Involvement of a sperm aminopeptidase in fertilization of the sea urchin

Summary Inhibitory efficiencies of bestatin methyl ester and its nine analogs for sea urchin sperm aminopeptidase activity were similar to the efficiency of the same compounds as inhibitors either of sperm respiration or of fertilization. This suggests that a sperm aminopeptidase plays a role in fertilization in the sea urchin, possibly through its involvement in sperm respiration.Acknowledgment. We are grateful to Dr Wataru Tanaka of Nippon kayaku co. for his gifts of bestatin and its analogs. We are also indebted to Mrs Akiko Tsuchida-Watanabe for her technical assitance, and to the staff of Asamushi Marine Biological Station, Tohoku University, where part of this work was carried out. This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.     

Timing of action of sperm proteases in ascidian fertilization

Summary The timing of action of three sperm proteases, acrosin, spermosin, and a chymotrypsin-like enzyme, in the fertilization of the ascidian,Halocynthia roretzi, was examined by adding specific protease inhibitors at various times after insemination. The results indicate that the last two enzymes both function at the early stage of the process of sperm penetration through the egg investment, while acrosin functions at the late stage.Acknowledgments. We are indebted to Dr M. Hoshi of Tokyo Institute of Technology for his helpful discussion, and to Dr T. Someno of Nippon Kayaku Kogyo Co. for his generous gifts of Z-Val-Pro-Arg-H and leupeptin. This work was supported by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan, and from Naito Research Foundation.     

Synthesis,crystallization and properties of acetyl phenylalanyl lysine chloromethyl ketone: A potential inhibitor of serine proteases

Summary A procedure for the synthesis of acetyl phenylalanyl lysine chloromethyl ketone is described. The chloromethyl ketone derivative is a noncompetitive inhibitor of trypsin (K₁=5.9×10^–3 M).Acknowledgments. The authors are grateful to Dr Arthur L. Bacon for his constant support and encouragement. Special thanks are due to Dr William Kray for initiating this research project. We thank Mrs Juliette Barclay, Ms Earlene Spratling and Ms Ollivette Hill for their technical assistance in conducting some of the experiments. This investigation was supported by MBS grant RR8105 from the General Research Support Branch, Division of Research Resources, National Institutes of Health.     

Involvement of prolyl endopeptidase in ascidian fertilization

Inhibitory effects on fertilization of the ascidian of three benzyloxycarbonyl(Z)-aminoacyl prolinals and Z-Gly-Pro-chloromethyl ketone added before and after insemination were examined. The results suggest that the prolyl endopeptidase is involved in the process of fertilization, especially in a process taking place between chorion elevation and cell cleavage.     

Evidence for the metabolism of phospholipids and triacylglycerols in humanAscaris lumbricoides

Summary HumanAscaris lumbricoides has the necessary mechanism for the biosynthesis and degradation of phospholipids and triacylglycerols, as in most other species. Piperazine decreases the level of triacylglycerols of this parasite by stimulating the activity of lipase and partially inhibiting the activity of phosphatidate phosphatase.Acknowledgments. Thanks are due to the University of Kerala for facilities provided, Dr T. Ramasarma of the I.I. Sc., Bangalore for help in the labelling studies, Dr. G. D. Cain of the University of Iowa for valuable comments and Dr P. Vijayagopal of ANU for the gift of CDP-choline.     

C-banding pattern on the chromosomes of the Japanese house shrew,Suncus murinus riukiuanus,and its implication

Summary The C-band on the chromosomes of the Japanese house shrew,Suncus murinus riukiuanus (Insectivora), was studied. Various types of C-banding pattern were found in the genome of this subspecies. Such banding patterns could be useful for an understanding of autosome and sex-chromosome polymorphisms withinS. murinus.Acknowledgment. The authors are very grateful to Prof. T. Wasano of Fukuoka University for his encouragement, Dr K. Ogawa of Fukuoka University, the Nohara family for help in collecting specimens, and Prof. E.W. Jameson, Jr, University of California, Dr J.W. Bickham and his students of Texas A & M University for comments.     

The occurrence of arachidonic acid in the venom duct of the marine snailConus textile

Summary A crude extract of the venom of the marine snailConus textile Linné caused a powerful contraction in the guinea-pig isolated ileum. The extract of the venom has been fractionated, and the fractions monitored by the contractile effect. Arachidonic acid was shown to be present as an active substance.Acknowledgment. We are grateful to Mr Z. Nagahama of Okinawa island for suppling specimens ofC. textile and to Dr T. Higashijima and Prof. T. Miyazawa (Department of Biophysics and Biochemistry, Tokyo University) for 270 MHz¹H-NMR-measurements.     

Purification of 20α-hydroxysteroid dehydrogenase from human erythrocytes

Summary 20-hydroxysteroid dehydrogenase was found in the supernatant after hemolysis of human erythrocytes, and the enzyme was isolated from the membrane-free hemolysate on a DEAE-cellulose column. The NADPH-generating system was essential for the reaction.Acknowledgments. We wish to thank Dr A. Tomoda, Department of Biochemistry, Kanazawa University School of Medicine, and Dr T. Yubisui, Department of Biochemistry, Medical College of Oita, for their advice and useful discussions.     

Sperm chymotrypsin-like enzymes of different inhibitor-susceptibility as lysins in ascidians

Summary Inhibitory effects of three peptidyl phenylalaninals on fertilization and on chymotrypsin-like enzyme activity of sperm in three species of ascidians were examined. The results suggest that a sperm chymotrypsin-like enzyme is indispensable for the fertilization in each of the ascidians, and that these enzymes have different susceptibilities to inhibitors.12 November 1986Acknowledgments. We are indebted to Dr M. Hoshi of Tokyo Institute of Technology for his helpful discussion. This work was supportd by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan.     

Chromosome variations inXenopsylla astia Rothschild, 1911 (Siphonaptera). A preliminary report

Summary 2 populations of rat fleas,Xenopsylla astia, collected from (A) Bombay and (B) Trivandrum, were found to differ in chromosome number as follows; (A) 2n=10 in both sexes; (B) 2n=18 (male) and 20 (female). Preliminary observations have further shown that the 2 populations differ in certain morphological characters and that their reciprocal hybrids are sterile in the male and partially sterile in the female sex.Acknowledgments. We wish to express our deepest gratitude to Dr Robert E. Lewis, Professor of Entomology, Iowa State University, USA, for confirming our identification of the 2 samples of rat fleas. We are indebted to Dr N.R. Prabhoo, Reader in Zoology, University of Kerala, Trivandrum, for his advice and helpful suggestions. We are also indebted to Prof. K.M. Alexander, for encouragement and providing facilities. Financial help of the University Grants Commission, India, to one of us (C.T.) is also gratefully acknowledged.     

Collagen synthesis of cultured fibroblast from Werner's syndromes of premature aging

Summary The difference of collagen producibility between 2 groups of skin fibroblasts from patients with Werner's syndrome with skin change and with normal skin, and the difference of collagen accumulation to cell layer between skin fibroblast from Werner's syndrome and controls were studied.Acknowledgments. We are grateful to Dr M. Ohkido and Dr I. Matsuo of the Department of Dermatology of Tokai University for their supply of materials and generous advice and Mr K. Takeichi of the Department of Pathology of Tokai University for his technical assistance.     

Isolation of isoflavonoid phytoalexins from seeds ofPhaseolus vulgaris L.

Summary Bean seeds challenged withRhizoctonia solani or exposed to natural microflora yielded the isoflavonoid phytoalexins: phaseollin, phaseollidin, phaseollinisoflavan, coumestrol and kievitone. The data suggest that seeds may have the major host-defence factors of a plant.The author thanks Dr Suresh S. Patil of University of Hawaii for his guidance and provision of laboratory facilities. Gift of phytoalexin samples by Dr S. Pueppke and Dr N.T. Keen is gratefully acknowledged. East-West Center, Honolulu, Hawaii, USA, is thanked for the financial assistance.     

Host-plant pyrrolizidine alkaloids inNyctemera annulata Boisduval: Their persistence through the life-cycle and transfer to a parasite

Summary Larvae ofNyctemera annulata Boisduval ingest pyrrolizidine alkaloids fromSenecio spathulatus A. Rich. which subsequently appear in the adult months and their eggs; the acquisition of the alkaloids by a parasite of theN. annulata larvae provides a further illustration of their ability to persist in insects.Acknowledgments. We thank Dr R. Yamdagni of the Chemistry Department, University of Calgary, Canada, and Dr O. Madsen of the Organic Chemistry Department, The Technical University of Denmark, Lyngby, Denmark, for much help with the MS data collection. We are also very grateful to Dr C.C.J. Culvenor, CSIRO Animal Health Lab., Parkville, Victoria, Australia, who most generously provided us with authentic reference specimens of pyrrolizidine alkaloids.     

The origin of the series elastic component in single crayfish muscle fibres

Summary High-speed cinematographic studies on the length changes of single tetanized crayfish muscle fibres during a step decrease in length revealed a fairly uniform distribution of a highly compliant series elastic component along the fibre length, suggesting that it mainly originates from some structures in each sarcomere other than the cross-bridges.We wish to thank Dr H. Hashizume of the Engineering Research Institute, Tokyo University for providing facilities to use the ultra high-speed cine-camera, and Mr T. Tsuno, Mr Y. Nakamura and Miss S. Gomi for their technical assistance.     

The origin of the instantaneous elasticity in single frog muscle fibres

Summary High-speed cinematographic recordings of single tetanized muscle fibres during a quick decrease in length revealed that the shortening was mostly localized at the fibre segment nearest the released end of the fibre, indicating that the instantaneous elasticity may not originate from the elasticity of the cross-bridges.We wish to thank Dr H. Hashizume of the Engineering Research Institute, Tokyo University for providing facilities to use the ultra high-speed cinecamera, and Mr T. Tsuno, Mr Y. Nakamura and Miss S. Gomi for their technical assistance.     

Evolutionary implications of ascorbic acid production in the Australian lungfish

Summary It is shown that the Australian lungfish (Neoceratodus forsteri) synthesizes ascorbic acid in its kidney, suggesting that ascorbic acid synthesis started before the origin of the tetrapods.Acknowledgments. We would like to thank Dr G.C. Grigg, University of Sydney, Mr R. B. Humphries, Dr P.A. Janssens and Dr C.H. Tyndale-Biscoe, Australian National University, and Dr. M. Griffiths and J.C. MacIlroy C.S.I.R.O., Division of Wildlife Research for supplying the tissue.     

An immunohistochemical study on the distribution of histiocytes containing S-100 protein-like antigen in cutaneous T-cell lymphoma/leukemia; a preliminary report

The immunohistochemical distribution of histiocytes containing S-100 protein-like antigen in the skin lesions of cutaneous T-cell lymphoma/leukemia is investigated. Marked hyperplasia of these histiocytes is found in two cases of mycosis fungoides.We thank Prof. Y. Ishida, Gunma University School of Medicine, Dr K. Iwata, Kochi Prefectural Central Hospital, Dr M. Motoi, Okayama University Medical School, Dr H. Enzan, Kochi Medical School, for supplying the material.     

Ca-antagonistic substance from soft coral of the genusSarcophyton

Summary A 14-membered ring diterpenoid named cembrane, possessing Ca-antagonistic action on the isolated rabbit aorta, has been isolated from a soft coral of the genusSarcophyton, and its structure established by X-ray and spectroscopic data.The authors thank Mr. Z. Nagahama (Okinawa) for collection of soft corals, Miss R. Abe (the present institute) for her skillful assistance, and Dr. T. Higashijima, Prof. T. Miyazawa (Tokyo University), Mr. T. Hayase and Dr. H. Ohtani (Mitsubishi Chemical Industries Ltd) for¹H and¹³C NMR-measurements.     

Anti-endotoxin (anti-lipid-A) antibodies

Summary Anti-lipid-A, anti-endotoxin, antibodies have been measured by a passive haemolysis test using antigen from anE. coli Re mutant. Titres in the normal population are low but do rise in situations in which there has been gram-negative sepsis. Absence of raised titres in other situations has profound implications.We are most grateful to Dr Günter Schmidt of Freiburg for his gift of Lipid-A antigen. We also thank Dr L. Monnens of Nijmegen and Dr T.M. Barratt of London for their sera.     

Lectin-induced lymphocyte stimulation in pigs — a kinetic study of NORs

Summary Early reactions of rRNA-cistrons in lectin-stimulated PBL cultures of domestic pigs are shown. Interphases showing 3 or more clusters of stained areas were attributed mainly to cells after first division. Clear differences are shown between PHA-and ConA-induced NOR-expression patterns compared to the patterns in PW-stimulated cultures.We thank Dr W. Gratze for valuable techniqual assistance, Dr M. Kundi from the Institute of Environmental Health of the University of Vienna for the statistical treatment of our data and the Ludwig Boltzmann Society for financial support.