Microsatellite instability and loss of heterozygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis

OBJECTIVES: The purpose of this study was to compare D-speed film, E-speed film, and the Soredex Digora system with respect to the detection of periradicular pathosis. STUDY DESIGN: Radiographic images of 100 cadaver jaws were made with E-speed film, D-speed film, and the Soredex Digora. Each set of 100 images was interpreted by four observers, with 30 days separating each of three viewing sessions from the next. The presence or absence of pathologic (inflammatory) periradicular bone resorption was determined by histologic examination of the samples. The observer performance was compared with the true histologic findings and evaluated with receiver operating characteristic and corrected receiver operating characteristic analysis. RESULTS: No statistically significant differences were found in diagnostic performance among the three radiographic techniques. In addition, no imaging technique was a good indicator of pathosis as determined by histologic analysis. CONCLUSION: Under the conditions of this study, it was determined that D-speed film, E-speed film, and the Soredex Digora were equivalent diagnostic imaging modalities with regard to the detection of pathologic periradicular bone resorption. No technique predictably indicated inflammatory resorption.     

Quantitative trait loci for murine growth

Body size is an archetypal quantitative trait with variation due to the segregation of many gene loci, each of relatively minor effect, and the environment. We examine the effects of quantitative trait loci (QTLs) on age-specific body weights and growth in the F2 intercross of the LG/J and SM/J strains of inbred mice. Weekly weights (1-10 wk) and 75 microsatellite genotypes were obtained for 535 mice. Interval mapping was used to locate and measure the genotypic effects of QTLs on body weight and growth. QTL effects were detected on 16 of the 19 autosomes with several chromosomes carrying more than one QTL. The number of QTLs for age-specific weights varied from seven at 1 week to 17 at 10 wk. The QTLs were each of relatively minor, subequal effect. QTLs affecting early and late growth were generally distinct, mapping to different chromosomal locations indicating separate genetic and physiological systems for early and later murine growth.     

Microsatellite variability and genetic distances

We analyze the within- and between-population dynamics of the distribution of the number of repeats at multiple microsatellite DNA loci subject to stepwise mutation. Analytical expressions for moments up to the fourth order within a locus and the variance of between-locus variance at mutation-drift equilibrium have been obtained. These statistics may be used to test the appropriateness of the one-step mutation model and to detect between-locus variation in the mutation rate. Published data are compatible with the one-step mutation model, although they do not reject the two-step model. Using both multinomial sampling and diffusion approximations for the analysis of the genetic distance introduced by Goldstein et al. [Goldstein, D. B., Linares, A. R., Cavalli-Sforza, L. L. & Feldman, M. W. (1995) Proc. Natl. Acad. Sci. USA 92, 6723-6727], we show that this distance follows a chi 2 distribution with degrees of freedom equal to the number of loci when there is no variation in mutation rates among the loci. In the presence of such variation, the variance of the distance is obtained. We conclude that the number of microsatellite loci required for the construction of phylogenetic trees with reliable branch lengths may be several hundred. Also, mutations that change repeat scores by several units, even though extremely rare, may dramatically influence estimates of population parameters.     

Microsatellite instability in colorectal adenomas

BACKGROUND & AIMS: Microsatellite instability in apparently sporadic, predominantly right-sided colon cancers seems to be the result of an acquired, rather than germline, genetic change that impairs mismatch repair. The timing of this change with respect to the adenomacarcinoma sequence has not been determined. The aim of this study was to evaluate colonic adenomatous polyps for microsatellite instability to determine whether instability reflects an early genetic change in colonic neoplasia. METHODS: Ninety-three sporadic colonic adenomas (44 right-sided and 49 left-sided) from 48 individuals were evaluated for microsatellite instability with a set of 10 polymerase chain reaction primer sets. RESULTS: Eighty percent of adenomatous polyps showed no instability. Ninety-eight percent showed instability with <30% of primer sets. Aside from one right-sided adenoma with 78% instability, there was no level of instability with a higher proportion of right-sided than left-sided adenomas. CONCLUSIONS: Colonic adenomas show far less microsatellite instability than carcinomas, and the marked right-sided predominance of instability observed in colon cancers was not observed. Instability is usually not an early event in the development of colonic neoplasia. A distinct pathway to sporadic colorectal cancer initiated by mismatch repair deficiency, although not excluded, is not suggested by these data.     

Microsatellite hybrid capture technique for simultaneous isolation of various STR markers

The microsatellite hybrid capture technique was designed to enrich simultaneously for various microsatellite repeats from a genomic clone. It is illustrated in this report that different repeat motifs, including polymorphic ones, can be efficiently isolated in a single experiment. In principle, this technique can be applied to any type of genomic clones to facilitate the isolation of informative markers for fine mapping of subchromosomal regions of interest, and for linkage and association studies of candidate genes for which no polymorphic markers are available yet.     

Identification of rat susceptibility loci for adjuvant-oil-induced arthritis

One intradermal injection of incomplete Freund's adjuvant-oil induces a T cell-mediated inflammatory joint disease in DA rats. Susceptibility genes for oil-induced arthritis (OIA) are located both within and outside the major histocompatibility complex (MHC, Oia1). We have searched for disease-linked non-MHC loci in an F2 intercross between DA rats and MHC-identical but arthritis-resistant LEW.1AV1 rats. A genome-wide scan with microsatellite markers revealed two major chromosome regions that control disease incidence and severity: Oia2 on chromosome 4 (P = 4 x 10(-13)) and Oia3 on chromosome 10 (P = 1 x 10(-6)). All animals homozygous for DA alleles at both loci developed severe arthritis, whereas all those homozygous for LEW.1AV1 alleles were resistant. These results have general implications for situations where nonspecific activation of the immune system (e.g., incomplete Freund's adjuvant-oil) causes inflammation and disease, either alone or in conjunction with specific antigens. They may also provide clues to the etiology of inflammatory diseases in humans, including rheumatoid arthritis.     

Editorial: Multiple loci for human globin genes

It is often suggested that when z-scores are used for linkage estimation, a maximum z-score of 3 (or maximum antilod score, M = 1000) is good evidence for the presence of linkage. It is pointed out that a better formula, applicable if nothing more is known about the loci concerned, would be that the probability that linkage is present is roughly 1--4/Msigma, whereas sigma is the standard error, i.e. square root(2-3 X downward curvature of z curve at peak). A consideration of typical cases suggests that a value M = 1000 would imply something like a 90% chance of linkage. It would seem simpler to use the formula, probability of linkage approximately to 1--H/20, where H is the average height of the antilod curve.     

Microsatellite instability in human solid tumors

BACKGROUND: Microsatellite instability (MIN) has been identified in a wide variety of human tumors, both familial and sporadic. In this study the authors attempted to correlate MIN with other biologic parameters to assess the significance of MIN in cancer. METHODS: The current literature up to May 1997 was reviewed critically. Comparative assessment and analysis of published MIN data in human solid tumors was addressed. RESULTS: Based on review of the current medical literature, the following conclusions can be drawn: 1) MIN associated with inherited mutations of the DNA mismatch repair genes (predominantly hMSH2/hMLH1) appears to characterize only the hereditary nonpolyposis colon carcinoma (HNPCC)/Muir-Torre family cancer syndrome category, and a subset of young colorectal carcinoma patients. Constitutional hMSH2/hMLH1 mutations rarely are reported in other than colon MIN+ tumor types; 2) MIN in non-HNPCC tumors generally is not associated with somatic mutations in the mismatch DNA repair genes most commonly involved in HNPCC; 3) loci of individual chromosomes containing microsatellite markers demonstrating high MIN frequency may be linked to particular tumor types (tumor specific MIN hot spots); 4) the gel banding patterns of MIN observed in noncolon tumors differ significantly from those reported previously in HNPCC; 5) although overall no association between MIN and histopathology is observed in the literature, a statistically higher MIN frequency has been noted in certain tumor subtypes; and 6) MIN in tumors can be associated with early or late stages of tumor progression, and also has been found in nontumor tissues. CONCLUSIONS: Molecular diagnosis using MIN analysis has been documented in at least two types of tumors (HNPCC and sporadic bladder carcinoma), suggesting a potential role of MIN in the diagnosis and/or prognosis of other solid human tumors as well.     

Microsatellite analysis of Duchenne muscular dystrophy

Duchenne muscular dystrophy is X-linked recessive neuromuscular disorders caused by mutations in the dystrophin gene. Prenatal diagnosis and carrier detection are usually performed using diallelic RFLP-markers which are not always informative. Now 30 of microsatellite marker have reported, these microsatellite polymorphism can easily be amplified using PCR technique. If mutations are known to localize in this region of the dystrophin gene or if routine RFLP-analysis is uninformative, the analysis of microsatellite markers is the preferable technique in prenatal diagnosis and carrier detection.     

Microsatellite instability in European hepatocellular carcinoma

A radioimmunoassay has been established to measure urinary aquaporin-2 excretion (u-AQP2). To elucidate how u-AQP2 changes when endogenous vasopressin is increased independently of plasma osmolality, we estimated u-AQP2 during general anesthesia for surgery. We collected urine and blood samples from 50 patients before and 90 and 180 min after anesthetic induction. Plasma (29.1+/-12.6 pg/mL) and urinary (565.1+/-207.0 ng/gCr) vasopressin levels were markedly increased after anesthetic induction. Although no significant alteration of plasma osmolality or serum sodium concentration was observed during 180 min, u-AQP2 was significantly increased (preinduction 224.5+/-24.2 fmol/ mgCr; 90 min 243.3+/-31.8; 180 min 331.4+/-45.9), paralleling an increase of plasma and urinary vasopressin. The plasma vasopressin concentration after anesthetic induction was far in excess of that expected based on plasma osmolality. Individual plasma and urinary vasopressin concentrations correlated significantly with u-AQP2. At 180 min after anesthesia, plasma osmolality did not change, but urine osmolality decreased despite increased u-AQP2, and a preanesthetic positive correlation between urine osmolality and u-AQP2 disappeared. Thus, although u-AQP2 correlates with increased intrinsic vasopressin levels, the increase in u-AQP2 did not directly contribute to urine concentration. Apparently, an escape from the physiologic effects of high vasopressin level occurs during anesthesia via a mechanism independent of aquaporin-2. We conclude that the anesthetic would interfere with the urinary concentrating capacity at the level of AQP2-action. IMPLICATIONS: The excessive increase of intrinsic vasopressin exactly augmented urinary aquaporin-2 excretion, resulting in urine concentration; however, anesthesia seemed to modify this process possibly by interfering with the aquaporin-2 action.     

Microsatellite instability in human atherosclerotic plaques

The aetiopathology of atherosclerosis remains obscure. Although histologically the accumulation of lipids and the proliferation of the smooth muscle cells represents the main feature of the disease, little is known as regards the molecular alterations associated with the atherosclerotic lesions. In the present study we investigated whether an elevated mutational rate is detectable in human atheromatous plaques. Thirty specimens were assessed for microsatellite instability (MI) by 7 microsatellite markers and MI, in at least one marker, was apparent in 6 (20%) cases. Our data suggest that decreased fidelity in DNA replication and repair may be associated with the development of the disease.     

Microsatellite instability analysis: a multicenter study for reliability and quality control

The molecular biology section of the Hereditary Non-Polyposis Colorectal Cancer study group-Germany, instituted a multicenter study to test the reliability and quality of microsatellite instability (MSI) analysis. Eight laboratories compared MSI analyses performed on 10 matched pairs of normal and tumor DNA from patients with colorectal carcinomas. A variety of techniques were applied to the detection of microsatellite changes: (a) silver and ethidium bromide staining of polyacrylamide gels; (b) radioactive labeling; and (c) automated fluorescence detection. The identification of highly unstable tumors and tumors without MSI was achieved in high concordance. However, the interpretation of the band patterns resulted in divergent classifications at several microsatellite marker loci for a large fraction of this tumor/normal panel. The data on more than 30 primers per case suggest that the enlargement of the microsatellite panel to more than 10 loci does not influence the results. In this study, cases with MSI in less than 10% of loci were classified as microsatellite stable, whereas MSI was diagnosed in cases with more than 40% of all markers unstable. We propose that a panel of five microsatellite loci consisting of repeats with different lengths should be analyzed in an initial analysis. When less than two marker loci display shifts in the microsatellite bands from tumor DNA, the panel should be enlarged to include an additional set of five marker loci. The number of marker loci analyzed as well as the number of unstable marker loci found should always be identified. These criteria should result in reports of MSI that are more comparable between studies.     

Microsatellite mutations in spontaneously aborted embryos

OBJECTIVE: To evaluate the incidence of microsatellite instability in spontaneously aborted embryos. DESIGN: Retrospective study. SETTING: Laboratory of Clinical Virology and Department of Obstetrics and Gynecology, University Hospital, Medical School, University of Crete. PATIENT(S): Thirty-five women in whom spontaneous abortions occurred between the 6th and 20th weeks of pregnancy. INTERVENTION(S): Thirty-five aborted embryonic tissues were analyzed with seven microsatellite markers, and their haplotypes were compared with the corresponding pattern of their parents. MAIN OUTCOME MEASURE(S): Microsatellite DNA. RESULT(S): Microsatellite instability was observed in 8 of 35 cases (23%). In 7 of 8 positive cases, microsatellite instability was restricted to one of the seven microsatellite markers, whereas in one case, three microsatellite markers were affected by instability. A statistically significant association was found between microsatellite instability and a previous normal childbirth. CONCLUSION(S): Genetic instability is a detectable phenomenon in spontaneous abortions, representing a significant increase in the mutational rate of the embryo and providing evidence for a mechanism associated with the phenomenon of spontaneous abortion. We conclude that this elevated mutational rate affects active genomic sequences that play a critical role in the viability of the embryo, leading to cell death and abortion.     

Microsatellite variation in the Australian dingo

Shoulder arthroplasty (SA) is commonly performed in patients with rheumatoid arthritis (RA) who have been treated with long-term immunosuppressive medication. RA is associated with an increased risk of neoplasms of the immune system. A case of non-Hodgkin's lymphoma as an unexpected diagnosis after the routine pathologic examination of the soft tissues after SA was detected in a 54-year-old woman with long-standing RA and prolonged immunosuppressive therapy. Although this case does not support the cost-effectiveness of routine specimen evaluation during SA, we suggest that histological analysis of the surgical tissues is appropriate and should be performed in all patients who have been treated with prolonged immunosuppressive medication, especially RA patients as well as patients who have suspicious surgical findings.     

Identifying loci required for follicular patterning using directed mosaics

We have developed a 'directed mosaic' system in Drosophila by using the GAL4 system to control the expression of the yeast recombinase, FLP, in a spatial and temporal fashion. By directing FLP expression, we show that it is possible to efficiently and specifically target loss-of-function studies for vital loci to the developmental pathway of interest. A simple F1 adult phenotypic screen demonstrated that most adult tissues can be analyzed with this approach. Using GAL4 lines expressed during oogenesis, we have refined the system to examine the roles of vital loci in the development of the follicular epithelium. We have identified essential genes involved in egg chamber organization, cell migration and cell shape. Further, we have used this technique to gain insights into the role of the Drosophila EGF receptor pathway in establishing the egg axes. Finally, using different UAS-FLP, GAL4 and existing FRT lines, we have built stocks that permit the analysis of approximately 95% of the genome in follicular mosaics.