Sesquiterpene lactones from Calea megacephala

Chemical analysis of Calea megacephala yielded two new sesquiterpene lactones. The new compounds represent a heliangolide, desacylisovalerylheliangine, and the ternifolin derivative 8β-methacryloyloxyternifolin. Their structures were established by spectroscopic methods.     

Effects of mutagenesis of residue 221 on the properties of bacterial and mitochondrial elongation factor EF-Tu

During protein biosynthesis, elongation factor Tu (EF-Tu) delivers aminoacyl-tRNA (aa-tRNA) to the A-site of ribosomes. This factor is highly conserved throughout evolution. However, several key residues differ between bacterial and mammalian mitochondrial EF-Tu (EF-Tu(mt)). One such residue is Ser221 (Escherichia coli numbering). This residue is conserved as a Ser or Thr in the bacterial factors but is present as Pro269 in EF-Tu(mt). Pro269 reorients the loop containing this residue and shifts the adjoining beta-strand in EF-Tu(mt) compared to that of E. coli EF-Tu potentially altering the binding pocket for the acceptor stem of the aa-tRNA. Pro269 was mutated to a serine residue (P269S) in EF-Tu(mt). For comparison, the complementary mutation was created at Ser221 in E. coli EF-Tu (S221P). The E. coli EF-Tu S221P variant is poorly expressed in E. coli and the majority of the molecules fail to fold into an active conformation. In contrast, EF-Tu(mt) P269S is expressed to a high level in E. coli. When corrected for the percentage of active molecules, both variants function as effectively as their respective wild-type factors in ternary complex formation using E. coli Phe-tRNA(Phe) and Cys-tRNA(Cys). They are also active in A-site binding and in vitro translation assays with E. coli Phe-tRNA(Phe). In addition, both variants are as active as their respective wild-type factors in ternary complex formation, A-site binding and in vitro translation assays using mitochondrial Phe-tRNA(Phe).     

中国11个双子叶植物原白中排印错误之更正

对我国11个双子叶植物（Dicotyledon）原白中模式标本引证的排印错误做了更正：砚山锥栗（壳斗科）原白中错误地将模式标本引证为王启无84116,实际应为王启无84416,前者属于菊科植物Inuna helianthus-aquatica C.Y.Wu ex Ling。长果柯（壳斗科）原白中错误地将模式标本引证为K.M.Feng 13012,实际应为K.M.Feng 13102,前者属于冬青科植物Ilex triflora Bl.。福建红小麻（荨麻科）原白中错误地将主模式标本引证为C.J.Chen&Z.Y.Li 109,实际应为C.J.Chen&Z.Y.Li 103,前者属于荨麻科植物Oreocnide frutescens（Thunb.）Miq.。少毛全缘叶紫麻（荨麻科）原白中错误地将主模式标本引证为N.K.Chun 44099,实际应为N.K.Chun 44033,前者属于杜鹃花科植物Lyonia ovalifolia（Wallich）Drude var.rubrovenia（Merr.）Judd.。甘南铁线莲（毛茛科）原白中错误地将主模式标本引证为Baishuijiang Exped.4490,实际应为Baishuijiang Exped.4990,前者属于卫矛科植物Euonymus alatus（Thunb.）Sieb.。矮粗距翠雀花（毛茛科）原白中错误地将模式标本引证为Sichuan Veg.Exped.3137,实际应为Sichuan Veg.Exped.3173,前者属于龙胆科植物Gentiana conduplicata T.N.Ho。镇康黄芪（豆科）原白中错误地将主模式标本引证为T.T.Yu 17255,实际应为T.T.Yu 17225,前者属于莎草科植物Scirpus lushanensis Ohwi。宽翼棘豆（豆科）原白中错误地将模式标本引证为Qinghai-Xizang Comp.Exped.9484,实际应为Qinghai-Xizang Comp.Exped.9485,前者属于石竹科植物Arenaria kansuensis Maxim.。肾瓣黄芪（豆科）原白中错误地将模式标本引证为Qinghai-Xizang Comp.Exped.3650,实际应为Qinghai-Xizang Comp.Exped.3605,前者属于麻黄科植物Ephedra gerardiana Wall.ex Mey.。湖南长柄槭（槭树科）原白中错误地将模式标本引证为李泽棠2944,实际应为李泽棠2994,前者属于杜鹃花科植物Pieris formosa D.Don。峨眉勾儿茶（鼠李科）原白中错误地将模式标本引证为杨光辉54729,实际应为杨光辉54723,前者属于山茱萸科植物Helwingia chinensis Batalin.。     

A revision of the genus Halectinosoma (Harpacticoida: Ectinosomatidae): a reappraisal of H. sarsi (Boeck) and related species

Ectinosomatid material was examined from extensive collections around the British Isles and from several additional world localities and museum collections, permitting a revision of the genus Halectinosoma. This paper describes 12 morphologically similar species and erects five new species. H. sarsi (Boeck) is regarded as species incertae sedis, while H. sarsi sensu Sars (1904) is redescribed as H. pseudosarsi sp. nov.; H. sarsi sensu T. & A. Scott (1894) is synonymized with H. canaliculatum (Por). H. propinquum is regarded as a junior subjective synonym of H. chrystalli (T. Scott). H. clavatum (Sars) is resurrected, having previously been regarded as a synonym of H. brunneum (Brady). A key for the identification of females belonging to this group of species is provided.     

Resonance Raman spectroscopy of polynucleotides

Resonance Raman Spectroscopy allows a selective study of the bases of DNA and therefore of the interactions of these bases with ligands. This technique is also sensitive to structural modifications. We show here that, first, the structures of native poly(dA-dT).poly(dA-dT) and poly(dA).poly(dT) are not the same and that, secondly, it is possible to characterize the B----Z transition of poly(dG-dC).poly(dG-dC). The study of the Raman hypochromism during the thermal denaturation of the polynucleotides reveals that the stacking of the adenines in poly(dA).poly(dT) is near that observed in poly(rA) but differs of this stacking in poly(dA-dT).poly(dA-dT). The enhancement of the intensity of the guanine line at 1193 cm-1 and of the cytosine lines at 780 cm-1, 1 242 cm-1 and 1268 cm-1 as well as the shift of the guanine line at low frequency should allow to characterize a small proportion of base pairs in Z form in any DNA.     

Flunitrazepam binding sites in rat diaphragm. Receptors for direct neuromuscular effects of benzodiazepines?

The evidence for direct muscle relaxant effects of benzodiazepines is controversial. We now show that a crude membrane preparation of rat diaphragm possesses binding sites for [3H]flunitrazepam (FNZ). Scatchard analysis gave a binding site density of 1689 +/- 143 fmol/mg protein (Kd = 25.6 +/- 2.6 nM). These sites are of the "peripheral" type since clonazepam fails to displace [3H]FNZ as effectively as R05-4864 (IC50 values: 7.5 x 10(-6) M and 8 x 10(-9) M, respectively). Diazepam is almost as effective as R05-4864 and potently displaces [3H]FNZ binding (IC50 = 3 x 10(-8) M). We propose that the previously described effects of diazepam on rat diaphragm are mediated through high-affinity binding sites.     

Conformational peculiarities of polynucleotides with a nonrandom base sequence according to the 1H----3H exchange rate in C8H groups of purinic residues

We have determined the 1H----3H exchange rate constants between water and C8H groups of purinic residues of alternating polynucleotides poly(dA-dT).poly(dA-dT), poly(dG-dC).poly(dG-dC) and poly(dA-dC).poly(dG-dT) as well as homopolynucleotides poly(dA).poly(dT) and poly(dG).poly(dC) in aqueous solutions with high-salt concentrations (3 M NaCl and 4-6 M CsF), in water-ethanol (60%) solution and in 0.15 M NaCl at 25 degrees C. The rate constants for adenine (kA) and guanine (kG) of polynucleotides were compared with corresponding constants for E. coli DNA. dGMP nd dAMP at the same conditions. The relation between exchange rates and conformations of polynucleotides permits the study of their conformational peculiarities in solution. Of three alternating polynucleotides examined in 0.15 M NaCl the exchange retardation was observed only for poly(dA-dT).poly(dA-dT) as compared with that in B-DNA, which is in good agreement with the B-alternating "wrinkled" DNA model. The conformations of poly(dG-dC).poly(dG-dC) and poly(dA-dC).poly(dG-dT), according to the exchange data obtained are within the B form. For homopolynucleotides in 0.15 M NaCl, the KA value for poly(dA).poly(dT) is nearly the same as kA for B-DNA, which indicates the similarity of their conformations, whereas the kG value for poly(dG).poly(dC) is 1.7-fold lower in comparison with the kG value in B-DNA. This seems to be connected with the existence of B = A conformation equilibrium for poly(dG).poly(dC) in solution. The increase of NaCl concentration to 3 M results in a B----Z transition in the case of poly(dG-dC).poly(dG-dC) and in the shift of B = A equilibrium towards the A-form in the case of poly(dG).poly(dC) as is evidenced by alterations of their KG values. Poly(dA-dT).poly(dA-dT) in 6 M CsF and poly(dA-dC).poly(dG-dT) in 4.3 M CsF maintain their inherent conformations in 0.15 M NaCl in spite of the fact that they are characterised by the "X-type" CD-spectrum at these conditions. According to the exchange data the conformation of poly(dA).poly(dT) in 6 M CsF corresponds to the "heteronomous" DNA model or some other structure with lower accessibility of C8H groups of adenylic residues.     

豌豆中脱落酸含量、叶片导性与土壤含水量之间的关系

脱落酸与植物水分胁迫的关系已进行了大量的研究,现已明确脱落酸可以抑制气孔的开放(Jones和 Mansfield 1970,Weyers和 Hillman 1979,Henson等 1989),但它的作用机制仍然很不清楚。已有报告指出,当土壤水分亏缺时,ABA可以作为根与地上部间的通讯信号,由很运到地上部并抑制气孔的开放(Blackman和 Davies1985,Gollan等 1986,Zhang和 Davies     

Monomeric sarcosine oxidase: evidence for an ionizable group in the E.S complex

Monomeric sarcosine oxidase (MSOX) contains covalently bound FAD and catalyzes the oxidation of sarcosine (N-methylglycine) and other secondary amino acids, including L-proline. The reductive half-reaction with L-proline proceeds via a rapidly attained equilibrium (K(d)) between free E(ox) and the E(ox).S complex, followed by a practically irreversible reduction step (E(ox).S --> E(red).P) associated with a rate constant, k(lim). The effect of pH on the reductive half-reaction shows that the K(d) for L-proline binding is pH-independent (pH 6.46-9.0). This indicates that MSOX binds the zwitterionic form of L-proline, the predominant species in solution at neutral pH (pK(a) = 10.6). Values for the limiting rate of reduction (k(lim)) are, however, strongly pH-dependent and indicate that an ionizable group in the E(ox).L-proline complex (pK(a) = 8.02) must be unprotonated for conversion to E(red).P. Charge-transfer interaction with L-proline as the donor and FAD as acceptor is possible only with the anionic form of L-proline. The ionizable group in the E(ox).L-proline complex is required for conversion of enzyme-bound L-proline from the zwitterionic to the reactive anionic form, as judged by the independently determined pK(a) for charge-transfer complex formation with the MSOX flavin (pK(a) = 7.94). The observation that the anionic form of L-proline with a neutral amino group is the reactive species in the reduction of MSOX is similar to that observed for other flavoenzymes that oxidize amines, including monoamine oxidase and trimethylamine dehydrogenase.     

Demonstration of two stable potential states in the squid giant axon under tetraethylammonium chloride

1. Intracellular injection of tetraethylammonium chloride (TEA) into a giant axon of the squid prolongs the duration of the action potential without changing the resting potential (Fig. 3). The prolongation is sometimes 100-fold or more. 2. The action potential of a giant axon treated with TEA has an initial peak followed by a plateau (Fig. 3). The membrane resistance during the plateau is practically normal (Fig. 4). Near the end of the action potential, there is an apparent increase in the membrane resistance (Fig. 5D and Fig. 6, right). 3. The phenomenon of abolition of action potentials was demonstrated in the squid giant axon treated with TEA (Fig. 7). Following an action potential abolished in its early phase, there is no refractoriness (Fig. 8). 4. By the method of voltage clamp, the voltage-current relation was investigated on normal squid axons as well as on axons treated with TEA (Figs. 9 and 10). 5. The presence of stable states of the membrane was demonstrated by clamping the membrane potential with two voltage steps (Fig. 11). Experimental evidence was presented showing that, in an "unstable" state, the membrane conductance is not uniquely determined by the membrane potential. 6. The effect of low sodium water was investigated in the axon treated with TEA (Fig. 12). 7. The similarity between the action potential of a squid axon under TEA and that of the vertebrate cardiac muscle was stressed. The experimental results were interpreted as supporting the view that there are two stable states in the membrane. Initiation and abolition of an action potential were explained as transitions between the two states.     

中国6个植物原白中排印错误之更正

对我国紫草科（Boraginaceae）的密花齿缘草（Eritrichium confetiflorum W. T. Wang）、锚刺果（Glochisocaryum kansuense W. T. Wang）和大花长叶微孔草（Microula trichocarpa（Maxim.） Johnst. var. macrantha W. T. Wang）,唇形科（Lamiaceae）的宽叶香茶菜（Rabdosia latifolia C. Y. Wu & H. W. Li）,苦苣苔科（Gesneriaceae）的矮直瓣苣苔（Ancylostemon humilis W. T. Wang）以及葫芦科（Cucurbitaceae）的黑子赤瓟（Thladiantha villosula Cogn. var. nigrita A. M. Lu & Z. Y. Zhang）原白中主模式标本引证的排印错误做了更正。密花齿缘草原白中错误地将主模式标本引证为杨昌友700422,实际应为杨昌友700442。锚刺果原白中用中文错误地将主模式标本引证为傅坤俊1989,实际应为傅坤俊1489,前者属于百合科（Liliaceae）的Asparagus sichuanicus S. C. Chen & D. Q. Liu。大花长叶微孔草原白中主模式标本引证为李馨70287,实际应为李馨72087,前者属于杜鹃花科（Ericaceae）的Rhododendron edgarianum Rehder & Wilson。宽叶香茶菜原白中主模式标本引证为曲桂龄2049,实际应为曲桂龄2046,前者属于蔷薇科（Rosaceae）的Rosa caudata Baker。矮直瓣苣苔原白中主模式标本引证为杨光辉59063,实际应为杨光辉59043,前者属于桦木科（Betulaceae）的Betula fargesii Franch.。黑子赤瓟原白中主模式标本引证为冯国楣3439,实际应为冯国楣3429,前者属于伞形科（Apiaceae）的Bupleurum rockii H. Wolff。     

Obesity-related changes in high-density lipoprotein metabolism

Obesity is associated with a 3-or-more-fold increase in the risk of fatal and nonfatal myocardial infarction (1,2,3,4,5,6). The American Heart Association has reclassified obesity as a major, modifiable risk factor for coronary heart disease (7). The increased prevalence of premature coronary heart disease in obesity is attributed to multiple factors (8,9,10). A principal contributor to this serious morbidity is the alterations in plasma lipid and lipoprotein levels. The dyslipidemia of obesity is commonly manifested as high plasma triglyceride levels, low high-density lipoprotein cholesterol (HDLc), and normal low-density lipoprotein cholesterol (LDLc) with preponderance of small dense LDL particles (7,8,9,10). However, there is a considerable heterogeneity of plasma lipid profile in overweight and obese people. The precise cause of this heterogeneity is not entirely clear but has been partly attributed to the degree of visceral adiposity and insulin resistance. The emergence of glucose intolerance or a genetic predisposition to familial combined hyperlipidemia will further modify the plasma lipid phenotype in obese people (11,12,13,14,15).     

Influence of Temperature on Embryogenesis in Meloidogyne javanica

The optimum temperature for embryogenesis in Meloidogyne javanica lies between 25 and 30 C. Embryogenesis is slightly more rapid at 30 C (9-10 days), but more eggs complete development at 25 C (11-13 days). At temperatures of 25, 27.5, and 30 C, embryogenesis is about twice as rapid as at 20 C (23-25 days), and about four times as rapid as at 15 C (46-48 days). Time-lapse studies showed that the thermal optimum is similar throughout the different stages of embryonic development.     

Manipulation of thiol contents in plants

As sulfur constitutes one of the macronutrients necessary for the plant life cycle, sulfur uptake and assimilation in higher plants is one of the crucial factors determining plant growth and vigour, crop yield and even resistance to pests and stresses. Inorganic sulfate is mostly taken up as sulfate from the soil through the root system or to a lesser extent as volatile sulfur compounds from the air. In a cascade of enzymatic steps inorganic sulfur is converted to the nutritionally important sulfur-containing amino acids cysteine and methionine (Hell, 1997; Hell and Rennenberg, 1998; Saito, 1999). Sulfate uptake and allocation between plant organs or within the cell is mediated by specific transporters localised in plant membranes. Several functionally different sulfate transporters have to be postulated and have been already cloned from a number of plant species (Clarkson et al., 1993; Hawkesford and Smith, 1997; Takahashi et al., 1997; Yamaguchi, 1997). Following import into the plant and transport to the final site of reduction, the plastid, the chemically relatively inert sulfate molecule is activated through binding to ATP forming adenosine-5'-phosphosulfate (APS). This enzymatic step is controlled through the enzyme ATP-sulfurylase (ATP-S). APS can be further phosphorylated to form 3'-phosphoadenosine-5'-phosphosulfate (PAPS) which serves as sulfate donor for the formation of sulfate esters such as the biosynthesis of sulfolipids (Schmidt and J?ger, 1992). However, most of the APS is reduced to sulfide through the enzymes APS-reductase (APR) and sulfite reductase (SIR). The carbon backbone of cysteine is provided through serine, thus directly coupling photosynthetic processes and nitrogen metabolism to sulfur assimilation. L-serine is activated by serine acetyltransferase (SAT) through the transfer to an acetyl-group from acetyl coenzyme A to form O-acetyl-L-serine (OAS) which is then sulhydrylated using sulfide through the enzyme O-acetyl-L-serine thiol lyase (OAS-TL) forming cysteine. Cysteine is the central precursor of all organic molecules containing reduced sulfur ranging from the amino acid methionine to peptides as glutathione or phytochelatines, proteines, vitamines, cofactors as SAM and hormones. Cysteine and derived metabolites display essential roles within plant metabolism such as protein stabilisation through disulfide bridges, stress tolerance to active oxygen species and metals, cofactors for enzymatic reactions as e.g. SAM as major methylgroup donor and plant development and signalling through the volatile hormone ethylene. Cysteine and other metabolites carrying free sulfhydryl groups are commonly termed thioles (confer Fig. 1). The physiological control of the sulfate reduction pathway in higher plants is still not completely understood in all details. The objective of this paper is to summarise the available data on the molecular analysis and control of cysteine biosynthesis in plants, and to discuss potentials for manipulating the pathway using transgenic approaches.     

Mammalian DNA methyltransferases prefer poly(dI-dC) as substrate

The synthetic duplex DNA, poly(dI-dC).poly(dI-dC), is methylated in vitro by human or murine DNA methyltransferases at 20-100 times the rate of other nonmethylated DNAs. Preparation of the hemimethylated derivative, poly(dI-dMeC).poly(dI-dC), of this polymer increases its effectiveness as a substrate by 2-fold, making it 4-10 times more effective as a substrate for mammalian DNA methyltransferases than any other hemimethylated DNA so far reported. However, the apparent slower rate of de novo methylation of poly(dI-dC).poly(dI-dC) as compared to the hemimethylated derivative is due to substrate inhibition, unique to the unmethylated polymer, as the rates of de novo and maintenance methylation are identical at low substrate concentrations.     

Learning and discrimination of coloured papers in the walking blowfly,Lucilia cuprina

Summary A new training and testing paradigm for walking sheep blowflies, Lucilia cuprina, is described. A fly is trained by presenting it with a droplet of sugar solution on a patch of coloured paper. After having consumed the sugar droplet, the fly starts a systematic search. While searching, it is confronted with an array of colour marks consisting of four colours displayed on the test cardboard (Fig. 1). Colours used for training and test include blue, green, yellow, orange, red, white and black.Before training, naive flies are tested for their spontaneous colour preferences on the test array. Yellow is visited most frequently, green least frequently (Table 2). Spontaneous colour preferences do not simply depend on subjective brightness (Table 1).The flies trained to one of the colours prefer this colour significantly (Figs. 5 and 9–11). This behaviour reflects true learning rather than sensitisation (Figs. 6–7). The blue and yellow marks are learned easily and discriminated well (Figs. 5, 9, 11). White is also discriminated well, although the response frequencies are lower than to blue and yellow (Fig. 11). Green is discriminated from blue but weakly from yellow and orange (Figs. 5, 9, 10). Red is a stimulus as weak as black (Figs. 8, 9). These features of colour discrimination reflect the spectral loci of colours in the colour triangle (Fig. 14).The coloured papers seem to be discriminated mainly by the hue of colours (Fig. 12), but brightness may also be used to discriminate colour stimuli (Fig. 13).     

DNA sequence has an effect on the extent and kinds of alkylation of DNA by a potent carcinogen

A system has been developed to study the effects of base sequence (neighboring bases) upon the alkylation of guanine (G) and adenine (A) bases in DNA. The study was performed on the synthetic polydeoxyribonucleotides, poly(dG).poly(dC), poly(dG-dC).poly(dG-dC), poly(dA).poly(dT), poly(dA-dT).poly(dA-dT), poly(dA-dC).poly(dG-dT), poly(dA-dG).poly(dC-dT), as well as calf thymus DNA. Each polynucleotide was treated with N-[3H]methyl-N-nitrosourea (MNU), depurinated, and the freed alkylpurines separated by HPLC and quantitated by liquid scintillation counting. The amounts of 3-methylguanine (3-MG), 7-MG, and O6-MG relative to guanine, and 3-methyladenine (3-MA) and 1-MA plus 7-MA relative to adenine, and also the O6-MG/7-MG ratios were highly reproducible for a given polynucleotide. Significant differences were found in the amounts of each of the methylpurines formed when compared among the six synthetic polynucleotides and DNA. This evidence is interpreted as an effect upon alkylation which is ultimately dependent upon the base sequence. These findings may have significance in defining the specificity of chemical carcinogens in terms of the susceptability to modification of nucleotide sequences such as those found in certain oncogenes.     

Apolipoprotein E and cholesterol metabolism in the pathogenesis and treatment of Alzheimer's disease

There is much evidence suggesting that there is a strong relationship between the deterioration of brain lipid homeostasis, vascular changes and the pathogenesis of Alzheimer's disease (AD). These associations include: (1). recognition that a key cholesterol transporter, apolipoprotein E type 4, acts a major genetic risk factor for both familial and sporadic AD; (2). epidemiological studies linking cardiovascular risk factors, such as hypertension and high plasma cholesterol, to dementia; (3). the discovery that small strokes can precipitate clinical dementia in cognitively normal elderly subjects; (4). the modulation of degradation of the amyloid precursor protein by cholesterol administration in cell culture and in animal models of beta-amyloid overproduction; and (5). the beneficial effect of cholesterol-lowering drugs, such as Probucol and statins, in combating common AD. The recent finding that there is a genetic association between the HMGR gene locus and sporadic AD further suggests that brain cholesterol metabolism is central to AD pathophysiology, and a potential therapeutic target for disease stabilization and primary disease prevention.     

Glomales SSUrRNA gene diversity

The arbuscular mycorrhiza (AM) is one of the most important symbioses on earth, formed between about 80% of vascular plants and AM fungi (Smith & Read, 1997), which are placed in the order Glomales (Morton & Benny, 1990). A discussion has recently arisen about the rDNA-gene and general genome organization of this fungal group, which, as far as is known, does not possess sexuality (Sanders, 1999). This discussion mainly arose as a result of the assumption that one spore of an AM fungus ( Scutellospora castanea ) contains small subunit (SSU) rRNA genes that belong, phylogenetically, to different families within the Glomales, namely the Gigasporaceae and Glomaceae (Hosny et al ., 1999). Questions about genetic drift, infrequent recombination, concerted evolution and the validity of phylogenetic techniques have arisen (Hosny et al ., 1999; Sanders, 1999). However, this assumption might well be based on the sequence of an Ascomycete contaminant.