The purification and immunocharacterisation of N-methylputrescine oxidase from transformed root cultures of Nicotiana tabacum L. cv SC58

The enzyme N-methylputrescine oxidase which catalyses the conversion of N-methylputrescine to N-methylpyrrolinium salt has been purified to homogeneity from transformed roots of Nicotiana tabacum L. cv SC58. The enzyme has an apparent sub-unit molecular weight of 53 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis with gel-filtration studies, indicating that the native form is a dimer. The K _m of the enzyme for N-methylputrescine has been estimated to be 0.1 mM. Polyclonal antibodies raised to the purified protein recognise one product in an immunoblot of a crude extract of transformed root tissue and will immunoprecipitate N-methylputrescine oxidase activity from such an extract. The antibodies also show a high degree of specificity in immunoblots of crude extracts of transformed root cultures from a range of other solanaceous and non-solanaceous species but do not cross-react with a partially purified preparation of pea-seedling diamine oxidase.Abbreviations MPO N-methylputrescine oxidase - PVDF polyvinylidene difluoride - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis We would like to thank members of the Plant Cell Biotechnology Group, Institute of Food Research, Norwich Laboratory, for their helpful discussions during the preparation of this paper.     

Octopamine receptor subtypes and their modes of action

Octopamine receptor subclasses were first proposed to explain differences in the pharmacological profiles of a range of physiological responses to octopamine obtained in the extensor-tibiae neuromuscular preparation of the locust. Thus, OCTOPAMINE₁ receptors which inhibit an endogenous myogenic rhythm, increase intracellular calcium levels. Also OCTOPAMINE₂ receptors which modulate neuromuscular transmission in this preparation, increase the level of adenylate cyclase activity. The current status of this classification is reviewed by examining the pharmacology of responses to octopamine in a range of preparations. It is concluded that the distinction between OCTOPAMINE₁ and OCTOPAMINE₂ receptor types is still valid, but that OCTOPAMINE₂ receptors exhibit some tissue specific variations. Studies on a clonedDrosophila octopamine/tyramine (phenolamine) receptor are discussed and illustrate many of the difficulties presently encountered in making a definitive classification of octopamine receptors. These include the possibilities that single receptors may activate multiple second messenger systems and that different agonists may differentially couple the same receptor to different second messenger systems.     

A study of the flights habits of the European chafer

By using a large cage in the field and a system of marking, the flight activities of 36 females and 49 males of the European chafer were followed during June and July, 1954.It was found that the beetles made one to 11 flights during their life span with males averaging five flights and females 4.5. More males made eight to 11 flights than did females, and males made more flights on consecutive evenings. The average longevity of the males was found to be six days while the females lived on the average for 6.5 days.The main flight period of the European chafer lasts about one month though some beetles can be observed flying in the evenings for about two months after the main flight has ended. The populations observed in the field are composed of successive groups of beetles which emerge, fly for several days then die. These groups overlap each other so that high numbers of beetles are present in the trees each evening during the main flight period. Variations in the physical characteristics of the soil habitat could be responsible for the differences in the time of maturity of the larvae and the subsequent first-time emergence of the adults.

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Zusammenfassung Im Juni und Juli 1954 wurde die Flugtätigkeit von 36 weiblichen und 49 männlichen Tieren von Amphimallon majalis (Razoumowsky) unter Zuhilfenahme eines grossen Käfigs im freien Felde und eines geeigneten Bezeichnungssystems der Tiere verfolgt.Es zeigte sich, dass die Käfer während ihrer Lebensdauer ein bis elf Flüge unternehmen und zwar betrug der Durchschnitt für männliche Tiere fünf, für weibliche 4,5 Flüge. Mehr Männchen als Weibchen flogen 8 bis 11 mal und die Männchen flogen häufiger an aufeinanderfolgenden Abenden. Für männliche Käfer war die durchschnittliche Lebensdauer 6 Tage, während die weiblichen einen Durchschnitt von 6,5 Tagen erreichten.Die Hauptflugperiode von Amphimallon majalis erstreckt sich über etwa einen Monat, obwohl man auch 2 Monate lang nach der Hauptflugzeit des abends noch einige Käfer fliegen sehen kann. Die Gesamtzahl der im Versuchsfeld beobachteten Insekten setzt sich aus auf einander folgenden Gruppen von Käfern zusammen, die nach einander ausschlüpfen, mehrere Tage fliegen und dann sterben. Diese Gruppen überschneiden sich, so dass während der Hauptflugzeit abends eine grosse Anzahl von Käfern in den Bäumen zu finden ist. Es ist sehr wohl möglich, dass Unterschiede in der physikalischen Beschaffenheit des Bodens für die Länge der Reifezeit der Larven verantwortlich sind und somit auch für den Zeitpunkt des Erstfluges der entwickelten Käfer.

     

Analysis of the proposed Fe-SX binding region of Photosystem 1 by site directed mutation of PsaA in Chlamydomonas reinhardtii

The psaA and psaB genes of the chloroplast genome in oxygenic photosynthetic organisms code for the major peptides of the Photosystem 1 reaction center. A heterodimer of the two polypeptides PsaA and PsaB is thought to bind the reaction center chlorophyll, P700, and the early electron acceptors A₀, A₁ and Fe-S_X. Fe-S_X is a 4Fe4S center requiring 4 cysteine residues as ligands from the protein. As PsaA and PsaB have only three and two conserved cysteine residues respectively, it has been proposed by several groups that Fe-S_X is an unusual inter-peptide center liganded by two cysteines from each peptide. This hypothesis has been tested by site directed mutagenesis of PsaA residue C575 and the adjacent D576. The C575D mutant does not assemble Photosystem 1. The C575H mutant contains a photoxidisable chlorophyll with EPR properties of P700, but no other Photosystem 1 function has been detected. The D576L mutant assembles a modified Photosystem 1 in which the EPR properties of the Fe-S_A/B centers are altered. The results confirm the importance of the conserved cysteine motif region in Photosystem 1 structure.Dedicated to the memory of Daniel I. Arnon.     

Is tuberculosis taken seriously in the United Kingdom?

Tuberculosis has been the subject of much concern in recent years. Notifications have increased, inadequacies in surveillance revealed, and policies for BCG immunisation and screening of immigrants questioned. Until recently the disease was given low priority in the United Kingdom. There is no overall strategic framework for tackling tuberculosis, and fears have been expressed about the future of local tuberculosis control programmes in the new market economy of the NHS. An action plan for tuberculosis within the context of a national programme is urgently required. Only then will a major impact on the incidence of the disease be seen.     

Preservation of Rhizobium viability and symbiotic infectivity by suspension in water

Three Rhizobium japonicum strains and two slow-growing cowpea-type Rhizobium strains were found to remain viable and able to rapidly modulate their respective hosts after being stored in purified water at ambient temperatures for periods of 1 year and longer. Three fast-growing Rhizobium species did not remain viable under the same water storage conditions. After dilution of slow-growing Rhizobium strains with water to 10(3) to 10(5) cells ml-1, the bacteria multiplied until the viable cell count reached levels of between 10(6) and 10(7) cells ml-1. The viable cell count subsequently remained fairly constant. When the rhizobia were diluted to 10(7) cells ml-1, they did not multiply, but full viability was maintained. If the rhizobia were washed and suspended at 10(9) cells ml-1, viability slowly declined to 10(7) cells ml-1 during 9 months of storage. Scanning electron microscopy showed that no major morphological changes took place during storage. Preservation of slow-growing rhizobia in water suspensions could provide a simple and inexpensive alternative to current methods for the preservation of rhizobia for legume inoculation.     

Discrimination by male ring-tailed Lemurs (Lemur catta) between the scent marks of male and those of female conspecifics

Olfaction plays an important role in the social communication of all prosimians. (The experiment reported in this paper forms part of an intensive chemobehavioral study of olfaction in Lemur catta (ring-tailed lemur) being carried out in this laboratory.) Five male Lemur cattawere tested on their behavioral responses to paired scent stimuli. Responses measured were (1) total investigation time, (2) arm-marking, (3) ABO/BO rubbing, and (4) flehmen. Males showed a strong discrimination between the scent stimuli,giving higher levels of response to female scent on measures 1, 3, and 4. This response suggests an olfactory-related preference by males for female scent under controlled conditions. This preference may be a consequence of the females’ dominance over males and the brevity of estrus in L. catta,both of which would favor such choice behavior.     

Sapatoxins,aliphatic ester tigliane diterpenes from Sapium indicum

From the unripe fruits of Sapium indicum, three aliphatic esters of the tigliane nucleus were isolated. These compounds were derivatives of 4-deoxyphorbol. Sapatoxin A was identified as 12-O-[n-deca-2,4,6-trienoyl]-4-deoxyphorbol-13-acetate, B as 12-O-[n-deca-2,4,6-trienoyl]-4-deoxy-5-hydroxyphorbol-13-acetate and C as 12-O-[n-deca-2,4,6-trienoyl]-4,20-dideoxy-5-hydroxyphorbol-13-acetate, by spectroscopic analysis and hydrolysis reactions.     

Biphasic effect of acute ethanol administration on rat liver tyrosine-2-oxoglutarate aminotransferase activity.

1. Acute ethanol administration causes a biphasic change in rat liver tyrosine aminotransferase activity. 2. The initial decrease is significant with a 200 mg/kg dose of ethanol, is prevented by adrenoceptor-blocking agnets and by reserpine, but not by inhibitors of ethanol metabolism, and exhibits many of the characteristics of the inhibition caused by noradrenaline. 3. The subsequent enhancement of the enzyme activity by ethanol is not associated with stabilization of the enzyme, but is sensitive to actinomycin D and cycloheximide. 4. It is suggested that the initial decrease in aminotransferase activity is caused by the release of catecholamines, whereas the subsequent enhancement may be related to the release of glucocorticoids.