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[Traditional radiology in the assessment of posttraumatic carpal instability].

INTRODUCTION: Carpal instability is a painful posttraumatic syndrome with early or late loss of the normal alignment of the carpal bones, which can be caused by a variety of injuries, from minor sprain to major fracture-dislocation of the carpal-wrist complex. If the trauma causing instability is a fracture, a severe dislocation or a fracture-dislocation, the radiographic diagnosis is not particularly difficult. The morphologic and dynamic complexity of the carpal region represents, instead, a major obstacle in the radiologic diagnosis of mild or moderate sprains because the morphologic alterations on standard static views are minimal or absent in these conditions. MATERIALS AND METHODS: We reviewed 214 injuries causing posttraumatic carpal instability including both the cases classified by the Data Analysis Center of the Istituto Ortopedico Rizzoli as carpal dislocations and fracture-dislocations from January, 1975, to July, 1996, and the more recent cases directly observed at our Casualty Clinic. In the former cases, we reviewed only the available images, while our patients were examined with comparative standard and under stress or dynamic views. RESULTS: Of 214 lesions causing posttraumatic carpal instability, 43 along the great arch were classified as severe because they were easily detectable on standard films and 171 along the small arch were classified as mild because slight/no abnormalities were detected on standard static views. Only dynamic imaging showed posttraumatic carpal instability demonstrating the integrity of the ligaments and of the carpal hinges, as well as gaps or asymmetry not detected on static views. CONCLUSIONS: We suggest the systematic use of dynamic imaging in the cases where static findings are negative or poor, in the patients with a painful wrist after an apparently minor sprain. Missed or delayed diagnoses are thus reduced, as well as the consequent joint incongruity and/or chronic subluxation which may severely impair these patients.

Carpal Bones↗

Genomic instability in colorectal carcinomas: comparison of different evaluation methods and their biological significance.

In order to demonstrate the relationship between microsatellite instability and other types of genomic instability, a series of 56 sporadic colorectal carcinomas was investigated by flow cytometrical ploidy analysis, oligonucleotide fingerprinting, and microsatellite polymerase chain reaction (PCR). Stabilization of the p53 gene product was analysed by immunohistochemistry and proliferative activity was determined flow cytometrically and by silver staining of nucleolar organizer regions (AgNORs). Of the 56 carcinomas, 11 (19 per cent) exhibited microsatellite instability; 33 of the cases were aneuploid (59 per cent) and 29 (52 per cent) showed alterations of the oligonucleotide fingerprints. There was a significant correlation of microsatellite instability with localization of these tumours proximal to the splenic flexure, diploid DNA content, and less frequent p53 stabilization. A solid growth pattern, mucinous differentiation, and a Crohn's-like lymphoid infiltrate were also characteristic for those tumours. The results demonstrate for the first time a significantly lower proliferative activity in tumours with microsatellite instability. Data obtained from DNA flow cytometry or from oligonucleotide fingerprinting did not correlate with such tumour characteristics. It is proposed that the use of microsatellite PCR facilitates specifically the detection of a group of colorectal cancers which may differ in pathogenesis and perhaps prognosis.

Adult↗

Microsatellite instability in precancerous lesions and adenocarcinomas of the stomach.

BACKGROUND: It is generally known that replication errors (RERs) at microsatellite loci detected in human malignancies reflect a genetic instability that is caused by abnormalities of DNA mismatch repair system and underlie human carcinogenesis. The authors analyzed RERs in precancerous lesions and adenocarcinomas of the stomach to learn when genetic instability occurs in stomach carcinogenesis. In addition, the authors examined genetic instability occurs in stomach carcinogenesis. In addition, the authors examined genetic alterations of the p53 and adenomatous polyposis coli (APC) genes to investigate the correlation between genetic instability and genetic alterations in these tumor suppressor genes. METHODS: The authors examined microsatellite assay at 9 microsatellite loci in 24 sporadic gastric cancers, 12 gastric adenomas, and 9 intestinal metaplasia mucosae of the stomach from patients with gastric cancers using fresh frozen or formalin fixed and paraffin embedded samples paired with normal mucosae. They also screened loss of heterozygosity (LOH) of the p53 and APC genes by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: In total, the RER(+) phenotypes were observed in 8 of 24 (33%) gastric cancers, 5 of 12 (42%) gastric adenomas, and 3 of 9 (33%) intestinal metaplasia mucosae of the stomach. Histology, RERs were detected in 3 of 9 (33%) well differentiated adenocarcinomas, 2 of 11 (18%) poorly differentiated adenocarcinomas, and 3 of 4 (75%) scirrhous type gastric cancers respectively. Several cases showed RERs at many microsatellite loci simultaneously. Some RER(+) phenotypes had genetic alterations of the p53 or APC genes detected by LOH using PCR-RFLP analysis. However, no significant correlation was found between RER(+) phenotypes and LOH in these genes. CONCLUSIONS: The frequency of RERs in detected gastric cancers were almost the same when compared with previously reported data. Interestingly, RERs were detected in greater than 30% of precancerous lesions, suggesting that genetic instability is an early somatic event of multistep stomach carcinogenesis. It also suggests that the adenoma-carcinoma sequence does exist in stomach carcinogenesis, especially in well differentiated adenocarcinomas. Moreover, alterations in the p53 and APC genes detected by PCR-RFLP analysis did not correlate with RER(+) phenotypes.

Adenocarcinoma↗

Microsatellite instability in transitional cell carcinoma of the urinary tract and its relationship to clinicopathological variables and smoking.

To determine whether microsatellite instability is involved in the development of transitional cell carcinoma (TCC) of the urinary tract, a microsatellite instability assay was carried out using PCR with 9 microsatellite loci. Thirty-eight TCC samples (30 patients with bladder cancer, 5 with renal pelvic tumors and 3 with ureteral tumors) and 1 lymph node with metastasis were examined. Microsatellite instability was found in 8 of 38 tumors examined, and 3 showed alterations in more than 2 microsatellite loci. All 8 tumors were beyond grade 2 and stage pT2 advanced tumors. Stages pT1-2 and pT3-4 patients differed significantly. Microsatellite instability was greater in smokers than non-smokers, but the differences were not significant. Microsatellite instability in TCC of the urinary tract is rare in superficial tumors but more common in invasive tumors. Microsatellite alterations would thus appear to occur, and possibly be importantly involved, in the tumorigenesis of urinary tract TCC.

Carcinoma, Transitional Cell↗

High frequency of genetic instability of microsatellites in human prostatic adenocarcinoma.

In order to investigate the genomic instability associated with prostate cancer, 36 microsatellite marker loci on chromosomes 1p, 3p, 5q, 8p, 8q, 9p, 11q and 13q were analyzed using microdissected samples from prostate cancer and adjoining microscopically normal tissues from the same slide. DNA was extracted from the normal and tumor cells of 40 microdissected prostate-cancer samples, amplified by PCR, and analyzed for microsatellite instability (MSI) using 36 different polymorphic DNA markers. In the present study, we have utilized a highly refined technique of PCR product separation on a sequencing gel, developed in our laboratory, which clearly shows high-quality results for the microsatellite instability in prostate cancer. The results of this study suggest that 45% (18 out of 40) showed genomic instability at a minimum of 1 locus; 4 cases each showed MSI at one and 2 loci, 4 cases had MSI at 3 loci, 3 cases showed MSI at 5 loci, while one case each showed MSI at 7, 8 and 15 loci. There was no significant correlation between the MSI and stage or grade of the tumors. This extensive study on genomic instability in prostate cancer found the occurrence of MSI to be very high, which suggests a role of MSI in the pathophysiology of prostate cancer.

Adenocarcinoma↗

Genetic instability and mutation of the TGF-beta-receptor-II gene in ampullary carcinomas.

Ampullary carcinomas are relatively rare cancers of which very little is known in terms of carcinogenetic mechanisms at the molecular level. Genetic instability caused by mutations of mismatch-repair genes has been demonstrated to be responsible for hereditary non-polyposis colorectal cancers and a sub-set of sporadic colorectal cancers. In some of those tumors showing genetic instability, the transforming-growth-factor-beta(TGF-beta)-receptor-II gene has been found to be mutated in repetitive sequences and considered to be a target of replication error. We studied the role of genetic instability and associated TGF-beta-receptor-II-gene mutations in a series of 18 sporadic cases by analyzing 5 microsatellite loci (D2S123, D3S1029, D5S409, TP53 and BAT26) and by sequencing a poly-A repeat (nucleotides 709-718) in the TGF-beta-receptor-II gene. Microsatellite instability was observed in 4 (22.2%) and gene mutations in 14 (77.8%) cases. These data indicate that the TGF-beta-receptor-II gene might be a preferential target of genetic instability whose alteration might be specifically advantageous and constitute a common step in the development of ampullary carcinomas.

Aged↗

Pronounced microsatellite instability in transitional cell carcinomas from young patients with bladder cancer.

Microsatellites may show loss of heterozygosity as well as instability of the repeats. We examined 22 different microsatellites in 14 bladder tumours (7 grade II non-invasive, 7 grade III/IV invasive) and found altered CA repeat length compared with leukocytes, indicating instability, in several microsatellites in all tumours. Instability was significantly more frequent in low stage tumours compared with high stage tumours. The number of new bands occuring was also significantly higher in low stage tumours (median 7.2) compared with high stage tumours (median 3.3). Furthermore, patients with a disease course > or = 1 year had significantly more unstable microsatellites (10.83) than those with a disease course < 1 year (mean 8.88). Examination of biopsies from normal bladder mucosa showed no instability. In 2 cases in which selected site biopsies were taken, alterations differed from the tumours, pointing at a different clonal development. LOH was most frequent in 9p markers in low stage tumours. In a group of markers located at 2p, 17p (p53), 9q, 5q and 10p, LOH was significantly more frequent in high stage tumours. Microsatellites placed at MSH2 and MLH1 loci showed LOH in several cases, indicating that the profound microsatellite instability could partly be an effect of damage to these genes.

Adult↗

Role of genomic instability in meningioma progression.

Microsatellite length instability, probably resulting from defective DNA mismatch repair mechanisms, has been described in a variety of cancers. Such genetic instability may play a significant role in tumor formation and progression. To investigate the role of microsatellite alterations in meningioma tumorigenesis and progression, we examined 33 microsatellite markers on nine chromosomes for abnormalities in 18 benign, 15 atypical, and 11 malignant meningiomas. In each tumor, at least 15 markers were investigated. Microsatellite instability was not detected in any of the cases examined. However, loss of heterozygosity for markers from various chromosomes was seen frequently among atypical and malignant meningiomas. Although some of these chromosomal losses might represent random events, our data also indicate a role for specific loci on chromosome arms 14q, 1p, 10q, and possibly 9p in the development of malignancy in meningiomas. Our results argue against a significant role for a generalized microsatellite instability phenotype in meningiomas, but they suggest that genomic instability resulting in frequent allelic deletions may contribute to meningioma progression.

Alleles↗

Perpetuating radiation-induced chromosomal instability.

Chromosomal instability is characterized by an increased rate of chromatid and chromosome rearrangements occurring multiple generations after exposure to ionizing radiation. Although chromosomal instability has been observed in many different cell types after exposure to ionizing radiation, the mechanisms perpetuating the manifestation of this instability phenotype are unknown. This review explores the current evidence concerning the role of dicentric-mediated bridge-breakage-refusion cycles and the role of recombination at interstitial telomere bands in mediating chromosomal instability. In addition, the contribution of genetic and epigenetic factors are discussed with regard to the predisposition of cells to chromosomal instability.

Animals↗

Microsatellite instability, MLH-1 promoter hypermethylation, and frameshift mutations at coding mononucleotide repeat microsatellites in ovarian tumors.

BACKGROUND: Microsatellite instability (MI) is frequent in endometrial carcinomas (ECs), but its occurrence in ovarian tumors is uncertain. Microsatellite instability positive ECs frequently are associated with frameshift mutations in coding mononucleotide tracts in IGFIIR, BAX, hMSH6, and hMSH3. METHODS: DNA from 52 consecutive patients with ovarian tumors (10 benign, 7 borderline, and 35 malignant) was obtained from neoplastic and normal tissue. After preliminary results, the series was expanded by including 41 additional, previously selected, endometrioid and clear cell carcinomas. Microsatellite instability analysis was assessed by evaluating three (CA)n dinucleotide repeats (D2S123, D5S346, D17S250) and two mononucleotide tracts (BAT 25 and BAT 26). Frameshift mutations at coding mononucleotide repeats (IGFIIR, TGF beta II, BAX, hMSH6, and hMSH3) were investigated by single-strand conformation polymorphism analysis and DNA sequencing. MLH-1 methylation was assessed by methylation specific PCR. RESULTS: Microsatellite instability was identified in only 2 of the 52 (3.8%) tumors of the initial series (1 endometrioid and 1 clear cell carcinoma). After expanding the initial series of 15 endometrioid and clear cell carcinomas with 41 additional endometrioid and clear cell carcinomas, MI was found in 7 of the total series of 56 endometrioid and clear cell carcinomas (12.5%). Frameshift mutations in coding mononucleotide tracts were detected in BAX (6 of 7), IGFIIR (1 of 7), and MSH3 (2 of 7). MLH-1 promoter hypermethylation was identified in three of six MI positive tumors. CONCLUSIONS: Microsatellite instability was infrequent in this series of ovarian tumors, and it was limited to endometrioid and clear cell carcinomas. Like EC, many ovarian carcinomas with MI follow the same process of MLH-1 promoter methylation and accumulation of mutations in coding mononucleotide tracts.

Adaptor Proteins, Signal Transducing↗

Dynamics of microtubules visualized by darkfield microscopy: treadmilling and dynamic instability.

Individual microtubules undergoing treadmilling in vitro were visualized by darkfield light microscopy, and the relationship between treadmilling and dynamic instability was studied as a function of microtubule-associated proteins (MAPs). In order to demonstrate treadmilling directly by real-time observation, we constructed three-block microtubules, the center-block of which was decorated with Tetrahymena dynein. The decorated block can easily be distinguished from undecorated blocks in the darkfield microscope because the decorated one appears much thicker. At steady-state conditions, the length of an undecorated block at one end increased and that at another end decreased, while the decorated center-block did not change in its length. The results from these direct observations show that calf brain 3X-microtubules exhibit a treadmilling flux of 0.9 micron/h. Using a similar microscopy technique, we previously demonstrated that phosphocellulose PC-microtubules existed in either the growing or the shortening phase and alternated quite frequently at steady-state conditions (dynamic instability). How does treadmilling relate to dynamic instability? An image recording of individual 3X-microtubules containing MAPs revealed that the microtubules undergo treadmilling and do not exhibit any dynamic instability. This evidence shows that MAPs suppress the dynamic instability of microtubules. That is, treadmilling can take place in the steady state only after microtubules have been stabilized by MAPs.

Animals↗

Quality assessment of confocal microscopy slide-based systems: instability.

BACKGROUND: All slide-based fluorescence cytometry detections systems basically include an excitation light source, intermediate optics, and a detection device (CCD or PMT). Occasionally, this equipment becomes unstable, generating unreliable and inferior data. METHODS: A number of tests have been devised to evaluate equipment performance and instability. The following four instability tests are described: galvanometer scanning, stage drift, correct wavelength spectral detection, and long-term laser power. RESULTS: Quality assurance tests revealed that a confocal microscope can become unstable in the following parameters, yielding inaccurate data: laser power, PMTs functionality, spectrophotometer accuracy, galvanometer scanning and laser stability, and stage drift. Long-term laser power stability has been observed to vary greatly. CONCLUSIONS: Confocal systems can become unstable in the following parameters: long-term laser power, galvanometer scanning, spectrophotometer accuracy, and stage stability. Instability in any of these parameters will affect image quality. Laser power fluctuations result from either a defective Acousto-optic tunable filter or improper heat dissipation. Spectrophotometer instability will generate unreliable spectra data, extra light reflections, and poor image quality. Galvanometer scanning instability yields poor image quality while microscope stage drift results in a sample going out of the plane of focus. With minor modifications, these tests may be applicable to other slide-based systems.

Animals↗

Tetra- and pentanucleotide short tandem repeat instability in gastric cancer.

The association between genetic instability in repetitive DNA domains and cancer has been reported in different types of malignancies. In this work we perform a comparative study of 29 gastric tumors with paired normal tissue using seven tetra-(FES/FPS, VWA31/A, HTPO, TH01, MBPB) and pentanucleotide (CD4, TP53) STR polymorphic markers regarding loss of heterozygosity and replication error status. Furthermore, we compare the gene frequencies obtained in normal tissue from patients with those of a normal control population from the same area, looking for allele associations between any of these polymorphic loci and gastric cancer risk. The results have shown that FES/FPS and TP53 present the higher rates of somatic instability. The observed results for TP53 are in accordance with those previously reported in gastric carcinogenesis, while instability of FES/FPS is for the first time reported in this tumor type. Our data suggest that different loci show different rates of instability and/or loss of heterozygosity and do not seem to consist of a result of an RER+ phenotype affecting several genomic repetitive domains. Furthermore, the instability in markers TH01, MBPB, TP53, and FES was generally detected in genotypes involving alleles with a high number of repeats. Comparing gene frequencies in patients and normal controls, no significant differences were found, although longer alleles are consistently more frequent in patients for the markers MBPB, TH01, and CD4.

Alleles↗

Genomic instability in head and neck cancer patients.

BACKGROUND: In the normal host, there are a variety of cellular systems that ensure the accurate replication and repair of DNA. Recent evidence suggests that there are individual variations in the ability to preserve the genome. Certain individuals have defects in these checkpoints and have an inherent genomic instability. They are susceptible to the accumulation of DNA damage and are prone to carcinogenesis. This article examines the role of genomic instability in the development of head and neck cancer. RESULTS: Patients with either the chromosomal instability syndromes or the Li-Fraumeni syndrome have marked defects in either DNA repair or apoptosis. These patients are prone to have head and neck neoplasms develop. Head and neck cancer patients also have a diminished ability to repair DNA damage compared with the "normal" population. Abnormalities have been identified in mutagen sensitivity, the expression of DNA mismatch repair enzymes, the expression of p53, and telomerase activity when head and neck cancer patients are compared with controls. CONCLUSION: Subpopulations exist who have increased genomic instability. These individuals are at an increased risk for the accumulation of DNA mutations and the development of head and neck cancer. More research is needed to identify specific mechanisms of genomic instability and to further define the importance of this phenomenon.

Carcinoma, Squamous Cell↗

Spindle assembly checkpoint defects and chromosomal instability in head and neck squamous cell carcinoma.

Alterations in chromosomal number and structure are found in most solid malignancies including head and neck squamous cell carcinoma (HNSC), however, the presence of ongoing, chromosomal instability in HNSC and its relation to spindle assembly checkpoint defects has not been formally demonstrated. We investigated the status of chromosomal instability (CIN) in HNSC primary tumors and cell lines as well as spindle assembly checkpoint integrity in HNSC cell lines. Centromeric fluorescence in situ hybridization (FISH) was carried out on expanded single cell-derived colonies from HNSC cell lines and primary HNSC touch preparations. The deviation of chromosomes from the modal number in single cell derived colonies was 18.4-27% in 6 HNSC cell lines, and 2-3% in a control cell line, HCT116. Twelve primary tumors and 4 normal controls were also studied; all primary tumors demonstrated significant deviation from the modal chromosomal number (average 33.7%, range = 29.9-43.9%), compared to normal controls (average 4.6%, range = 3.6-5.6%). Additional characterization of the rate of chromosomal breakage was carried out by dual color FISH simultaneously using centromeric and telomeric probes for individual chromosomes on expanded singe cell-derived colonies and primary HNSC. Control HCT 116 colonies demonstrated a mean discordance between number of centromeric and telomeric hybridization signals in 21% (range = 19-23%) of cells, whereas HNSC cell line colonies demonstrated a mean discordance of 50% (range = 38-55%), with the majority of instances of discordant signal indicating telomeric loss. Similarly, touch preparations from primary HNSC demonstrated discordance in hybridization signal of centromeric vs. telomeric signal of 26.3% (range = 18.5-42%), with normal controls showing a rate of discordance of 6.4% (range = 4-8%). Finally, all 6 HNSC cell lines demonstrated partial impairment of mitotic arrest in response to nocodazole, indicating that impairment of the spindle assembly checkpoint may contribute to chromosomal instability in HNSC. Ongoing instability in chromosomal number and structure are consistent features of primary HNSC and cell lines. Spindle assembly checkpoint impairment occurs in HNSC cell lines and may contribute to chromosomal instability in HNSC.

Carcinoma, Squamous Cell↗

Multiple manifestations of X-ray-induced genomic instability in Chinese hamster ovary (CHO) cells.

Carcinogenesis is postulated to follow a multistep cascade in which the first genetic event may destabilize cells and thereby facilitate the induction of subsequent mutations within the same cell. It has recently been shown that exposure to ionizing radiation can in itself induce a persistent, heritable genetic instability in cells. To further investigate this phenomenon, we utilized a mutationally unstable population derived from a single Chinese hamster ovary (CHO) cell that survived X irradiation. We exposed these cells to a second dose of radiation, selected hypoxanthine phosphoribosyl transferase (HPRT) mutant subclones, and identified the type of mutations involved. We found complete deletions, continuous tract partial deletions, single-exon deletions, discontinuous-exon deletions ("skip mutations"), and point mutations (changes of less than 100 bp) among the isolated HPRT mutants. We hypothesized that the skip mutation clones might be more likely to demonstrate genomic instability. To test this hypothesis, mutant subclones were screened for three markers of genetic instability: alteration of minisatellite sequences, change in telomere length, and induction of chromosomal aberrations. Clones with skip mutations and single-exon deletions possessed elevated frequencies of minisatellite alterations and chromosomal aberrations, particularly rings and dicentrics. All mutant clones showed longer telomere terminal restriction fragment lengths than did wild-type cells. These results are consistent with the hypothesis that irradiation may induce a global instability phenotype, since the multiple alterations observed are mechanistically distinct, heritable cellular modifications that arose in the clonogenic progeny of the irradiated cells. Skip mutations may be one manifestation of this instability, but their presence was not specifically associated with the other genetic alterations.

Animals↗

Urethral instability: clinical and urodynamic characteristics.

Filling cystometry was performed on 397 patients with lower urinary tract complaints. The overall prevalence of urethral instability was 12.6% (50/397). Of these 187 patients had detrusor instability (DI). Within the DI group, clinical and urodynamic characteristics were compared. Patients with an unstable urethra had a shorter functional urethral length (P = 0.005). For those with DI, 28 of 187 (15%) had urethral instability. Forty (21%) women had a decrease in maximal urethral pressure that preceded the detrusor contraction (type II DI). When those with type II DI were excluded, the difference in functional urethral length was not observed. Patients with type II DI experienced an earlier sensation of fullness of 228 mL compared with 283 mL for the other patients with DI (P = 0.001). There was a positive association between urethral instability and type II DI. Four different patterns of urethral pressure changes were observed. Based on these findings, it appears that a urethral abnormality may be the primary disorder in patients with type II DI. The differentiation of the various subtypes of urethral instability and DI might be important for directing therapy.

Female↗

Effect of transurethral resection of the prostate on detrusor instability and urge incontinence in elderly males.

Detrusor instability is common in men with evidence of outflow obstruction due to benign prostatic hypertrophy and typically reverses in about two thirds of patients after transurethral resection of the prostate (TURP). It is also common among the elderly without outflow obstruction and may lead to urge incontinence. To determine whether TURP has an effect on detrusor instability and urge incontinence in elderly men, or whether these abnormalities are due to other age-associated changes, 12 males (mean age 80 years) with urge incontinence or frequency and urgency of micturition, and symptomatic benign prostatic hypertrophy, were studied by 24-hour monitoring of incontinence and videourodynamic examination, before and after TURP; 7/12 patients were significantly cognitively impaired. Preoperatively, all patients showed detrusor instability, which reversed postoperatively in only one patient, a significantly smaller proportion than that consistently reported in younger patients. Preoperatively, 11/12 patients were incontinent. After TURP, 8/11 patients had an improvement in the amount of incontinence, by up to 458 g in 24 hours. Those who improved had been urodynamically more severely obstructed preoperatively. Those with the most improvement were also cognitively impaired. We conclude that, in the geriatric population, detrusor instability and urge incontinence may be the result of age-associated changes and not secondary to obstruction. Detrusor instability is likely to persist following TURP. Preoperative urodynamic assessment of obstruction in the incontinent male with benign prostatic hypertrophy may be useful since the severity of incontinence responds well to TURP if there is marked obstruction. Cognitive impairment should not be a deterrent to operation.

Aged↗