Oligosecretory biclonal multiple myeloma.

Among the plasma cell dyscrasias, non-secretory myeloma is one of the rarest. This diagnosis is based on the absence of monoclonal proteins in the serum and urine. When serum free light chains are trace and the kappa: lambda ratio normal, clonality may however be established by PCR. We present a case of an oligosecretory myeloma confirmed by PCR, which would have hitherto been classified as non-secretory.

A model for cutaneous squamous cell carcinoma in vemurafenib therapy.

Vemurafenib is a chemotherapeutic BRAF inhibitor, or dabrafenib, that has been FDA-approved for treatment in metastatic melanoma positive for the V600E mutation. BRAF inhibitors, including vemurafenib, are linked to the development of cutaneous squamous cell carcinoma and keratoacanthoma. Furthermore, pathological analysis has shown these secondary tumors do not harbor the same mutations as the primary cancer, suggesting de novo pathogenesis. In accordance, patients require close dermatological follow-up due to the high prevalence rates of these tumors. This paper takes the form of an extensive case-and-review article exploring the development of these tumors and their management.

Karyopyknotic cytoplasmic inclusions in neutrophils.

Karyopyknotic cytoplasmic inclusions in neutrophils (KPCI) have been previously called "Howell-Jolly" like inclusions and identified in immunosuppressed patients with human immunodeficiency virus (HIV) and in patients with various malignancies who have undergone chemotherapy. Attempts to characterize these inclusions have included the Grocott's methenamine silver (GMS), periodic acid Schiff (PAS), Gram, and Feulgen stains. Previous authors have concluded that these inclusions are of deoxyribonucleic acid (DNA) origin. We present a case describing an additional method to confirm this observation and to document previously undescribed curvilinear forms.

Molecular pathology--translating research into clinical practice: an expanding frontier in surgical oncology.

Molecular assays have now become essential to the pathologist and clinician alike in diagnosing and managing disease. This article highlights the techniques and molecular targets no longer ancillary to basic research. Ripe for discussion are the likely future impact of genetics on clinical care, the potential models for service provision, and the broader ethical, legal, and social issues related to the use of genetic information for nonmedical purposes. Molecular methods are forecasted to increase in assisting in the diagnosis of human diseases. The author's mission is to embrace this discipline and use these technologies in clinical practice.

Glioblastoma multiforme in two non-nuclear family members.

BACKGROUND: In spite of traditional and current epidemiological research, there have been few environmental risk factors identified for malignant brain tumors. It has been an equally difficult challenge to identify genetic causes for brain tumors because of the rarity of families with multiple affected individuals, which prevents the use of traditional methods of genetic analysis such as genetic linkage, sib-pair, or even population-based association studies. Thus, it is important to take advantage of rare occasions of familial brain tumors. METHODS: Identification and careful study of such families may provide important clues about the etiology of brain malignancies. We studied one family of which two nonnuclear family members were affected with pathologically diagnosed glioblastoma multiforme. Fluorescence in situ hybridization (FISH) assays were used on archival sections from each patient's tumor to investigate the loss and/or gain of important allelic endpoints. Tissue sections were prepared and processed for FISH. DNA probes for targeted gene loci were used to assess allelic gain/loss. FISH probes targeted regions including 19q13, 1p36, 10q/phosphate and tensin homolog (PTEN), chromosome 3, chromosome 7, chromosome 17/17q and p53/17p. RESULTS: FISH analyses identified distinct abnormalities in the two patients, suggesting that despite the familial connections and histologically similar tumors, genetic abnormalities are abundant and heterogeneous among these malignancies. CONCLUSION: These abnormalities, however, serve to contribute to valuable information regarding patient outcomes, albeit their precise roles in the etiology of this malignancy are yet to be determined.

Characterization of a new myeloid leukemia cell line with normal cytogenetics (CG-SH).

A new myeloid leukemia cell line (CG-SH) with normal cytogenetics was established from a patient with acute myelogenous leukemia (AML) following myelodysplastic syndrome (MDS). The cells of CG-SH are immature blasts and have an immature myeloid phenotype (positive for myeloperoxidase, CD7, CD34, CD38, CD117, HLA-DR, negative for CD10, CD19, CD20, CD41, CD42). A partial expression of CD13, CD15, CD65 and a weak expression of CD33 and CD133 was noted. The cells are negative for EBER. By molecular analysis, a mutation of NRAS and heterozygous mutations of RUNX1 were detected. No mutations were detected in FLT3-ITD, MLL-PTD or NPM1. By real-time PCR, a series of 19 microRNAs was identified which are strongly expressed in CG-SH. In conclusion, a new cell line was established which will be useful for the study of AML with normal cytogenetics and mutations in NRAS and/or RUNX1.

Comparison of FISH and quantitative RT-PCR for the diagnosis and follow-up of BCR-ABL-positive leukemias.

BACKGROUND: For Philadelphia chromosome positive (Ph+) leukemias (chronic myelogenous leukemia [CML], acute lymphoblastic leukemia [ALL], and rare other leukemias), both allogeneic transplantation and treatment with tyrosine kinase inhibitors offer chances of molecular remission (the molecular marker being consistently undetectable). Molecular remission is defined as a reduction in the quantification of BCR-ABL transcripts to an undetectable level by molecular diagnostic methods, and is considered as a surrogate marker for cure or long-term disease control. The molecular diagnostic methods including fluorescence in situ hybridization (FISH) and quantitative reverse transcription-polymerase chain reaction (QRT-PCR) are more sensitive than classical cytogenetic analysis for the detection of BCR-ABL positive cells. QRT-PCR, due to its superior sensitivity, is considered the gold standard for the follow-up of Ph+ leukemias treated with imatinib. AIM: The objective of our study was to compare the diagnostic and clinical usefulness of FISH and QRT-PCR at different timepoints for Ph+ leukemias. PATIENTS AND METHODS: We investigated 23 unselected patients with Ph+ CML (n = 21) or Ph+ ALL (n = 2) at 77 different timepoints in a comparative study with both FISH and QRT-PCR using commercially available reagents in a routine laboratory. RESULTS: Our study demonstrated a good correlation of QRT-PCR with FISH in detecting the BCR-ABL fusion gene among patients with CML or ALL (coefficient of correlation = 0.77493, p < 0.0001, using Spearman's correlation procedure). All newly diagnosed or untreated cases were positive with both methods. Lower coefficients of correlation were found when FISH and QRT-PCR were correlated with the white blood cell count (WBC). An overall concordance of FISH and QRT-PCR (being either negative or positive in both tests) was found in 65 cases (84.4%) and a discrepancy identified in 12 cases (15.6%). CONCLUSIONS: We confirm that QRT-PCR allows precise measurement of low levels of BCR-ABL transcripts and can serve as a sensitive indicator for minimal residual disease. In addition, we demonstrate in most cases a good correlation of QRT-PCR with FISH in detecting the BCR-ABL fusion gene among patients with CML or Ph+ ALL. FISH is not suitable for monitoring minimal residual disease.

Fluorescence in situ hybridization: method of choice for a definitive diagnosis of mantle cell lymphoma.

Fluorescence in situ hybridization (FISH) using IGH/CCND1 probes was used to analyze 35 specimens including 27 paraffin sections, 3 bone marrow aspirates, and 5 peripheral blood smears. The 27 paraffin sections included 7 bone marrows, 10 lymph nodes, 3 spleens, 3 tonsils, 3 gastrointestinal biopsies, and 1 skin biopsy. Among these cases, 23 specimens were from 20 patients with mantle cell lymphoma (MCL) and 12 specimens were from 12 patients with non-MCL lymphomas/lymphoid hyperplasia. Specimens from all MCL patients showed positive results with FISH. In one patient, the archived paraffin sections were negative with FISH, but a fresh peripheral blood specimen showed a positive result. Negative results were obtained in all specimens from non-MCL cases. Flow cytometric analysis revealed that all cases of MCL showed CD19/CD5 staining, but the percentages of cells positive for CD23 and FMC-7 were variable, thus they cannot be depended upon for a definitive diagnosis of MCL. Immunohistochemical stains demonstrated positive staining for CD5 and CD20 and negative staining for CD23 in MCL cases but cyclin D1 was positive in only 10 of 13 MCL cases studied. Therefore, it appears that immunophenotyping alone is not sufficient to establish a definitive diagnosis of MCL. FISH should be routinely used when the diagnosis needs confirmation. FISH can be performed in a routine clinical laboratory, and it is applicable to archived material for retrospective studies. Other molecular cytogenetic techniques in comparison with FISH are discussed.