RAPID HLA-DRB1 GENERIC TYPING BY PCR-SSP METHOD

We report a simple and rapid HLA-DRBI generic typing method, PCRSSP, which is practical and inexpensive. We use 9 sequence-specific primers and 2 group specific Primers to define the HI-ADRB1 specificities DR1,DR2, DR3, DR4,DR5,DR6, DR7, DR8, DR9 and DR10. The HLA DR3, DR5,DR6 and DR8 can be amplified by the two primers of DR3568 and DRB1. The DR6 specificity can be identified by excluding the DR3, DR5 and DR8 when the DR3568 are positive. Any individuals can be typed with some exception: the three pairs of phenotype DR3/DR3 and DR3/DR6, DR5/DRS and DR5/DR6,DR8/DR8 and DR8/DR6 cannot be discriminated from each other. We typed 106 unrelated healthy people from Beijing locations in two weeks. We think this typing method is suitable to replace the error-prone serologic HLA-DR tests in routine clinical practice, including the Prospective typing of cadaveric organ donors.

Fuzzy Extensive Economical Control

ln this paper, FEEC theory is studied. An economical space is a dynamic model for any scope of certain economy and market . ln an economical space, any point represents some economical area or entity and is assigned an variable economic condition Any subspace is assigned an variable economic evaluation. FEEC controller operates oz. economic evaluation and the public parts in economic condition. Any adjustment done by FEEC controller asks the point itself to improve other members to meet the requirements. ln this paper, the template for economic condition and economic evaluation is defined , and then the generic type of economical space and FEE. controller is also given.

AN ASSOCIATION STUDY BETWEEN ESSENTIAL HYPERTENSION AND HLA-DRB1 ALLELES

It is well established that genetic and environmental factors are involved in the etiology of essential hypertension(EH). Previous studies have suggested that at least one of the HLA genes is responsible for the genetic susceptibility to EH. Our aim in the present study was to investigate this issue in China by the PCR-SSP HLA-DRB1 typing method. The results showed an increased frequency of HLA-DR2 and a decreased frequency of HLA-DR7 with EH patients compared with controls. We consider that HLA-DR2 may represent a marker for susceptibility to FH in the North Chinese population.

Insights into the genus Diaporthe:phylogenetic species delimitation in the D.eres species complex

The genus Diaporthe comprises pathogenic,endophytic and saprobic species with both temperate and tropical distributions.Cryptic diversification,phenotypic plasticity and extensive host associations have long complicated accurate identifications of species in this genus.The delimitation of the generic type species Diaporthe eres has been uncertain due to the lack of ex-type cultures.Species limits of D.eres and closely related species were evaluated using molecular phylogenetic analysis of eight genes including nuclear ribosomal internal transcribed spacer(ITS),partial sequences of actin(ACT),DNA-lyase(Apn2),translation elongation factor 1-α(EF1-α),beta-tubulin(TUB),calmodulin(CAL),60s ribosomal protein L37(FG1093)and histone-3(HIS).The occurrence of sequence heterogeneity of ITS within D.eres is observed,which complicates the analysis and may lead to overestimation of the species diversity.The strict criteria of Genealogical Concordance Phylogenetic Species Recognition(GCPSR)were applied to resolve species boundaries based on individual and combined analyses of other seven genes except the ITS.We accept nine distinct phylogenetic species including Diaporthe alleghaniensis,D.alnea,D.bicincta,D.celastrina,D.eres,D.helicis,D.neilliae,D.pulla and D.vaccinii.Epitypes are designated for D.alnea,D.bicincta,D.celastrina,D.eres,D.helicis and D.pulla.Modern descriptions and illustrations are provided for these species.Newly designed primers are introduced to amplify and sequence the Apn2(DNA-lyase)gene in Diaporthe.Based on phylogenetic informativeness profiles,EF1-α,Apn2 and HIS genes are recognised as the best markers for defining species in the D.eres complex.

Pleosporales

One hundred and five generic types of Pleosporales are described and illustrated.A brief introduction and detailed history with short notes on morphology,molecular phylogeny as well as a general conclusion of each genus are provided.For those genera where the type or a representative specimen is unavailable,a brief note is given.Altogether 174 genera of Pleosporales are treated.Phaeotrichaceae as well as Kriegeriella,Zeuctomorpha and Muroia are excluded from Pleosporales.Based on the multigene phylogenetic analysis,the suborder Massarineae is emended to accommodate five families,viz.Lentitheciaceae,Massarinaceae,Montagnulaceae,Morosphaeriaceae and Trematosphaeriaceae.

Towards a natural classification of Botryosphaeriales

The type specimens of Auerswaldia,Auerswaldiella,Barriopsis,Botryosphaeria,Leptoguignardia,Melanops,Neodeightonia,Phaeobotryon,Phaeobotryosphaeria,Phyllachorella,Pyrenostigme,Saccharata,Sivanesania,Spencermartinsia and Vestergrenia were examined and fresh specimens of Botryosphaeriales were collected from Thailand.This material is used to provide a systematic treatment of Botryosphaeriales based on morphology and phylogeny.Two new genera,Botryobambusa and Cophinforma are introduced and comparedwith existing genera.Four species newto science,Auerswaldia dothiorella,A.lignicola,Botryosphaeria fusispora and Phaeobotryosphaeria eucalypti,are also described and justified.We accept 29 genera in Botryosphaeriales,with Macrovalsaria being newly placed.In the phylogenetic tree,the 114 strains of Botyrosphaeriales included in the analysis cluster into two major clades with 80%,96%and 1.00(MP,ML and BY)support,with Clade A containing the family type of Botryosphaeriaceae,and Clade B containing Phyllosticta,Saccharata and Melanops species.This group may represent Phyllostictaceae.In Clade A the taxa analyzed cluster in eight sub-clades(Clades A1-8).Clade A1 comprises three distinct subclusters corresponding to the genera Diplodia(Diplodia Clade),Neodeightonia(Neodeightonia Clade)and Lasiodiplodia(Lasiodiplodia Clade).Clade A2 clusters into three groups representing Phaeobotryosphaeria(100%),Phaeobotryon(100%)and Barriopsis(94%).Clade A3 incorporates 17 strains that cluster into three well-supported genera(Dothiorella(86%),Spencermartinsia(100%)and Auerswaldia(63%);the position of Macrophomina is not stable.Clade A4 is a single lineage(100%)representing the new genus Botryobambusa.Clade A5 is a wellsupported subclade incorporating Neofussicoccum.Clade A6 represents the type species of Botryosphaeria,three other Botryosphaeria species and two other genera,Neoscytalidium and Cophinforma gen.nov.Clade A7 comprises two Pseudofusicoccum species and Clade A8 has two Aplosporella species.These sub-clades may eventually require separate families but this requires analysis of a much larger dataset.Our data advances the understanding of Botryosphaeriales,there is,however,still much research to be carried out with resolution of families and genera,linkage of sexual and asexual morphs and differentiation of cryptic species.

Epitypification and neotypification:guidelines with appropriate and inappropriate examples

review of phylogenetic studies carried out together with morphological ones shows that a major problem with most early studies is that they concentrated on techniques and used material or strains of fungi that in most cases were not carefully reference,and in a worrying number of cases wrongly named.Most classical species,particularly of microfungi,are not represented by adequate type material,or other authoritatively identified cultures or specimens,that can serve as DNA sources for phylogenetic study,or for developing robust identification systems.Natural classifications of fungi therefore suffer fromthe lack of reference strains in resultant phylogenetic trees.In some cases,epitypification and neotypification can solve this problem and these tools are increasingly used to resolve taxonomic confusion and stabilize the understanding of species,genera,families,or orders of fungi.This manuscript discusses epitypification and neotypification,describes how to epitypify or neotypify species and examines the importance of this process.A set of guidelines for epitypification is presented.Examples where taxa have been epitypified are presented and the benefits and problems of epitypification are discussed.As examples of epitypification,or to provide reference specimens,a new epitype is designated for Paraphaeosphaeria michotii and reference specimens are provided for Astrosphaeriella stellata,A.bakeriana,Phaeosphaeria elongata,Ophiobolus cirsii,and O.erythrosporus.In this way we demonstrate how to epitypify taxa and its importance,and also illustrate the value of proposing reference specimens if epitypification is not advisable.Although we provided guidelines for epitypification,the decision to epitypify or not lies with the author,who should have experience of the fungus concerned.This responsibility is to be taken seriously,as once a later typification is made,it may not be possible to undo that,particularly in the case of epitypes,without using the lengthy and tedious formal conservation and rejection processes.