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PubMedDate not supplied

Pathogenesis of varicocele: experimental study using flow cytometric DNA analysis.

Despite extensive investigation, a cause-and-effect relationship between varicocele and male infertility has not been fully proven. We investigated the effect of a unilateral varicocele on both testes. Spermatogenesis in the rats with a surgically induced left varicocele was evaluated 5 weeks postoperatively by measurement of testicular weight, flow cytometric DNA analysis and evaluation of the mean seminiferous tubular diameter (MSTD). In rats with varicoceles, the testicular weight, the percentage of haploid cells and the MSTD were decreased in both testes in contrast to sham-operated rats. There was a greater decrease on the left side. These results suggest that unilateral varicocele may impair spermatogenesis in both testes, with the impairment being greater on the ipsilateral side.

Animals↗
PubMed1997

BOX-polymerase chain reaction-based DNA analysis of nonserotypeable Streptococcus pneumoniae implicated in outbreaks of conjunctivitis.

Nonserotypeable isolates predominate in epidemic conjunctivitis caused by Streptococcus pneumoniae. Previous evaluations of outbreaks of pneumococcal conjunctivitis have relied on epidemiologic factors and the nontypeability of the isolates to infer that a single clone was involved. In the present study, BOX-polymerase chain reaction DNA analysis was used to characterize nonserotypeable S. pneumoniae isolated by conjunctival culture during a recent conjunctivitis outbreak and to compare these isolates with those from outbreaks described earlier. The recent outbreak was caused by a single pneumococcal clone. Outbreaks in separate parts of the United States in 1980-1981 were all caused by the same clone. Cluster analysis revealed a high degree of genetic relatedness among isolates causing conjunctivitis compared with that among other nonserotypeable S. pneumoniae, with the closest relatedness being found among the 1996 and 1980-1981 conjunctival isolates.

Conjunctivitis↗
PubMed2002

Dogs as alternative intermediate hosts of Taenia solium in Papua (Irian Jaya), Indonesia confirmed by highly specific ELISA and immunoblot using native and recombinant antigens and mitochondrial DNA analysis.

Serology (ELISA and immunoblot) using native glycoproteins, affinity purified glycoproteins, and a recombinant antigen is known to be highly specific to Taenia solium cysticercosis in humans and pigs. These techniques were applied for dogs in the highly endemic area of cysticercosis in Papua (Irian Jaya), Indonesia. Analysis of dog sera by both ELISA and immunoblot revealed 7 of 64 dogs were highly positive. Examination of two sero-positive dogs revealed cysticerci of T. solium in the brain and heart of these dogs. Mitochondrial DNA analysis confirmed that they were the same as T. solium previously confirmed from pigs and biopsies from local people from Irian Jaya. It is suggested that the life cycle of T. solium may be completed not only between humans and pigs but also between humans and dogs.

Animals↗
PubMed2001

[Morphological changes and quantitative DNA analysis of Schwann cells in peripheral nerves after high voltage electrical injury].

OBJECTIVE: To explore the characteristics and the mechanism of the injury of Schwann cells and nerve fibres of the peripheral nerves inflicted by high voltage electricity. METHODS: Rabbits injured by high voltage electricity were employed as the model. Thirty rabbits were randomly divided into control (9) and experimental (21) groups. The dynamic quantitative DNA analysis and the change of the morphology and structure of the sciatic nerve were observed with LM and EM on 0, 3, 7, 10, 14 and 21 postburn days (PBDs), respectively. RESULTS: There was continuous degeneration of the axon and medullary sheath of sciatic nerve tissue and its neogenesis was inhibited. The DNA synthesis in Schwann cell initiated on 3 postburn day (PBD). The distribution of the current was heterogeneous on the cross section of peripheral nerve fibres. CONCLUSION: The obvious features of peripheral nerve injured by high voltage electricity included delayed initiation of the DNA synthesis of Schwann cell and the secondary necrosis and inhibition of neogenesis process of Schwann cell.

Animals↗
PubMed2001

[Advances in immunoserology tests in clinical medicine: autoantibodies, immune complexes and DNA analysis].

I reviewed first the history of detection for autoantibodies and the methods to detect the circulating immune complexes. Then I presented several unusual tests for detecting autoantibodies which I had experienced. They are agglutination tests in agarose gel using the particulate antigens of human tissues or lipid antigens and the mixed agglutination test using the cultured cells. The characteristics of rheumatoid factor(RF) were analyzed by the following methods: 1) solid-phase radioimmunoassay for IgG-RF using the formalinized sheep RBC, 2) mixed agglutination test for IgG-RF on the slide glass smeared with the sensitized sheep RBC, 3) hemolysis in agarose gel for competitive reaction of RF with complement to the IgG hemolysin and 4) ELISA for detecting the complement-fixation of the monoclonal IgM-RF. The IgG-Fc and C3b receptors on the tissue sections of the mammalian aorta were detected by an adhesion of the IgG-sensitized or C3b-bound RBC. DNA-analysis studies using the molecular biology techniques which were done in the Department of Internal Medicine II, Fukushima Medical University were presented. 1) Nucleic acid sequences of the cloned DNA polypeptide fragments in the serum of patients with systemic lupus erythematosus. 2) SSCP analysis for clonality of Vb repertoires in T cell receptors in the patients with rheumatoid arthritis, primary biliary cirrhosis or CREST syndrome. 3) Detection of mRNA of TNFa and Fas ligand in the mononuclear cells(FICL-PCR) in the synovial fluids of RA patients.

Agglutination Tests↗
PubMed1999

DNA analysis of hepatocytes: a novel method for estimating the value of liver tissue vaccine (LTV).

Because of the socio-economic condition of African countries, the traditional types of vaccines should be encouraged. LTV in a preliminary study, proved to be of value in reducing morbidity and mortality in Schistosoma mansoni infection. In the present study and because of the vaccine contains liver tissue homogonate, the present authors analyed the DNA content of the hepatocytes of the vaccinated and control groups of animals to know any effects of the vaccine on the hepatocytes ploidy. The DNA content of vaccinated animals was higher. The results were discussed. It was concluded that for testing S. mansoni vaccine, DNA analysis of heptocytes for detecting proliferative S-phase is an important as a positive immune reaction.

Animals↗
PubMed2003

Phylogenetic diversity of lactic acid bacteria associated with paddy rice silage as determined by 16S ribosomal DNA analysis.

A total of 161 low-G+C-content gram-positive bacteria isolated from whole-crop paddy rice silage were classified and subjected to phenotypic and genetic analyses. Based on morphological and biochemical characters, these presumptive lactic acid bacterium (LAB) isolates were divided into 10 groups that included members of the genera Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, and WEISSELLA: Analysis of the 16S ribosomal DNA (rDNA) was used to confirm the presence of the predominant groups indicated by phenotypic analysis and to determine the phylogenetic affiliation of representative strains. The virtually complete 16S rRNA gene was PCR amplified and sequenced. The sequences from the various LAB isolates showed high degrees of similarity to those of the GenBank reference strains (between 98.7 and 99.8%). Phylogenetic trees based on the 16S rDNA sequence displayed high consistency, with nodes supported by high bootstrap values. With the exception of one species, the genetic data was in agreement with the phenotypic identification. The prevalent LAB, predominantly homofermentative (66%), consisted of Lactobacillus plantarum (24%), Lactococcus lactis (22%), Leuconostoc pseudomesenteroides (20%), Pediococcus acidilactici (11%), Lactobacillus brevis (11%), Enterococcus faecalis (7%), Weissella kimchii (3%), and Pediococcus pentosaceus (2%). The present study, the first to fully document rice-associated LAB, showed a very diverse community of LAB with a relatively high number of species involved in the fermentation process of paddy rice silage. The comprehensive 16S rDNA-based approach to describing LAB community structure was valuable in revealing the large diversity of bacteria inhabiting paddy rice silage and enabling the future design of appropriate inoculants aimed at improving its fermentation quality.

DNA, Ribosomal↗
PubMed1988

Flow cytometric DNA analysis of colon adenocarcinomas: a comparative study of preparatory techniques.

DNA quantitation of solid tumors requires either nuclear extraction or disaggregation into single cells for flow cytometric (FCM) analysis. We investigated methods for disaggregating colon carcinomas comparing mechanical and enzymatic techniques in fresh tumors and compared these to extraction of nuclei from paraffin blocks. Mechanical dissociation resulted in the highest yield of tumor cells, especially tumor cells with abnormal DNA content. In addition, mechanically disaggregated intact cells, particularly when dual labeled with cytokeratin, produced optimum DNA histograms. Cytokeratin staining allowed gating on epithelial cell populations and excluded contaminating diploid inflammatory and stromal cells, improving our ability to calculate synthesis phase fractions (SpF). Human colon tumor nuclei extracted from paraffin-embedded tissue resulted in greater variation of DNA histograms and less sensitivity in identifying aneuploid cells compared to mechanically dissociated intact cells from fresh tumors. The quality of the histograms and the ability to identify aneuploid cell populations were improved by modifying the standard "Hedley" technique by additional enzyme incubation. Mechanical dissociation of fresh tumors was the preferred technique for FCM DNA analysis. Nuclear extraction from paraffin blocks resulted in some loss of sensitivity in identifying aneuploid cell lines and decreased ability to measure SpF.

Adenocarcinoma↗
PubMed1990

Prognostic value of pathologic features and DNA analysis in endometrial carcinoma.

In a series of 38 nonmetastasizing and 29 metastasizing endometrial cancers, histologic subtype, aneuploidy, nuclear grade, and depth of myometrial invasion were the features most distinctly separating these two groups of patients. The most important factor was histologic type: papillary, papillary serous, adenosquamous, clear cell, and diffusely infiltrating carcinomas were strongly correlated with metastasis using chi square analysis. Nuclear grade 3 and deep myometrial invasion were similar to each other in sensitivity and predictive value, and both were also strongly correlated with metastatic risk. Aneuploidy, also correlated with metastasis, was similar to nuclear grade 3 and deep myometrial invasion in predictive value but was more sensitive. DNA analysis appeared to be most useful when histologic studies were inconclusive in assigning a high degree of risk. Twice as many aneuploid carcinomas having nuclear grade 2 (intermediate risk) morphology proved to be metastatic. A similar relationship was found with deep myometrial invasion. A quantitative correlation between the DNA index and the risk of metastasis was found; a DNA index above 1.5 was a strong attribute of the metastatic group.

Aneuploidy↗
PubMed1988

X/Y translocation in a family with X-linked ichthyosis, chondrodysplasia punctata, and mental retardation: DNA analysis reveals deletion of the steroid sulphatase gene and translocation of its Y pseudogene.

We describe a family with two male members showing an X/Y translocation (karyotype: 46,Y,der(X)t(X;Y)(p22;q11]. At physical examination both patients showed ichthyosis, mental retardation and dysmorphic features. Chondrodysplasia punctata and short stature were present in one case. Direct DNA analysis, using a steroid sulphatase cDNA probe, was performed in one patient, his mother and sister, both carriers of the translocation. We found that the translocated region of the Y chromosome includes the steroid sulphatase pseudogene. These results suggest that in our patients the X/Y translocation may be derived from a recombinational event between homologous regions located on the short arm of the X chromosome and the long arm of the Y chromosome. Clinical and molecular studies on the present family add further information for the construction of a tentative physical map of the distal Xp.

Child↗
PubMed1997

Treatment of minute medullary thyroid carcinoma in multiple endocrine neoplasia 2A families first diagnosed by DNA analysis of RET proto-oncogene mutations: a case report.

Multiple endocrine neoplasia (MEN) 2A is an inherited disease characterized by the development of medullary thyroid carcinoma, pheochromocytoma and hyperparathyroidism. It has recently been shown to be associated with germ-line mutations in the RET proto-oncogene. We describe a 21-year-old man from a MEN 2A family who was found by DNA analysis to be a gene carrier of MEN 2A and then was diagnosed, using a stimulated calcitonin test, as having presymptomatic medullary thyroid carcinoma. His morbidity seems to have been cured by total thyroidectomy as postoperative calcitonin levels after stimulation are normal. It is concluded that direct genetic analysis for mutations in the RET proto-oncogene should be the diagnostic test of choice for identifying family members at risk for MEN 2A.

Adrenal Gland Neoplasms↗
PubMed1992

Carrier detection in X-linked ocular albinism of the Nettleship-Falls type by DNA analysis.

X-linked ocular albinism (XOA) is characterized by anomalies of the eyes and hypopigmentation or absence of pigment in skin, hair and eyes due to a hereditary inborn error of metabolism affecting the pigment cells. The gene of XOA of the Nettleship-Falls type (OA1) has been mapped to Xp22.3, and several closely linked RFLP loci have been identified. Linkage analysis and deletion mapping have established the marker gene order Xpter-STS-DX237-(OA1,DXS143,DXS85)-DXS1 6-DXS43-Xcen. Although the position of OA1 has yet not been fully resolved, we report on the first carrier detections in OXA of the Nettleship-Falls type by DNA analysis using markers which unquestionably flank OA1.

Albinism, Ocular↗
PubMed2003

Pulmonary lymphangioleiomyomatosis: a case report with immunohistochemical details and DNA analysis.

A 47-year-old woman is presented with pulmonary lymphangioleiomyomatosis (PLAM) involving the bilateral lung and slight pulmonary function abnormality. Computed tomography scan showed bilateral microcyst formation in the lung. Histologically, proliferating spindle shaped cells with centrilobular emphysema were main findings. Immunohistochemically, these proliferating spindle shaped cells were positive for alpha-smooth muscle actin, desmin, vimentin, HMB45, estrogen receptor and progesterone receptor, but negative for S-100, cytokeratin. Single strand conformation polymorphism (SSCP) and DNA analysis for tuberous sclerosis 1 and 2 showed no significant abnormality.

Actins↗
PubMed2001

Random amplified polymorphic DNA analysis of Acinetobacter species isolated from worn contact lenses.

The purpose of this study was to explore the genotypes of Acinetobacter species and to compare the genotypes and phenotypes of the Acinetobacter isolated from contact lens wearers. Nineteen strains were used in the study, 13 were isolated from patients experiencing an adverse response event, and six strains were isolated from asymptomatic contact lens wearers. Random amplified polymorphic DNA and phenotypic analyses were carried out using commercially available kits. Random amplified polymorphic DNA analysis showed a higher discrimination power compared with the phenotypic analysis. The test strains were loosely clustered into six genotypic groups with no definite relation to any clinical events. These results indicate that many genotypes of Acinetobacter can cause adverse responses, and the initial source of the microorganisms rather than their clinical sequelae may determine classification.

Acinetobacter↗
PubMed2003

DNA analysis servers: plot.it, bend.it, model.it and IS.

The WWW servers at http://www.icgeb.trieste.it/dna/ are dedicated to the analysis of user-submitted DNA sequences; plot.it creates parametric plots of 45 physicochemical, as well as statistical, parameters; bend.it calculates DNA curvature according to various methods. Both programs provide 1D as well as 2D plots that allow localisation of peculiar segments within the query. The server model.it creates 3D models of canonical or bent DNA starting from sequence data and presents the results in the form of a standard PDB file, directly viewable on the user's PC using any molecule manipulation program. The recently established introns server allows statistical evaluation of introns in various taxonomic groups and the comparison of taxonomic groups in terms of length, base composition, intron type etc. The options include the analysis of splice sites and a probability test for exon-shuffling.

Computer Graphics↗