Five New Phenolic Glycosides from Viburnum luzonicum

Viburnum luzonicum is widely distributed in China. Its branch extracts showed potential α-amylase and α-glucosidase inhibitory activities. In order to discover new bioactive constituents, five undescribed phenolic glycosides, viburozosides A−E ( 1 – 5 ), were obtained by bioassay-guided isolation coupled with HPLC-QTOF-MS/MS analysis. Their structures were elucidated by spectroscopic analyses, including 1D NMR, 2D NMR, ECD, and ORD. All compounds were tested for their α-amylase and α-glucosidase inhibitory potency. Compound 1 showed significantly competitive inhibition against α-amylase (IC₅₀=17.5 μM) and α-glucosidase (IC₅₀=13.6 μM).     

Light, temperature and nutrients as factors in photosynthetic adjustment to an elevated concentration of carbon dioxide

The short-term stimulation of the net rate of carbon dioxide exchange of leaves by elevated concentrations of CO₂ usually observed in C₃ plants sometimes does not persist. Experiments were conducted to test whether the patterns of response to the environment during growth were consistent with the hypotheses that photosynthetic adjustment to elevated CO₂ concentration is due to (1) feedback inhibition or (2) nutrient stress. Soybean [Glycine max (L.) Merr. cv. Williams] and sugar beet (Best vulgaris L. cv. Mono Hye-4) were grown from seed at 350 and 700 μl^? CO₂, at 20 and 25°C, at a photon flux density of 0.5 and 1.0 mmol m^?2 S^?1 and with three nutrient regimes until the third trifoliolate leaf of soybean or the sixth leaf of sugar beet had finished expanding. Net rates of CO₂ exchange of the most recently expanded leaves were then measured at both 350 and 700 μl 1^?1 CO₂. Plants grown at the elevated CO₂ concentration had net rates of leaf CO₂ exchange which were reduced by 33% in sugar beet and 23% in soybean when measured at 350 μl 1^?1 CO₂ and when averaged over all treatments. Negative photosynthetic adjustment to elevated CO₂ concentration was not greater at 20 than at 25°C, was not greater at a photon flux density of 1.0 than at 0.5 mmol m^?2 S^?1 and was not greater with limiting nutrients. Furthermore, in soybean, negative photosynthetic adjustment could be induced by a single night at elevated CO₂ concentration, with net rates of CO₂ exchange the next day equal to those of leaves of plants grown from seed at the elevated concentration of CO₂. These patterns do not support either the feedback-inhibition or the nutrient-stress hypothesis of photosynthetic adjustment to elevated concentrations of CO₂.     

Effects of temperature and photoperiod on flowering in Hyptis brevipes

Few tropical species have been tested for their flowering response under controlled conditions. Hyptis brevipes Poit, is an annual herb, commonly found in wet margins of streams and ponds, being considered a weed for some perennial plantations in Brazil. Under experimental glasshouse conditions, this species proved to be an obligate short-day plant. Flowering was delayed when photoperiods longer than 8 h were given, the critical photoperiod being between 12 and 13 h. When both temperature and photoperiod were controlled, at 20°C a longer protoperiod (by almost 1 h) is still inductive compared to 25 and 30°C. The number of short-day cycles required for full induction is relatively high and dependent upon temperature; at 20°C or above, 10 cycles are adequate, but at 15°C, more short-day cycles are needed. The number of inflorescences formed as well as the floral index vary according to daylength × temperature × inductive cycle number, allowing flowering to be assessed quantitatively. Long days are inhibitory to flowering, either suppressing it completely (when symmetrically intercalated among 24 inductive cycles) or preventing the floral index from increasing.     

Chilling stress and oxygen metabolizing enzymes in Zea mays and Zea diploperennis*

Abstract. The activities of five active-oxygen scavenging enzymes were compared for cold-lability and three were compared for chilling induction in two Zea genotypes of contrasting susceptibility to photoinhibition during chilling. Activities of superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), dehydroascorbate reductase (DHAR, EC 1.8.5.1), and glutathione reductase (GTR, EC 1.6.4.2) in leaf extracts from plants grown without chilling stress were assayed at 19°C and 5°C. Enzymes from the chilling-susceptible Z. Mays cv. LG11 had lower specific activities at 5°C than did enzymes from the chilling-tolerant Z. diploperennis, except for MDHAR where no significant differences were observed. The activities of SOD and APX from Z. diploperennis were double those of Z. mays at both assay temperatures. Monodehydroa-scrobate reductase and glutathione reductase activities in both species were reduced by 63–78% at a 5°C assay temperature. The dehydroascorbate reductase (DHAR) showed the greatest low-temperature lability losing 96% (Z. diploperennis) and 100% (Z. mays) of its activity at 5°C. To examine possible chilling-induced changes in levels of enzyme activity, plants of both Zea genotypes were transferred to growth chambers at 10°C at moderate light intensities. Glutathione reductase activity was found to increase within 24h in Z. diploperennis, but it decreased slightly in Z. mays. MDHAR activity decreased by 50% in Z. diploperennis but showed only a transient increase in activity in Z. mays.     

Alpha-aminoxy acids as building blocks for the oxime and hydroxylamine pseudopeptide links. Application to the synthesis of human elastase inhibitors.

The aminoxy acids NH2-O-C(alpha)HR-CO2H are much more easily obtained in the enantiomerically pure form than the analogous hydrazino acids NH2-NH-C(alpha)HR-CO2H, and it has been shown that the isosteric amidoxy psi[CO-NH-O] and hydrazide psi[CO-NH-NH] amide surrogates Induce two quite similar gamma-like folded structures. An aminoxy acid can also be N-coupled to a peptide aldehyde to give the aldoxime psi[CH = N-O] link or to a peptide ketone to form the ketoxime psi[CR= N-O] link. The former can be further reduced into the hydroxylamine psi[CH2-NH-O] link which gives rise to reduced amidoxy peptides. The structural properties Induced by these amide surrogates were studied, using IR and NMR spectroscopy, paying particular attention to the Z/E-isomerism of the oxime link. In order to investigate their inhibitory potency, the three amide surrogates were introduced in the Pro3-Val4 and Val4-Ala5 position of Z-Ala1-Ala2-Pro3-Val4-Ala5-Ala6-NHiPr, a substrate which is cleaved in the Val4-Ala5 position by human leukocyte elastase (HLE). The [Val4psi[CO-NH-O]Ala5] analogue was still a substrate, while the [Pro3psi[CO-NH-O]Val4] and [Val4psi[CH = N-O]Ala5] pseudopeptides acted as HLE competitive inhibitors.     

Mutual support and selection between branches of damaged plants

Snow (1931) showed that the fate of a branch of a plant that had suffered local herbivory could be determined by correlative effects of other branches. This neglected work was continued in the current study, herbivory being simulated by the removal of leaves of different ages from pea plants with two branches. A damaged branch was suppressed when an undamaged alternative branch was present; otherwise the damage never prevented continued development. The removal of mature leaves had a smaller effect than the removal of immature, expanding leaves. When leaves were removed from both branches it was the branches that suffered less damage to their immature leaves that continued shoot development. Branches from which all photosynthetic leaves were repeatedly removed developed only when they retained their immature leaves and remained dominant, inhibiting the development — but not the photosynthesis — of the other branch on the same plant. Accounting for these results requires mechanisms that compare the different branches of the plant and select for development the ones that have the greatest potential for future, rather than present, photosynthesis. It is concluded that a compartmentalized or sectorial distribution of essential substrates can be modified by correlative relations that are probably mediated by hormones.     

酵母醇脱氢酶ADHI的纯化及动力学研究

本文报道了啤酒酵母醇脱氢酶组成型同工酶ＡＤＨＩ的快速高效的纯化方法。通过活性蓝色染料柱亲和层析的方法将该酶纯化至电泳均一,收率达４７％,对该酶的产物抑制及端点抑制动力学研究结果支持Ｗｒａｔｔｅｎ,Ｃｌｅｌａｎｄ提出的序列有序机制。     

Modeling of the H-reflex facilitation during ramp and hold contractions

Healthy subjects were asked to make a voluntary ramp and hold contraction. The duration of the ramp stage was 500 ms, and the torque increment in this period was set to 15 Nm. The contraction was made from a relaxed and from a 5 Nm background torque situation. Hoffmann (H-) reflexes were elicited during the voluntary contraction, mostly with 100 ms intervals. These experiments showed an increase (facilitation) in the H-reflex before the torque or the EMG started to increase. This facilitation of the H-reflex remained during all the stages of the voluntary movement and declined to normal levels again only at the very end of the hold phase, which lasted for one second. This specific pattern of facilitation during a voluntary contraction was modeled using a modeling language, that is specifically designed to calculate neuronal systems with a high degree of reality (Ekeberg et al., 1991). Our model consisted of a motoneuron pool with 200 neurons connected to an EMG-model of the human soleus muscle and an extra group of higher-level neurons for controlling the amount of decrease of presynaptic inhibition. The model was used to simulate the observed modulation of the H-reflex with both a presynaptic and a postsynaptic mechanism. Simulations showed that a continuous change in the descending control signals is needed to make the model based on postsynaptic mechanism fit with the experimental data, whereas no extra control from the CNS over the excitatory drive to the motoneuron pool is needed when the decrease of presynaptic inhibition mechanism is applied.     

CaATP inhibition of the MgATP-dependent proton pump (H+-ATPase) in bacterial photosynthetic membranes with a mechanism of alternative substrate inhibition

 The membrane-bound F1 sector of the H⁺–ATPase complex (F-type ATPase) in dark-adapted photosynthetic chromatophores is endowed with MgATP- and CaATP-dependent ATPase activities, both sensitive to inhibitors such as oligomycin and venturicidin. Because of contatamination of free Mg^{2 +} and Ca²⁺ ions in chromatophore preparations, kinetic characterization of the two hydrolitic reactions can be performed only in the presence of both substrates, using a model for two alternative substrates. The two activities are characterized by similar maximal rates and affinity constants [V_MgATP and V_CaATP: 13±1 and 10±1 nmol s^–1 ATP hydrolyzed (μmol BChl)^–1; K_MgATP and K_CaATP: 0.22±0.06 and 0.20±0.05 mm]. However, only the MgATP-dependent ATPase is coupled to Δ*_{H +} generation. In this process CaATP acts as an alternative substrate and a competitive inhibitor of the proton pump, with a K_I coincident with K_CaATP for the hydrolytic activity. This finding highlights the central role that the coordination chemistry of the ion-nucleotide complex plays in determining the proton gating mechanism at the catalytic site(s) of the enzyme complex. These results are discussed on the basis of the coordination properties of the ions and of the available information on the protein structure. Received: 5 December 1995 / Accepted: 7 March 1996     

水稻巯基蛋白酶抑制剂的分子修饰与其对稻瘟病菌的抑制作用

水稻巯基蛋白酶抑制剂（ＣＰＩ）经用二硫苏糖醇,对氯汞苯甲酸和碘乙酸修饰后,对木瓜蛋白酶的抑制活性并无改变;用Ｎ－乙基顺丁烯二酰亚胺与ＣＰＩ反应,可以测出ＣＰＩ分子内有１９个巯基被修饰,被修饰后,抑制活性仍无改变,表明水稻ＣＰＩ的抑制活性不需要巯基参与;应用Ｎ－溴代丁二酰亚胺与ＣＰＩ反应,可测出ＣＰＩ分子内有２个Ｔｒｐ被修饰,修饰后,抑制活性全部丧失,表明Ｔｒｐ是保持抑制活性所必需的基团。水稻巯基蛋白酶抑制剂和丝氨酸蛋白酶抑制剂对稻瘟病菌丝体的生长均有抑制作用,但后者的抑制作用比前者更强,若将两种抑制剂混合使用,则对稻瘟病菌丝体的抑制作用非常强烈;当抑制剂加入量达７２μｇ时,即可产生明显的抑制作用。