Peptide des β-Alanins als Ersatz für die freie Aminosäure bei pantothensäurebedürftigen Hefezellen und ihr Verhalten gegenüber dem β-Alanin-Antagonisten Asparagin

Zusammenfassung Pantothensäurebedürftige Hefezellen können ihren Bedarf an diesem Vitamin nicht allein aus -Alanin decken, sondern auch aus Benzoyl--Alanin, -Alanyl-d,l-Norleucin und -Alanyl-l-Histidin. Der Antagonist Asparagin hemmt die Verwertung dieser Peptide genauso wie diejenige der freien Aminosäure. Durch höhere Konzentrationen an -Alanin oder -Alanyl-d,l-Norleucin läßt sich die Hemmwirkung nicht allein kompensieren, es kommt sogar zu einer Förderung des Hefewachstums. Der Antagonist wird dann zum Synergisten.

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Summary The -alanine containing peptides benzoyl--alanine, -alanyl-d,l-norleucine and -alanyl-l-histidine can substitute for the amino acid -alanine in a pantothenic acid requiring yeast. Asparagine, an antagonist of -alanine, affects these peptides in a similar manner. In combination with an overdose of -alanine or -alanyl-d,l-norleucine, asparagine is no longer an antagonist but becomes a synergist.

     

No linkage to the 3β-HSD gene cluster in a kindred affected with 3β-hydroxy-Δ5-C27steroid dehydrogenase deficiency and early onset hepatic failure

We studied the segregation of the genes for 3-hydroxy-C_19/21-steroid dehydrogenase types I and II (3-HSD I and II) in a consanguineous family affected with 3-hydroxy-⁵-C₂₇steroid dehydrogenase (3-OH-C₂₇-SD) deficiency. The results show that the C₂₇ and C_19/21 steroid dehydrogenase activities are encoded by distinct genes that are not in genetic linkage. Further kindreds would assist in screening for linkage of 3-OH-C₂₇-SD to other members of the 3-hydroxysteroid dehydrogenase gene family.     

Prediction of the Secondary Structure Contents of Globular Proteins Based on Three Structural Classes

The prediction of the secondary structural contents (those of -helix and -strand) of a globular protein is of great use in the prediction of protein structure. In this paper, a new prediction algorithm has been proposed based on Chou's database [Chou (1995), Proteins 21, 319–344]. The new algorithm is an improved multiple linear regression method, taking into account the nonlinear and coupling terms of the frequencies of different amino acids and the length of the protein. The prediction is also based on the structural classes of proteins, but instead of four classes, only three classes are considered, the class, class, and the mixed + and / class or simply the class. Thus the ambiguity that usually occurs between + proteins and / proteins is eliminated. A resubstitution examination for the algorithm shows that the average absolute errors are 0.040 and 0.035 for the prediction of -helix content and -strand content, respectively. An examination of cross-validation, the jackknife analysis, shows that the average absolute errors are 0.051 and 0.045 for the prediction of -helix content and -strand content, respectively. Both examinations indicate the self-consistency and the extrapolating effectiveness of the new algorithm. Compared with other methods, ours has the merits of simplicity and convenience for use, as well as high prediction accuracy. By incorporating the prediction of the structural classes, the only input of our method is the amino acid composition and the length of the protein to be predicted.     

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.     

The glycolipid specificity ofErythrina cristagalli agglutinin

The interaction of¹²⁵I-labeledErythrina cristagalli agglutinin (ECA) with neutral glycosphingolipids on thin layer chromatograms was examined by the overlay technique followed by radioautography. The lectin bound topara-globoside with a sensitivity about 10 times higher than to lactosylceramide or globoside, in agreement with the specificity of the lectin forN-acetyllactosamine. The lower limit of detection ofpara-globoside was about 0.66 nmol. The specific binding of ECA to this glycolipid was confirmed by a highly sensitive enzyme-linked lectin assay (ELLA), utilizing the horseradish peroxidase-avidin-biotin system for detection of bound lectin. Overlays of neutral glycosphingolipid extracts from human erythrocyte membranes and from human granulocytes with ECA demonstrated that the lectin can be employed for the detection of small amounts ofpara-globoside in biological materials also in the presence of excess globoside. No staining was obtained when thin layer chromatograms of neutral glycosphingolipid extracts from rabbit erythrocyte membranes were overlayed with¹²⁵I-ECA. Afterin situ treatment of the chromatograms with -galactosidase, the lectin bound to several components, one of which had a mobility corresponding to that of the pentahexosylceramide Gal3Gal4GlcNAc3Gal4Glc1Cer, the major neutral glycosphingolipid of rabbit erythrocytes, thus providing further evidence for the specificity of ECA forpara-globoside.Abbreviations GSL glycosphingolipid(s) - CDH lactosylceramide, Gal4Glc1Cer - CTH trihexosylceramide, Gal4Gal4Glc1Cer - GLOB globoside, GalNac3Gal4Gal4Glc1Cer - PG para-globoside, Gal4GlcNAc3Gal4Glc1Cer - AsG_M1 asialo-G_M1, Gal3GalNAc4Gal4Glc1Cer - FORS Forsmann antigen, GalNAc3GalNAc3Gal4Gal4Glc1Cer - CPH pentahexosylceramide, Gal3Gal4GlcNAc3Gal4Glc1Cer - ECA Erythrina cristagalli agglutinin - SBA soybean agglutinin - PBS phosphate-buffered saline - PVP-40 polyvinylpyrrolidone M.W. 40000 - BSA bovine serum albumin - HRP-avidin horseradish peroxidase conjugated to avidin - ELLA enzyme-linked lectin assay - ELISA enzyme-linked immunosorbent assay - PMNL polymorphonuclear leukocytes - HPTLC high performance thin layer chromatography     

Comparative studies of the development of endosperm-degrading enzymes in malting sorghum and barley

The endosperm cell walls of barley are degraded extensively during malting whilst those of sorghum are not. Malting barley produced endo--1,3:1,4-glucanase, endo--1,3-glucanase and pentosanase in large quantities. In contrast, malting sorghum developed mainly endo--1,3-glucanase and pentosanase. Although the limited break-down of the endosperm cell walls of sorghum may reflect sub-optimal activities of -glucanases, such as endo--1,3:1,4-glucanases, it is possible that the highly intractable nature of the cell walls and their high protein content (approx. 60%) may contribute to the low susceptibility of sorghum endosperm cell walls to enzymic degradation during malting.

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Résumé Les parois cellulaires endospermiques de l'orge sont fortement dégradées pendant le maltage, tandis que celles du sorghum ne le sont pas. L'orge en maltage produit l'endo--1,3:1,4-glucanase, l'endo--1,3-glucanase et la pentosanase en grandes quantités. Par contre, le sorghum en maltage dévéloppe principalement l'endo--1,3-glucanase et la pentosanase. Bien que la destruction limitée des parois cellulaires endospermiques puisse réflecter des activités sub-optimales des -glucanases, comme l'endo--1,3:1,4-glucanase, il n'en est pas molns possible que la nature hautement intractile des parols cellulaires et leur contenu élevé en protéines (approximativement 60%) pulsse contribuer à la faible susceptibilite des parois cellulaires endospermiques du sorghum à la dégradation enzymatique durant le maltage.

     

Model of neural visual system with self-organizing cells

This paper describes a model of a neural visual system of a higher animal, in which the capability of pattern recognition develops adaptively. To produce the adaptability, we adopted self-organizing cells, and with them modeled feature-detecting cells which were discovered by Hubel and Wiesel and whose plasticity was found by Blakemore and Cooper. Combining the self-organizing cells and the learning principle of a Perceptron-type system, we constructed a model of the whole visual system. The model is also equipped with an eye movement control mechanism for gazing, which reduces the number of selforganizing cells required for pattern recognition, thus contributing to their quick self-organization. Computer simulation and an experiment using a hardware simulator showed that self-organizing cells quickly become sensitive to the features often seen and that the resulted system can classify patterns with a rather small number of feature-detecting cells.     

Analysis of responses to human synthetic adrenomedullin and calcitonin gene-related peptides in the hindlimb vascular bed of the cat

Vasodilator responses to human adrenomedullin (hADM), a newly discovered hypotensive peptide, human calcitonin gene-related peptide- (hCGRP-) and hCGRP-, which share structural homology with hADM, were compared in the hindlimb vascular bed of the cat under constant flow conditions. Injections of hADM (0.003-1 nmol), hCGRP-, and hCGRP- (0.003-0.3 nmol) into the perfusion circuit caused dose-related decreases in hindlimb perfusion pressure. Vasodilator responses to hCGRP- and hCGRP- were similar in potency and duration, and the doses of hCGRP- and hCGRP- required to reduce hindlimb perfusion pressure 40 mm Hg (ED40 mm Hg) were significantly lower than the ED40 mm Hg for hADM. The duration of the hindlimb vasodilator responses to hCGRP- and hCGRP- were significantly longer than the duration of the response to hADM. Amylin, a peptide that shares structural homology with ADM and with CGRP, had no significant effect on hindlimb perfusion pressure when injected in doses up to 1 nmol. Decreases in hindlimb perfusion pressure in response to hADM, hCGRP-, and hCGRP- were not altered by L-N5-(1-iminoethyl)-ornithine (L-NIO) in a dose of the nitric oxide synthase inhibitor that decreased the vasodilator response to acetylcholine or by the cyclooxygenase inhibitor, meclofenamate, in a dose that decreased the vasodilator response to archidonic acid. The present data demonstrate that hADM, hCGRP-, and hCGRP- have potent, but relatively short-lasting, vasodilator activity, and that vasodilator responses are not dependent on the release of nitric oxide or vasodilator prostaglandins in the hindlimb vascular bed of the cat.     

Somatic embryogenesis in callus cultures of Rosa hybrida L. cv. Landora

Callus was initiated from immature leaf and stem segments of rose (Rosa hybrida cv. Landora) and subcultured every four weeks on a basal medium of half-strength Murashige & Skoog (1962) salts plus 30 g l^-1 sucrose (1/2 MS) and supplemented with 2.2 M BA, 5.4 M NAA and 2.2–9.0 M 2,4-D. Embryogenic callus and subsequently somatic embryos were obtained from 8-week-old callus culture on 1/2 MS+2.2 M BA+0.05 M NAA+0.3 M GA₃+200–800 mg l^-1 L-proline. Long-term cultures were established and maintained for up to 16 months by repeated subculture of embryogenic callus on L-proline deficient medium. About 12% of cotyledonary stage embryos taken from cultures cold-stored at 8±1°C for 4 days germinated on 1/2 MS+2.2 M BA+0.3 M GA₃+24.7 M adenine sulphate.Abbreviations BA benzyladenine - NAA -naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid     

Incorporation of 14CO2 in prenylquinones of Chlorella pyrenoidosa

Incorporation and release of ¹⁴C-label in prenylquinones of Chlorella was investigated under steady state conditions. After one hour of ¹⁴CO₂-photosynthesis all plastid quinones investigated were labeled. The highest label was found in phylloquinone (18%) while -tocopherol exhibits the lowest label (0.38%). Among the plastoquinones, plastohydroquinone-9 shows a higher labeling degree (5.1%) and a faster labeling kinetic than plastoquinone-9 (1.6%). After replacement of ¹⁴CO₂ against ¹²CO₂ the total radioactivity in plastohydroquinone-9, -tocopherol and phylloquinone decreases but in -tocoquinone and plastoquinone-9 proceeds further. From this labeling kinetic we conclude, that newly synthesized [¹⁴C]-tocopherol molecules are converted to [¹⁴C]-tocoquinone and [¹⁴C]plastohydroquinone-9 molecules to [¹⁴C]plastoquinone-9. From their ¹⁴C-incorporation kinetic half-lives could be calculated for all prenylquinones in the same ranges as previously found for the chlorophylls and carotenoids (Grumbach et al., 1978). Half-lives are shorter in plastohydroquinone-9 (30 min) and plastoquinone-9 (40 min) than in phylloquinone (55 min), -tocoquinone (50 min) and -tocopherol (220 min). This means that all prenyl-lipids such as chlorophyll a, -and -carotene, plastohydroquinone-9 and plastoquinone-9 which are more directly involved in the process of photosynthesis are subject to a continuous and higher turnover than the xanthophyll and -tocopherol. From the fast labeling kinetic and short half-lives of -tocoquinone and especially phylloquinone with a labeling degree of 12% after one hour of ¹⁴CO₂ photosynthesis we suppose that perhaps these two prenylquinones are also involved in the photosynthetic activity of chloroplasts.     

Evolutionary origin of the class A and class C β-lactamases

Summary The protein sequences of 18 class A -lactamases and 2 class C -lactamases were analyzed to produce a rooted phylogenetic tree using the DD peptidase of Streptomyces R61 as an outgroup. This tree supports the penicillin-binding proteins as the most likely candidate for the ancestoral origin of the class A and class C -lactamases, these proteins diverging from a common evolutionary origin close to the DD peptidase. The actinomycetes are clearly shown as the origin of the class A -lactamases found in other non-actinomycete species. The tree also divides the -lactamases from the Streptomyces into two subgroups. One subgroup is closer to the DD peptidase root. The other Streptomyces subgroup shares a common branch point with the rest of the class A -lactamases, showing this subgroup as the origin of the non-actinomycete class A -lactamases. The non-actinomycete class A -lactamase phylogenetic tree suggests a spread of these -lactamases by horizontal transfer from the Streptomyces into the non-actinomycete gram-positive bacteria and thence into the gram-negative bacteria. The phylogenetic tree of the Streptomyces class A -lactamases supports the possibility that horizontal transfer of class A -lactamases occurred within the Streptomyces.     

K-current fluctuations in inward-rectifying channels of frog skeletal muscle

Summary K currents and K-current fluctuations were recorded in inwardly rectifying K channels of frog skeletal muscle under voltage-clamp conditions. External application of 0.2 to 10mm Cs reduces the inward mean K current but produces a distinct increase of the spectral density of K-current fluctuations. The additional fluctuations arise from the random blocking by Cs ions. From the variance of current fluctuations, the steady-state current and the probability of the open unblocked channel an effective single-channel conductance ^* was calculated. ^* strongly depends on the external Cs concentration (7.8 pS at 0.2mm Cs, 2.1 pS at 10mm Cs). This dependence is interpreted in terms of a two-step blocking process: (1) a fast exchange of Cs ions between the external solution and a first binding site inside the channel which leads to the Cs-modulated effective single-channel conductance, and (2) a slow Cs binding to a second site deeper in the channel which produces the observed current fluctuations. With this hypothesis we obtained a real single-channel conductance of 10 pS and a real density ofn4 inwardly rectifying channels per m² of muscle surface area.     

Is There a Field-Theoretic Explanation For Precursor Biopolymers?

A Hu-Barkana-Gruzinov cold dark matter scalarfield may enter a weak isospin invariant derivative interactionthat causes the flow of right-handed electrons to align parallelto (). Hence, in the outer regions of galaxies where () islarge, as in galactic halos, the derivative interaction mayinduce a chirality-imbued quantum chemistry. Such a chirality-imbued chemistry would in turn be conducive to the formation ofabundant precursor biopolymers on interstellar dust grains,comets and meteors in galactic halo regions, with subsequentdelivery to planets in the inner galactic regions where and() are concomitantly near zero and left-right symmetricterrestrial quantum chemistry prevails.     

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.     

A new enzymatic route to the synthesis of 12-ketoursodeoxycholic acid

Summary Cholic acid (3,7,12-trihydroxy-5-cholanoic acid) was completely and selectively transformed into 12-ketoursodeoxycholic acid (3,7-dihydroxy-12-oxo-5-cholanoic acid) by means of two consecutive enzymatic steps catalyzed, the first, by 7- and 12-hydroxysteroid dehydrogenase and, the second, by 7-hydroxysteroid dehydrogenase. Coenzyme regeneration was carried out with -ketoglutarate-glutamate dehydrogenase and glucose-glucose dehydrogenase, respectively.     

Restriction fragment analysis of ‘null’ forms at the Gli-1 loci of bread and durum wheats

Summary Wheat accessions lacking some of the - and -gliadin components encoded by the Gli-1 loci on the short arm of chromosome 1D in bread wheat and chromosome 1A in durum wheat were studied by two-dimensional polyacrylamide gel electrophoresis and restriction fragment analysis. Digested genomic DNAs of normal and null forms were probed with a cDNA clone related to -/-gliadins and with a genomic clone encoding an LMW subunit of glutenin. The hybridisation patterns with the -/-gliadin probe were similar to those of cvs Chinese Spring and Langdon used as standards for bread and durum wheats, respectively, but several restriction fragments located on the 1D chromosome of bread wheat and the 1A chromosome of durum wheat were absent in the null forms. In addition, specific LMW glutenin fragments encoded by the same chromosomes were also absent in the null forms, suggesting that simultaneous deletions of blocks of genes for both -/-gliadins and LMW glutenins had occurred. Comparisons of the protein and RFLP patterns enabled some proteins to be mapped to specific restriction fragments.     

Abbau der 4-Chlorbenzoesäure durch eineArthrobacter-Species

Zusammenfassung EineArthrobacter-Species, die 4-Chlorobenzoesäure als einzige Kohlenstoffquelle verwerten kann, gibt beim Wachstum auf dieser Verbindung 4-Hydroxybenzoesäure und Protocatechusäure ins Medium ab. Der weiter Abbau des aromatischen Ringes erfolgt durch meta-Spaltung. Beim Wachstum derArthrobacter-Species auf Benzoesäure trit im Medium cis,cis-Muconsäure auf. In diesem Fallewird also der ortho-Weg eingeschlagen. Die Enzyme für beide Abbauwege sind induzierbar.

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Degradation of 4-Chlorobenzoic acid by anArthrobacter species

AnArthrobacter sp. growing on 4-Chlorobenzoic acid as its sole source of carbon excretes 4-hydroxybenzoic acid and protocatechuic acid into the culture medium. Protocatechuic acid is further attacked by meta-cleavage. During growth of theArthrobacter sp. on benzoic acid cis-cis muconic acid can be isolated from the medium, suggesting the involvement of the ortho-cleavage pathway. The enzymes both for the meta- and the ortho-cleavage pathway are inducible.

Dem Andenken an Professor Bernhauer gewidmet     

Idioms of distress: Kinship and sickness among the people of the Kingdom of Tonga

Idioms of distress refers to the popular expression of emotional tension that arises in the relationship between sickness and kinship. By reference to case studies and discussions among the Polynesian people of Tonga, the author shows where such tension arises and how it influences the sickness process. Sickness is necessarily a collective phenomenon which can best be understood not simply as a clinical event, but as an experience that is part of the experience of family. Various ways of expressing distress as a reflexive encounter between personal and cultural meaning systems are reviewed, as are several new concepts such as doing sickness as kinship, and turning in the process of decision making in the kinship management of sickness. The explanatory models of sickness in Tonga are shown to encompass culturally sanctioned expressions of distress as part of the adaptive coping mechanisms in that society. Distress frequently emerges in somatic form, as a number of studies have shown. However, the author emphasizes the kinship meaning of sickness, kinship management and sickness therapy, the adaptive process of idiomatic expressions of distress, which are expanded here and offered as potential avenues for elaboration in other cultural milieu. Two aspects of the notion idioms of distress are noted, and the phenomenon is understood as a process which acts as a prime mover in social change.     

The structure of the incompatibility factors of Schizophyllum commune: Constitution of the three classes of B factors

Summary The B factors of Schizophyllum commune are of 3 classes: The high recombining class I has 7 alleles and 7 alleles; the low recombining classes are class II with 7 allels and probably 2 alleles and class III with probably 2 (or also 2) alleles and 7 allels. A fourth hypothetical class (-) was not found and either does not exist or is indistinguishable from class III by the tests employed. The and alleles differ from and by either (a) mutations affecting both mating specificity and recombination frequency, or (b) deletions involving most of the B region.The research was supported by a grant from the Atomic Energy Commission of the U.S. No. (30-1)-3875 and was performed at the Biological Laboratories, Harvard University, Cambridge, Mass., U.S.A.     

Calpain-PKC Inter-Relations in Mouse Hippocampus: A Biochemical Approach

In previous studies, we isolated and identified a -calpain/PKC complex from rabbit skeletal muscle. Here, we have used specific purification procedures in order to study the interactions between -calpain and PKC in mouse hippocampus, a brain structure implicated in memory processes. We observed that -calpain and conventional PKCs (, II and ) are co-eluted after anion exchange chromatography. In contrast to our previous results obtained on skeletal muscle, -calpain and PKC isoenzymes were dissociated after gel filtration chromatography. Furthermore, -calpain induced the proteolytic conversion of PKC, II, and into PKM, II, and with a preferential hydrolysis of PKC, a specific isoenzyme of the nervous system. Although the -calpain/PKC interactions in the hippocampus are quite different from skeletal muscle, our results however, point out the functional importance of these inter-relations. Moreover, as PKC has been involved in the biochemical events underlying learning and memory, the preferential relationship between -calpain and PKC promotes the importance of the role that -calpain could play in the cellular mechanisms of memory formation.