Evolutionary aspects of “chloroplast-like” trnN and trnH expression in higher-plant mitochondria

Two identical “chloroplast-like” tRNA^Asn genes, trnN1 and trnN2, have been identified in the potato (Solanum tuberosum) mitochondrial genome. The flanking sequences of trnN1 are unrelated to the corresponding authentic potato chloroplast regions, whilst those of trnN2 are very similar to the chloroplast sequences. The trnN1 copy is present in the mitochondrial genome of various plants whereas the second copy, trnN2, is absent from all the other plant genomes studied so far. Interestingly, both trnN copies are expressed in potato mitochondria. Sequences flanking the chloroplast-like tRNA^His gene (trnH), present as a single copy in the potato mitochondrial DNA, are unrelated to the corresponding chloroplast sequences, whereas chloroplast-derived sequences have been maintained in the vicinity of the maize chloroplast-like mitochondrial trnH gene. However, both the potato and the maize trnH are expressed in mitochondria. Received: 10 April / 1 August 1997     

Study on the Relationship between Cytogenetics and Phenotypic Effect in Turner＇s Syndrome

ＳｔｕｄｙｏｎｔｈｅＲｅｌａｔｉｏｎｓｈｉｐｂｅｔｗｅｅｎＣｙｔｏｇｅｎｅｔｉｃｓａｎｄＰｈｅｎｏｔｙｐｉｃＥｆｆｅｃｔｉｎＴｕｒｎｅｒ＇ｓＳｙｎｄｒｏｍｅＨＵＸｉａｏｆｅｎｇ（胡晓峰）；ＺＨＵＢａｏｓｈｅｎｇ（朱宝生）；ＬＩＮＨａｎｈｕａ（林汉华）...     

人乳头瘤病毒16型DNA转化人胚肺细胞的实验研究

人乳头瘤病毒(HPV)的16,18,31,33型等与宫颈癌的发病有关,其中HPV16与宫颈癌关系密切。为进一步研究HPV16的致癌性,我们用克隆的HPV16 DNA(2μg/10~5细胞)转染体外培养的人胚肺细胞,并进行了细胞存活时间、血清依赖性、着壁依赖性、间接免疫酶检测、HPV16 DNA、同源序列检测、染色体核型等生物学的研究。结果表明,转染细胞存活时间延长、在软琼脂培养基中形成集落、HPV16特异抗原得以表达、HPV16 DNA的同源序列存在于细胞中。表明本实验用HPV16DNA转染的人胚肺细胞具备转化细胞的某些特征,HPV16有使人胚肺细胞转化的作用。     

Genomic DNA fingerprints and phenotypic characteristics of serotype B Haemophilus influenzae isolates from Italy

Three different restriction enzymes (PstI, EcoRI, SspI) were used to analyze the total genomic DNA fingerprints of 52 Haemophilus influenzae type b (Hib) isolates collected between 1982 and 1992 from patients and carriers in central-northern Italy. The same isolates were also characterized by biotyping and antimicrobial agent susceptibility typing. In addition, 13 Hib reference strains from Sweden and the Netherlands were subjected to DNA fingerprinting and compared to Italian isolates.Both genotypic and phenotypic analysis revealed low variability among the Italian study isolates. Most were biotype I and all study isolates but one were susceptible to ampicillin, chloramphenicol, rifampin, third-generation cephalosporins and cotrimoxazole. Among the 52 Italian isolates, 3 distinct DNA patterns were identified, and 88.5% of study strains belonged to the same DNA group. There was sharing of the predominant DNA profile among isolates cultured in different years from different geographical areas and different invasive, respiratory and surface infections. However, another DNA pattern was only found in carrier isolates and in one surface infection isolate.Comparison by DNA fingerprinting showed that the majority of Italian isolates were closely related to most of the analyzed Swedish and Dutch reference strains, previously shown by other techniques to be predominant in those areas. This finding provides additional support for the hypothesis that there may be a dominant European Hib clone.The results show that DNA fingerprinting is a reliable method for Hib characterization and may be a useful additional epidemiological tool for this microorganism.     

Suppression of Gl Arrest and Enhancement of G2 Arrest by Inhibitors of Poly(ADP-ribose) Polymerase: Possible Involvement of Poly(ADP-ribosyl)ation in Cell Cycle Arrest Following γ-Irradiation

Low-dose γ-irradiation of mouse embryonic fibroblast C3D2F1 3T3-a cells caused Gl arrest along with G2 arrest and inhibition of replicative DNA synthesis. When the cells were cultured in the presence of inhibitors of poly(ADP-ribose) polymerase [EC 2.4.2.30], such as 3-aminobenzamide, benzamide and luminol, Gl arrest of C3D2F1 3T3-a cells was suppressed and enhancement of G2 arrest was observed. In contrast, 3-aminobenzoic acid, a non-inhibitory analog of 3-aminobenzamide, did not suppress Gl arrest following γ-irradiation. These results suggest that the poly(ADP-ribosyDation reaction is critical for the pathway of Gl arrest and is also involved in the pathway of G2 arrest.     

Nephron Sparing Surgery for De Novo Renal Cell Carcinoma in an Allograft Kidney: A Case Report

De novo renal cell carcinoma in a renal allograft is rare and has special implications in renal transplant recipients. We describe a patient with a renal allograft who developed a de novo renal cell carcinoma in the functioning renal allograft 258 months after transplantation. The patient underwent enucleation of the tumor because preoperative MRI showed it was well-encapsulated. A DNA banding study showed that the tumor originated from the donor. Indications for conservative renal surgery in renal cell carcinoma have been increasing. Accordingly, 1 option in the treatment of de novo renal cell carcinoma in a functioning renal allograft is enucleation as a method of nephron sparing surgery.     

Unusual presentation and clinical variability in Belgian pedigrees with progressive external ophthalmoplegia and multiple deletions of mitochondrial DNA

We studied 14 patients from three unrelated Belgian pedigrees with a familial mitochondrial disorder and multiple deletions of mitochondrial DNA (mtDNA). In one family with an oculopharyngeal presentation there is a clear autosomal dominant inheritance. Progressive external ophthalmoplegia (PEO), “ragged red fibres” (RRF) and multiple deletions of mtDNA are common to all three families. Therefore a diagnosis of autosomal dominant progressive ophthalmoplegia with multiple deletions of mtDNA (adPEO) was made in one family at least. Our data confirm the previous observations that adPEO is a systemic disorder rather than a pure myopathy. In our pedigrees frequently associated features include axonal peripheral neuropathy, dysphagia, psychiatric illness, and sudden death. Mild ataxia, pes cavus and mitral valve prolapse with associated mitral insufficiency also occur. In some cases onset is atypical with neuropathy, adolescent onset myopathy or psychiatric illness. In such cases the common features of PEO and muscle weakness always complete the clinical phenotype later during the course of the disease. Biochemical studies on mitochondrial fractions prepared from one patient's muscle, revealed no abnormalities of respiratory chain enzyme activities.     

环氧化酶-2基因启动子甲基化及表达与胃黏膜病变的关系

目的 探讨环氧化酶2(COX-2)基因启动子区甲基化水平和蛋白表达与胃黏膜病变的关系,并对其相关的影响因素进行研究.方法 以1201例患有不同胃黏膜病变的高危人群为研究对象,采用免疫组织化学方法榆测COX-2表达,用亚硫酸氢钠-变性高效液相色谱(DHPLC)对COX-2启动子甲基化率进行定量分析,采用13C尿素呼气实验(13C-UBT)对幽门螺旋杆菌(H priori)感染状况进行测定.结果 COX-2甲基化率中位数随胃黏膜病变的加重逐渐升高,在浅表性胃炎和慢性萎缩性胃炎(SG/CAG)、肠上皮化生(IM)及不确定性异型增生和异璎增生(Ind DYS/DYS)病变中分别为10.6%、11.8%、13.8%,各病变组之间差异有统计学意义(X2=8.312,P=0.016).分层分析显示,在H pylori感染阴性病例中,COX-2甲基化率仍随病变加重明显升高,在SG/CAG、IM、Ind DYS/DYS病变中其中位数分别为8.8%、10.6%、14.1%(X2=6.629,P=0.036).进一步分析发现,COX-2甲基化率随着COX-2表达强度的增强而降低,由COX-2弱阳性表达的13.3%降至强阳性表达的7.6%(X2=10.400,P=0.015).结论 COX-2启动子甲基化水平与胃黏膜病变程度及H pylori感染状况密切相关,并与COX-2表达强度呈负相关,说明COX-2启动子区异常甲基化可能在胃黏膜病变的演变过程中起重要作用.     

A nuclear mutant of Chlamydomonas that exhibits increased sensitivity to UV irradiation,reduced recombination of nuclear genes,and altered transmission of chloroplast genes

Summary Meiotic progeny of Chlamydomonas reinhardtii normally receive chloroplast genomes only from the mt ⁺ parent. However, exceptional zygotes, which transmit the chloroplast genomes of both parents or, more rarely, only those of the mt ^- parent, arise at a low frequency. Mutations at the mt ⁺-linked mat-3 locus were found previously to elevate the transmission of chloroplast genomes from the mt-parent, resulting in a much higher than normal frequency of exceptional zygotes. In this paper we demonstrate that an ultraviolet-sensitive nuclear mutation mapping at the uvsE1 locus, which is unlinked to mating type, also promotes chloroplast genome transmission from the mt ^- parent. This mutant, which was previously shown to reduce recombination of nuclear genes in meiosis, acts synergistically which the mat3-3 mutation to produce an extremely high frequency of exceptional zygotes. Through the use of restriction fragment length polymorphisms existing in the chloroplast genomes of C. reinhardtii and the interfertile strain C. smithii, we show that chloroplast DNA fragments from the mt ^- parent normally begin to disappear shortly after zygote formation. However, this process appears to be blocked totally in the absence of wild-type uvsE1 and mat-3 gene products. Our findings are consistent with the hypothesis that both gene products contribute to the mechanism responsible for uniparental inheritance of the chloroplast genome from the mt ⁺ parent.     

赖型钩体flaB2与VR1012中的CpG基序分析

目的：对问号赖型钩端螺旋体（赖型钩体）DNA疫苗[包括内鞭毛蛋白基因（flaB2)和质粒DNA表达载体（VR1012）]的CpG基序（CpG motifs)进行分析，为DNA疫苗免疫机制的阐明和提高DNA疫苗的效能奠定基础。方法：以flaB2与VR1012构建重组DNA的免疫原，对flaB2及VR1012全核苷酸序列进行计算机分析（分类、计数和定位）。结果：CpG的“C”的侧翼为两个嘌呤，“G”的侧翼为两个嘧啶，在flaB2中共3个，分别为GACGCT，GACGTC和GACGCC；在VR1012中共11个，分别为GACGTC1个，GACGCT2个，GACGCC1个，GACGTT1个，GGCGTT2个，GGCGCT2个，GGCGCC1个，AACGCT1个，其中特别重要的TGACGTCA4个和TAACGCCA有1个，位于5'端456－463；509－516；592－599；778－785和486－493；4个TGACGTCA和1个TAACGCCA均位于5'端且相对集中。结论：赖型钩体flaB2与VR1012构成的DNA疫苗含有TGACGTCA等CpG，这些基序又称免疫刺激序列，构成了DNA疫苗中的佐剂。