Possible bioactive conformations of alpha-melanotropin

Rat brain microtubules were prepared at the adult stage and from immature (i.e., 4-day-old) animals. At an early stage of development, the composition of microtubule-associated proteins is qualitatively different from that found at the adult stage [(1982) Eur. J. Biochem. 129, 465-471]. The influence of calmodulin on the time course of assembly of second cycle microtubules was compared at both stages of brain development (i.e., microtubules originating from 4-day-old and adult animals). In the presence of Ca2+ the inhibition of microtubule assembly was more pronounced at a young stage of brain development than at the adult stage. Cross-linking studies with 125I-labeled calmodulin further established that the two major microtubule-associated proteins, MAP2 and TAU were able to bind to calmodulin at both stages of brain development but with different intensities. The labeling with 125I-labeled calmodulin was Ca2+-dependent, specific, displaced by unlabeled calmodulin and trifluoperazine.     

Modification of isolated α and β chains of human hemoglobin by the metal tetrasulfonated phthalocyanines. Studies on hybrid phthalocyanine-heme intermediates

α and β chains of hemoglobin have been modified with cobalt(II) tetrasulfonated phthalocyanine in place of heme. They display properties very similar to those of iron(II) phthalocyanine modified α and β chains. Mixed together they form tetrameric cobalt(II) phthalocyanine hemoglobin.Incorporation of Co(II)L into α and β globins results in stabilization of the protein structure, which is shown by a marked increase in its helicity content. Cobalt phthalocyanine substituted α and β chains are able to combine reversibly with oxygen giving more stable oxygenated species than their native analogues. The rate of both processes is lower in the case of the modified α chain. Recombination of the phthalocyanine α and β chains with the alternate heme containing chains give tetrameric hybrid hemoglobins. These comprise two phthalocyanine modified subunits and two heme containing subunits. The helicity content of the tetrameric hybrid hemoglobin calculated for one subunit is lower that the arithmetic mean of helicities for its isolated subunits. This suggests a destabilizing chain-chain interaction within the tetramer. Unlike in the separated subunits, oxygen binding by hybrid hemoglobins is irreversible. Deoxygenation by argon bubbling leads to the formation of inactive species which in oxygen atmosphere undergo irreversible oxidation with destruction of the complex.     

Synthesis, characterization, DFT studies and DNA binding of mixed platinum (II) complexes containing quinoxaline and 1,2-dithiolate ligands

The complexes Pt(pq)Cl2(1) and Pt(pq)(bdt) (2) (where pq = 2-(2'pyridyl)quinoxaline and bdt=benzene-1,2-dithiolate) have been synthesized and fully characterized by UV-visible (UV-Vis), Fourier Transformer Infrared Spectra (FTIR), 1 and 2D NMR and cyclic voltammetry (CV). Interactions of the tested systems (the aforementioned complexes 1 and 2) and the free ligands pq and bdt with double stranded calf thymus DNA (CT-DNA) were studied by UV-spectrophotometric (melting curves) and circular dichroism (CD) measurements. The results suggest that both complexes 1 and 2, are able to form adducts with DNA and to distort the double helix by changing the base stacking. Complex 2 forms stronger adducts to CT-DNA than complex 1 and this is probably due to the substitution of the chlorine atoms of 1 by the 1,2-dithiolate ligand (bdt) in 2. The latter induces an extensive distortion in the planarity of 2 as density functional theory (DFT) calculations reveal. Besides, the light absorbing complex 2 possess intense mixed metal ligand to ligand charge transfer (MM'LLCT) transition in the visible region of the spectrum and could act as photoluminescent metal-based probe for the study of DNA binding. Thus, the photocleavage of DNA by 2 has been studied by UV-Vis and CD spectra and monitored by agarose gel electrophoresis. Under our experimental conditions, it is unclear that complex 2 can photocleave DNA. Furthermore, the ability of 2 to inhibit proliferation of human tumor cell lines was tested and the results indicate some cytoxytic effect on the SF-286 cells.     

Effect of spermine on interaction of DNA polymerase α from the loach (Misgurnus fossilis) eggs with DNA

Polyamines (putrescine, spermidine and spermine) cause a marked increase in the activity of the loach Misgurnus fossilis DNA polymerase α on activated (gapped) DNA. The stimulatory effect increases in the order: putrescine, spermidine, spermine. Kinetic analysis shows that spermine does not change the affinity of the polymerase for dTTP, but it decreases the enzyme affinity for DNA. The apparent K_m of the polymerase for activated DNA progressively increases from 14 to 1200 μM (nucleotide), if the concentration of spermine rises up to 2 mM, while V_max reaches a maximum at 0.5 mM spermine and then drops at higher polyamine concentrations. Native calf thymus DNA and especially single-stranded DNA from phage M13 appear to be inhibitors of α-polymerase activity on gapped DNA. Dixon plots suggest simple competitive inhibition of the polymerase activity by single- or double-stranded DNA and absence of cooperativity in the interaction of the polymerase with DNA. Hill-plot analysis is compatible with the interpretation that there is only one DNA binding site on each DNA polymerase α molecule. Spermine, even at low concentrations, decreases sharply the affinity of the enzyme for double-stranded DNA, while the enzyme affinity for single-stranded DNA changes insignificantly. Another result of spermine action is the destabilization of the polymerase-DNA complex. The ratio of the ‘static affinity’ of the enzyme to its ‘kinetic affinity’ decreases 2.2-fold in the presence of 0.5 mM spermine. As a result, the sensitivity of DNA synthesis to 3′-deoxy-3′-aminothymidine 5′-triphosphate and to 1-β-d-arabinofuranosylcytidine 5′-triphosphate decreases in the presence of the polyamine. Both spermine effects, the decrease in the ‘nonproductive binding’ of the polymerase to double-stranded regions in DNA and the destabilization of the polymerase-DNA complex, presumably account for the increase in the activity of the loach α-polymerase on activated DNA.     

Application of reversed-phase liquid chromatography and prepacked C18 cartridges for the analysis of oxytetracycline and related compounds

The reversed-phase (RP) chromatographic separation of oxytetracycline (OTC) 4-epioxytetracycline (4-epiOTC), α-apooxytetracycline (α-apoOTC), and β-apooxytetracycline (β-apoOTC) has been accomplished on an Inertsil C₈ column at ambient temperature. Using the simplex method of solvent optimization, a 0.1 M ammonium acetate buffer (pH 3.0)-acetonitrile-tetrahydrofuran (72.5:12.5:15, v/v/v) mobile phase was found to give excellent separation of the compounds. OTC, 4-epiOTC, α-apoOTC and β-apoOTC were resolved in 35 min with calculated detection limits of 40, 20, 50 and 140 ng/ml, respectively. Solid-phase extraction (using RP C₁₈ cartridges) of OTC and OTC degradation compounds from distilled water and porcine muscle was tested at four concentration levels ranging from 200 to 2000 ng/ml (g); overall mean recovery of OTC from distilled water and porcine tissue was greater than 90% and 70%, respectively.     

氧离子辐射对体外人精子自发化学发光、运动性、顶体反应和存活率的影响(英文)

我们研究了不同剂量(0,0.25,0.5,1,2,4,8,16,32,64Gy)3.17MeV/u氧离子对体外人精子自发化学发光(SCL)、运动性,顶体反应(AR)和存活率的影响。结果表明,精子SCL随辐射剂量明显增强,最低有效剂量为0.5Gy;精子运动性在0.5—2Gy,AR在0.5—4Gy辐射组均明显高于其对照组,但均随辐射剂量的进一步增加而明显降低;在0.25—8Gy剂量之间,精子存活率与其对照组相比无明显变化,但当剂量进一步增加至16—64Gy时,存活率急剧下降。这些结果提示,低剂量重离子辐射能对精子运动性及AR等功能产生兴奋效应。而大剂量则明显抑制这些功能,甚至造成精子死亡。其作用机制可能与重离子辐射诱导的自由基反应有关。     

Binding of collagen to group A, B, C, D and G streptococci

Abstract Binding of ¹²⁵I-labelled collagen type II to group A, B, C, D and G streptococci was studied. Strains of all five serogroups were found to bind. Binding to one high-binding strain (group G, strain 12127) was characterised. This was reversible, saturable with time and inhibited by unlabelled type II collagen, but not by other proteins such as fibronectin and ovalbumin. However, binding was inhibited by unlabelled type I, II and III collagens and gelatin, suggesting that a common structure of various collagens is involved in binding.     

Membrane damage by staphylococcal α-toxin to different types of cultured mammalian cell

Staphylococcal α-toxin was shown to be more membrane-damaging to epithelial-like cells than to neuroblasts or normal fibroblasts. Mouse adrenal cortex tumor (Y₁Ac) epithelioid cells and human embryonal lung (MRC-5) fibroblasts were used for further comparison. Alpha-toxin was considerably more cytotoxic to adrenal cells than to fibroblasts. This difference did not depend on the presence of fibronectin on the fibroblast surface, or on a general difference in the response to other membrane-damaging hemolytic toxins and detergents. Incubation of adrenal cells at 0°C with α-toxin induced some irreversible change, and membrane damage and a cytotoxic effect developed upon further incubation in toxin-free growth medium. In fibroblasts the membrane damage progressed slowly and only in the continued presence of the toxin. Toxin-induced damage to transport and synthetic functions in fibroblasts was reversible upon removal of the toxin after prolonged exposure. It is proposed that adrenal cells may carry a cell-surface receptor to which α-toxin binds specifically, thereby allowing the toxin to exert its cell damaging effect.     

Vegetation succession in lakes of West Connemara,Ireland: comparing predicted and actual changes

Abstract. Changes in the vegetation of lakes and wetlands were investigated over a period of 18 years. It was assumed that changes in vegetation were related to changes in agricultural land use resulting in increased phosphate levels in surface waters. Data were collected in 1975, 1988 and 1993. Multivariate techniques were used to relate changes in vegetation to changes in environmental factors. With the use of a Markovian chain model, vegetation development was projected into the future. Projections based on vegetation dynamics between 1975 and 1988 were compared with actual changes in the vegetation. The vegetation dynamics appeared stable on a regional scale but quite dynamic on a local scale. A continuous decline in species diversity was noted as well as an overall increase of phosphate level. However, only minor changes in vegetation could be attributed to this increase of phosphate. Major changes were a result of fluctuations in water level. These changes coincide with periods of drier and wetter climate. Because of the fluctuating nature of these changes, predicted vegetation change did deviate from the observed change.     

柚的起源、演化及分布初探

柚 (Ｃｉｔｒｕｓｇｒａｎｄｉｓ (Ｌ .)Ｏｓｂｅｃｋ)是桔属的三个基本种之一 ,是一类重要的种质资源 ,经济价值高 ,近年来国内外的开发利用方兴未艾。柚果以其果形美观 ,味道鲜美 ,营养丰富 ,且耐储藏 ,易运输 ,素有“天然罐头”之称。我国是柑桔的起源中心和遗传变异中心之一 ,资源极其丰富。中国柚的人工栽培最早 ,夏书《禹贡》就有“扬州厥包橘柚锡贡”的记载 ;《吕氏春秋》有“果之美者 ,云梦之柚”之说 ,证明柚的栽培至少有三千多年的历史。我国目前在广东、广西、福建、四川、台湾、湖南、江西、浙江、江苏、安徽、云南、贵州、湖北…     

Phytogeographical and community similarities of alpine tundras of Changbaishan Summit,China, and Indian Peaks,USA

Abstract. We compared the diversity, phytogeography, and plant communities in two mid-latitude alpine tundras with comparable aerial and elevational extents: Changbaishan Summit in eastern Asia and Indian Peaks in western North America. Despite wide separation, the two areas shared 72 species. In all, 43% of the species on Changbaishan Summit are also distributed in the alpine zones of western North America, while 22% of the species on Indian Peaks are also distributed in the alpine zones of eastern Asia. Almost all the shared species also occur in the Beringian region. Phytogeographical profiles of species and genera showed that 69% of species and over 90% of genera in both alpine tundras belong to the three phytogeographical categories: cosmopolitan, circumpolar, and Asian-North American. We attributed the current floristic relationship between these widely separated areas to the periodic past land connection between the two continents during the Tertiary and Pleistocene. Indian Peaks has a closer floristic relationship with the Arctic tundra than does Changbaishan Summit. Indian Peaks also has 45% higher species richness and lower vegetation cover than Changbaishan Summit. Plant communities from the two areas were completely separated in the two-way indicator species analysis and non-metric multidimensional scaling on floristic data at both species and generic levels, whereas ordination of communities by soil data produced a greater overlap. The plant communities on Changbaishan Summit in general have lower alpha diversity, higher beta diversity (lower between-community floristic similarity), and more rare species than does Indian Peaks. Mosaic diversity does not differ in the two alpine tundras, although the analysis suggests that Changbaishan Summit communities are more widely spaced on gradients than the Indian Peaks communities.     

Arabinosyl nucleosides. XIII. Influence of arabinofuranosylthymine on DNA-, RNA- and protein-synthesizing systems in vitro

The natural metabolite of the sponge Cryptotethya crypta, arabinofuranosylthymine (araThd), is intracellularly phosphorylated to araTTP. The present study demonstrates that araTTP inhibits both isolated DNA polymerases α and the DNA polymerase β from L5178y cells competitively with respect to the analogous substrate dTTP. The affinity of araTTP is higher to the DNA polymerase α than to the DNA polymerase β.The activity of mammalian DNA-dependent RNA polymerases I, II and III as well as the incorporation rate of a protein cellfree system is not affected by high doses of araTTP.     

A revised structure for the triterpene rigidenol

The structure of the triterpene rigidenol has been revised to 11α-hydroxy-lup-20(30)-en-3-one.     

Structure-function studies on the cyclic peptide MT-II, lactam derivative of alpha-melanotropin.

The alanine-substituted and the retro, enantio, and retro-enantio analogs of MT-II, a potent agonist at melanocortin (MC) receptors, were prepared by solid-phase synthesis and evaluated for their ability to bind and activate human MC3, MC4, and MC5 receptors. Replacement of His with Ala resulted in [Ala6]-MT-II with affinity and agonist potency at human MC3, MC4, and MC5 receptors similar to MT-II. Substitution of Arg with Ala gave compound 100-fold less potent than MT-II, but replacement of Phe or Trp with Ala led to inactive compounds (at the micromolar concentrations). The significant drop of potency of the retro, enantio, and retro-enantio analogs of MT-II, demonstrated a crucial role of side-chain topology, and to a lesser degree, of peptide backbone in interactions of MT-II with the melanocortin receptors. The nuclear magnetic resonance analysis of MT-II suggested involvement of Phe and Arg residues in H-bonds stabilizing the bent conformations of the peptide backbone.     

Anti-Tumor Activity of an Enzymatically Synthesized α-1,6 Branched α-1,4-Glucan,Glycogen

Oral administration of an enzymatically synthesized α-1,4:1,6-glycogen (ESG) at a dose of 50 μg/ml significantly prolonged the survival time of Meth A tumor-bearing mice. ESG also significantly stimulated macrophage-like cells (J774.1), leading to augmented production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α). The weight-average degree of polymerization (DPw) and the ratio of branch linkage (BL) of ESG were 149,000 and 8.1% respectively. β-Amylase-treated ESG, however, lost J774.1-activating activity although inhibited subcutaneous growth of Meth A tumor cells admixed with it. Its DPw and BL changed to 126,000 and 20% respectively. Partially degraded amylopectin [(AP), DPw: 110,000, BL; 5.1] was also effective at stimulating J774.1, but its activity was lower than that of ESG. Other α-glucans [cycloamylose (CA), enzymatically synthesized amylose (ESA), highly branched cyclic dextrin (HBCD), and β-amylase-treated HBCD], of which DPw was lower than that of ESG, showed no J774.1-activating activity and weaker anti-tumor activity.     

云南省(虫齿)目二新种和灰背蛇(虫齿)的记述((虫齿)目:厚(虫齿)科、叉(虫齿)科、围(虫齿)科)

本文记述了云南省(虫齿)目二新种,Tapinella bannana sp.n.和Peripsocus plurimaculatus sp.n.及一新种记录种Ophiodopelma semicets Lee and Thornton,其雄虫为首次记载。     

(-)-Epigallocatechin-3-gallate prevents oxidative damage in both the aqueous and lipid compartments of human plasma

When human plasma was exposed to the hydrophilic radical initiator, AAPH, (-)-epigallocatechin-(3)-gallate (EGCG) dose-dependently inhibited the aqueous compartment oxidation (IC(50)=0.72 microM) (monitored by DCFH oxidation) and spared the lipophilic antioxidants, alpha-tocopherol, and carotenoids, but not ascorbic acid. When radicals were selectively induced in the lipid compartment by the lipophilic radical initiator, MeO-AMVN, EGCG spared alpha-tocopherol, but not carotenoids and inhibited the lipid compartment oxidation (monitored by BODIPY 581/591) with a potency lower than that found in the aqueous compartment (IC(50)=4.37 microM). Our results indicate that EGCG, mainly localized in the aqueous compartment, effectively quenches aqueous radical species, thus limiting their diffusion into the lipid compartment and preventing lipid-soluble antioxidant depletion. Further, ESR experiments confirmed that EGCG recycled alpha-tocopherol through a H-transfer mechanism at the aqueous/lipid interface affording an additional protective mechanism to the lipid compartment of plasma.     

The folding pathway of a single domain in a multidomain protein is not affected by its neighbouring domain

Domains are the structural, functional, and evolutionary components of proteins. Most folding studies to date have concentrated on the folding of single domains, but more than 70% of human proteins contain more than one domain, and interdomain interactions can affect both the stability and the folding kinetics. Whether the folding pathway is altered by interdomain interactions is not yet known. Here we investigated the effect of a folded neighbouring domain on the folding pathway of spectrin R16 (the 16th α-helical repeat from chicken brain α-spectrin) by using the two-domain construct R1516. The R16 folds faster and unfolds more slowly in the presence of its folded neighbour R15 (the 15th α-helical repeat from chicken brain α-spectrin). An extensive Φ-value analysis of the R16 domain in R1516 was completed to compare the transition state of the R16 domain alone with that of the R16 domain in a multidomain construct. The results indicate that the folding pathways are the same. This result validates the current approach of breaking up larger proteins into domains for the study of protein folding pathways.     

Synthesis,characterization, DNA interaction,and cytotoxicity of novel Pd(II) and Pt(II) complexes

Four complexes [Pd(L)(bipy)Cl]·4H₂O (1), [Pd(L)(phen)Cl]·4H₂O (2), [Pt(L)(bipy)Cl]·4H₂O (3), and [Pt(L)(phen)Cl]·4H₂O (4), where L = quinolinic acid, bipy = 2,2’-bipyridyl, and phen = 1,10-phenanthroline, have been synthesized and characterized using IR, ¹H NMR, elemental analysis, and single-crystal X-ray diffractometry. The binding of the complexes to FS-DNA was investigated by electronic absorption titration and fluorescence spectroscopy. The results indicate that the complexes bind to FS-DNA in an intercalative mode and the intrinsic binding constants K of the title complexes with FS-DNA are about 3.5?×?10⁴ M^?1, 3.9?×?10⁴ M^?1, 6.1?×?10⁴ M^?1, and 1.4?×?10⁵ M^?1, respectively. Also, the four complexes bind to DNA with different binding affinities, in descending order: complex 4, complex 3, complex 2, complex 1. Gel electrophoresis assay demonstrated the ability of the Pt(II) complexes to cleave pBR322 plasmid DNA.