Ancestral origins of the Machado-Joseph disease mutation: a worldwide haplotype study

Machado-Joseph disease (MJD) is an autosomal dominant neurodegenerative disorder originally described in families of Portuguese-Azorean ancestry. The cloning of the MJD1 gene allowed identification of the disease in many other populations, and MJD is now known to be the most common cause of dominant spinocerebellar ataxia. The hypothesis that its present world distribution could result from the spread of an original founder mutation has been raised, both at historical and molecular levels. In the present study, we tested this hypothesis by linkage-disequilibrium analysis of tightly linked polymorphisms and by haplotype comparison, in 249 families from different countries. We typed five microsatellite markers surrounding the MJD1 locus (D14S1015, D14S995, D14S973, D14S1016, and D14S977), and three intragenic single-base-pair polymorphisms (A(669)TG/G(669)TG, C(987)GG/G(987)GG, and TAA(1118)/TAC(1118)). The results show two different haplotypes, specific to the island of origin, in families of Azorean extraction. In families from mainland Portugal, both Azorean haplotypes can be found. The majority of the non-Portuguese families also share the same intragenic haplotype seen in the families coming from the island of Flores, but at least three other haplotypes were seen. These findings suggest two introductions of the mutation into the Portuguese population. Worldwide, the sharing of one intragenic haplotype by the majority of the families studied implies a founder mutation in MJD.     

Tyr-716 in the platelet-derived growth factor beta-receptor kinase insert is involved in GRB2 binding and Ras activation.

Ligand stimulation of the platelet-derived growth factor (PDGF) beta-receptor leads to activation of its intrinsic tyrosine kinase and autophosphorylation of the intracellular part of the receptor. The autophosphorylated tyrosine residues mediate interactions with downstream signal transduction molecules and thereby initiate different signalling pathways. A pathway leading to activation of the GTP-binding protein Ras involves the adaptor molecule GRB2. Here we show that Tyr-716, a novel autophosphorylation site in the PDGF beta-receptor kinase insert, mediates direct binding of GRB2 in vitro and in vivo. In a panel of mutant PDGF beta-receptors, in which Tyr-716 and the previously known autophosphorylation sites were individually mutated, only PDGFR beta Y716F failed to bind GRB2. Furthermore, a synthetic phosphorylated peptide containing Tyr-716 bound GRB2, and this peptide specifically interrupted the interaction between GRB2 and the wild-type receptor. In addition, the Y716(P) peptide significantly decreased the amount of GTP bound to Ras in response to PDGF in permeabilized fibroblasts as well as in porcine aortic endothelial cells expressing transfected PDGF beta-receptors. The mutant PDGFR beta Y716F still mediated activation of mitogen-activated protein kinases and an increased DNA synthesis in response to PDGF, indicating that multiple signal transduction pathways transduce mitogenic signals from the activated PDGF beta-receptor.     

Determination of free concentration of sameridine in blood plasma by ultrafiltration and coupled-column liquid chromatography

Sameridine is a new candidate drug with both local anaesthetic and analgesic properties. The free concentration of sameridine in blood plasma was determined by coupled-column liquid chromatography. Following adjustment of the pH and the temperature of the plasma samples, the free fraction was prepared by ultrafiltration. The coupled-column liquid chromatographic system consisted of a reversed-phase column, a cation-exchange extraction column and a cation-exchange analytical column. Sameridine was detected by UV determination at 205 nm and the system showed high selectivity. The limit of quantification was 1 nM and the within-day precision was 4.6% (R.S.D., n=10).     

Effect of microwave radiation on permeability of liposomes. Evidence against non-thermal leakage

The effect of 2.45 GHz microwave radiation on the permeability of unilamellar phosphatidylcholine liposomes has been studied. Leakage of 5(6)-carboxyfluorescein from the liposomes was measured using spectrofluorimetry after exposure to either microwaves or thermal heating for 5–20 min intervals. The exposure temperature, 37.6 ± 0.5°C, was well above the phase transition temperature of the lipid membrane. The microwave exposure did not result in any non-thermal increase in permeability above that produced by thermal heating. This study refutes the results reported by Saalman et al. [1] in which an increased liposome permeability due to microwave exposure was reported. The refined analysis in the present study shows that this increased liposome permeability was not a non-thermal microwave effect.     

High-performance liquid chromatography–tandem electrospray mass spectrometry for the determination of lidocaine and its metabolites in human plasma and urine

A sensitive, selective and accurate high-performance liquid chromatographic–tandem mass spectrometric assay was developed and validated for the determination of lidocaine and its metabolites 2,6-dimethylaniline (2,6-xylidine), monoethylglycinexylidide and glycinexylidide in human plasma and urine. A simple sample preparation technique was used for plasma samples. The plasma samples were ultrafiltered after acidification with phosphoric acid and the ultrafiltrate was directly injected into the LC system. For urine samples, solid-phase extraction discs (C₁₈) were used as sample preparation. The limit of quantification (LOQ) was improved by at least 10 times compared to the methods described in the literature. The LOQ was in the range 1.6–5 nmol/l for the studied compounds in plasma samples.     

湖南苗族、侗族5项人类群体遗传学特征的调查

对湖南苗族、侗族共648例的5项人类群体遗传学特征(利眼、足趾长、起步类型、耳耵聍、指甲型)进行了调查。结果显示:1)两个民族的多数人属于右利眼、拇趾长于二趾、左起步、干型耵聍和长形指甲;2)两个民族的左利眼出现率和拇趾长于二趾的出现率高于国内多数人群;3)足趾长出现率存在明显的民族间差异;苗族利眼和方形指甲出现率均表现出明显的民族内性别差异;4)5项特征间的相关关系不明显。     

蛇菰的化学成分研究

从湖北恩施地区产蛇菰(Balanophora japonicaMakino)中得到4个成分,分别鉴定为蛇麻脂醇乙酯(lupeolacetate,1)、β-香树脂醇乙酯(-βamyrin acetate,2)、没食子酸(gallic acid,3)和β-谷甾醇(-βsitosterol,4),其中化合物1、2和3为首次从该植物中得到。     

一个人类新基因ZNF322的研究初报

利用果蝇心脏发育基因zfh-1的cDNA序列和计算机克隆方法,在9号染色体上发现了一个新的人类同源基因,经国际基因全程委员会批准,命名为ZNF322。该基因总长度约为2.8kb,为一个连续的序列,编码一个402个氨基酸的蛋白质,与小鼠zfp-25编码的蛋白质最相似（同源度为51.4%）。该基因在人体肾、脑、结肠、胚胎心脏及黑色素瘤等多种组织中广泛表达,其功能有等深入研究。     

遮荫对半夏叶片光合色素与保护酶活性的影响

对半夏进行不同程度遮荫处理,研究了其生长过程中叶组织的光合色素含量、叶绿素a/b值、保护酶体系(SOD、POD、CAT)活性和丙二醛(MDA)含量的变化及其生理特性.结果表明:不同处理的半夏在生长过程中,光合色素的含量呈先升后降的趋势;与全光照相比,遮荫处理的叶绿素含量较高,叶绿素a/b值较低,类胡萝卜素含量在生长发育中早期较低而后期较高;SOD、POD、CAT活性呈由低到高再降低的趋势,MDA含量持续升高;遮荫处理的SOD、POD活性和MDA含量均低于全光照处理,CAT活性则表现为先高于后低于全光照处理.适度的遮荫可以提高半夏叶绿素的含量,降低膜系统受到的伤害,改善半夏的倒苗状况,为半夏叶片的生长提供较好的环境.