Metabolic disorders in patients operated for pancreatic cancer.

Adenocarcinoma of the pancreas presents a major threat with a 5-years survival rate of 5%. Whipple pancreaticoduodenectomy (PD) is the standard procedure for cephalo-pancreatic neoplasm. After an extended resection and reconstruction of superior gastrointestinal tract the digestive physiology might be heavily disrupted. A literature review of metabolic alterations of patients who suffered a major pancreatic resection is performed, regarding micronutrients, lipid absorption and pancreatogenic diabetes. Long-term survivors following PD generally have a satisfactory nutritional status although with subclinical iron, vitamin D and selenium deficiency. These patients should be followed-up also regarding these micronutrients and properly dietary supplemented when necessary, also considering the increased life expectancy. Approximately 17-25% of patients will develop insulin-dependent diabetes but pancreatogenic diabetics have elevated levels of serum insulin and minimal or absent response to food intake, as opposed to a type I diabetics, where insulin serum is normal or elevated and there is an exaggerated response to ingestion of sugar.

Hematologic masquerade of rhabdomyosarcoma.

We present the case of a 12-year-old boy admitted with the diagnosis of acute leukemia, but found to have an infiltration of the marrow by an alveolar rhabdomyosarcoma (RMS) as determined by the cytogenetic demonstration of a t(2;13)(q25;q14). A primary tumor could not be found. With this case as a basis, we have tabulated features of all similar cases in the literature and discuss the possible optimal approaches to establishing the correct diagnosis in such patients.

FISH Analysis.

In situ hybridization of specific DNA or RNA sequences to cellular targets was developed over 20 yr ago (1,2). The early techniques employed isotopically labeled probes and subsequent autoradiographic detection using a photographic emulsion overlying the metaphase chromosomes, nuclei, or whole cells. However, autoradiography requires long exposure periods, and is not practical for clinical application. In the late 1970s, nonisotopic methods of nucleic acid labeling were developed. The subsequent improvements in the detection of reporter molecules using immunocytochemistry and immunofluorescence, in conjunction with advances in fluorescence microscopy and image analysis, have made the technique safer, faster and reliable.

Cytogenetics analysis.

The establishment of reliable and meaningful chromosomal (cytogenetic, karyotypic) changes in hematological disorders, primarily the leukemias and lymphomas, must be based on the examination of the involved cells or tissues. Thus, in the case of the leukemias bone marrow (BM) aspirations yield optimal results in the preponderant number of patients, whereas in the lymphomas affected tissues, usually lymph nodes, are the best source of cells carrying cytogenetic anomalies. Generally, BM is not a good source of cells for cytogenetic analysis in lymphoma. Not only is the marrow often not affected by the lymphoma, but also when it is, the number of abnormal cells is relatively small and/or the abnormal cells are not in division and, hence, do not yield a sufficient number of metaphases for cytogenetic analysis. In some situations, blood cells can be utilized as a source of metaphases affected by karyotypic changes, e.g., in cases with about 10% immature cells in the peripheral blood (PB), in chronic lymphocytic leukemia (CLL), in cases where the marrow is fibrotic or extremely hypocellular, or in determining the presence of Ph+cells in established cases of chronic myelocytic leukemia (CML) (1,2).

Cytogenetic and molecular genetic techniques as adjunctive approaches in the diagnosis of bone and soft tissue tumors.

Important and meaningful advances have been made in mesenchymal tumor cytogenetics during the last two decades. A number of bone and soft tissue tumors have been shown to have recurrent, if not specific, chromosomal changes, particularly translocations. These changes not only serve as aids in the diagnosis and classification of bone and soft tissue tumors, especially in the differential diagnosis of those of confusing nature, but have also guided molecular studies in establishing the underlying genes involved. To date, a number of tumor-specific gene fusions have been identified and many have been shown to encode aberrant transcription factors. These key biological events in bone and soft tissue tumors are crucial not only to our understanding of the sarcomagenetic processes leading to the various tumors, but also ultimately in the design of specific therapies tailored to the genetic events in mesenchymal neoplasms.

Evidence by spectral karyotyping that 8q11.2 is nonrandomly involved in lipoblastoma.

We report two cases of lipoblastoma with chromosome 8-related aberrations, ie, a 92,XXYY,t(7;8Xp22;q11.2)x2 [8]/46,XY[16] in Case 1 and a 46,XY,-8,-13,add(16) (q22),+mar, +r [cp13]/46,XY[7] in Case 2. Using spectral karyotyping and fluorescence in situ hybridization techniques, the karyotype of Case 2 was redesignated as 46,XY, r(8), del(13)(q12), der(16)ins(16;8)(q22; q24q11.2)[cp13]/46,XY[7]. This report delineates a new chromosome rearrangement, ie, der(16)ins(16;8)(q22; q24q11.2) in lipoblastoma, and also confirms the t(7; 8)(p22;q11.2), reported only once previously, as a recurrent translocation involved in such a tumor. These findings provide valuable information for clinical molecular cytogenetic diagnosis of lipoblastoma. Furthermore, this report highlights the value of cytogenetic and molecular cytogenetic analysis in differential diagnosis of childhood adipose tissue tumors and adds to the number of lipoblastomas reported with chromosomal abnormalities at 8q11.2.

Development of a disease specific quality of life (QoL) questionnaire module to supplement the EORTC core cancer QoL questionnaire, the QLQ-C30 in patients with pancreatic cancer. EORTC Study Group on Quality of Life.

There is overwhelming consensus that quality of life assessment is urgently required in pancreatic cancer, yet little research has been conducted. We report on the development of a disease specific questionnaire module to supplement the EORTC core cancer module, the QLQ-C30 in patients with pancreatic cancer, using EORTC quality of life study group guidelines for module development. Relevant QoL issues were generated from literature searches and interviews with health professionals and patients with pancreatic cancer. Issues were constructed into items and provisionally translated. The provisional module was pretested in patients in 8 European centres. The resulting module the QLQ-PAN26 includes 26 items related to disease symptoms, treatment side-effects and emotional issues specific to pancreatic cancer. This should ensure that the module will be sensitive to assess the small but important disease and treatment related QoL changes in pancreatic cancer. The use of the QLQ-C30 and QLQ-PAN26 will provide a comprehensive system of QoL assessment in international trials of pancreatic cancer.

Identification of a novel amplicon at 1q31 in pancreatic cancer cell lines.

Pancreatic adenocarcinoma is a highly lethal malignant tumor that is increasing in frequency, now ranking fifth in the United States as a cause of death attributed to cancer. Patients with pancreatic carcinoma have one of the poorest prognoses of all cancer patients, with the number of deaths being approximately 75% of the total number of cases. The use of comparative genomic hybridization (CGH) has gained widespread use in the study of some types of solid tumors, and it seems to be a very good approach in pancreatic adenocarcinomas, in which just a few cases have been studied cytogenetically, mostly owing to the fact that these tumors are very difficult to grow in culture. Fourteen pancreatic cancer lines were examined with CGH. In 11 of these lines, we found an amplicon at 1q31, not previously reported in pancreatic cancer. More studies need to be done in primary tumors to determine the involvement of 1q31 in this type of tumor.

A group of previously not recognized cytogenetic abnormalities in myeloid hematological malignancies.

We have identified a group of previously not reported chromosome abnormalities related to myeloid hematological malignancies. Cases 1 and 2 were observed to have an additional i(4)(p10) as the sole anomaly with similar clinical features of myeloid disorders; that is, acute nonlymphocytic leukemia (ANLL-M2) and myelodysplastic syndrome (MDS)-refractory anemia with an excess of blasts in transformation, respectively. Fluorescence in situ hybridization studies with the use of a 4p-specific microdissection probe further confirmed the presence of an i(4)(p10) in these patients. Case 3 was diagnosed with ANLL-M1 and had an additional i(8)(p10) as the only change, also confirmed by a whole-chromosome painting procedure. In cases 4-6, deletions of 18q at breakpoints q12, q23, and q21 were identified as the sole anomaly in a myeloproliferative disorder (MPD), MPD, and MDS, respectively. X-autosome translocations other than t(X;10)(p11;p11) and t(X;11)(q13;q23) have not been reported as recurrent or primary changes in hematological disorders. In the present study, a t(X;9)(q26;q22) and t(X;5)(q13;q33) as the sole anomaly were found in cases 7 and 8, respectively. Both cases had the same diagnosis of MDS. Considering that trisomies 4 (+4) and 8 (+8) are common anomalies in MDS and ANLL, our findings strongly indicate that amplification of genes on 4p and 8p, but not on 4q and 8q, may play a crucial role in the pathogenesis of MDS and ANLL. In addition, genes on 18q12-23 and on Xq13-26 may be involved in the pathogenesis of myeloid disorders.

PHA/IL2: an efficient mitogen cocktail for cytogenetic studies of non-Hodgkin lymphoma and chronic lymphocytic leukemia.

This study documents the utility of a mitogen/cytokine cocktail composed of phytohemagglutinin and Interleukin 2 (PHA/IL2) used to stimulate cultures from patients with chronic lymphoproliferative disorders. We report the results of a selected series of 57 patients with non-Hodgkin lymphoma (NHL) or chronic lymphocytic leukemia (CLL), in which only the culture stimulated with PHA/IL2 demonstrated the presence of an abnormal clone. On average, cells in the abnormal clone comprised 40% of the mitotic cells in this culture. The most common abnormalities observed in these patients were trisomy 12, present in 39% of the cases, and t(14;18), seen in 14% of cases.

Identification of a ring chromosome in a myxoid malignant fibrous histiocytoma with chromosome microdissection and fluorescence in situ hybridization.

We investigated the origin of a ring chromosome in a myxoid malignant fibrous histiocytoma (MFH) by microdissection and fluorescence in situ hybridization (FISH) analyses. Cytogenetically, only two ring chromosomes were observed; the smaller ring was seen more frequently. The latter was microdissected, and the material used for FISH. Hybridization of the microdissected labeled DNA to normal metaphase cells revealed that the signal localized only to 20q. Three signals were seen in the tumor cells using either the microdissected 20q probe or chromosome 20 centromeric probe, indicating the involvement of both the long arm and the centromere in the ring chromosome. The short arm of chromosome 20 did not appear to be involved in the formation of the ring chromosome.

Clear-cell and papillary carcinoma of the kidney: an analysis of chromosome 3, 7, and 17 abnormalities by microsatellite amplification, cytogenetics, and fluorescence in situ hybridization.

Clear-cell and papillary renal cell carcinomas (RCCs) have specific genetic changes that allow them to be classified on the basis of histopathology and on the basis of cytogenetic and molecular genetic findings. Clear-cell carcinomas are characterized by a deletion of gene sequences on the short arm of chromosome 3 (3p). Papillary RCCs do not have 3p deletions but have an increase in chromosomal number that usually includes trisomies of chromosomes 7 and 17. This study was undertaken to determine whether PCR-amplified DNA microsatellites can be used to detect numerical abnormalities of chromosomes 7 and 17 and whether the numerical abnormalities and 3p deletions that are detected by microsatellite analysis can be correlated with histopathologic tumor types. A series of histologically unambiguous RCCs consisting of three papillary and ten clear-cell RCCs were studied by cytogenetics and by fluorescence in situ hybridization (FISH) with chromosome 7 and 17 centromeric probes. Microsatellites on the long and short arms of chromosomes 3, 7, and 17 were amplified in paired normal tissue and tumor samples, and the reaction products were analyzed for differences between the normal and the tumor allele ratios. Clear-cell carcinomas showed loss of heterozygosity (LOH) of 3p but not 3q alleles in eight of ten cases. LOH of 3p and 3q was seen in one case of papillary RCC that cytogenetically had two normal chromosomes 3. This indicated a nondisjunction duplication that could be confused with monosomy 3 if only microsatellite studies were performed. Differences in microsatellite allele ratios between normal tissue and tumor correlated with the presence of trisomy 7 that was identified in clear-cell and papillary RCCs by cytogenetics and by FISH. Microsatellite analysis did not detect numerical chromosome 17 abnormalities in the papillary RCCs but did show an abnormality in one clear-cell carcinoma that was markedly aneusomic for chromosomes 7 and 17 by FISH. In this collection of cases, microsatellite amplification genetically distinguished only clear-cell RCCs showing 3p but not 3q LOH as a separate class of tumors. The method detected abnormalities in chromosome number that were found in both clear-cell and papillary RCCs.