Two sulfur-containing ansamycin antibiotics from Streptomyces albolongus

Two sulfur-containing ansamycin antibiotics were isolated from the culture broth of Streptomyces albolongus C-46366; the major one was identical with awamycin and the minor one was a new ansamycin antibiotic, ansathiazin. Their structures were elucidated from their reactions and spectroscopic analyses. These antibiotics were active against gram-positive bacteria, acid-fast bacteria and a protozoan.     

Lipopurealins,novel bromotyrosine derivatives with long chain acyl groups,from the marine spongePsammaplysilla purea

Summary Lipopurealin-A,-B and-C have been isolated from the Okinawan marine spongePsammaplysilla purea. They are inhibitors of Na, K-ATPase, and the structures have been elucidated on the basis of spectral data. They are bromotyrosine derived structures with saturated long chain acyl parts.Physiologically active marine natural products from Porifera X. Preceding paper: Nakamura, H., Wu, H., Kobayashi, J., Nakamura, Y., Ohizumi, Y., and Hirata, Y., Tetrahedron Lett.26 (1985) 4517.     

Bombykol biosynthesis from deuterium-labeled (Z)-11-hexadecenoic acid

Summary (Z)-[11,12d ₂]-11-Hexadecenoic acid was applied topically to the pheromone gland of silkworm pupa. After eclosion, the gland components were analyzed by capillary GC-MS. (Z)-11-Hexadecenoic acid, a characteristic fatty acid of the gland, was found to be a precursor of bombykol, (Z)-11-hexadecenol and bombyk acid.We are greatful to Dr Y. Tamaki and Mr H. Sugie of the National Institute of Agricultural Science for mass spectrometric analysis and Dr M. Noda of Kyoto Prefectural University for valuable discussion. This work was supported by a Grant-in-aid for Scientific Research (to K.H. No. 56112011) from the Ministry of Education, Science and Culture of Japan.     

A testis-specific lactate dehydrogenase in the pipid frog,Hymenochirus boettgeri

Summary Lactate dehydrogenase zymograms of mature testes ofHymenochirus boettgeri show in addition to the five isozymes composed of LDH-A and LDH-B subunits, a second 5-band system which is due to isozymes formed between LDH-A and a third subunit, LDH-C. These testis-specific LDH-C isozymes appear around 6 months after metamorphosis indicating that their expression is correlated with sexual maturity as is generally the case in mammals. This is the first report of a testis-specific LDH isozyme in a lower tetrapod; such isozymes have hitherto only been reported in mammals and in the pigeon.This study was supported by a grant from the Fond National Suisse de la Recherche Scientifique to H.R.K. We thank M. Sateur and F. Bourquin for their technical help.     

Hydroxylation of menthols and cineoles withm-chloroperbenzoic acid

Summary Reaction of menthols and cineoles withm-chloroperbenzoic acid afforded tertiary, secondary, and primary alcohols, some of which were natural products having potent plant growth regulatory activity or were mammlianm metabolies.We thank Nippon Terpene Co. Ltd for their generous gift of the compounds used in this work and Drs M. Kido and Y. Fukuyama, Otsuka Pharmaceutical Co. Ltd, for measurement of high resolution mass spectra. The present work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Health and Welfare.     

Origin and identification of bacteria which produce kairomones in the frass ofAcrolepiopsis assectella (Lep., Hyponomeutoidea)

The volatiles used by the parasitoidDiadromus pulchellus to find its host, the leek moth, are produced by the bacteria developing in the frass of the host larvae. The origin and the nature of these bacteria were investigated. Samples were taken from healthy leeks and from infested leeks in the field, as well as from the frass of larvae reared in the laboratory either on the host plant or on an artificial diet. The various species of bacteria identified were cultured in the presence of precursors of leek sulphur volatiles and their volatile emissions were analysed.Klebsiella oxytoca and variousBacillus, common decomposers of plant matter, were the principal species producing active volatiles which were alkyl disulphides.     

A new selective insecticidal uncoupler of oxidative phosphorylation

Summary A new aryl hydrazone structure with high insecticidal activity against the Australian sheep blowfly,Lucilia cuprina, was shown to have a higher activity as an uncoupler of oxidative phosphorylation in insect compared to mammalian mitochondrial preparations. This compound possesses the requirements of other uncouplers in its measured pKa and lipid solubility. However, when compared to a closely related structure with similar physicochemical properties, its insecticidal and insect mitochondrial uncoupling activities are greater and it exhibits decreased mammalian toxicity corresponding to this differential biochemical selectivity.Acknowledgment. We thank Mr K. Rihs for preparation of the hydrazones and Prof. Dr K.H. Büchel for supply of hydrazone III.     

Inhibitory effect of bacterial ubiquinones on the settling of barnacle,Balanus amphitrite

In an attempt to clarify the influence of marine bacteria on the settling of fouling invertebrate larvae, we screened for inhibitors, produced by marine bacteria, of settling by cyprids of the barnacle,Balamus amphitrite. We found that the culture broth ofAlteromonas sp. strain number KK10304, which was associated with the marine sponge,Halichondria okadai, effectively inhibited settling of the cyprids. Bioassay-guided isolation indicated ubiquinone-8 (1) as an effective inhibitor of cyprid settling. As ubiquinones are widely distributed in bacteria, several related compounds were also tested.     

Insect antifeedant terpenes,hot-tasting to humans

Summary Four dialdehyde sesquiterpenoids (1–4) isolated from East AfricanWarburgia plants exhibit powerful antifeeding activity for larvae of the armyworms,Spodoptera exempta andS. littoralis. The hotness of these sesquiterpenes is associated with this activity. The specific absolute stereochemistry of these antifeedants appears to govern the hotness of the taste.The authors are grateful to Proff. K. Nakanishi, A.S. Kende and T. Kubota for their interest and valuable discussion. Succulatal was provided by Dr. Y. Asakawa. This work was initiated during the stay of I.K. at the International Center of Insect Physiology and Ecology, Nairobi, Kenya. We also thank Mr A. Chapya for technical assistance.     

Allozymes and the hybrid origin of the parthenogenetic lizardCnemidophorus exsanguis

Summary Allozyme electrophoresis was used to determine the hybrid parentage of the triploid parthenogenetic speciesCnemidophorus exsanguis. 19 of 33 loci were polymorphic inC. exsanguis and/or the 7 potential bisexual progenitors. It was determined from allele distributions thatC. exsanguis containes genomes form 3 different species, probablyC. costatus, C. inornatus andC. septemvittatus.Acknowledgments. We thank R.L. Bezy, W.M. Brown, A.E. Greer, C.S. Lieb, J.L. Patton and K.L. Wright for assistance in the field; the Departments of Game and Fish of Arizona and New Mexico and the Direccion General de la Fauna Silvestre, Mexico for providing collecting permits; and R.L. Bezy, H.W. Greene and D.B. Wake for discussions and criticisms. This research was supported in part by NSF Grants DEB 76-20599 and DEB 81-05615 and by a Research Fellowship in Herpetology from the Natural History Museum of Los Angeles County to DAG.     

Competitiveness and communication for effective inoculation byRhizobium,Bradyrhizobium and vesicular-arbuscular mycorrhiza fungi

After a short summary on the ecology and rhizosphere biology of symbiotic bacteria and vesicular-arbuscular (VA) mycorrhiza fungi and their application as microbial inocula, results on competitiveness and communication are summarized. Stress factors such as high temperature, low soil pH, aluminium concentrations and phytoalexins produced by the host plants were studied withRhizobium leguminosarum bv.phaseoli andRhizobium tropici onPhaseolus beans. Quantitative data for competitiveness were obtained by usinggus ⁺ (glucoronidase) labelled strains, which produce blue-coloured nodules. ForPhaseolus-nodulating rhizobia, a group specific DNA probe was also developed, which did not hybridize with more than 20 other common soil and rhizosphere bacteria. Results from several laboratories contributing to knowledge of signal exchange and communication in theRhizobium/Bradyrhizobium legume system are summarized in a new scheme, including also defense reactions at the early stages of legume nodule initiation. Stimulating effects of flavonoids on germination and growth of VA mycorrhiza fungi were also found. A constitutive antifungal compound in pea roots, -isoxazolinonyl-alanine, was characterized.     

β-Lactam resistance in Staphylococcus aureus: the adaptive resistance of a plastic genome

Staphylococci have two mechanisms for resistance to β-lactam antibiotics. One is the production of β-lactamases, enzymes that hydrolytically destroy β-lactams. The other is the expression of penicillin-binding protein 2a (PBP 2a), which is not susceptible to inhibition by β-lactam antibiotics. Strains of S. aureus exhibiting either β-lactamase or PBP 2a-directed resistance (or both) have established a considerable ecological niche among human pathogens. The emergence and subsequent spread of bacterial strains designated as methicillin-resistant S. aureus (MRSA), from the 1960s to the present, has created clinical difficulties for nosocomial treatment on a global scale. The recent variants of MRSA that are resistant to glycopeptide antibiotics (such as vancomycin) have ushered in a new and disconcerting chapter in the evolution of this organism. Received 2 April 2005; received after revision 15 July 2005; accepted 25 July 2005     

(Z)-1,17-diaminooctadec-9-ene,a novel aliphatic diamine from coccinellidae

Summary The isolation and structure determination of (Z)-1,17-diaminooctadec-9-ene from several species of Coccinellidae is reported.Maître de Recherches du Fonds National de la Recherche Scientifique.We gratefully acknowledge the Fonds National de la Recherche Scientifique for financial support. We express our sincere thanks to Professor K. S. Hagen for a generous gift ofH. leis conformis.     

Development of DNA probes for cytotoxin and enterotoxin genes in enteric bacteria

Summary DNA probes to identify the genes encoding toxins in enteric bacteria have been developed. Use of these probes reduces the number of animals required for toxicity testing, as suspect bacteria can be directly tested for the presence of toxin. We have augmented the gene probes available by developing probes against theEscherichia coli enterotoxin LTII and shiga toxin fromShigella dysenteriae 1.The LTII gene fromE. coli 357900 was identified and characterised and a suitable internal probe was obtained. The LTII gene was found not to be common among enterobacteriae from various geographical locations. Isolates predominately of animal origin from Nigeria and Thailand hybridized with the probe.The shiga toxin gene was isolated fromS. dysenteriae 1 by a combination of in vivo and in vitro methods. An internal probe was identified and used against different serogroups ofShigella andE. coli isolated. The probe was found to hybridize withS. dysenteriae 1 isolates and also someS. flexneri andS. sonnei strains. Representatives were tested for toxin production and found to produce toxin at low levels.     

A rat mutant unable to synthesize vitamin C

Summary A colony of Wistar rats with a hereditary defect in L-ascorbic acid-synthesizing ability was established. This rat, like primates and guinea pigs, lacks L-gulonolactone oxidase (EC 1.1.3.8) which catalyzes the last step of L-ascorbic acid biosynthesis. When L-ascorbic acid was added to their drinking water, the rats grew almost normally and were fertile. These mutant rats should be useful not only for nutritional and parmacological studies on vitamin C but also for genetic studies on the lack of this enzyme.Acknowledgments. We are indebted to K. Katagiri, the previous director of Aburahi Laboratories, for his encouragement during the experiment. We are also grateful to H. Nishimura, Professor Emeritus of Kyoto University, and Y. Hasegawa of our laboratory for their helpful suggestions.     

Bactericidal activity againstPseudomonas aeruginosa is acquired by cultured human monocyte-derived macrophages after uptake of myeloperoxidase

Myeloperoxidase (MPO) is an enzyme located within polymorphonuclear neutrophils capable of producting cytotoxic oxidant species that are particularly active against bacteria with polysaccharide capsules.Pseudomonas aeruginosa (10⁶ bacteria per 1 ml) are killed within 1 h in vitro by a MPO/H₂O₂/Cl^– system (48 mU=132 ng of MPO). The question arose as to whether human macrophages would acquire cytotoxic activity when loaded with this enzyme. Monocytes were therefore isolated from human blood and cultured for up to ten days to induce maturation to macrophages. These cells lost endogenous MPO within five days while H₂O₂ production in response to stimulation by phorbol myristate acetate (10^–6M) decreased to 23% within ten days. On the other hand, their capacity to take up exogenous MPO increased fourfold from day three to day ten. Human macrophages cultured from eight days (when both H₂O₂ production and MPO uptake were sufficient) were therefore used to study the effects of MPO uptake on cytocidal activity againstPseudomonas aeruginosa. After a 1 h MPO loading period, macrophages (5×10⁵ cells per ml) were incubated in the presence of bacteria (0.5 to 2×10⁶ bacteria per ml) for 2 h at 37°C. At a bacteria/macrophage ratio of 1, only 34.8±7.0% of bacteria survived (compared to killing by non-loaded macrophages), while 74.4±9.3% survived at a ratio of 4. From these results, we conclude that loading macrophages with exogenous MPO could enhance their microbicidal activity, suggesting a potentially useful therapeutic application.     

The mating behavior of individuals ofDrosophila pseudoobscura from New Zealand

Summary Recently discovered individuals in New Zealand of the North American speciesDrosophila pseudoobscura were analyzed for any divergence in mating behavior. Cultures collected from five localities in North American were mated with a stock from Rotorua, New Zealand. No significant divergence was obtained in any of the within North Americ, and more importantly, between North American and New Zealand mating experiments. Further analyses also showed no development of sterility between recently caught New Zealand and North American flies. We discuss our results in relation to others of this type.Publication No. 5 from the Evolutionary Genetics Laboratory, University of Auckland.Acknowledgements. We thank the University Grants Committee, grant No. 140Z88, for financial assistance. We thank D.G. Futch., L. Barr, A. Beckenach and H. Spencer for their kind assistance in sending us wild caught cultures.     

Karyotypes of several frogs from Korea,Taiwan and the Philippines

Summary Rana amurensis coreana, R. plancyi chosenica, R. latouchii, R. narina andOoeidozyga laevis have 2n=26 chromosomes,R. kuhlii has 2n=22, andKaloula picta has 2n=28. Males ofR. narina have a conspicuous heteromorphic pair, No. 8, which might play a rôle in sex-determination.Supported in part by a grant from the Japan-American Scholarship Foundation.I wish to thank Dr H.K. Kim, Dr Y.-S. Liang, Dr V. Samson-Carino, Mr C.-S. Wang, Mr P.S. Lin and Mr B. Durusan for their aid in every aspect during the collecting trips.     

One-sided sexual isolation betweenDrosophila takahashii andDrosophila pseudotakahashii

Summary In order to elucidate the phylogenetic relationships betweenD. takahashii andD. pseudotakahashii, 2 closely related allopatric species, sexual isolation was studied by the male-choice method. The present data indicate that there is a one-sided mating preference between these species. On a basis of the results, their evolutionary sequence is discussed.We are indebted to Dr I. R. Bock, Australia, and Dr O. Kitagawa, Japan, for kindly providing the live cultures. Thanks are due to the Head of the Zoology Department, B.H.U. for the provision of laboratory facilities and to the C.S.I.R., New Delhi, India, for awarding a Post-doctoral fellowship to Y.N.D.     

Sex determination in mammals — Before and after the evolution of SRY

Therian mammals (marsupials and placentals) have an XX female: XY male sex chromosome system, which is homologous to autosomes in other vertebrates. The testis-determining gene, SRY, is conserved on the Y throughout therians, but is absent in other vertebrates, suggesting that the mammal system evolved about 310 million years ago (MYA). However, recent work on the basal monotreme mammals has completely changed our conception of how and when this change occurred. Platypus and echidna lack SRY, and the therian X and Y are represented by autosomes, implying that SRY evolved in therians after their divergence from monotremes only 166 MYA. Clues to the ancestral mechanism usurped by SRY in therians are provided by the monotremes, whose sex chromosomes are homologous to the ZW of birds. This suggests that the therian X and Y, and the SRY gene, evolved from an ancient bird-like sex chromosome system which predates the divergence of mammals and reptiles 310 MYA. Received 4 March 2008; received after revision 22 April 2008; accepted 3 June 2008