Chronic hepatitis C. Advances in diagnostic testing and therapy.

The methods for diagnosing hepatitis C virus infection have been evolving since the first-generation enzyme-linked immunosorbent assay antibody test was devised in 1989. In addition to assaying for serum antibodies against viral proteins, serum and liver tissue can be tested for viral RNA, evidence of ongoing viral replication. The improving ability to diagnose hepatitis C has furthered the understanding of the natural history of this infection. Acute hepatitis C results in chronic elevations of serum transaminase levels following nearly one half of cases. Cirrhosis complicates approximately 20% of chronic infections. Long-standing chronic hepatitis C may play a role in the pathogenesis of hepatocellular carcinoma. Sustained normalization of serum transaminase levels, often accompanied by a decrease in or disappearance of viral RNA, occurs in approximately 25% of patients with chronic hepatitis C who are treated with a 6-month course of recombinant interferon alfa. This treatment can occasionally be complicated by hematologic, endocrinologic, and psychiatric adverse effects but is usually fairly well tolerated. Whether interferon therapy will diminish the risk of cirrhosis or carcinoma is not yet known. This article reviews the diagnosis of chronic hepatitis C infection as well as the mechanisms of action, efficacy, and adverse effects associated with interferon alfa therapy.

Revised estimates of diagnostic test sensitivity and specificity in suspected biliary tract disease.

BACKGROUND: The purpose of this study was to estimate the sensitivity and specificity of diagnostic tests for gallstones and acute cholecystitis. METHODS: All English-language articles published from 1966 through 1992 about tests used in the diagnosis of biliary tract disease were identified through MEDLINE. From 1614 titles, 666 abstracts were examined and 322 articles were read to identify 61 articles with information about sensitivity and specificity. Application of exclusion criteria based on clinical and methodologic criteria left 30 articles for analysis. Cluster-sampling methods were adapted to obtain combined estimates of sensitivities and specificities. Adjustments were made to estimates that were biased because the gold standard was applied preferentially to patients with positive test results. RESULTS: Ultrasound has the best unadjusted sensitivity (0.97; 95% confidence interval, 0.95 to 0.99) and specificity (0.95; 95% confidence interval, 0.88 to 1.00) for evaluating patients with suspected gallstones. Adjusted values are 0.84 (0.76 to 0.92) and 0.99 (0.97 to 1.00), respectively. Adjusted and unadjusted results for oral cholecystogram were lower. Radionuclide scanning has the best sensitivity (0.97; 95% confidence interval, 0.96 to 0.98) and specificity (0.90; 95% confidence interval, 0.86 to 0.95) for evaluating patients with suspected acute cholecystitis; test performance is unaffected by delayed imaging. Unadjusted sensitivity and specificity of ultrasound in evaluating patients with suspected acute cholecystitis are 0.94 (0.92 to 0.96) and 0.78 (0.61 to 0.96); adjusted values are 0.88 (0.74 to 1.00) and 0.80 (0.62 to 0.98). CONCLUSIONS: Ultrasound is superior to oral cholecystogram for diagnosing cholelithiasis, and radionuclide scanning is the test of choice for acute cholecystitis. However, sensitivities and specificities are somewhat lower than commonly reported. We recommend estimates that are midway between the adjusted and unadjusted values.

Vitamin A hepatotoxicity: a cautionary note regarding 25,000 IU supplements.

Vitamin A hepatotoxicity has been reported at doses exceeding 50,000 IU/day. At 25,000 IU vitamin A per day, although elevated liver enzymes may be seen, hepatotoxicity is rare. We report a case of severe hepatotoxicity associated with the habitual daily ingestion of 25,000 IU of vitamin A bought as an over-the-counter dietary supplement. With the general availability of high-dose supplements and recent literature emphasizing the importance of vitamin A adequacy, the potential for vitamin A hepatotoxicity may increase. Health professionals should remain aware of the potential for vitamin A hepatotoxicity and elicit a vitamin A history in all patients being evaluated for liver dysfunction.

Nonalcoholic steatohepatitis.

Nonalcoholic steatohepatitis (NASH) is a liver disease that, until recently, has been underrecognized as a common cause of elevated liver enzymes. This distinct clinical entity is characterized by liver biopsy findings similar to those seen in alcoholic hepatitis but in the absence of alcohol consumption sufficient to cause such changes. Patients with NASH are often middle-aged and obese, with coexisting diabetes or hyperlipidemia, but NASH also occurs in younger lean, otherwise healthy individuals and even in children. Although NASH is generally a benign disorder, it may be progressive, leading to cirrhosis and complications of portal hypertension. Liver biopsy remains the gold standard for diagnosis. Therapy for NASH remains poorly defined, although weight reduction and ursodeoxycholic acid may have a beneficial effect.

A tumor profile in Down syndrome.

We conducted a review of cancers in Down syndrome (DS), because solid tumors are poorly understood in DS. Cancers are in excess in this condition because of the 20-fold excess of leukemias, whereas malignant solid tumors seem to be globally underrepresented as compared with those in the general population. However, among these tumors, some tumors are in excess: lymphomas, gonadal and extragonadal germ cell tumors, and possibly retinoblastomas and pancreatic and bone tumors. Neoplasms in excess are seen earlier, sometimes in fetal life (leukemias and testicular germ cell tumors) or neonatally (leukemias and lymphoma) and affect mainly male subjects. There seems to exist an excess of rare karyotypes. Other tumors are underrepresented, particularly neuroblastomas and nephroblastomas, in young children, and perhaps common epithelial tumors in adults. These observations suggest that DS has a particular tumor profile, with some tissues more affected by malignant diseases (hematopoietic tissue and germ cells) and others that seem to be protected (central and peripheral nervous system, renal tissue, and epithelial tissues). The mechanism is mainly genetic, but differences in exposure to exogenous agents compared with the general population must be kept in mind. These findings are of interest for the management of these patients and early detection of cancers. Better knowledge of this tumor profile could help us to understand the mechanisms of carcinogenesis and should be compared to the current knowledge of genes on chromosome 21.

Keratosis pilaris and ulerythema ophryogenes associated with an 18p deletion caused by a Y/18 translocation.

We present a patient with partial monosomy of the short arm of chromosome 18 caused by de novo translocation t(Y;18) and a generalized form of keratosis pilaris (keratosis pilaris affecting the skin follicles of the trunk, limbs and face-ulerythema ophryogenes). Two-color FISH with centromere-specific Y and 18 DNA probes identified the derivative chromosome 18 as a dicentric with breakpoints in p11.2 on both involved chromosomes. The patient had another normal Y chromosome. This is a third report the presence of a chromosome 18p deletion (and first case of a translocation involving 18p and a sex chromosome) with this genodermatosis. Our data suggest that the short arm of chromosome 18 is a candidate region for a gene causing keratosis pilaris. Unmasking of a recessive mutation at the disease locus by deletion of the wild type allele could be the cause of the recessive genodermatosis.

Correlation of percutaneous liver biopsy fragmentation with the degree of fibrosis.

BACKGROUND: Although fragmentation of a liver biopsy specimen has been considered to be suggestive of cirrhosis, the evidence for this is difficult to find in the published literature. AIM: To determine whether fragmentation of percutaneous liver biopsy specimens correlates with the degree of fibrosis. METHODS: One hundred and eighty-six patients underwent percutaneous liver biopsy prospectively. The specimens were measured for the length and number of fragments. The extent of fibrosis was scored by a pathologist blind to the clinical data. Length and fragmentation data were compared between the different stages. RESULTS: The overall median fragment length was 1.85 cm and the median fragment number was four. Specimens with advanced fibrosis (stages III-IV) had more fragments than those with no or mild fibrosis (stages 0-II) (P < 0.0001). The aggregate fragment length decreased with increasing stage of fibrosis (P < 0.0001). Specimens with greater than 12 fragments were seen only with advanced fibrosis. CONCLUSIONS: Fragmentation of percutaneous liver biopsy specimens is common and increases with progression from early to advanced fibrosis. Fibrotic specimens fragment more often and more extensively.

A randomized trial of high-dose interferon alpha-2b, with or without ribavirin, in chronic hepatitis C patients who have not responded to standard dose interferon.

BACKGROUND: Conventional interferon monotherapy fails to achieve virological clearance in most hepatitis C-infected patients. The use of high-dose induction regimens may improve the initial clearance of virus, while the addition of ribavirin appears to improve the rates of sustained response once clearance is achieved. AIM: To compare the efficacy and safety of re-treatment with an induction regimen of high-dose interferon alpha-2b, with or without ribavirin, in chronic hepatitis C patients who have not responded to standard dose interferon monotherapy. METHODS: Previous virological non-responders to standard dose interferon (3-5 MU three times weekly for > or = 12 weeks) were randomized to receive, unblind, either 10 MU interferon alpha-2b daily for 10 days, then 5 MU daily for 74 days, then 5 MU three times weekly for 24 weeks (total 36 weeks) (group A), or the above regimen with the addition of ribavirin, 1000-1200 mg/day, at day 11 (group B). All patients were followed up for 24 weeks after completion of therapy. RESULTS: End of treatment virological response was noted in one of 10 (10%) patients in group A and in eight of 15 (54%) patients in group B (P=0.04). The sole end treatment responder in group A and three in group B relapsed on follow-up. The apparent improvement in response in group B compared to group A nearly reached statistical significance (group B 5/15 vs. group A 0/10; P=0.06). CONCLUSIONS: In this small pilot study, a 36-week high-dose induction interferon monotherapy protocol did not yield sustained responses in previous non-responders to standard dose interferon. However, the same regimen with ribavirin yielded a 33% sustained response rate, nearly reaching statistical significance. The therapy was well tolerated, despite the higher doses of interferon used and the addition of ribavirin. High-dose interferon with ribavirin appears to be a therapeutic option for non-responders to conventional interferon monotherapy.

Effects of low-dose neutrons applied at reduced dose rate on human melanoma cells.

Human melanoma cells that are resistant to gamma rays were irradiated with 14 MeV neutrons given at low doses ranging from 5 cGy to 1.12 Gy at a very low dose rate of 0.8 mGy min(-1) or a moderate dose rate of 40 mGy min(-1). The biological effects of neutrons were studied by two different methods: a cell survival assay after a 14-day incubation and an analysis of chromosomal aberrations in metaphases collected 20 h after irradiation. Unusual features of the survival curve at very low dose rate were a marked increase in cell killing at 5 cGy followed by a plateau for survival from 10 to 32.5 cGy. The levels of induced chromosomal aberrations showed a similar increase for both dose rates at 7.5 cGy and the existence of a plateau at the very low dose rate from 15 to 30 cGy. The existence of a plateau suggests that a repair process after low-dose neutrons might be induced after a threshold dose of 5-7.5 cGy which compensates for induced damage from doses as high as 32.5 cGy. These findings may be of interest for understanding the relative biological effectiveness of neutrons and the effects of environmental low-dose irradiation.

Cytogenetic characterization of a bladder carcinoma cell line by fluorescence in situ hybridization.

A bladder carcinoma cell line, RT112/84, has been cytogenetically characterized by classical techniques and fluorescence in situ hybridization. The RHG banding and FISH analysis revealed a mixture of two clones and multiple chromosome rearrangements.

Rehybridization on metaphases studied previously by FISH. An approach to analyze chromosome aberrations.

A new protocol for fluorescence in situ rehybridization is described. Biotin-labeled chromosome-specific DNA probes were hybridized onto metaphases which previously had been studied by FISH. This method makes it possible to reexamine the same metaphase spreads with different DNA probes. It allows for precise characterization of cytogenetic translocations and for detection of multiple aberrations presented in a karyotype. It is especially useful in cases where a limited number of cytogenetic preparations are available.

Cytogenetic studies in a case of T-cell prolymphocytic leukemia.

The authors describe cytogenetic aberrations observed in a case of T prolymphocytic leukemia. C11 deletion (q14) B5 deletion (pter), D14q +, E20 trisomy, and two markers are the main anomalies of the complement.

Chronic myelogenous leukemia (CML) with translocation (8;22): a new variant.

A case of chronic myelogenous leukemia (CML) in a young woman with a new variant Ph1-translocation--i.e., t(8;22) (q24;q12)--is described. The clinical and biological aspects of the disease did not seem to differ from those of the usual cases of CML.

Karyotype and FISH analysis of a newly established cell line derived from a human bladder carcinoma.

A new human malignant urologic cell line was established in vitro from a moderately differentiated transitional cell carcinoma of the bladder and cytogenetically characterized. Repeated chromosome analyses of the cell line using conventional RHG and GTG banding and non-radioactive in situ hybridization showed a stable karyotype with a modal number of 48 and chromosomal rearrangements, some of which have not been previously described. Numerical deviation included three trisomies (+7, +8, +9) and one nullisomy (-19, -19). Structural changes involved a balanced translocation (1;5)(q12;q12), an isochromosome 3q, a 14p+, and two markers. Fluorescence in situ hybridization (FISH), using biotin-labeled alpha satellite probes for chromosome 9 or painting for chromosomes 1 and 8, applied to interphase nuclei or metaphases showed similar results to those found by conventional cytogenetic study. This cell line may be an interesting model for fuller characterization by molecular biology studies and for testing anti-cancer drugs in vitro.

Translocation t(3;20) associated with thrombocythemia in Ph-positive CML.

A patient with Philadelphia (Ph) chromosome positive chronic myelocytic leukemia is described who also developed an abnormality of chromosome #3, i.e., t(3;20)(p21;p13), in blast crisis. This abnormality may be connected with the advent thrombocythemia. The disease was a thrombopenia in the initial phase.

Structural and numerical aberrations of chromosome 22 in a case of follicular variant of papillary thyroid carcinoma revealed by conventional and molecular cytogenetics.

This study reports a case of papillary carcinoma with vesicular components showing multiclonal aberrations of chromosome 22 as revealed by RHG-banding cytogenetics and by fluorescence in situ hybridization (FISH; whole chromosome 22 and BCR-ABL-specific locus probes, multi-FISH). Four clones with chromosome 22 changes as the sole abnormality were seen. The main abnormal clone lacked the whole chromosome 22. A del(22)(q11) was observed in a second group of cells. The third clone had an idic(22). Finally, FISH revealed a fourth abnormal cell population with a der(17)t(?17;22). Some of these chromosome 22 alterations have been described in other solid tumors such as meningiomas and neurinomas, suggesting a common genetic pathway of tumor progression occurring in a multistep process. Chromosome 22 changes do not seem to be involved in pure papillary thyroid tumors and therefore could be related to the maintenance of a follicular-type histological pattern.

Nonrandom changes of chromosome 10 in bladder cancer. Detection by FISH to interphase nuclei.

Fluorescence in situ hybridization (FISH) to interphase nuclei has been a valuable method for examining the chromosome copies in tumor cells in clinical practice. Twelve cases of transitional cell carcinoma (TCC) of the bladder were investigated with a biotin-labeled repetitive DNA probe to detect numerical aberrations of chromosome 10 in interphase nuclei. The cells containing one fluorescent signal were screened in two of seven non-invasive tumors and in four of five invasive tumors. Two patients presented two FISH spots of different sizes. More than two signals were seen in one invasive tumor. The findings suggest that partial or complete loss of a chromosome 10 is a nonrandom aberration in bladder cancer.

Synthesis and Characterization of Hydrotalcite-like Compounds Containing V(3+) in the Layers and of Their Calcination Products.

The synthesis and full characterization of a new hydrotalcite-like compound with the formula [Mg(0.71)V(0.29)(OH)(2)](CO(3))(0.145).0.72H(2)O and with V(3+) in the layers are described. The influence of hydrothermal treatment and drying rate on the crystallinity of the materials obtained is discussed. The evolution to mixed oxides upon calcination at different temperatures (448, 548, 773, 1023, and 1273 K) under different atmosphere environments (air or nitrogen) for 2 h has been studied. Characterization of the original layered materials and of the calcination products has been carried out by X-ray diffraction, thermal analysis, Fourier-transform infrared spectroscopy, BET specific surface area determination, temperature-programmed reduction, and transmission electron microscopy. X-ray absorption spectroscopies (XANES and EXAFS) have also been used to assess the local geometry of vanadium ions in the different compounds prepared. All experimental data agree with a well-crystallized hydrotalcite-like compound after thermal treatment, and also a minor effect of the drying rate on the crystallinity has been found. Thermal decomposition yields poorly crystalline layered compound at 448 K that undergoes transformation to mostly amorphous materials when calcined at 548-773 K, finally leading to a mixture of MgO and Mg(3)V(2)O(8), which has increasing crystallinity as the calcination temperature increases. XAS results indicate the presence of V(3+) ions in an octahedral coordination in the parent sample calcined at 448 K and tetrahedrally coordinated V(5+) species for samples calcined at higher temperatures, calcination giving rise to a better ordering of the second coordination sphere. Similar results were found when calcination was performed in nitrogen, although higher temperatures were needed to achieve the same result.

Preparation and Study of Decavanadate-Pillared Hydrotalcite-like Anionic Clays Containing Cobalt and Chromium.

Hydrotalcite-like compounds containing Co(II) and Cr(III) in the brucite-like layers have been prepared. The interlayer anion was carbonate or decavanadate. The chemical formulas of the samples are [Co(0.65)Cr(0.35)(OH)(2)] (CO(3))(0.175).1.22H(2)O (sample CoCrC) and [Co(0.61)Cr(0.39)(OH)(2)](V(10)O(28))(0.065).1.35H(2)O (sample CoCrV). The compounds have been characterized by X-ray diffraction, XAS, vis-UV, FT-IR, and Raman spectroscopies, while the surface textures were assessed by nitrogen adsorption. Reducibility has been studied by temperature-programmed reduction. A similar characterization study has been carried out on samples obtained after calcination of the parent samples in air at increasing temperatures. Results indicate an ordered structure, with Co(II) and Cr(III) ions in octahedral holes of the brucite-like layers and, in CoCrV, decavanadate species with its main C(2) axis parallel to the layers. Thermal decomposition at increasing temperatures takes place, for CoCrC, through intermediate formation of Cr(VI) species, which are again reduced to Cr(III) at higher temperatures; simultaneously, Co(II) is oxidized to Co(III) (even at 673 K), thus leading to formation of Co(II)Co(III)Cr(III)O(4). However, decomposition of CoCrV takes place through partial depolymerization of decavanadate species and formation of Co(II)Cr(2)O(4) and Co(II)(2)V(2)O(7), without intermediate formation of Cr(VI) species.

Chromosome analysis by image processing in a computerized environment. Clinical applications.

Dealing with a routine regional cytogenetic activity, we have developed and adapted to clinical work a semi automatic karyotyping machine. Attempts for an accurate automated chromosome classification using a neural network have led to partial results. A specific adaptation to cancer cytogenetics is under development (determination of the modal number, translocations analysis with densitometric curves, automatic identification of markers). A specific program allows quantification of chromosome labelling with radioactive probes. Exchanges of digitized karyotypes are feasible with labs using automated karyotyping machines. A local network connects several karyotyping and metaphase finding stations. Guidelines for an international data bank concerning abnormal chromosome images have been elaborated. On the other hand the ISH techniques have been applied to the following topics: identification of human chromosome aberrations in amniotic and chorionic cells, chromosome studies of human gametes and embryos (including sex determination), identification of markers in cancer cells.