Use of 5,6-dichloro-l-β-D-ribofuranosylbenzimidazole to distinguish light stimulation from cytokinin stimulation of amaranthin synthesis in Amaranthus tricolor

In Amaranthus tricolor seedlings, amaranthin synthesis can be induced by cytokinins and (or) by light. It was found that 5,6-dichloro-1--D-ribofuranosylbenzimidazole (DRB) was a specific inhibitor of the cytokinin response whereas the light response was unchanged. DRB also inhibited the negative response of pigment production to an anticytokinin, 7-(pentylamino)-3-methylpyrazolo(4,3-d) pyrimidine (PAMPP).DRB can be considered as a specific inhibitor of the physiological responses to cytokinins. The evolution of the inhibition percentage versus cytokinin or anticytokinin concentration shows that DRB is not a `competitor' of cytokinins.The experiments suggest that cytokinins or anticytokinins act as modulators of a fundamental light activated reaction of amaranthin synthesis.Address where proofs should be sent: Laboratoire de Physiologie végétale, Faculté des Sciences et des Techniques, Université de Nice, F-06034 Nice Cedex, France     

HLA-DRB1 * 03, but not the TNFA -308 promoter gene polymorphism, confers protection against fistulising Crohn’s disease

 Crohn’s disease (CD) appears in forms so diverse that it has been hypothesized CD might be a syndrome, with different pathogenic mechanisms leading to the various clinical phenotypes. This may plausibly explain the conflicting and inconclusive results with regard to HLA associations in unselected groups of patients. The power of these association studies may increase when disease heterogeneity is taken into account. As fistulising CD has been proposed as a separate subgroup of patients with CD, we studied the carrier frequencies (CF) of the DRB1 alleles in 35 unrelated Caucasian Dutch CD patients with proven peri-anal fistulas. A striking decrease in the frequency of the DRB1 ^* 03 allele was found in those patients with peri-anal fistulas when compared with a panel of 2400 healthy controls (HC) (3% vs 25%; P = 0.005; Odds Ratio [OR] = 0.09). The DRB1 ^* 03 allele is in strong linkage disequilibrium with a polymorphism at position –308 in the promoter region of the gene encoding TNFα (TNFA-308 ^* 2). We investigated whether this allele frequency was decreased as well. Surprisingly, the CF of TNFA-308 ^* 2 was 29%, not different from the CF of 98 HC (34%; P = 0.7; OR = 0.8). This study is the first showing a significant negative association between DRB1 ^* 03 and a particular subgroup of CD patients. Thus, patient selection may largely determine the outcome of genetic association studies in CD, as we previously observed no association with this allele in an unselected population of CD patients. As DRB1 ^* 03 frequency, but not the closely linked TNFA-308 ^* 2, was decreased, this suggests recombination between the DRB1 and TNFA loci in this group of patients, and may help to define the biological basis of fistula formation. Received: 29 September 1997 / Revised: 3 December 1997     

用聚合酶链反应-指纹图法分析HLA-DRB基因型

将不同个体的DNA经聚合酶链反应(PCR)扩增其HLA-DRB基因的高度多态区域,扩增产物经12％的聚丙烯酰胺凝胶电泳,照像后分析不同个体的指纹图型。PCR-指纹图法(PCR-Fingerprinting)为一种简便,快速的基因分析方法,可加速无关骨髓供者的筛选,它在器官移植及法医的个体识别中有一定实用价值。     

Full-length sequence analysis of the HLA-DRB1 locus suggests a recent origin of alleles

The HLA region harbors some of the most polymorphic loci in the human genome. Among them is the class II locus HLA-DRB1, with more than 400 known alleles. The age of the polymorphism and the rate at which new alleles are generated at HLA loci has caused much controversy over the years. Previous studies have mostly been restricted to the 270 base pairs that constitute the second exon and represent the most variable part of the gene. Here, we investigate the evolutionary history of the HLA-DRB1 locus on the basis of an analysis of 15 genomic full-length alleles (10-15 kb). In addition, the variation in 49 complete coding sequences and 322 exon 2 sequences were analyzed. When excluding exon 2 from the analysis, the diversity at the synonymous sites was found to be similar to the intron diversity. The overall diversity in noncoding region was also similar to the genome average. The DRB1*03 lineage has been found in human, chimpanzee, bonobo, gorilla, and orangutan. An ancestral "proto HLA-DRB1*03 lineage" appeared to have diverged in the last 5 million years into the human-specific lineages *08, *11, *13, and *14. With exception to exon 2, both the coding- and the noncoding diversity suggests a recent origin (<1 million years ago) for most of the alleles at the HLA-DRB1 locus. Sites encoding for amino acids involved in antigen binding [antigen recognizing sites (ARS)] appear to have a more ancient origin. Taken together, the recent origin of most alleles, the high diversity between allelic lineages, and the ancient origin of sequence motifs in exon 2, is consistent with a relatively rapid generation of novel alleles by gene conversion like events.     

Ovar-MHC Polymorphism in Malpura and Avikalin Sheep Vaccinated for Peste des Petits Ruminants (PPR) Virus

India harbors a vast diversity of sheep (40 breeds). The study was carried out to assess the genetic diversity of DRB1 and DQA2 locus of the ovar-MHC and their possible association with Peste des petits ruminants (PPR) virus vaccine response in Malpura and Avikalin sheep breeds maintained at an organized institute flock in the semi-arid region of India. Genetic analysis revealed the rich diversity of DRB1 locus with 23 alleles in Malpura and 21 alleles in Avikalin sheep that included 9 new alleles. DQA2 locus also had rich diversity with 19 alleles in Malpura and 20 alleles in Avikalin sheep that included 7 new alleles. At the protein level, high variability alike at the nucleotide level was observed. A marker for footrot susceptibility, DQA2*1101 was absent in both breeds. Genotypic association of DRB1 and DQA2 with PPR vaccine response was statistically non-significant. Vaccine response being a multifactorial (polygenic and influenced by environment) variable, could not show statistically significant association with MHC genotypes in the present study. However, rich genetic diversity of DRB1 and DQA2 gene reflects the importance of this locus for future selection programs.     

中华菊头蝠MHCⅡ-DRB 基因遗传多态性及进化

采样自云南同一种群的中华菊头蝠共16 个个体,用于DRB 基因的分子进化和多态性研究。利用翼膜组织提取DNA 基因组,并PCR 克隆测序分析。获得了相差3 bp 的两种不同长度序列类型,A 序列类型263 bp,在研究群体中有15 个等位基因;B 序列类型260 bp,在研究群体中有8 个等位基因。在分析的74 个氨基酸变异位点上检测到12 个正向选择位点。在9 个个体中检测到分布频率最高的等位基因,也有多个等位基因只存在一个个体中。单个个体中最多存在6 个等位基因。遗传多态性分析表明中华菊头蝠DRB 基因具有较高的多态性。中华菊头蝠DRB 基因可能至少存在3 个重复座位。利用已发表的翼手目DRB 第二外显子序列构建的系统进化树表明中华菊头蝠MHCⅡ-DRB 基因处于独立进化支。     

应用SSP-PCR/SSO方法进行中国辽宁汉族人HLA-DRB1基因的遗传多态性研究

对１５９名中国辽宁汉族个体的基因组ＤＮＡ进行分析,共检出４２种等位基因,其中以ＤＲＢ１０９０１２（１２．８％）、０７０１（１０．７％）、１５０１（１０．４％）最为常见,其次为ＤＲＢ１１２０１（７９％）、１２０２（７５％）、１１０１（６６％）、０３０１（５．０％）。并发现辽宁汉族人ＤＲＢ１等位基因频率与白种人间存在明显差异,揭示不同人种有其自己的主要等位基因。同时对本技术在ＨＬＡ－ＤＲＢ１分型应用中的优点进行了讨论     

Description of bovine major histocompatibility complex class IIa haplotypes using parthenogenetic embryo‐derived cells

In this study, we report an approach to characterize individual BoLA haplotypes using cells from parthenogenetic bovine embryos derived from slaughterhouse ovaries. Eight of the 15 parthenogenetic embryos so obtained had not undergone meiotic recombination on the BoLA region and were suitable to describe BoLA haplotypes. Detailed analysis of the BoLA class IIa region identified seven different class IIa haplotypes, including six not previously described and two new alleles of BoLA‐DQA and one BoLA‐DQB. Our method provided reliable sources of homozygous DNA to describe BoLA haplotypes.     

JUNB promotes the survival of Flavopiridol treated human breast cancer cells

Chemotherapy resistance is a major obstacle to achieving durable progression-free-survival in breast cancer patients. Identifying resistance mechanisms is crucial to the development of effective breast cancer therapies. Immediate early genes (IEGs) function in the initial cellular reprogramming response to alterations in the extracellular environment and IEGs have been implicated in cancer cell development and progression. The purpose of this study was to investigate the influence of kinase inhibitors on IEG expression in breast cancer cells. The results demonstrated that Flavopiridol (FP), a CDK9 inhibitor, effectively reduced gene expression. FP treatment, however, consistently produced a delayed induction of JUNB gene expression in multiple breast cancer cell lines. Similar results were obtained with Sorafenib, a multi-kinase inhibitor and U0126, a MEK1 inhibitor. Functional studies revealed that JUNB plays a pro-survival role in kinase inhibitor treated breast cancer cells. These results demonstrate a unique induction of JUNB in response to kinase inhibitor therapies that may be among the earliest events in the progression to treatment resistance.