Specific inhibition of phosphate transport in mitochondria by N-ethylmaleimide

N-ethylmaleimide (NEM), a reagent that alkylates free sulfhydryl groups, was shown to be a highly effective inhibitor of the following coupled mitochondrial processes: oxidative phosphorylation, ATP-³²Pi exchange, Pi-induced light scattering and configurational changes, State III respiration, valinomycin-induced translocation of potassium with Pi as the anion, and calcium accumulation in presence of Pi. However, NEM was less effective or ineffective in inhibiting some processes that do not require inorganic Pi, namely electron transfer and ATPase activity, ADP binding, energized light scattering changes induced by arsenate and nonenergized light scattering changes induced by acetate. The rate of oxidative phosphorylation and of ATP-³²Pi exchange was normal in ETP_H particles prepared from NEM-treated mitochondria. Also NEM, even et levels 2–3 times greater than those required to inhibit oxidative phosphorylation in intact mitochondria, did not inhibit coupled processes in submitochondrial particles. We are proposing that NEM alkylates sulfhydryl groups in the mitochondrion that modulate Pi translocation, and that the suppression of Pi translocation blocks oxidative phosphorylation, the Pi-dependent energized configurational change in mitochondria and Pi-dependent transport processes.On leave of absence from the Department of Biochemistry, Cancer Institute Okayama University Medical School, Okayama, Japan.On leave of absence from the Department of Pathology, Nagoya University Medical School, Nagoya, Japan.     

Studies on anaerobic biphasic growth of a denitrifying bacterium, Pseudomonas stutzeri

Growth of Pseudomonas stutzeri(VAN NIEL strain) in the presenceof a limiting amount of nitrate under anaerobic conditions ischaracterized by 2 logarithmic phases separated distinctly byan intermediate phase where the growth rate is very low. Inthe first logarithmic phase nitrate is reduced stoichiometricallyto nitrite stage, and in the second phase nitrite is reducedto nitrogen gas. The nitrite reducing activity of cells in the second growthphase is 3–4 times higher than that of cells in the firstphase. The rise in nitrite reducing activity is correlated witha remarkable increase in the content of cytochromes a₂ and c-552. ¹Present address: Department of Biochemistry, Hiroshima UniversitySchool of Dentistry, Hiroshima, Japan. ²Present address: Institute of Molecular Biology, Faculty ofScience, Nagoya University, Nagoya, Japan. (Received June 16, 1969; )     

Biochemistry,Coming Along with Melodies: Instructional Design and Teaching Tips for Introduction of Biochemistry Course

The first class of a higher education course, in most cases, is an introduction that covers the learning goals, course overview, syllabus, and evaluation mode. The effectiveness of an introduction lecture is vital for students to develop an interest in the course. Biochemistry is the foundation of a vast array of scientific disciplines; therefore, it is a core course required for medical schools and life science majors. However, many students are often intimidated by the complex, intertwined metabolic pathways composed of a great variety of structurally diverse biomolecules. Well begun is half done: a good introduction is essential for the success of a course and this is particularly true for teaching of biochemistry. As an educator with over twenty years of teaching experience in biochemistry to students of biology, medicine, pharmacy, and nursing at Peking University, I have successfully enriched the introduction class with delightful biochemistry songs, multiple sources of educational materials, and biochemistry-related knowledge in medical humanities. These strategies and tips have proven effective, and I hope it can be enlightening and helpful to the teaching of biochemistry.     

Properties, development and cellular-localization of cinnamic acid 4-hydroxylase in cut-injured sweet potato

In sweet potato root tissue, cinnamic acid 4-hydroxylase activityincreased markedly in response to cut injury, and reached amaximum after 1 day of incubation. The patterns of developmentand successive decline were similar to those for phenylalanineammonia-lyase activity. The development of both enzyme activitieswas inhibited by cycloheximide. The activity was strictly dependenton pH of the homogenizing and reaction media. The optimum pHof the reaction was 8.0. The respective Km values for trans-cinnamicacid and NADPH were 2.6?10^-5 and 1.8?10^-6M. The activity wasnot affected by ß-mercaptoethanol and the intermediatesand product of the polyphenol biosynthetic pathway. Carbon monoxideinhibited strongly the activity and its inhibition was partiallyprevented by light. Thus, the enzyme may be involved in thecytochrome P-450 mediated electron transport system. Studiesusing differential centrifugation and sucrose density gradientcentrifugation, showed that the intracellular distribution of4-hydroxylase activity differed distinctly from that of themitochondrial marker enzyme and was not in accord with thatof NADPH-cytochrome c reductase activity. ¹This paper constitutes part 114 of the Phytopathological Chemistryof Sweet Potato with Black Rot and Injury. ²Present address: Laboratory of Biochemistry, Faculty of Agriculture,Nagoya University, Chikusa, Nagoya 464, Japan. (Received May 20, 1974; )     

Association of biochemistry grades with performance in pharmacology and anatomy in a Saudi Arabian medical college

The College of Medicine, King Khalid University of Abha, Saudi Arabia currently accepts 100 students a year, up from 50 to 70 students four years ago. The first-year students take four science courses (Biology,Chemistry, Physics, and Statistics) in addition to some general university requirement courses. Biochemistry is offered in the second year, Anatomy in the second and third year and Pharmacology in the fourth year. This study was carried out to determine the correlation between the performance of medical students in Biochemistry and their performance in Anatomy and Pharmacology. Data were obtained from two groups of students (Group 22 and 23) of the years 1995 and 1996 who had already taken Biochemistry, Anatomy and Pharmacology. Performance was equal in Pharmacology but in biochemistry performance of group 23 was clearly lower than group 22. Scores of students in Biochemistry course strongly correlated with the basic Pharmacology course (r=0.714, P<0.0001). Scores in Anatomy also correlated with those in Biochemistry (r=0.616, P<0.001) but much less with scores in Pharmacology (r=0.345, P<0.01).     

A festschrift for J. Martyn Bailey, a biochemist extraordinaire

A festschrift for Dr. John Martyn Bailey, Professor of Biochemistry and Molecular Biology was organized by the Biochemistry department of the George Washington University School of Medicine and Health Sciences on December 4-5, 2006 to honor his 48 years of contributions. He made important contributions in the areas of essential fatty acids, prostaglandins, thromboxanes and lipoxygenase metabolites.     

Faunal trophic interaction in an oligotrophic-dystrophic lake (Shirakoma-ike, Japan)

 The trophic interaction in the pelagic food web of oligotrophic-dystrophic Lake Shirakoma-ike was investigated by a stable isotope technique. Carbon and nitrogen isotope analyses revealed the trophic interaction among Piona carnea (a water mite), Chaoborus sp. (a phantom midge), Daphnia longispina (a cladoceran), and Acanthodiaptomus pacificus (a calanoida copepoda). The δ¹³C values for estimating the relative contributions of D. longispina and A. pacificus to carnivores were used, and they were also used for the constraint of the trophic enrichment factor of δ¹⁵N. Both Chaoborus sp. and P. carnea were suggested to consume D. longispina and A. pacificus at almost the same ratio (1 : 1). However, both carnivores preferred A. pacificus to D. longispina, because D. longispina was much more abundant in the lake than A. pacificus. Omnivory of P. carnea was also analyzed by the mixing model. Received: February 28, 2002 / Accepted: July 10, 2002 Present address: Hydrospheric Atmospheric Research Center, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan Tel. +81-52-789-3434; Fax +81-52-789-3436 e-mail: lee@ihas.nagoya-u.ac.jp Present address: Research Institute for Humanity and Nauture, Kyoto, Japan Acknowledgments We would like to thank Professor Kikuo Kato for providing useful comments and colleagues in the Laboratory of Biogeochemical Data Analysis, Nagoya University, for their support with the sample collection. We are in debt to Dr. R. Coucett for the kind suggestion on the trophic enrichment factors for blood-sucking insects. We also thank the members of the Suwa Hydrobiological Station, Shinshu University, who kindly allowed us to use facilities for sampling in Lake Shirakoma-ike. The zooplankton samples were partly provided by Mr. Masataka Yoshida. Correspondence to:J.-Y. Lee     

Immunoelectronmicroscopic localization of phenylethanolamine-N-methyltransferase in the bovine adrenal medulla

Summary The subcellular localization of phenylethanolamine-N-methyltransferase (PNMT) in the bovine adrenal medulla has been examined by a peroxidase-labelled immunoelectronmicroscopic method. The enzyme is localized in the ground substance of the epinephrine-containing cells, but not in the norepinephrine-containing cells. PNMT is not shown in the nucleus, mitochondria, Golgi complex, ribosomes and norepinephrine granules. There are some observations which indicate that PNMT might be also localized in epinephrine granules.The author wishes to thank Dr. M. Hoshino (Research Institute, Aichi Cancer Center, Nagoya) for his help in the immunological aspect of this work, Dr. T. Nagatsu and Mrs. Y. Numata (Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya) for their help in purification of PNMT and in preparation of the antibody, and Miss S. Iida (Aichi Prefectural College of Nursing, Nagoya) for her valuable assistance.     

Development of biochemistry in the Biology Department of Kharkhov State University. Formation of a new direction--physiology and biochemistry of aging

The article presents some data on teaching Biochemistry at the Nature Department of Kharkiv Imperial University Physico-Mathematical Faculty, as well as restoration of biochemical education and scientific researches as a result of the University reorganization in the Soviet period and foundation of the Biochemistry Chair at the Biological Faculty in the renovated Kharkiv State University. The analysis of scientific biochemical investigations conducted in the Kharkiv University till now is revealed. The special attention is paid to development of such a scientific trend as age physiology and biochemistry. The article deals with the comprehensive information on scientific and pedagogical activity of the outstanding scientists such as O. V. Nagorny and I. M. Bulankin as founders of the Kharkiv school of age physiology and biochemistry. The work has utilized some archive data.     

Orthovanadate Suppresses Accumulation of Phenylalanine Ammonia-Lyase mRNA and Chalcone Synthase mRNA in Pea Epicotyls Induced by Elicitor from Mycosphaerella pinodes

Orthovanadate delayed accumulation of mRNAs encoding phenylalanineammonia-lyase and chalcone synthase in pea epicotyls inducedby an elicitor from Mycosphaerella pinodes. However, accumulationof mRNA for a putative P-type ATPase was not affected. The relationshipbetweenthe ATPase and defense responses is discussed. ³Present address: Plant Pathology Laboratory, School of Agriculture,Nagoya University, Chikusa, Nagoya, 464-01 Japan.     

The 2009 Nobel Prize in Chemistry: Thomas A. Steitz and the structure of the ribosome

Over the past 200 years, there have been countless groundbreaking discoveries in biology and medicine at Yale University. However, one particularly noteworthy discovery with profoundly important and broad consequences happened here in just the past two decades. In 2009, Thomas Steitz, the Sterling Professor of Molecular Biophysics & Biochemistry, was awarded the Nobel Prize in Chemistry for "studies of the structure and function of the ribosome," along with Venkatraman Ramakrishnan of the MRC Laboratory of Molecular Biology and Ada E. Yonath of the Weizmann Institute of Science. This article covers the historical context of Steitz's important discovery, the techniques his laboratory used to study the ribosome, and the impact that this research has had, and will have, on the future of biological and medical research.     

Reminiscences,collaborations and reflections

This is a personal account by a semi old-timer who completed his official term as a professor of plant biochemistry at Nagoya University in Japan in 1992. My university student life began soon after the World War II (1948). I shared the hardships of many in my age group, in that life was difficult during my college years. I was fortunate to have the opportunity of studying in the USA on a Fulbright scholarship first at Purdue University (1955–1956), and then at the University of California, Berkeley (1956–1957). My graduate study and postdoctoral training in the new world were vitally refreshing and stimulating, which gave me the impetus for becoming a natural scientist associated with academic institutions. Consciously and subconsciously I was impressed by the friendly and liberal atmosphere surrounding young students as well as senior scholars in the United States. But more importantly, I was inspired by the critical and competitive minds prevailing among these people.The appointment as a biochemist at the International Rice Research Institute (IRRI) in the Philippines (1962–1964) was the real start of my professional career. The work was continued upon my return to Nagoya to become a staff member of the Research Institute for Biochemical Regulation (1964–1992). Throughout the years, my major research interest has covered photosynthesis as a whole, involving photosynthetic CO₂-fixation (RuBisCO), carbohydrate metabolism, e.g. starch biosynthesis and breakdown (-amylase), and metabolic regulation, which are interrelated in the basic metabolism of plant cells.I shall briefly describe in this article highlights from my studies and discoveries made and I shall also discuss their possible significance in plant metabolism, with the hope that it does not contradict my sense of humility: They are (a) discovery of ADPG in plants and its role in starch biosynthesis; (b) structure-function relationship of RuBisCO proteins, in particular on heterologous recombination of their subunits of plant-type enzyme molecules derived from the prokaryotic photosynthetic bacteria; (c) molecular evolution of RuBisCO genes; (d) mode of actions (formation, intracellular transport and secretion) of rice seed -amylase and its structural characteristics (distinctive glycosylation), and (e) DNA methylation and regulatory mechanism of photosynthesis gene expression in plastids (amyloplasts). In each step of my research, I shared joy, excitement, disappointment, and agony with my colleagues, an experience that may be common to all researchers. Although it is now becoming well recognized among the scientific community in Japan, I want to point out that interaction of multinational scientific minds in the laboratory produces a vital and creative atmosphere for performance of successful research. I experienced and realized this important fact in my earlier days in the USA and the Philippines. Inasmuch as I believe that this is the most crucial element for any research laboratory to possess, I fondly remember the friendships gained with numerous overseas visitors and collaborators who have contributed immensely to our work.Written at the invitation of Govindjee.     

A paradigm for axonal transport

Axonal transport has been extensively studied for a period of 20–30 years, but there is still no general consensus concerning the mechanism by which this transport process operates. An important development in this regard is the recent studies in the physical biochemistry group in the Department of Biochemistry at Monash University where it has been demonstrated that ordered flows may be generated spontaneously in polymer systems under non-equilibeium conditions. The new phenomenon exhibits many novel features, particularly with respect to polymer transport, which bear marked similarity to the behaviour of components in axonal transport. This article sets out to essentiallybring to the attention of those in the neurosciences some of the properties of ordered structured flows in polymer solutions. These properties may generate a different view in the understanding of the mechanism of axonal transport.     

Stimulation by Fungal Elicitor of Inositol Phospholipid Turnover in Tobacco Suspension Culture Cells

The effects of fungal elicitor on inositol phospholipid turnoverand induction of phenylalanine ammonia-lyase (PAL) activityin tobacco suspension culture cells were investigated. Tobaccocells labeled by [³H]inositol in vivo were treated with Phytophthoranicotianae elicitor and [³H]metabolites of inositol phospholipidturnover were examined. Stimulation of inositol phospholipidturnover was observed preceding the induction of PAL activity;inositol 1,4-bisphosphate increased 15 times over the control10 min after the elicitor treatment. Increase of inositol 1,4,5-trisphosphatewas only 38% of the control. Phosphatidylinositol and phosphatidylinositol4-phosphate transiently decreased by 21 and 35%, respectively.Phosphatidylinositol 4,5-bisphosphate was not affected significantlyby the elicitor. Inositol 1,4-bisphosphate was preferentiallyelevated by elicitation than 1,4,5-trisphosphate suggestingthat the regulatory mechanism of inositol phospholipid turnoverin tobacco cells is different from that in animal cells. Phosphatidylinositolkinase but not phospholipase C was activated by the elicitorin vitro. Elicitor-dose dependency curves in the induction ofPAL activity and in the stimulation of inositol phospholipidturnover showed a similar feature suggesting that inositol phospholipidturnover is involved in the elicitor-signal transduction intobacco cells. ¹Present address: The Johns Hopkins University School of Hygieneand Public Health, 615 N. Wolfe St., Baltimore, Maryland 21205,U.S.A. ²Present address: Nagoya University BioScience Center and GraduateSchool of Agricultural Sciences, Nagoya University, Chikusa-ku,Nagoya, 464-01 Japan.     

Protein Kinase Inhibitors Inhibit Stimulation of Inositol Phospholipid Turnover and Induction of Phenylalanine Ammonia-Lyase in Fungal Elicitor-Treated Tobacco Suspension Culture Cells

Elicitor prepared from Phytophthora nicotianae stimulated inositolphospholipid turnover and induced phenylalanine ammonia-lyaseactivity in tobacco suspension culture cells [Kamada and Muto(1994) Plant Cell Physiol. 35: 397]. Protein kinase inhibitors,K252a and staurosporine inhibited both responses. These resultssuggest that inositol phospholipid turnover plays an importantrole in PAL induction through protein kinases. In addition,their mode of inhibition were different, proposing that severaltypes of protein kinases are involved in these elicitor-inducedresponses. ¹Present address: The Johns Hopkins University School of Hygieneand Public Health, 615 N. Wolfe St., Baltimore, Maryland 21205,U.S.A. ²Present address: Nagoya University BioScience Center and GraduateSchool of Agricultural Sciences, Nagoya University, Chikusa-ku,Nagoya, 464-01 Japan.     

Bacterial rhodopsins: evolution of a mechanistic model for the ion pumps

Walther Stoeckenius received a MD degree at the University of Hamburg, Germany in 1950. After 18 months of clinical work as an intern, he began postdoctoral work on the development of pox viruses at the Institute for Tropical Medicine in Hamburg using mainly electron microscopy techniques. After two years he moved as Assistant Professor to the Department of Pathology at the University of Hamburg and became Docent for Pathology in 1958. In addition to teaching and routine pathology work, he continued to use electron microscopy to explore the fine structure of cells and developed an interpretation of the triple-layered appearance of membranes in electron micrographs in terms of molecular structure and the chemistry of osmium tetroxide fixation. In 1959 he obtained a position as Research Associate in Keith Porter's laboratory at Rockefeller University. This was changed after a few months to Assistant Professor and he stayed there, later as Associate Professor, for eight years. The work on membrane structure continued, and a model was developed that described the membrane as a lipid bilayer with embedded protein domains. In efforts to isolate such domains, the purple membrane and bacteriorhodopsin were discovered. In 1966, the lure of California became irresistible and Dr. Stoeckenius accepted a professorship at the University of California at San Francisco. The work on bacteriorhodopsin continued there with the emphasis changing from electron microscopy to spectroscopy and biochemical techniques. He is now Professor Emeritus there in the Department of Biochemistry and Biophysics and the Cardiovascular Research Institute.     

Biochemical education in Romania: Report on a workshop held in Bucharest, 16–20 September 1991

The Workshop Team from IUBMB consisted of Professor W Rombauts (Leuven, Belgium), Professor F Vella (Saskatoon, Canada), and Dr E J Wood (Leeds, UK), with local arrangements being in the hands of Dr S A Hulea (Secretary of the Romanian Biochemical Society). The Workshop was held at the Institute of Biochemistry, Romanian Academy of Sciences (Head: Professor Dr Cecilia Motas) and there were participants from Bucharest, Constantin, Cluj-Napoca, Galati, Tasi and Sibiu. Support was kindly provided by the Romanian Biochemical Society, World Vision International, The Commission of Biochemistry, Romanian Academy of Sciences, Institute of Biochemistry, University of Medicine and Pharmacy “Carol Davilla”, University of Bucharest, and IUBMB     

The inheritance of progressive muscular dystrophy in Japan

Summary In conclusion, there were no apparent differences between Japanese and foreign cases in clinical types, the frequency of each type, sex ratio, the age of onset, modes of the inheritance etc., except for a peculiar type of the disorder found in the South-Western Islands. However, sporadic cases were seem to be comparatively often encountered in each type especially in the Duchenne and limb-girdle type. Further, large families with as many victims as in foreign cases have not yet been reported in Japan.Professor of Nagoya University, M. D., D. Med. Sc.     

Diurnal differences in the supply of glucomannans and xylans to innermost surface of cell walls at various developmental stages from cambium to mature xylem in Cryptomeria japonica

Summary. We investigated the diurnal differences in the innermost surface of tracheid cell walls at various developmental stages from cambium to mature xylem. Cryptomeria japonica saplings were cultivated in a growth chamber with a light cycle set at 14 h of light and 10 h of darkness. Samples were collected from the saplings during both the light and dark periods. The innermost surface of cell walls was immunogold-labeled with anti-glucomannan or anti-xylan antiserum and was observed by field emission scanning electron microscopy. Diurnal differences in the aspect of the innermost surface of cell walls were seen only in S₂-layer-forming tracheids; cellulose microfibrils were clearly evident during the light period, and amorphous material containing glucomannans and xylans was prevalent during the dark period. Cellulose microfibrils were present at the primary-wall formation and S₁-layer-forming stages, and many warts were observed in the mature tracheids, regardless of the time of sampling. The densities of labeled glucomannans on the innermost surface of cell walls in S₁- and S₂-forming tracheids and of labeled xylans in S₂-forming tracheids during the dark period were significantly higher than those during the light period. These results suggest a diurnal periodicity in the supply of cell wall matrix containing hemicellulose to the innermost surface of developing secondary walls. Correspondence and reprints: Laboratory of Bio-material Physics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan. Present address: Chair of Climate Change Science for Forestry and Water Resources, Graduate School of Science and Technology, Niigata University, Niigata, Japan.