Chronic hepatitis B virus infection in patients with “Normal” ALT levels

Not many data exist to guide us in the management of patients with chronic hepatitis B virus infection and “normal” alanine aminotransferase levels. Many of these patients may not have normal levels on long-term follow-up or when the upper limit of normal is determined from a truly healthy reference population. These patients may have significant histologic disease and benefit from further investigation or treatment. This article focuses on the disease course of such patients.     

A case of type I Gaucher disease with cardiopulmonary amyloidosis and chitotriosidase deficiency

We present a case of Merkel cell carcinoma of the thigh diagnosed by conventional histology, immunohistochemistry, electron microscopy and cytogenetics. A unique chromosome 6 trisomy characterized this primary neoplasm, as confirmed by FISH study. The role of chromosome analysis and interphase cytogenetics is emphasized as an adjunct in the subtyping of tumours and their prognostic evaluation.     

Quantitation of Ergosterol Content: Novel Method for Determination of Fluconazole Susceptibility of Candida albicans

MIC end points for the most commonly prescribed azole antifungal drug, fluconazole, can be difficult to determine because its fungistatic nature can lead to excessive "trailing" of growth during susceptibility testing by National Committee for Clinical Laboratory Standards broth macrodilution and microdilution methods. To overcome this ambiguity, and because fluconazole acts by inhibiting ergosterol biosynthesis, we developed a novel method to differentiate fluconazole-susceptible from fluconazole-resistant isolates by quantitating ergosterol production in cells grown in 0, 1, 4, 16, or 64 microg of fluconazole per ml. Ergosterol was isolated from whole yeast cells by saponification, followed by extraction of nonsaponifiable lipids with heptane. Ergosterol was identified by its unique spectrophotometric absorbance profile between 240 and 300 nm. We used this sterol quantitation method (SQM) to test 38 isolates with broth microdilution end points of /=64 microg/ml (resistant) and 10 isolates with trailing end points by the broth microdilution method. No significant differences in mean ergosterol content were observed between any of the isolates grown in the absence of fluconazole. However, 18 susceptible isolates showed a mean reduction in ergosterol content of 72% after exposure to 1 microg of fluconazole/ml, an 84% reduction after exposure to 4 microg/ml, and 95 and 100% reductions after exposure to 16 and 64 microg of fluconazole/ml, respectively. Ten SDD isolates showed mean ergosterol reductions of 38, 57, 73, and 99% after exposure to 1, 4, 16, and 64 microg of fluconazole/ml, respectively. In contrast, 10 resistant isolates showed mean reductions in ergosterol content of only 25, 38, 53, and 84% after exposure to the same concentrations of fluconazole. The MIC of fluconazole, by using the SQM, was defined as the lowest concentration of the drug which resulted in 80% or greater inhibition of overall mean ergosterol biosynthesis compared to that in the drug-free control. Of 38 isolates which gave clear end points by the broth microdilution method, the SQM MIC was within 2 dilutions of the broth microdilution MIC for 33 (87%). The SQM also discriminated between resistant and highly resistant isolates and was particularly useful for discerning the fluconazole susceptibilities of 10 additional isolates which gave equivocal end points by the broth microdilution method due to trailing growth. In contrast to the broth microdilution method, the SQM determined trailing isolates to be susceptible rather than resistant, indicating that the SQM may predict clinical outcome more accurately. The SQM may provide a means to enhance current methods of fluconazole susceptibility testing and may provide a better correlation of in vitro with in vivo results, particularly for isolates with trailing end points.     

Controversies in surgical pathology: minimal involvement of sentinel lymph node in breast carcinoma: prevailing concepts and challenging problems

Sentinel lymph node biopsy has attained "standard of care' status in the management of breast carcinoma. However, the pathological interpretation and clinical consequences of "minimally involved' sentinel lymph nodes remain controversial. Herein, we present some of the complex and challenging pathological problems inherent in this evolving setting. Clearly, at least some of our current concepts regarding "minimally involved' sentinel lymph nodes need reappraisal.     

Fine-needle aspiration of a primary mediastinal large B-cell lymphoma: a case report with cytologic, histologic, and flow cytometric considerations

Fine-needle aspiration (FNA) cytology and immunophenotyping by flow cytometry (FCM) are increasingly being used for diagnosing and subclassifying lymphoma in the REAL/WHO classification. Herein, we report a case of primary mediastinal large B-cell lymphoma (PMBL), a subtype of diffuse large B-cell lymphoma in the WHO classification, diagnosed by FNA cytology in conjunction with FCM. This, to our knowledge, has not previously been reported. A 57-yr-old woman presented with bilateral axillary lymphadenopathy and intermittent shortness of breath. CT scan revealed a 5-cm anterior mediastinal mass and mediastinal lymphadenopathy. Endoscopic ultrasound-guided FNA of a 4.5-cm subcarinal lymph node showed medium to large atypical lymphocytes with scant to moderate finely vacuolated cytoplasm. Nuclei were enlarged, cleaved, noncleaved, lobulated, and hyperchromatic. The background showed lymphoglandular bodies. Malignant large cell lymphoma was cytologically diagnosed. FCM, performed on a portion of the FNA specimen, demonstrated large B cells devoid of surface immunoglobulin expression, the characteristic immunophenotype of PMBL. The histologic diagnosis was PMBL. Touch-imprint cytology of the histologic specimen showed large cells with a narrow rim of clear cytoplasm and prominent outer cell border. Nuclear features were similar to the FNA specimen. In the presence of a mediastinal mass, FNA cytology in conjunction with FCM can effectively diagnose PMBL in the appropriate clinical setting.     

Comparison of DNA- and RNA-based methods for detection of truncating BRCA1 mutations

A number of methods are used for mutational analysis of BRCA1, a large multi-exon gene. A comparison was made of five methods to detect mutations generating premature stop codons that are predicted to result in synthesis of a truncated protein in BRCA1. These included four DNA-based methods: two-dimensional gene scanning (TDGS), denaturing high performance liquid chromatography (DHPLC), enzymatic mutation detection (EMD), and single strand conformation polymorphism analysis (SSCP) and an RNA/DNA-based protein truncation test (PTT) with and without complementary 5' sequencing. DNA and RNA samples isolated from 21 coded lymphoblastoid cell line samples were tested. These specimens had previously been analyzed by direct automated DNA sequencing, considered to be the optimum method for mutation detection. The set of 21 cell lines included 14 samples with 13 unique frameshift or nonsense mutations, three samples with two unique splice site mutations, and four samples without deleterious mutations. The present study focused on the detection of protein-truncating mutations, those that have been reported most often to be disease-causing alterations that segregate with cancer in families. PTT with complementary 5' sequencing correctly identified all 15 deleterious mutations. Not surprisingly, the DNA-based techniques did not detect a deletion of exon 22. EMD and DHPLC identified all of the mutations with the exception of the exon 22 deletion. Two mutations were initially missed by TDGS, but could be detected after slight changes in the test design, and five truncating mutations were missed by SSCP. It will continue to be important to use complementary methods for mutational analysis.     

Flow-cytometry detection of minimal DNA aneuploidy in mature lymphoid leukemias - comparison with metaphase and interphase cytogenetics

The relative DNA content of peripheral blood cells from 79 cases with lymphoid leukemias was analyzed by a dual-parameter flow cytometric analysis. The leukemia samples corresponded to: chronic lymphocytic leukemia (CLL) 40, CLL with more than 10% prolymphocytes (CLL/PL) 12, CLL mixed 9, prolymphocytic leukemia (PLL) 5, and B-cell lymphoma in leukemic phase 13. DNA aneuploidy was found overall in 26 (32.9%) of the cases and these corresponded to: 7 (17.5%) with CLL, 7 (58.3%) with CLL/PL, 4 (44.4%) with CLL mixed, 2 (40%) with PLL and 6 (46.2%) with B-cell lymphoma. There was a good correlation between DNA content and cytogenetics/fluorescent in situ hybridization in all but 2 cases as follows: 6 of 7 cases with diploid DNA had normal karyotype and only one had trisomy 12: 4 of 6 cases with hyperdiploid DNA had trisomy 12, one had tetraploidy and only one had a normal karyotype. Two cases were hypodiploid both by DNA and cytogenetic analysis. Our findings demonstrate a higher incidence of DNA aneuploidy in B-cell lymphoma in leukemic phase, PLL, and atypical CLL in comparison with typical CLL and a good correlation with cytogenetics. We conclude that flow cytometric DNA analysis represents a useful, sensitive, and rapid method to detect and monitor minimal changes of DNA content in leukemic lymphocytes without the need of short-term cultures.     

Cytologic diagnosis of metastatic ovarian adenocarcinoma in the urinary bladder: a case report and review of the literature

A 53-yr-old woman with a 13-mo history of recurrent ovarian papillary serous adenocarcinoma presented with persistent microscopic hematuria. The patient was undergoing chemotherapy for her recurrent ovarian tumor when she was referred to the urology service for microscopic hematuria. An intravenous pyelogram was normal. Cystoscopy was performed, as well as a urinary bladder washing and mucosal biopsies for examination. Adenocarcinoma similar to the patient's primary ovarian tumor was detected in both cytology and histopathology specimens. Ovarian carcinoma comprises 1.3-4.0% of all metastatic neoplasms to the urinary bladder and is an important consideration in the differential diagnosis of a cytologic finding of adenocarcinoma in urine specimens of female patients, where it accounts for an even higher percentage of cases (1 of 3 adenocarcinoma diagnoses in a series of 4,677 urine specimens from female patients).