Cloning, sequencing and characterization of a chitin synthase gene fragment from the fungus Phascolomyces articulosus /

Phascolomyces articulosus genomic DNA was isolated from 48 h old hyphae and was used for amplification of a chitin synthase fragment by the polymerase chain reaction method. The primers used in the amplification corresponded to two widely conserved amino acid regions found in chitin synthases of many fimgi. Amphfication resulted in four bands (820, 900, 1000 and 1500 bp, approximately) as visualized in a 1.2% agarose gel. The lowest band (820 bp) was selected as a candidate for chitin synthase because most amplified regions from other fimgi so far exhibited similar sizes (600-750 bp). The selected fragment was extracted from the gel and cloned in the Hinc n site of pUC19. The derived plasmid and insert were designated ^\5C\9'PaCHS and PaCHS respectively. The plasmid pUC19-PaC/fS was digested by several restriction enzymes and was found to contain BamHl and HincU sites. Sequencing of PaCHS revealed two intron sequences and a total open reading frame of 200 amino acids. The derived polypeptide was compared with other related sequences from the EMBL database (Heidelberg, Germany) and was matched to 36 other fiilly or partially sequenced fimgal chitin synthase genes. The closest resemblance was with two genes (74.5% and 73.1% identity) from Rhizopus oligosporus. Southern hybridization with the cloned fragment as a probe to the PCR reaction showed a strong signal at the fragment selected for cloning and weaker signals at the other two fragments. Southern hybridization with partially digested Phascolomyces articulosus genomic DNA showed a single band. The amino acid sequence was compared with sequences from other chitin synthase gene classes using the CLUSTALW program. The chitin synthase fragment from Phascolomyces articulosus was initially grouped in class n along with chitin synthase fragments from Rhizopus oligosporus and Phycomyces blakesleeanus which also belong to the same class, Zygomycetes. Bootstrap analysis using the neighbor-joining method available by CLUSTALW verified such classification. Comparison of PaCHS revealed conservation of intron positions that are characteristic of chitin synthase gene fragments of zygomycetous fungi.

An international investigation on the validity of the CSAPPA scale in screening for developmental coordination disorder /

The main objective of the present investigation was to continue the research initiated by Hay and colleagues (2004) in examining the efficacy of the Children's Self-Perceptions of Adequacy in and Predilection for Physical Activity (CSAPPA) scale as a proxy for the short form of the Bruininks-Oseretsky Test of Motor Proficiency (BOTMP-SF) in screening for Developmental Coordination Disorder (DCD) in children. To better appreciate DCD knowledge outside Canada, the measurements of this investigation were expanded in Greece. A translated Greek CSAPP A scale and the BOTMP-SF were administered for the first time in Greek children. A second objective was to investigate the relationship between DCD and various risk factors of coronary artery disease (CAD) in Canadian and Greek children. A sample of 591 (Ms=322; Fs=269) Canadian and 392 (Ms=211; Fs=181) Greek children, aged 9 to 13 years, consented to the BOTMP-SF, CSAPP A Scale, participation in physical activity questionnaire, Leger 20-meter Multistage Shuttle Run test, and body fat using bioelectric impedance. Prevalence of DCD in Canada and Greece was 8% and 19%, respectively. Significant agreement (p

The People's Republic of China and her relations with the countries of the association of Southeast Asian nations : 1969-1975

Phe Ihesis examines the evolution of the -policies of the People fs Jtenublie of China towards J?hail°nd, PTal ysia, Singapore, Iidonesia pad the Philip-pines, organised in the Association of Southeast Asian Nations from 1969 to 1975• 2ze first central point of this study is an *ir sumption that the foreign relations of The People's Tepublic of Chi la Towards Southeast ^sia have been motivated by a dynamic interplay of t^o main factors: (1) Farxist-Leninist ideology and ICao J^e-tung Ph^ught, which dictate to China to behave as a revolutionary Dover vhich must assist the insurgent movements in the area in their strug fle to overthrow the local governments; (2) national interest, vhich demands of China to safeguard the southern flank of her territory bordering on Southerst 'sia through Friendly relations, trade and ot*»er conventional inrtniments of diplomacy. hile the tvo main motive factors are nuTually antagonistic and exclusivet the Chinere leaders are nevertheless at te mi ting to oring them iirco a coherent policy under Mao's theory of tve {hniity of op-nosites," vhich believes that it is -possible to reconcile these co-posing tendencies into a dynamic enuibrium through vhich both opnosites could be promoted at the same time although not to the same extent* la other words, the Chinese leaders conceive the dynamic equilibrium as a continuum between them in a mix in which one or the other orientation predominates in different •periods* Bins we might see China1 s conduct motivated in one period by mostly ideological considerations at the expense of the staire-to-state relations, then ve might see her policy in the middle of the continuum and suf ering from immo bill sine and just muddling through, or finally ?fe might see her emphasising friendly ties at the expense of support of revolutionary movements at the other extreme -point of the spectrum* !fhe mechanism vhich enables Peking to move from one pole to the other of the spectrum is activated by the following elementsJ (1) the result of an internal power struggle within the leadership in Peking between ideologically radical and moderate elements, which enables the victorious faction to initiate nev policies; (2) Peking's assessment of the changing intentions and capabilities of the major powers in the area; (3) internal changes within the countries of the area and the changing attitudes of their governments towards China; (4) changing fortunes of revolutionary movements operating in the area* 'Phe second major point of this study is an assertion that while China's conduct toward Southeast *lsia after her foundation in 1949 was primarily based upon ideological considerations, the beginning of the seventies saw the national interest reasserting itself as the leading motive factor* Thus China talks with her neighbours in Southeast asia in terms of relevance of fllong historical ties," casting herself into the role of a benevolent "older brother11 who is entitled to reopect and deference in exchange for patronage and protection* Hence the traditional echoes of the past are emerging ever stronger and influencing her postures towards the region, while the open support to revolutionary moevments is underplayed at the moment.