Studies on the Active Principles of Leucothoe grayana MAX.

It was established by the authors that Grayanotoxin-II has a novel tetracyclic structure (I) containing one exocyclic methylene group and five hydroxyl groups. The relations among these functional groups are described in this report.     

Evolution of Active Neurohypophysial Principles Among the Vertebrates

Nine active neurohypophysial peptides have been identified amongthe vertebrates. Arginine-vasotocin appears to be the most primitive.It occurs in cyclostomes and among representative species fromall major branches of vertebrate phylogeny. The eight otherneurohypophysial principles are presumed to have evolved fromarginine-vasotocin after one or more reduplications of the primitivearginine-vasotocin gene. Argininevasotocin cannot be detectedin the adult mammalian neurohypophysis but it has been foundin the pineal and subcommissural organs of adult mammals. Arginine-vasotocinmay be released into the cerebrospinal fluid and influence functionsmediated by hypothalamic hypophysiotropic factors. The primitivefunctions of arginine-vasotocin are not clear. They may relateto cardiovascular regulation since vascular responses to arginine-vasotocincan be demonstrated in species representing all major branchesof vertebrate evolution.     

Studies on the Active Site of Dihydroxy-acid Dehydratase

Several classes of substrate analogs of dihydroxy-acid dehydratase have been tested as inhibitors of this enzyme in an attempt to characterize its binding site and find what modifications in substrate structure lead to an affinity higher than that of the natural substrates. The substrate analogs were tested on dihydroxy-acid dehydratase from both spinach and Escherichia coli. One modification of the substrate that led to as much as a 1000-fold increase in binding affinity was replacement of the 3-hydroxyl group with a thiol. It has been shown previously that the 3-hydroxyl group of the substrate becomes a ligand for one Fe of the Fe-S clusters of these enzymes on binding to their active sites. It seems likely then that the tighter binding of the thiol containing analogs is due to the thiol group becoming a ligand to an iron of the Fe-S clusters of these enzymes. A second modification in substrate that led to as much as 1000-fold increase in binding affinity was the addition of a large lipophilic group. This suggests there is a large hydrophobic pocket or hydrophobic surface near the active site of dihydroxy-acid dehydratase. A modification in substrate that led to as much as a 50-fold increase in binding was the replacement of the carboxyl group of the substrate with phosphonate; however, this increase was limited to substrate analogs without a polar functionality on the carbon β to the phosphonate group. Bromopyruvate was found to irreversibly inactivate dihydroxy-acid dehydratase. Each good inhibitor we found was active on spinach dihydroxy-acid dehydratase and E. coli dihydroxy-acid dehydratase to a similar extent suggesting the active sites of the enzymes from these two organisms are similar. Some of the better inhibitors described in this report have mild herbicidal activity.     

Studies on Substances Active on the Behavior of Planarian

Substances which acted on the behavior of planarian were searched for, and two active components were isolated from culture broth of Streptomyces sp. 340. They were identified as trans-3-methylthioacrylic acid (MTAA) and 3-methylthiopropionic acid (MTPA).

It was found that many Streptomyces and some fungi accumulated MTAA, and many Streptomyces, fungi, bacteria and yeasts accumulated MTPA when the microorganisms were grown in the medium containing methionine.     

Studies on air-ion-enhanced iron chlorosis I. Active and residual iron

The effects of positive and negative air ions on the active and residual iron fractions of barley seedlings were studied during the course of iron chlorosis. Active iron is that fraction localized in the chloroplasts which dissolves in 1.0 N HC1 and participates in the biosynthesis of chlorophyll. Residual iron is not soluble in 1.0 N HC1 and is not concerned with the biosynthesis of chlorophyll. Air ions of either charge induced a significant decrease in active iron content which was associated with a decrease in chlorophyll content. Concomitantly there occurred an increase in both the residual iron and the cytochrome c fractions of the seedlings. There is evidence that the rise in residual iron content may involve not only cytochrome c but also other cytochromes and iron-containing enzymes as well. We have theorized that the site of air ion action in the experiments reported may be the regulatory systems controlling iron metabolism in the seed and young seedling. Through this action air ions apparently divert endogenous free-state iron from conversion to active iron and make it available for the production of a number of ironcontaining compounds which are components of the residual iron fraction.

---

Zusammenfassung Die Wirkung von positiven und negativen Luftionen auf die Aktiv- und Rest-EisenfraktÏon in Gerstenkeimlingen wurde im Verlauf der Eisenchlorose untersucht. Aktiv-Eisen ist die Fraktion in dem FarbstofftrÄger, die in 1.0 N HC1 lösbar und an der Chlorophyllsynthese beteiligt ist. Rest-Eisen istnicht löslich in 1.0 N HC1 und an der Chlorophyllsynthese unbeteiligt. Positive und negative Luftionen bewirkten eine signifikante Verminderung des Gehalts an Aktiv-Fe und Chlorophyll. Gleichzeitig wurden höhere Anteile an Rest-Fe und Cytochrom c gefunden. Es sind Hinweise dafür vorhanden, dass die Zunahme des Rest-Fe nicht nur das Cytochrom c,sondern auch andere Fe-haltige Enzyme betrifft. Die Autoren nehmen an, dass die Luftionen an dem Regulationssystem angreifen, das den Eisenstoffwechsel der Samen und Keimlinge kontrolliert. Luftionen verhindern scheinbar die Umbildung von endogenem Fe-freier Form zu Aktiv-Fe und machen es für die Bildung einer Reihe von Fe-haltigen Verbindungen zugÄnglich, die in der Rest-Fe Fraktion enthalten sind.

---

Resume Les effets de l'ionisation positive ou négative de l'air ont été étudiés fraction active et sur la fraction résiduelle du Fer contenu dans des germes d'orge au cours de la sidérochlorose. Le Fer actif est la fraction des chloroplastes soluble dans l'Acide Chlorhydrique 1,0 N et participant à la biosynthèse de la chlorophylle. Le Fer résiduel n'est pas soluble dans H C1 1,0 N et n'entre pas dans la biosynthèse de la chlorophylle. Les ions positifs et négatifs de l'air ont provoqué une réduction significative de la teneur en Fer actif et en chlorophylle. Simultanément, il est apparu un accroissement de Fer résiduel et du Cytochrome c. Certains indices suggèrent que l'accroissement du Fer actif porte non seulement sur le Cytochrome c mais aussi sur d'autres enzymes contenant du Fer. Les auteurs pensent que les ions de l'air agissent au niveau des systèmes de régulation du métabolisme du Fer dans les graines et dans les germes. Les ions de l'air détournent apparemment le Fer libre endogène de la conversion en Fer actif et le rendent disponible pour la formation d'un certain nombre de composés contenant du Fer qui entrent dans la fraction du Fer résiduel.

     

三颗针活性成分的研究

以~3H-硫氮草酮受体结合分析法为指标,从三颗针中分离得到四个生物碱活性单体,经光谱鉴定为小檗碱、小檗胺、巴马汀及药根碱。其中小檗胺为钙拮抗主要活性成分,并在红细胞过氧化氢实验中表现有较强的抗脂质过氧化活性,这些作用为其防治心肌缺血、抗心律不齐的机制提供有益的信息。     

Studies on the Active Transport of Calcium in Human Red Cells

The Ca⁺⁺ transport mechanism in the red cell membrane was studied in resealed ghost cells. It was found that the red cell membrane can transport Ca⁺⁺ from inside the cell into the medium against great concentration gradient ratios. Tracing the movement of ⁴⁵Ca infused inside red cells indicated that over 95% of all Ca⁺⁺ in the cells was transported into media in 20 min incubation under the optimum experimental conditions. The influence of temperature on the rate constant of transport indicated an activation energy of 13,500 cal per mole. The optimum pH range of media for the transport was between 7.5 and 8.5. As energy sources, ATP¹, CTP, and UTP were about equally effective, GTP somewhat less effective, and ITP least effective among the nucleotides tested. The Ca⁺⁺ transport does not appear to involve exchange of Ca⁺⁺ with any monovalent or divalent cations. Also, it is not influenced by oligomycin, sodium azide, or ouabain in high concentrations, which inhibit the Ca⁺⁺ transport in mitochondria or in sarcoplasmic reticulum. In these respects, the Ca⁺⁺ transport mechanism in the red cell membrane is different from those of mitochondria and the sarcoplasmic reticulum.     

生姜辛辣部位的高效液相色谱(HPLC)指纹图谱研究

采用HPLC法分析了4个居群生姜丙酮回流-乙醚萃取物中的多酚类化合物,建立了有较好专属性的HPLC指纹图谱,比较了姜辣素含量及其得率。     

Studies on Biologically Active Substances Formed by Bacilli

The products of several Bacillus strains were investigated on rabbit serum calcium decreasing, oxytocic and toad heart function promoting activities. These products were obtained from the clear supernatant fluid of the culture medium after the cells were removed by centrifugation.

For the production of rabbit serum calcium decreasing substance, Bacillus subtilis K and Bacillus natto No. 8 were found to be usefull, Bacillus megaterium KM, Bacillus cereus var. mycoides and Bacillus subtilis K produced oxytocic principle. Bacillus subtilis K, Bacillus brevis and Bacillus megaterium KM also produced toad heart function promoting factor.

A procedure was developed to obtain the electrophoretically homogenous rabbit serum calcium decreasing substance from culture filtrate of Bacillus subtilis K. The crude substance was obtained as isoelectric precipitate by adjusting the culture filtrate to pH 3.0. The crude substance was purified by gel filtration on a Sephadex G-75 column, isoelectric fractionation and chromatography on DEAE-cellulose column. The purified preparation was shown to be homogenous by Tiselius electrophoresis but was separated into two bands by polyacrylamide electrophoresis. The chemical analysis of this biologically active substance indicated this substance to be a lipoprotein. The substance decreased rabbit serum calcium level about 12% at 6~8hr after intravenous injection (dose; 0.5 mg/kg body weight).