Propionibacterium acnes GehA lipase, an enzyme involved in acne development, can be successfully inhibited by defined natural substances

Propionibacterium acnes, a usual inhabitant of human skin, plays an important role in acne development, related to the production of numerous enzymatic activities involved in the degradation of host molecules. Among these enzymes, P. acnes lipase (GehA, glycerol-ester hydrolase A) has been recognized as one of the major factors in the pathogenesis of acne, being responsible for the hydrolysis of sebum and the release of inflammatory compounds. Anti-acne treatments are based on the use of retinoids or benzoyl peroxide, frequently in combination with antibiotics. However, the low effectiveness of such treatments and the increasing antibiotic resistance has led to the development of alternative therapies such as Kampo formulations, containing traditional herbal drugs. Search for new anti-acne treatments led us to perform the cloning, characterization and inhibition of P. acnes GehA, considered an interesting pharmaceutical target for anti-acne therapies. The genetic, molecular and biochemical properties of the cloned lipase were analysed, and several inhibitor agents were tested, including natural substances like saponins, alkaloids or flavonoids. Among these, the flavonoids (±)-catechin and kaempferol were the most promising candidates for acne treatment, whereas saponins like glycyrrhicic acid and digitonin produced a lower inhibition of the enzyme. No inhibition by alkaloids was found. Therefore, the inhibition caused by (±)-catechin and kaempferol on GehA, together with their wide anti-acne properties and low toxicity, make them very suitable candidates for the treatment of acne and other P. acnes-related diseases.     

Salivary gland hyaluronidase in various species of phlebotomine sand flies (Diptera: psychodidae)

Hyaluronidase activity was detected and partially characterized in salivary gland extracts of females of six sand fly species. In Phlebotomus papatasi and Lutzomyia longipalpis the enzyme was active over a broad pH range; the pH optimum was 5.0. Besides high cleaving activity towards hyaluronic acid, it hydrolyzed chondroitin sulfates A and C. Hyaluronidases of various sand fly species differed in structure and sensitivity to reducing conditions. In the subgenera Phlebotomus (P. papatasi and P. duboscqi) and Adlerius (P. halepensis) the predominant active form of the enzyme was monomeric with the same apparent molecular weight under nonreducing and reducing conditions (around 65 kDa for P. papatasi and P. duboscqi and 110 kDa for P. halepensis). In P. sergenti the enzyme occurred as a putative homodimer but remained active under reducing conditions when separated into 60 kDa subunits. In L. longipalpis and P. perniciosus the activity was detectable under non-reducing conditions only. In P. duboscqi, low enzyme activity was found also in males. Salivary gland hyaluronidases of sand flies share characteristics with endo-N-acetyl-hexosaminidases of mammalian sperm cells and corresponding venom enzymes of Hymenoptera. Hypothetically, they facilitate blood meal acquisition but also may modulate immune reactions of the host and promote pathogen transmission.     

Yeasts as biological agents to control Botrytis cinerea

Yeasts, isolated from different sources, were identified and tested for inhibition using YMA-MB plates seeded with Botrytis cinerea strains. A total of 42 yeast strains of 20 different species were tested in vitro for antagonism against 18 pathogenic B. cinerea strains. Pichia membranifaciens, P. anomala and Debaryomyces hansenii displayed the most important inhibitory effect against Botrytis strains. In small-scale trials, post-harvest application of P. membranifaciens CYC 1106 to apple wounds inhibited B. cinerea CYC 20010. Purified killer toxin from P. membranifaciens CYC 1106 inhibited B. cinerea CYC 20010. Results indicated that certain yeasts, or their toxins such us P. membranifaciens CYC 1106 killer toxin, might have potential as novel agents to control B. cinerea.     

Anti-plasmodial and anti-trypanosomal activity of synthetic naphtho[2,3-b]thiophen-4,9-quinones

Naphtho[2,3-b]thiophen-4,9-quinone and five derivatives were prepared using the Friedel-Crafts reaction and tandem-lithiation of aromatic diethylamides. These quinones were evaluated for their trypanocidal and anti-plasmodial activities by their effects on: (1) growth of epimastigote forms of Trypanosoma cruzi in vitro, (2) lysis of trypomastigote forms of T. cruzi in murine blood, (3) growth of Plasmodium falciparum in vitro, and (4) inhibition of the recombinant enzyme trypanothione reductase. The parent compound, naphtho[2,3-b]thiophen-4,9-quinone (3a), was among the most active quinone tested in vitro against P. falciparum at 0.2 μM. However, it was inactive against P. berghei-infected mice treated with 2.3 mmol/kg daily for 5 days. Most of the quinones prepared were active against T. cruzi epimastigotes in culture but exhibited weak activity at 4 °C against trypomastigotes in murine blood as well against the enzyme trypanothione reductase. Further structural modifications will be necessary to improve the in vivo activity of the naphthothiophenquinones.     

Plant growth-promoting Pseudomonas bearing catabolic plasmids: Naphthalene degradation and effect on plants

Two IncP-9 naphthalene degradative plasmids pOV17 and pBS216 were transferred into plant growth-promoting Pseudomonas which were represented by species P. aureofaciens, P. chlororaphis, P. fluorescens, and P. putida. The strains with the same plasmid differed significantly by their growth parameters, stability of the plasmid and plant protective effect from naphthalene action. Strains P. putida 53a(pOV17) and P. chlororaphis PCL1391(pOV17) demonstrated higher population number in the rhizosphere. Moreover, they protected the mustard plants from naphthalene toxic influence more effectively than the wild type strain P. aureofaciens OV17(pOV17). The activity of catechol-2,3-dioxygenase in the strains with the plasmid pOV17 was higher than that in strains with the plasmid pBS216. The strain P. putida 53a(pOV17) with high catechol-2,3-dioxygenase activity has been demonstrated to have the best protective effect. The strain P. putida 53a(pBS216) without catechol dioxygenases activities did not have protective effect but suppressed the plant germination.     

Monoquaternary ammonium derivatives inhibit growth of protozoan parasites

The phospholipid metabolism of Plasmodium falciparum-infected erythrocytes has been shown to be an effective pharmacological target for novel chemotherapy. Thirty-seven monoquaternary ammonium derivatives analogous to choline were screened for their potential antiprotozoal activity against P. falciparum and Leishmania braziliensis. Twenty-three compounds inhibited chloroquine resistant and sensitive P. falciparum strains with inhibitory concentrations ranging from 0.001 μM to 47 μM. Among the inhibitors were six compounds with nanomolar activity containing at least one ethyl group in the polar head and a hydrophobic alkyl chain with 10 to 14 methylene groups. Four compounds also exhibited in vitro antileishmanial properties in the micromolar range.     

生长阶段及环境因子对东海原甲藻氨基酸氧化酶活性的影响

以中国沿海典型赤潮藻东海原甲藻为实验材料, 研究了不同生长阶段以及温度、光照和氮源对其氨基酸氧化酶活性的影响。结果表明, 在缺氮条件下东海原甲藻显示较高的氨基酸氧化酶活性。在15-30℃温度范围均检测出氨基酸氧化酶活性, 较低温度下(15-20℃)的氨基酸氧化酶活性显著高于高温处理组(25-30℃)(p<0.05), 其中20℃时的酶活最高。在50-100 μmol/(m²·s)的光强下, 氨基酸氧化酶活性较高(0.38-0.47 fmol/(h·cell)), 而在2 μmol/(m²·s)的低光强下, 虽然酶活受到显著抑制, 但仍达到0.28 fmol/(h·cell)。氮源组成对氨基酸氧化酶活性具有显著影响, 以丙氨酸为唯一氮源时的酶活最高(0.44 fmol/(h·cell)), NH₄⁺+丙氨酸为氮源时的酶活最低(0.22 fmol/(h·cell))。研究显示, 光照、温度和氮源是东海原甲藻氨基酸氧化酶活性的关键调控因子。东海原甲藻不仅能够有效利用游离氨基酸, 而且适应较广的温度和较低的光照条件, 这可能是东海原甲藻赤潮形成和持续的重要原因。     

3种速生树种落叶水浸提液对走马胎幼苗生长的化感效应

林药复合种植的关键在于科学理解造林树种与药材之间是否具有增益效应,其中避免化感抑制是重要方面之一。本研究选择以民族药用植物—走马胎（Ardisia gigantifolia Stapf）为研究对象,探讨走马胎与3种速生树种尾叶桉（Eualyptus urophylla S.T.Blakely）、湿地松（Pinus elliottii Engelm.）和杉木（Cunninghamia lanceolata（Lamb.）Hook.）复合种植中存在的化感作用。结果显示：与对照组CK相比,桉树落叶水浸提液降低了走马胎株高至少2.59%,对走马胎幼苗的株高有显著性抑制作用（P<0.05）,对走马胎幼苗的根长、地下部分干重以及生物量有极显著性抑制作用（P<0.01）,且随着浓度的增高,抑制作用增强。湿地松落叶水浸提液对走马胎幼苗的株高、根长以及地上部分干重出现低浓度促进高浓度没有显著性影响的双浓度效应,而对走马胎幼苗的基径、地下部分干重以及生物量呈现低浓度促进高浓度抑制的双浓度效应;杉木落叶水浸提液对走马胎幼苗生长的影响呈现低浓度促进高浓度抑制的双浓度效应现象。综合各项指标得出：走马胎与桉树之间存在较强的负化感效应,而走马胎与湿地松或杉木之间存在“低促高抑”的化感效应;     

Diurnal variations in physiology and behaviour of the noble crayfish Astacus astacus and the signal crayfish Pacifastacus leniusculus

Heart rate, locomotor activity, and oxygen consumption were recorded simultaneously and continuously in seven individuals of the noble crayfish Astacus astacus (Linneus 1758) and seven individuals of the signal crayfish Pacifastacus leniusculus (Dana 1852). The recordings were made in the laboratory over 7 days at 15°C under a 12 : 12 h dark : light regime. Circadian rhythms in heart rate, locomotor activity and oxygen consumption were found both in A. astacus and P. leniusculus. Increased heart rate, locomotor activity, and oxygen consumption levels during night time in both A. astacus and P. leniusculus illustrated expression of nocturnal behaviour. No differences in oxygen consumption levels were observed between A. astacus and P. leniusculus. Also, no significant difference between heart rate levels or heart rate variances was found in A. astacus and P. leniusculus at night. During day, however, heart rate levels, heart rate variances and locomotor activity were higher in P. leniusculus than in A. astacus. The higher activity level in P. leniusculus than in A. astacus during daytime indicates that P. leniusculus is less strictly nocturnal than is A. astacus.     

Attack, escape and predation rates of larvae of two aphidophagous ladybirds during conspecific and heterospecific interactions

The attack, escape and predation rates for larvae of aphidophagous ladybird Propylea dissecta (Mulsant) and Coccinella transversalis Fabricius were quantified as a potential mechanism leading to the differences in the incidence of cannibalism and intraguild predation. These rates were compared at four larval instars within and between the species. The attack rates of larvae of C. transversalis were significantly higher than those of P. dissecta towards conspecific and heterospecific victims. For both species, third instars exhibited maximum tendency to attack. Escape rates in C. transversalis were higher than P. dissecta. In P. dissecta, the second instars made a greater number of escapes than other conspecific instars after being attacked by same stage cannibal or heterospecific predator. In P. dissecta, first instars suffered maximum mortality due to cannibalism and intraguild predation by conspecifics and heterospecifics of the same and older developmental stage. No larvae of C. transversalis were eaten by P. dissecta of the same stage. These results suggest that the larvae of P. dissecta were more often potential cannibals than intraguild predators, while the reverse was the case in C. transversalis. Based on this finding, it could be predicted that in patchy prey habitats, high rates of larval cannibalism in P. dissecta would occur with a high risk of cannibalism of first instars. Larvae of C. transversalis would respond as intraguild predators, while those of P. dissecta as intraguild prey. The greater size and walking activity of C. transversalis could be possible reason for this tendency.     

Parasitism activity of Peristenus spp. (Hymenoptera: Braconidae) on Lygus lineolaris (Hemiptera: Miridae) nymphs prior to the establishment of P. digoneutis in southwestern Quebec

Parasitism of tarnished plant bug nymphs, Lygus lineolaris (Palisot de Beauvois) was investigated on different weeds and on lettuce, celery and alfalfa-clover, in a muck soil area of southwestern Quebec. The phenology of parasitism, defined as the time during which female parasitoids are active in the field, was evaluated. Two nymphal parasitoid species were found: Peristenus pallipes (Curtis) and P. pseudopallipes (Loan). Lygus lineolaris was most abundant on stinging-nettle with levels of parasitism of 17% (P. pallipes) and 27% (P. pseudopallipes) and on alfalfa-clover with levels of parasitism of 25% (P. pallipes) and 8% (P. pseudopallipes). On lettuce, parasitism by P. pseudopallipes was below 4% while no parasitism was observed on celery. The dissection and rearing methods were used to estimate parasitism, and dissection usually yielded parasitism levels higher than rearing. Degree-days requirements of both host and parasitoids were used to calculate the period of activity of parasitoid adults. The estimates of adult activity were 47 days from late May to early July for P. pallipes, and 42 days from mid-July to late August for P. pseudopallipes.     

基于DNA条形码技术对浙江省外来入侵福寿螺进行分子鉴定

外来入侵福寿螺对我国农业生产和水生生态系统平衡等造成严重危害。2010年, 种类鉴定研究首次揭示我国外来入侵福寿螺包括Pomacea canaliculata和P. maculata两个种, 而浙江省仅见P. canaliculata一种报道。P. canaliculata和P. maculata种间形态近似, 且受环境、食物源等因素影响, 同种内外壳形态特征多样, 因而基于形态特征进行种类的准确鉴定极为困难。本研究在采集浙江省7个区县的福寿螺样本的基础上, 利用DNA条形码技术扩增了101个不同个体的COI序列, 并从BOLD数据库下载了“P. canaliculata种团”的5个近缘种的55条COI序列用于分析, 其中包括P. lineata, P. dolioides和P. paludosa所有已发表序列, 以及P. canaliculata和P. maculata的南美洲样品的序列等。序列相似度比对、DNA条形码间隙和系统发育树等分析表明, COI序列可以实现近缘福寿螺的有效鉴别。待测的浙江省福寿螺样品中, 杭州江干区检测到P. canaliculata和P. maculata两种, 而舟山普陀区、绍兴上虞区和新昌县、温州瓯海区及杭州西湖区仅检测到P. canaliculata, 表明P. canaliculata在浙江省具有更广的分布范围。P. canaliculata和P. maculata分别形成4种和2种单倍型, 各区县样点分别包含1-3种单倍型, 浙江省各发生地呈现较低的遗传多样性。依据系统发育关系推测, 浙江省分布的P. canaliculata和P. maculata分别可能来源于阿根廷和巴西。     

松属针叶角质层微形态特征在分类学中的应用

角质层是表皮细胞壁表面的一层不透水的脂肪性物质。角质层与表皮细胞紧密结合,植物表皮细胞形态和排列方式、气孔器的形态结构等微形态特征均能在角质层上反映出来。利用光学显微镜和扫描电镜对松属（Pinus）12种植物针叶角质层微形态特征进行观察和比较,详细描述20个性状,其中12个性状来自角质层内表面,8个性状来自角质层外表面。结果表明,这些特征可为该属属下分类和相似种的鉴别提供有用信息,具有重要的分类学意义：①表皮细胞长度、表皮毛长度、角质层外表面起伏程度、表皮细胞轮廓、有无气孔塞和针絮状物质等角质层微形态特征具有自身特异性,在属下可作为松属组级水平上的分类依据。角质层微形态 特征不支持将五针松组（P. Section Cembra）和白皮松组（P. Section Parrya）合并为P. Section Quinquefolius的观点,亦不支持将油松组（P. Section Pinus）分成P. Section Pinus和P. Section Trifolius的看法。②白皮松（P. bungeana）针叶角质层微形态特征既与五针松组有相同之处,又与油松组有相似之处,还有部分特征显示出不同于松属其他种类的独特性,可为白皮松亚属（P. Subgenus Parrya）的建立提供新依据。③扫描电镜下表皮细胞垂周壁纹路,气孔塞有无和外表面气孔形状等特征可为形态相似种火炬松（P. taeda）和湿地松（P. elliottii）提供种间界定依据。     

Methionine analogues as inhibitors of methionyl-tRNA synthetase

A series of methionine analogues have been synthesized as inhibitors of methionyl-tRNA synthetase and evaluated for their inhibitory activities of E. coli methionyl-tRNA synthetase and bacterial growth. Among them, -methionine hydroxamate 20 has proved to be the best inhibitor of the enzyme with K_i = 19 μM and showed a growth inhibition against E.coli JM 109, P. vulganis 6059 and C. freundii 8090.     

光照、温度和氮对竹叶眼子菜生长的影响

实验选择长江中下游常见沉水植物竹叶眼子菜(Potamogeton malaianus Mig.),利用正交试验设计,通过室内静态模拟实验研究3种主要环境因子光照强度、温度、总氮浓度及其互作对竹叶眼子菜分苗生长的影响。结果表明:光照强度和温度为影响竹叶眼子菜生长的主效环境因子,且它们的交互作用对竹叶眼子菜生长有较为显著的影响;在2～8mg·L^-1的总氮浓度下,竹叶眼子菜均可以正常生长。观察结果可知,竹叶眼子菜在5320～12000lx光照强度、30℃、4～8mg·L^-1水体总氮浓度的条件下生长良好,竹叶眼子菜生长虽然对光照强度的要求较高,但其叶往往集中在水体表面形成冠层,能在透明度较低的水体中生长,加上对高温及高营养盐的耐受性较好,故推测竹叶眼子菜可作为夏季富营养化水体中恢复和重建沉水植被的先锋工具种。     

Drought stress induced changes in some organic substances in nodules and other plant parts of two potential legumes differing in salt tolerance

A greenhouse experiment was conducted to assess the effect of water stress on growth and metabolic changes in nodules and other plant parts of two leguminous species, Phaseolus vulgaris and Sesbania aculeata, with the major objective that nodules play a vital role in drought tolerance. Imposition of water deficit conditions for 45 days to 15-day-old plants of P. vulgaris and S. aculeata reduced shoot mass and nodule mass of both species, but the reduction was more pronounced in P. vulgaris than in S. aculeata. Nitrate reductase (NR) activity was reduced more in the leaves and nodules of P. vulgaris than in S. aculeata. Soluble proteins in the nodules of S. aculeata were more decreased as compared to that in P. vulgaris. Free amino acids increased in all parts of both species due to water deficit, but a higher increase was observed in leaf and nodules of P. vulgaris than in S. aculeata. Osmoprotectants such as proline and glycine betaine increased more in the nodules and other parts of S. aculeata under drought stress. In conclusion, S. aculeata (salt tolerant) showed a higher degree of drought tolerance than P. vulgaris (salt sensitive). Drought tolerance of S. aculeata was found to be associated with a smaller reduction in number and mass of root nodules, a high activity of nitrate reductase in leaves and nodules, high accumulation of free proline in roots and nodules, and high glycine betaine content in nodules.     

基于叶绿体DNA atpB-rbcL片段的典型樱亚属部分种的亲缘关系及分类地位探讨

蔷薇科典型樱亚属植物（Subg.Cerasus Koehne）（樱花类）具有重要的观赏价值,深入研究其种间亲缘关系及分类地位,对于国产樱属植物系统发育与种质资源创新具有重要的理论指导意义。基于PCR扩增与测序技术研究了从云南、江西、福建采集的典型樱亚属3个种（含变种）的叶绿体DNA非编码区atpB-rbcL片段,结合GenBank数据库已经提交的其它4个种的同源序列,共获得7个种的30条序列矩阵,以此构建了单倍型中介网络图与系统发育树,分析和探讨了它们的亲缘关系与分类地位。（1）在atpB-rbcL序列矩阵中,774个有效位点中共发现15个多态性位点,占分析序列的1.94%,（A+T）含量为70.50%,存在明显的碱基偏好;（2）DNASP 5共检测到9个单倍型,物种间平均单倍型多样性（H_d=0.880 5±0.026）,平均核苷酸多态性（π=0.007 11±0.000 54）,除尾叶樱桃（Prunus dielsiana）外（Hap5~Hap7）,其余物种拥有各自独特的1个单倍型,具有较丰富的遗传多样性;（3）综合Network中介图与系统发育重建结果：推测磐安樱桃（P.pananensis）处于较原始的进化阶段,相比尾叶樱桃,与浙闽樱桃（P.schneideriana）亲缘关系较近;长柱尾叶樱桃（P.dielsiana var.longistyla）与尾叶樱桃互为姊妹支,并共同构成独立进化单元。综上,叶绿体atpB-rbcL片段在典型樱亚属下种间的适用效果较好,可以优先选为DNA条形码;分子证据结合形态学特征支持将长柱尾叶樱桃作为尾叶樱桃变种的处理意见,而磐安樱桃的分类地位则需要更多的证据。     

Cloning and sequence analysis of the lipase and lipase chaperone-encoding genes from Acinetobacter calcoaceticus RAG-1, and redefinition of a Proteobacterial lipase family and an analogous lipase chaperone family

The genes encoding the lipase (LipA) and lipase chaperone (LipB) from Acinetobacter calcoaceticus RAG-1 were cloned and sequenced. The genes were isolated from a genomic DNA library by complementation of a lipase-deficient transposon mutant of the same strain. Transposon insertion in this mutant and three others was mapped to a single site in the chaperone gene. The deduced amino acid (aa) sequences for the lipase and its chaperone were found to encode mature proteins of 313 aa (32.5 kDa) and 347 aa (38.6 kDa), respectively. The lipase contained a putative leader sequence, as well as the conserved Ser, His, and Asp residues which are known to function as the catalytic triad in other lipases. A possible trans-membrane hydrophobic helix was identified in the N-terminal region of the chaperone. Phylogenetic comparisons showed that LipA, together with the lipases of A. calcoaceticus BD413, Vibrio cholerae El Tor, and Proteus vulgaris K80, were members of a previously described family of Pseudomonas and Burkholderia lipases. This new family, which we redefine as the Group I Proteobacterial lipases, was subdivided into four subfamilies on the basis of overall sequence homology and conservation of residues which are unique to the subfamilies. LipB, moreover, was found to be a member of an analogous family of lipase chaperones. We propose that the lipases produced by P. fluorescens and Serratia marcescens, which comprise a second sequence family, be referred to as the Group II Proteobacterial lipases. Evidence is provided to support the hypothesis that both the Group I and Group II families have evolved from a combination of common descent and lateral gene transfer.