Human autoantibodies to poly(adenosine diphosphate-ribose) polymerase.

The chromatin-bound enzyme poly(ADP-ribose) polymerase (ADPRP) is strongly stimulated by DNA with single- or double-stranded breaks, and transfers the ADP-ribose moiety of NAD to nuclear proteins. The activation of ADPRP is important for DNA repair and replication, and also has been postulated to play a role in the pathogenesis of lymphocyte dysfunction associated with chronic inflammatory diseases, and inborn errors of nucleoside metabolism. We have detected high titers of IgG autoantibodies to the ADPRP protein in six patients with rheumatic complaints. No other autoantibodies were detected in any of the six sera. The specificity of the anti-enzyme antibodies was established by (a) immunoprecipitation of ADPRP activity, (b) immunoprecipitation and immunoblotting of both the native 116-kD enzyme and its proteolytic digestion products. ADPRP was purified from human thymus and calf thymus. The autoantibodies reacted equivalently with both enzymes. The anti-ADPRP antibodies had a distinctive immunofluorescent pattern with HEp-2 cells, reacting intensely with nucleoli and metaphase chromosomes, and diffusely with the nucleus. Autoantibodies to ADPRP have not been described previously. The presence of a specific immune response against an enzyme that has been associated with various immunodeficiency syndromes raises intriguing possibilities concerning the relationship between DNA damage, immunodeficiency, and autoimmunity.     

Punch technique--an alternative approach to the repair of pierced earlobe deformities

The punch technique of repairing pierced earlobe deformities is a practical and simplified alternative to the usual method of repair. The procedure can easily be performed in any dermatology office since the instruments required are very basic. The technique is presented in a series of illustrations, and the results are cosmetically acceptable.     

Production and characterization of genetically engineered antibody molecules

Expression of antibody heavy- and light-chain genes by transfection permits the production of monoclonal antibodies with improved biological and antigen-binding properties. The immunoglobulin genes are placed in vectors containing a gene for encoding a protein that provides a biochemically selectable function in eukaryotic cells; these vectors are transfected into myeloma and hybridoma cells. Selection of drug-resistant cells permits the efficient isolation of the rare cells that express the transfected DNA. By placing heavy and light chains on plasmids with different selectable markers, one can deliver heavy- and light-chain genes simultaneously to the same cell. The transfected immunoglobulin genes are efficiently expressed and the proteins produced are a faithful mirror of the genes that were introduced. Using the standard techniques of genetic engineering and gene transfection, we can now produce antibodies of widely varying structures, including chimeric antibodies with segments derived from different species. These antibodies provide useful reagents to study structure-function relationships within the antibody molecule. Ultimately it will be possible to produce a new generation of antibody molecules with improved antigen-binding properties and effector functions.     

In vitro synthesis of antimycobacterial antibodies with different specificities in various tissues of leprosy patients

For the detection of the synthesis in vitro of anti-Mycobacterium leprae antibodies in various tissues of leprosy patients, biopsy specimens of skin lesions, nasal mucosa, larynx, lymph nodes, and bone marrow were cultured in a medium containing 14C-labeled lysine and isoleucine. The culture fluids were analyzed by crossed immunoelectrophoresis with intermediate gel and autoradiography. The results show that synthesis of anti-M. leprae antibodies occurs at the investigated sites of leprosy patients and that the specificities of the synthesized antibodies differ between sites in individual patients. It is conceivable that these antibodies play a role in the local defense against M. leprae.     

Effect of exercise on repetitive nerve stimulation studies: new appraisal of an old technique.

Repetitive nerve stimulation (RNS) is a simple and rapid method for evaluation of neuromuscular transmission defects. Although the effect of exercise in conjunction with RNS is well recognized, it has not been standardized in actual patient and control groups. In a prospective study over a period of 1 year, the authors evaluated the effect of exercise in conjunction with RNS in comparison with conventional 3-Hz RNS at rest in the clinical setting. Fifty-four patients who were referred for possible neuromuscular transmission disorders, in addition to 35 healthy control subjects, were studied. Amplitude and area decremental responses with RNS at rest and after 20 seconds of maximal exercise at 1-minute intervals up to 3 minutes were evaluated. The use of RNS with exercise resulted in additional diagnostic yield of up to 36.4% compared with conventional 3-Hz RNS at rest. The standardized use of exercise with RNS is advocated for increasing its diagnostic yield in the neurophysiologic laboratory.     

菠菜乙醇酸氧化酶编码区cDNA在酵母表达载体中的克隆

目的：克隆菠菜乙醇酸氧化酶cDNA并构建其酵母表达载体。方法：从新鲜菠菜叶子中提取总RNA，用RT－PCR法扩增菠菜乙醇酸氧化酶编码区cDNA并插入pMD-T载体中进行序列测定；将此cDNA构建入P.Pastoris酵母表达体系中的pPIC3.5k上的SnaBI/NotI位点，进行PCR筛选和限制性酶切鉴定。结果：测序表明获得的基因为乙醇酸氧化酶cDNA序列，与国外文献报道的序列相比有约98％的同源性；重组质粒的SnaBI/NotI双酶切证明构建正确。结论：菠菜乙醇酸氧化酶cDNA克隆及重组质粒pPIC3.5K-GO的获得，为酵母表达菠菜乙醇酸氧化酶提供了前题条件。     

宫颈上皮内瘤变108例临床分析

目的通过总结108例宫颈上皮内瘤变（CIN）的临床与预后，探讨分析CIN的诊断与治疗。方法归纳总结共108例各级CIN的临床特点及治疗方法，术前肉眼多点活检及阴道镜下活检与术后病理诊断的符合率，分析其预后。结果所有病人均有性生活史。除45／108（417％）无症状外，63／108（58．3％）有分泌物增多，或同房出血或阴道不规则流血。术前阴道镜下活检与术后病理诊断的符合率高于肉眼多．最活检符合率，但统计学差异无显著性，（P〉0．05）。根据病人要求、CIN级别及病变部位、范围采取不同的手术方式，其中14例随访超过5a，全部病人无1例复发，3、5a存活率均为100％。结论临床上对有性生活史的生育年龄妇女要警惕CIN，肉眼多点活检或阴道镜下活检可作为确诊手段，对不同的病人，依据病灶的性质，采用不同的手术方式，可避免复发，获100％的治愈率。     

Nucleotide polymorphism in colicin E1 and Ia plasmids from natural isolates of Escherichia coli.

We examined DNA sequence polymorphism for the colicin gene clusters of seven ColE1 and six ColIa plasmids obtained from natural isolates of Escherichia coli. These gene clusters harbor levels of nucleotide diversity ranging from 0.006 (ColIa) to 0.054 (ColE1). This level of diversity is similar to that observed for chromosomally encoded E. coli genes. However, the variance associated with these estimates is severalfold higher for the plasmid-encoded genes. This increased variance may be due to the differing plasmid population sizes. The pattern of colicin gene cluster polymorphism suggests that the two colicins are evolving in different fashions. ColE1 accumulates polymorphism at an elevated rate in the central domain of the colicin protein, while ColIa polymorphism is distributed evenly along the gene cluster. Comparison of the patterns of divergence between colicin and related proteins of ColIa and Ib and patterns of polymorphism within ColIa suggest that this gene cluster is not evolving in a neutral fashion. These data lend support to the hypothesis that colicin gene clusters may evolve under the influence of diversifying selection.