Nucleotide sequence of dengue type 3 virus genomic RNA encoding viral structural proteins

Complementary DNAs to the 5 proximal region of the dengue virus type 3 RNA were cloned into bacterial plasmids and the nucleotide sequence of 3,000 bases from the 5 terminus of the genome were determined by DNA and RNA sequencing methods using dideoxy chain-termination reactions. Comparison of the nucleotide sequence thus obtained with those of other flavivirus genomes revealed significant homology existing in nucleotide sequence of the flavivirus genomes. When we compared amino acid sequence deduced from the nucleotide sequence with those of other flaviviruses, this genome region was found to include sequences encoding three viral structural proteins C, M, and E and a part of the viral nonstructural protein NS1 in this order in addition to the 5-noncoding sequence. The characteristics and functions of these proteins were discussed based on the deduced amino acid sequences and their hydrophobic profiles. The genetic relationship of flaviviruses was also discussed based on the genetic variation observed in their genomes.     

Human growth hormone 1 (GH1) gene expression: complex haplotype-dependent influence of polymorphic variation in the proximal promoter and locus control region

The proximal promoter region of the human pituitary expressed growth hormone (GH1) gene is highly polymorphic, containing at least 15 single nucleotide polymorphisms (SNPs). This variation is manifest in 40 different haplotypes, the high diversity being explicable in terms of gene conversion, recurrent mutation, and selection. Functional analysis showed that 12 haplotypes were associated with a significantly reduced level of reporter gene expression whereas 10 haplotypes were associated with a significantly increased level. The former tend to be more prevalent in the general population than the latter (p<0.01), possibly as a consequence of selection. Although individual SNPs contributed to promoter strength in a highly interactive and non-additive fashion, haplotype partitioning was successful in identifying six SNPs as major determinants of GH1 gene expression. The prediction and functional testing of hitherto unobserved super-maximal and sub-minimal promoter haplotypes was then used to test the efficacy of the haplotype partitioning approach. Electrophoretic mobility shift assays demonstrated that five SNP sites exhibit allele-specific protein binding. An association was noted between adult height and the mean in vitro expression value corresponding to an individual's GH1 promoter haplotype combination (p=0.028) although only 3.3% of the variance of adult height was found to be explicable by reference to this parameter. Three additional SNPs, identified within sites I and II of the upstream locus control region (LCR), were ascribed to three distinct LCR haplotypes. A series of LCR-GH1 proximal promoter constructs were used to demonstrate that 1) the LCR enhanced proximal promoter activity by up to 2.8-fold depending upon proximal promoter haplotype, and that 2) the activity of a given proximal promoter haplotype was also differentially enhanced by different LCR haplotypes. The genetic basis of inter-individual differences in GH1 gene expression thus appears to be extremely complex.     

天然免疫与获得性免疫的进化关系

免疫有天然免疫和获得性免疫两种类型,它们有不同的机制和起源.天然免疫可识别某些"非己”细胞或分子并加以清除;获得性免疫则对分子抗原表位进行识别,按抗原提呈细胞等有无协同信号(发育阶段/类型)而有所区别.两者有不同的生物学起源与意义;天然免疫源于防御入侵者的需求,获得性免疫则源于系统及个体自身发育中调节细胞发育的需求.两者嫁接性混合进化形成了复杂的可识别"自己/非己”的免疫系统,并留下了神奇的机制.     

Optimizing vaccine design for cellular processing,MHC binding and TCR recognition

In this review we describe the methods and processes that our group have developed while aiming to test and design multiepitope vaccines for infectious diseases and cancer. Testing the performance of vaccines composed of epitopes restricted by human leukocyte antigen (HLA) molecules is accomplished by in vitro antigenicity assays, as well as in vivo immunogenicity assays in HLA transgenics. The efficiency by which multiepitope vaccines are processed is optimized by spacer residues, which are designed to facilitate generation by natural processing of the various class I- and class II-restricted epitopes. Methods and strategies to test and optimize HLA binding affinity, patient coverage from the vaccine construct, and TCR recognition of HLA/epitope complexes are also discussed.     

Evolution of age at menarche and at onset of regular cycling in a large cohort of French women

BACKGROUND: Early exposure to ovarian hormones is considered to increase breast cancer incidence. The age at which the ovaries become functional is thus important. METHODS: We explored the evolution of age at first menstruation and at onset of regular cycling in 86 031 women participating in the E3N-EPIC cohort study, part of the European Prospective Investigation into Cancer. RESULTS: We observed an increase in mean age at menarche among women born between 1925 and 1930, followed by a steady decrease in the youngest birth cohorts. In contrast, age at onset of regular cycling increased gradually from 1925 onwards. There was thus a steady increase in the interval between age at menarche and at onset of regular cycling, mainly due to an increase in the percentage of women in whom regular cycling started at least 5 years after menarche (from 9.0% among women born in 1925-1929 to 20.8% in those born in 1945-1950). The increase in the interval between menarche and onset of regular cycling was even greater among women with a late menarche. CONCLUSIONS: This increase might be due to a change in dietary intake and/or physical exercise aimed at achieving the slim silhouette desired by the younger generations.     

Physical mapping of the plastid genome from the chlorophyll c-containing alga,Cryptomonas Φ

Summary A physical map of the circular plastid genome of Cryptomonas has been constructed using the enzymes SacI, BamHI, SmaI, SalI, PstI and XhoI. In addition, fine-structural mapping of the inverted repeat region has been performed using AvaI, BglII, EcoRI and XbaI. The inverted repeat is very small, encompassing no more than 6 kb and containing only genes for the rRNAs. It divides the plastid genome into a small singlecopy region of 12–13 kb which contains genes for phycoerythrin and the 32 kd photosystem II polypeptide, and a large single-copy region of 93–94 kb, giving a total size of 118 kb. The genes for the large subunit of ribulose-1,5-bisphosphate carboxylase (Rubisco) and the beta subunit of ATP synthase CF₁ are encoded in the large single-copy region. The evolutionary significance of the organization of this plastid genome, the first presented from the chlorophyll c-, phycobiliprotein-containing group of algae, is discussed.     

Genome analysis of dengue type-1 virus isolated between 1990 and 2001 in Brazil reveals a remarkable conservation of the structural proteins but amino acid differences in the non-structural proteins

We have investigated the genetic diversity of dengue type-1 (DEN-1) virus in Brazil. The full nucleotide sequences of three DEN-1 virus isolated from DEN fever (DF) and DEN hemorrhagic fever patients in northeastern Brazil in 1997 (BR/97) and one from a DF patient in the south of Brazil in 2001 (BR/01) were compared to that of the reference strain BR/90 obtained in the city of Rio de Janeiro in 1990. Sequence analysis showed that the structural proteins were remarkably conserved between all isolates. A total of 27 amino acid changes occurred throughout the non-structural proteins. Among them, nine amino acid substitutions were specific of BR/97 and BR/01 isolates, indicating that in situ evolution of these strains had occurred. Within the BR/97 and BR/01 samples, some amino acid substitutions have been previously identified in DEN-1 virus strains sequenced so far, suggesting that recombination events might have occurred.     

Cytogenetics of a new cytotype of African Mus (subgenus Nannomys) minutoides (Rodentia, Muridae) from Kenya: C- and G- banding and distribution of (TTAGGG) n telomeric sequences

We present the results of a cytogenetic study on Mus (Nannomys) minutoides from Kenya by means of C- and G- banding and in-situ fluorescence hybridization (FISH) to localize the telomeric sequences. The karyotype is characterized by the occurrence of several Rb chromosomes Rb(1.X), Rb(1.Y). Rb(2.17), Rb(3.13), Rb(4.10), Rb(5.11), Rb(6.7), Rb(8.12), not previously described for this species. This finding suggests a high level of chromosomal diversification, which means it is possible to consider this cytotype as a new, well-differentiated, chromosomal lineage within the subgenus. The C-banding of the metaphases illustrated conspicuous blocks of centromeric heterochromatin at the paracentromeric regions of all telocentric chromosomes. Centromeric heterochromatin is not visible on all biarmed chromosomes. Following hybridization with telomeric probes, bright interstitial telomeric sequence (ITS) fluorescence signals are evident at the pericentromeric area of all Rb chromosomes, with the exception of Rb(2.17). Considering the localization of the C-positive heterochromatin and of the telomeric sequences, the events leading to the Kenyan cytotype from an all-telocentric condition probably included two steps: first, fusion without loss of heterochromatin and pericentromeric telomeric sequences; second, the reduction of the C-positive satellite DNA followed by the amplification of telomeric sequences in the C-negative paracentromeric region of Rb chromosomes. The presence of a single Rb(2.17) without ITS indicates possible variations of this mechanism.     

An influenza A(H3) reassortant was epidemic in Australia and New Zealand in 2003

During 2003, Australia and New Zealand experienced substantial outbreaks of influenza. The strain responsible was an A(H3N2) influenza virus described as A/Fujian/411/2002-like, which had circulated as a minor variant in the previous Northern Hemisphere (NH) winter, mainly in Korea and Japan. Early in the year the isolates were very similar to those that had been previously isolated in the NH, however, a reassortant strain emerged early in the New Zealand winter, followed by the appearance of similar viruses in Australia and other regional areas. While the hemagglutinin HA1 sequence of these viruses demonstrated only minor differences from the A/Fujian/411/2002 reference strain, the neuraminidase gene was clearly different from that of other recently circulating H3 viruses and most closely matched an earlier reference strain A/Chile/6416/2001. Three internal genes (NS, NP, M) in the reassortant viruses were also more closely related to the A/Chile/6416/2001 lineage. This reassortant A(H3) virus predominated in Australia and New Zealand in 2003 was also seen in Brazil and Malaysia during 2003 and was widespread in the United States and Europe during their 2003-04 winter. Interestingly most of the strains of A(H3) that were isolated at the beginning of the 2004 winter in Australia, did not have this earlier A/Chile/6416/2001-like neuraminidase but had a neuraminidase that was similar to that of the reference strain A/Fujian/411/2002. This was suggestive of the re-introduction of influenza A(H3) from other countries, however, there was still low level circulation of the reassortant virus in 2004 with isolates detected in Australia and Singapore.     

The chromosome complement ofAcomys spp. (Rodentia,Muridae) from Oursi,Burkina Faso—the ancestral karyotype of thecahirinus-dimidiatus group?

We present here data on chromosome banding analysis (R- and C-bands) ofAcomys sp. (Rodentia, Muridae) from Oursi, Burkina Faso, characterized by 2n=FN=68 and comparison of its banding patterns with those ofAcomys dimidiatus from Saudi Arabia (2n=38, FN=70), studied previously. The study revealed complete homology between acrocentric chromosomes ofAcomys sp. and chromosome arms of 16 pairs of metacentric and two pairs of acrocentric chromosomes ofA. dimidiatus. In addition to monobrachial homology, one tandem translocation accompanied by a centromeric shift was identified in the karyotype of the latter species. The data obtained show that karyotypes of all the species of theAcomys cahirinus-dimidiatus group studied previously may be derived from that ofAcomys sp. from Oursi by means of numerous non-homologous Rb translocations and 1–2 tandem translocations, and thus its karyotype may be considered as ancestral for thecahirinus-dimidiatus group.accepted for publication by M. Schmid