. The myofibrils are significantly thinner than in m. parietalis fibers, run as quite independent well separated units, possess thicker Z-disks, and lack M-lines. Large amounts of myosatellite cells are associated with these red fibers.Triads are located near A/I-junctions in all four fiber types and occur irregularly, the density of triads being different in the various fiber types.We are indebted to Dr. Finn Walvig, Biological Station, University of Oslo, Drøbak, for supply of hagfishes, and we also wish to thank Dr. Jan K. S. Jansen, Institute of Physiology, University of Oslo, for valuable suggestions during this study.     

Nicotiana chloroplast genome

Summary Using the existing restriction map and probes from wheat and pea ct-DNA, seven protein genes have been localized in the chloroplast genome of N. tabacum. On the clock-like map, the location of each gene is indicated by its time zone: the 15.2 kD polypeptide of the cytochrome b/f complex at 315, cytochrome f at 430, LS of RuBPCase at 450, both and subunits of ATP synthase at or near 500, proton-translocating subunit of ATP synthase at 820, subunit of ATP synthase at 840 and the 32 kD protein at 930. The genome organization of Nicotiana chloroplast DNA is similar to spinach.     

Habituation rules for a theory of the cerebellar cortex

A quantitative model of cerebellar cortical function is described with a complete formalization based on (i) the topology of cerebellar cortical neuronal network, (ii) some particular synaptic properties of cell classes in cerebellum cortex, and (iii) the dynamics of excitation in this network. For (i), a construction of functional classes around one Purkinje cell is given and their existence is discussed. For (ii), as in Marr-Albus model, the modifiability of synapses between parallel fibres and Purkinje cell is assumed. But the formalization permits to consider the consequences of such a property at the level of glomerulus (with granule cells) which is known as a complex transformation system. For (iii) habituation rules are assumed. It is shown that this method leads to some interesting properties in the functioning of cerebellar cortex. Particularly, emitting frequency along a Purkinje cell axon results from a discrimination by the system between transformed input signals and an external noise due to all other contexts, and learning could be considered as the result of a conflict between a set of patterns and the transformed input signals. This model could be a basis for future numerical simulations.     

Protoplasmic streaming in the giant unicellular green algaAcetabularia mediterranea

Summary Protoplasmic Streaming inAcetabularia mediteranea has been studied by microcinematography in 1. germinating zygotes, 2. germlings before the differentiation of rhizoids and apices, 3. young cells with rhizoids and apices, 4. vegetative cells-several centimeters in length, 5. cells with a maximum sized cap, containing secondary nuclei, and 6. cells after cyst formation. Intracellular transport is found to occur at a network-system of thin filaments and at a different system of headed streaming bands. At the network of filaments chloroplasts are found to move at a velocity of 1–2 m/sec. Headed streaming bands move along the filaments and may lead without interruption from the rhizoid to the apex of the cell andvice versa. The front zone of the streaming bands is occupied by a leading cytoplasmic head-structure. Small vesicles, polyphosphate granula and secondary nuclei are the predominant moving structures in headed streaming bands. The velocity of these particles is found to be 3–11 m/sec. The filament system is found during all developmental stages. Headed streaming bands are undetectable in germinating zygotes and develop from small cytoplasmic droplets in germlings to broad heavily loaded bands in the huge vegetative cell.Transport of secondary nuclei by headed streaming bands is not observed during mitotic divisions and after cyst formation, though moving bands are still present for several weeks after cyst formation.