Identification of regulatory regions and regulatory protein complexes of the Spirulina desD gene under temperature stress conditions: Role of thioredoxin as an inactivator of a transcriptional repressor GntR under low-temperature stress

In the present study, electrophoretic mobility shift assays were used to identify temperature responsive elements in the 5' upstream region (5' UTR) of the Spirulina desD gene. Overlapping, synthetic oligonucleotides of both sense and anti-sense strands that spanned the entire 5' UTR of the gene were analyzed. The responsive DNA-binding protein complexes were identified using liquid chromatography-tandem mass spectrometry. The results indicated that the cold-responsive elements were located at -453 to -247, -197 to -151, -105 to -76, and -50 to -1, whereas the low-temperature specific regulatory regions were located at -372 to -352. Moreover, the heat-responsive elements were located at -347 to -243, -197 to -151, and -124 to -1, whereas the high-temperature specific elements were located between -130 to -101 and -30 to -1. In terms of regulatory protein complexes under the two stress conditions, Trx was only detected in the low-temperature responsive protein complex, and divalent cations were essential for the binding of the protein complex to the regulatory elements. Furthermore, Trx was shown to play a critical role as a reducing agent that inactivates the Spirulina desD repressor, GntR. Consequently, the desD gene expression is induced under the low-temperature condition.     

A study of 3891 cases of mycoses in the tropics.

4103 cases suspected of mycoses were analysed as to sex, age and site of disease and 3891 were proved cases. This group formed 50% of total mycoses or 13-93% of all dermatoses recorded in the Government General Hospital, Madras, during the period of study. There were 66-26% adult female, 27-6% adult male and 6-14% were below 13 years. Dermatophytoses were found in 73-5%; the other common diseases were tinea versicolor (17-68%) and candidiasis (12-43%). Multiple sites of involvement or more than 1 disease in the same individual were mostly observed. The incidence of piedra (0-1%) and deep mycoses (0-156%) was very low. Mycetoma was the common disease (5/6) in deep mycoses. In dermatophytoses, tinea corporis (49-71%) and tinea cruris (47-85%) commonest; tinea axillaris (3-42%), tinea capitis (1-72%) and tinea barbae (1-29%) were less common. The incidence of tinea manuum, tinea pedis and tinea unguium was similar (4-97%-6-38%). High temperature and humidity were related to the higher incidence of tinea corporis, tinea cruris and tinea versicolor. Mainly children suffered from tinea capitis. All other mycoses were commonly found in adults between 2nd and 3rd decades. In all mycoses but candidiasis, female predominated. Cutaneous candidiasis was mainly a problem of housewives. Among the dermatophytes Trichophyton violaceum was predominant (33-7%) followed by T. rubrum (32-6%). Trichophyton schoenleinii and M. gypseum were rarely isolated. From mycetoma, Madurella mycetomii, Nocardia braziliensis, N. asteroides and Actinomadura spp. were isolated. Demonstration of Cryptococcus laurentii in 1 case is reported in this area for the first time.     

花椒挥发油的超临界CO2萃取法与水蒸气蒸馏法提取的比较

采用超临界萃取法与水蒸气蒸馏法从花椒中提取花椒挥发油,用GC—MS方法测定各种提取方法得到挥发油的化学成分及相对含量。结果表明,水蒸气提取挥发得率为3．8％（W／W）,主要成分为柠檬烯（22．75％）、沉香醇（21．70％）、罗勒烯（3,7-二甲基-1,3,7-辛三烯）（14．27％）等。超临界CO2萃取法挥发油得率为5．9％（W／W）,主要成分为柠檬烯（14．01％）、沉香醇（13．18％）、罗勒烯（3,7-二甲基-1,3,7-辛三烯）（26．23％）等。两种方法所得到的挥发进行比较,水蒸气蒸馏法分离法得到的挥发油中分离出78个峰,其中含量和相似度高的成分有31种,占86．31％。超临界CO2萃取法提取的花椒挥发油中分离出了152个峰,含量和相似度高的成分有36种,占74．72％。两种提取方法所得到的主要挥发油中有27种成分相同。     

葡萄无核基因定位与作图的研究

以UBC 269484和GSLP1569的序列为支点,设计合成了包括UBC 269和GSLP1在内的9条引物,以葡萄 有核亲本红地球和无核亲本红光无核的DNA为模板,对这9个引物进行筛选,结果GSLP1、39970524 5号引物和 39970524 6号引物在无核亲本红光无核上扩增出了特异标记GSLP1569、39970524 5 564、39970524 6 1538和 39970524 6 1200。用这3个特异引物在红地球、红光无核、无核白和红地球×红光无核杂交组合F1代163株杂 种的DNA样上进行PCR扩增,结果4个特异标记在F1群体中与无核主效基因共分离。4个特异标记也出现于所 用组合中无核基因原始供给者无核白上。这些标记和葡萄无核主效基因相连锁。用QTXb17遗传作图软件,对葡 萄无核主效基因S定位与作图,当P=0.01时,LOD值在32.7～46.4之间,置信界限在0.2～9.9之间。这4个 特异标记和无核主效基因S处于在同一连锁群,位于无核主效基因S的两侧,覆盖基因组12.3cM。特异标记 39970524 5 564、GSLP1569、39970524 6 1538、39970524 6 1200距S基因的遗传距离分别为0.6cM、1.2cM、 4.9cM和11.1cM。     

Characteristic beta-globin gene cluster haplotypes of Evenkis and Oroqens in north China

Haplotype frequencies of the beta-globin gene cluster were estimated for 114 Evenkis and 81 Oroqens from northeast China, and their characteristics were compared with those in Japanese, Koreans, and three Colombian Amerindian groups of South America (Wayuu, Kamsa, and Inga tribes). A major 5' subhaplotype (5' to the delta-globin gene) was + - - - - in Evenkis, whereas + - - - -, - + + - +, and - + - + + were the major subhaplotypes in Oroqens. One possible candidate for an ancestral 5' subhaplotype, - - - - -, was found in one Evenki (0.5%) and three Oroqen chromosomes (2.0%). They were observed as heterozygous forms for + ---- and -----. Major haplotypes were +-----+, + -----+-, and + - - - - + + in Evenkis, whereas they were +-----+,-++-+-+, +----+-, and -+-++-+ in Oroqens. The lowest Nei's genetic distance values of Evenkis or Oroqens based on the 5' subhaplotype frequency distributions were observed in relation to the Wayuu or Koreans, respectively, but those of Evenkis and Oroqens based on the haplotype frequency distributions were found in relation to Koreans.     

Changes of action potential and L-type calcium channel current of Sprague-Dawley rat ventricular myocytes by different amlodipine isomers

To investigate the effects of S- and R-amlodipine (Aml) on action potential (AP) and L-type calcium channel current (ICa-L), the whole-cell patch-clamp technique was used on rat ventricular myocytes to record AP, ICa-L, peak currents, steady-state activation currents, steady-state inactivation currents, and recovery currents from inactivation with S-Aml and R-Aml at various concentrations. Increasing concentrations of S-Aml gradually shortened AP durations (APDs). At concentrations of 0.1, 0.5, 1, 5, and 10 micromol/L, S-Aml blocked 1.5% +/- 0.2%, 25.4% +/- 5.3%, 65.2% +/- 7.3%, 78.4% +/- 8.1%, and 94.2% +/- 5.0% of ICa-L, respectively (p < 0.05), and the half-inhibited concentration was 0.62 +/- 0.12 micromol/L. Current-voltage curves were shifted upward; steady-state activation and inactivation curves were shifted to the left. At these concentrations of S-Aml, the half-activation voltages were -16.01 +/- 1.65, -17.61 +/- 1.60, -20.17 +/- 1.46, -21.87 +/- 1.69, and -24.09 +/- 1.87 mV, respectively, and the slope factors were increased (p < 0.05). The half-inactivation voltages were -27.16 +/- 4.48, -28.69 +/- 4.52, -31.19 +/- 4.17, -32.63 +/- 4.34, and -35.16 +/- 4.46 mV, respectively, and the slope factors were increased (p < 0.05). The recovery times from inactivation of S-Aml were prolonged (p < 0.05). In contrast, R-Aml had no effect on AP and ICa-L (p > 0.05) at the concentrations tested. Thus, only S-Aml has calcium channel blockade activity, whereas R-Aml has none of the pharmacologic actions associated with calcium channel blockers.     

Genetic mapping and characterization of Pseudomonas aeruginosa mutants defective in the formation of extracellular proteins.

We isolated 15 mutants of Pseudomonas aeruginosa PAO which were defective in the formation of certain extracellular proteins, such as elastase, staphylolytic enzyme, and lipase ( Xcp mutants). The mutations were mapped on the chromosome by conjugation and transduction. The locations were xcp -1 near 0', with the gene order cys-59- xcp -1- proB , and loci xcp -2, xcp -3, and xcp -31 at 35', with the gene order trpC , D- xcp -3/ xcp -31- xcp -2- argC . Loci xcp -4 and xcp -41 through xcp -44 were cotransducible with proA at 40'; loci xcp -5, xcp -51, xcp -52, and xcp53 were located at 55', with the gene order leu-10- trpF -met-9010- xcp -53- xcp -5/ xcp -51/ xcp+ ++-52, and xcp -6 was located at 65' to 70', between catA and mtu-9002. Nine mutations ( xcp -2, xcp -3, xcp -31, xcp -4, and xcp -41 through xcp -45) caused decreased production of extracellular enzymes. Six strains with mutations xcp -1, xcp -5, xcp -51, xcp -52, xcp -53, and xcp -6 produced cell-bound exoproteins and had defective release mechanisms. The regulation of production of alkaline phosphatase and phospholipase C is different from other exoproteins , such as elastase, but they all seem to share a common release mechanism. Alkaline protease had separate mechanisms for regulation and release, since this protease was found in culture supernatants of all but one of the mutants, and none of the strains had cell-bound enzyme.     

Ionic composition of endolymph and perilymph in the inner ear of the oyster toadfish, Opsanus tau

The concentrations of free Na+, K+, Ca(+, and Cl(-)in endolymph and perilymph from the inner ear of the oyster toadfish, Opsanus tau, were measured in vivo using double-barreled ion-selective electrodes. Perilymph concentrations were similar to those measured in other species, while endolymph concentrations were similar to those measured previously in elasmobranch fish, though significantly different from concentrations reported in mammals. Perilymph concentrations (mean +/- std. dev.) were as follows: Na+, 129 mmol l(-1) +/- 20; K+, 4.96 mmol l(-1) +/- 2.67; Ca2+, 1.83 mmol l(-1) +/- 0.27; and Cl(-), 171 mmol l(-1) +/- 20. Saccular endolymph concentrations were Na+, 166 mmol l(-1) +/- 22; K+, 51.4 mmol l(-1) +/- 16.7; Ca2+, 2.88 mmol l(-1) +/- 0.27; and Cl(-), 170 mmol l(-1) +/- 12; and semicircular canal (utricular vestibule) endolymph concentrations were Na+, 122 mmol l(-1) +/- 15; K+, 47.7 mmol l(-1) +/- 13.2; Ca2+, 1.78 mmol l(-1) +/- 0.48; Cl(-), 176 mmol l(-1) +/- 27. The relatively high concentrations of Ca2+ and Na+ in the endolymph may have significant implications for the physiological function of the mechanoelectrical transduction channels in the vestibular hair cells of fish compared to those of their mammalian counterparts.     

Synthesis of poly-O-sulfated glycosides of 2,5-anhydro-D-mannitol

2,5-Anhydro-3-O-beta-D-glucopyranosyl-; -3-O-alpha-L-idopyranosyl-; -3-O-alpha-D-arabinopyranosyl-; -3-O-alpha-L-arabinopyranosyl-; -3-O-beta-D-maltopyranosyl-; -3-O-beta-D-gentiobiopyranosyl-; -1,6-di-O-beta-D-glucopyranosyl-; -1,6-di-O-alpha-L-idopyranosyl-; -1-O-beta-D-maltopyranosyl-; -1,3,6-tri-O-beta-D-glucopyranosyl-; -1,6-di-O-beta-maltopyranosyl- and -1,6-di-O-beta-D-gentiobiopyranosyl-2,5-anhydro-D-mannitol as well as their poly-O-sulfated derivatives were synthesized. The IP3-IC50 values of their sodium and/or potassium salts were determined for structure-activity studies aiming at the synthesis of new, orally active antiasthmatic compounds.     

三棱的化学成分研究

目的：研究三棱Sparganium stoloniferum Buch．-Ham．的化学成分。方法：采用硅胶柱层析,Sephadex LH-20,ODSC-18,重结晶等方法,根据理化性质和光谱鉴定化合物的结构。结果：从三棱中得到8个化合物,分别为棕榈酸（1）,5-羟甲基糠醛（2）,6,7,10-三羟基-8-十八烯酸（3）,棕榈酸单甘油酯（4）,5-Hydroxy-2-（hydroxymethyl）pyridine（5）,β-谷甾醇（β-sitosterot）（6）,β-胡萝卜苷（daucosterol）（7）,Betulinic acid（8）。结论：化合物5,8为首次从该植物中分离得到。     

Mass spectra of methyl-branched hydrocarbons from eggs of the tobacco hornworm

Hydrocarbons from three homologous series of branched alkanes from the eggs of the tobacco hornworm, Manduca sexta (L.), were identified by mass spectrometry. Gas-liquid chromatography (GLC) peaks 37-A (equivalent chain length of 37.2) and 39-A (equivalent chain length of 39.2) were mixtures of 13-, 15-, 17-, and 19-methylheptatriacontane and 13-, 15-, 17-, and 19-methylnonatriacontane, respectively. GLC peaks 33-B, 37-B, and 39-B with equivalent chain lengths of 33.4, 37.4, and 39.4, respectively, were mixtures of 13,17- and 15,19-dimethyltritriacontane, 13,17-, 15,19-, and 17,21-dimethylheptatriacontane, and 13,17-, 15,19-, and 17,21-dimethylnonatriacontane, respectively. GLC peak 37-C (equivalent chain length of 37.6) was a mixture of 11,15,19-, 13,17,21-, and 15,19,23-trimethylheptatriacontane.     

Identification of major glycoconjugates from Mycobacterium bovis culture filtrate by biotin-hydrazide labeling

To identify Mycobacterium bovis glycoproteins, carbohydrates present in a delipidized M. bovis culture filtrate protein extract were biotin-hydrazide labeled. 11 carbohydrate-containing protein with a moleculra weight of 15-, 19-, 25-, 32-, 35-, 39-, 42-, 48-, 52-, 58-, and 62-kDa were detected. The 52- and 32-kDa protein were deglycosylated by endoglycosidase-F.     

The three-dimensional folding of ribosomal RNA; localization of a series of intra-RNA cross-links in 23S RNA induced by treatment of Escherichia coli 50S ribosomal subunits with bis-(2-chloroethyl)-methylamine.

Intact 50S ribosomal subunits from E.coli were cross-linked with the symmetrical bifunctional reagent bis-(2-chloroethyl)-methylamine. After deproteinization, selected regions of the 23S RNA were excised by treatment with ribonuclease H in the presence of appropriate complementary decadeoxynucleotides, and screened for the presence of intra-RNA cross-links by two-dimensional gel electrophoresis. Individual isolated cross-linked RNA fragments were analysed by our established procedures. Sixteen intra-RNA cross-links were identified, three of which corresponded to those previously published. The thirteen 'new' cross-links were localized in the 23S RNA at positions 774-78 linked to 792-94, 876-79 linked to 899-900, 979-81 or 983-84 to 2029, 1715 to 1743-46, 1911-21 to 1964, 1933 to 1966, 2032 to 2054-55, 2112 to 2169-71, 2116-17 to 2163-67, 2128-32 to 2156-59, 2392-93 to 2422-23, 2737-38 to 2763-66, and 2791 to 2890. These results are discussed in the context of three-dimensional model-building studies with the 23S RNA, with particular reference to the environment of the 'active centre' of the 50S subunit.     

Synthesis of peptides of the preS1-region of the viral hepatitis B envelope protein and localization of antigenic determinants]

We synthesized the 24-41, 30-36, 31-36, 24-30 fragments of the preS1-region of the hepatitis B (subtype ayw) envelope. The peptides were prepared by the solid phase synthesis on perfluorpolyethylene polymer grafted with polystyrene. The peptide chains were elongated from C-terminus using activated esters and symmetrical anhydrides of Boc-amino acids, cleaved off the solid phase by HBr or TFMSA in TFA, purified by gel filtration, and, after conjugation with protein carriers, inoculated into test animals. The resultant antibodies were shown to react with peptides. The blood sera from patients with acute hepatitis B reacted with the conjugates of peptides 24-41, 30-36, 31-36 in the immunoenzymic solid phase assay. The monoclonal antibodies for the preS1-polypeptide were shown to react with peptides 24-41, 30-36, 31-36 and with their conjugates. The results obtained were proved by the data of the epitope-mapping with overlapping hexapeptides.     

Localisation of a series of intra-RNA cross-links in the secondary and tertiary structure of 23S RNA, induced by ultraviolet irradiation of Escherichia coli 50S ribosomal subunits.

Intra-RNA cross-links were introduced into E. coli 50S ribosomal subunits by mild ultraviolet irradiation. The subunits were partially digested with cobra venom nuclease, and the cross-linked RNA complexes were isolated by two-dimensional electrophoresis. Many of the complexes were submitted to a second partial digestion procedure. Oligonucleotide analysis of the RNA fragments obtained in this manner enabled cross-links between the following ribonuclease T1 oligonucleotides in the 23S RNA to be established: positions 292-296 and 339-350; 601-604 and 652-656; 1018-1022 and 1140-1149; 1433-1435 and 1556-1560; 1836-1839 and 1898-1903; 2832-2834 (tentative) and 2878-2885; 2849-2852 and 2865-2867 (tentative); 739-748 and 2609-2618; 571-577 and 2030-2032; 1777-1792 (tentative) and 2584-2588. The first seven of these cross-links lie within the secondary structure of the 23S RNA, whereas the last three are tertiary structural cross-links. The degree of precision of the individual determinations was variable, depending on the nucleotide sequence in the vicinity of the cross-link site concerned.     

Combining ability of summer-squash lines with different degrees of parthenocarpy and PRSV-W resistance

The aim was to assess heterosis in a set of 16 summer-squash hybrids, and evaluate the combining capacity of the respective parental lines, which differed as to the degree of parthenocarpy and resistance to PRSV-W (Papaya Ringspot Virus-Watermelon strain). The hybrids were obtained using a partial diallel cross design (4 × 4). The lines of parental group I were 1 = ABX-037G-77-03-05-01-01-bulk, 2 = ABX-037G-77-03-05-03-10-bulk, 3 = ABX-037G-77-03-05-01-04-bulk and 4 = ABX-037G-77-03-05-05-01-bulk, and of group II, 1' = ABX-037G-77-03-05-04-08-bulk, 2' = ABX-037G-77-03-05-02-11-bulk, 3' = Clarice and 4' = Caserta. The 16 hybrids and eight parental lines were evaluated for PRSV-W resistance, parthenocarpic expression and yield in randomized complete-block designs, with three replications. Parthenocarpy and the resistance to PRSV-W were rated by means of a scale from 1 to 5, where 1 = non-parthenocarpic or high resistance to PRSV-W, and 5 = parthenocarpic or high susceptibility to PRSV-W. Both additive and non-additive gene effects were important in the expression of parthenocarpy and resistance to PRSV-W. Whereas estimates of heterosis in parthenocarpy usually tended towards a higher degree, resistance to PRSV-W was towards higher susceptibility. At least one F(1) hybrid was identified with a satisfactory degree of parthenocarpy, resistance to PRSV-W and high fruit-yield.     

Processing pathway deduced from the structures of N-glycans in Carica papaya

A processing The processing pathway of N-glycans in Carica papaya was deduced from the structures of N-glycans. The N-glycans were liberated by hydrazinolysis followed by N-acetylation. Their reducing-end sugar residues were tagged with 2-aminopyridine and the pyridylamino (PA-) sugar chains thus obtained were purified by HPLC. Eleven PA-sugar chains were found, and their structures were analyzed by two-dimensional sugar mapping combined with partial acid hydrolysis and exoglycosidase digestion. The structures of the N-glycans were of the highmannose types with xylose and fucose; however, among them two new N-glycans, Manalpha1-6(Manalpha1-3)Manalpha1-6(Xylbeta1-2)+ ++Manbeta1-4GlcNAcbeta1- 4(Fucalpha1-3)GlcNAc and Manalpha1-3Manalpha1-6(Xylbeta1-2)Manbeta1-4G lcNAcbeta1-4(Fucalpha1-3 )GlcNAc, were found. Judging from these structures together with Manalpha1-6(Manalpha1-3)Manalpha1-6(Manalpha1-3) (Xylbeta1-2)Manbeta1- 4GlcNAcbeta1-4(Fucalpha1-3)GlcNAc reported previously [Shimazaki, A., Makino, Y., Omichi, K., Odani, S., and Hase, S. (1999) J. Biochem. 125, 560- 565], a processing pathway for N-glycans in C. papaya is inferred in which the activity of Golgi alpha-mannosidase II is incomplete.     

Synthesis and immunogenic properties of peptides--fragments of the immunodominant regions of the VP1 protein of the Asia-1 type of foot- and-mouth disease virus

Potential immunodominant epitopes were predicted on the basis of a theoretical analysis of the antigenic structure of the VP1 protein of the type Asia-1 foot-and-mouth disease virus. Peptides corresponding to the 140-153, 136-153, 132-153, 143-157, 137-157, and 193-208 fragments of the VP1 protein sequence were synthesized by the solid phase method, and the immunogenic properties of the peptides were studied on guinea pigs. The shortest peptide exhibiting the protective effect was found to correspond to the, 140-153 fragment of the VP1 sequence. The Plm-(Gly)3-(140-153)-(Gly)2-Lys(Plm)-Leu and [Ac-(140-153)-(Gly)3]8-(Lys)7-Gly synthetic constructions in combination with adjuvants provided up to 80% protection of immunized animals against infection with the foot-and-mouth disease virus.     

利用GIS和RS确定长白山自然保护区森林景观分布的环境范围

在对遥感数据进行景观分类和对环境因子进行空间表达基础上,在地理信息系统的支持下,确定长白山自然保护区森林景观分布的环境(包括年均温、年降水量、坡度和坡向)范围．结果表明。从苔原、岳桦、云冷杉到阔叶红松林,最适海拔高度范围依次为1780-2212、1705-1956、1042-1625、823-1184m;最适温度范围分别为-4．75～-2．40℃、-3．42～-2．07℃、-1．49-1．39℃、0．71-2．37℃;最适降水范围分别为1034～1110、1014-1060、883-1017、824-925mm;长白山自然保护区的森林景观主要分布在平、缓坡地上,并与坡向关系密切。苔原在各个坡向上均有分布。且在各个坡向上分布面积的变化不大;岳样、云冷杉林、阔叶红松林、山杨白样林主要呈现北、西北向分布,其次为东北、西向分布;落叶松林主要为东北向分布。其次为东和北向分布;疏林主要为西向分布,其次为西南、西北和南向分布;风倒区主要为西、西南、西北向分布。