Cancer Genetics

We report the progression of splenic marginal zone lymphoma (SMZL) with circulating villous lymphocytes to Burkitt lymphoma with the presence of a t(8;14)(q24;q32) followed by a highly aggressive course. While the initial indolent lymphoma had an IgM λ immunophenotype the Burkitt lymphoma was IgM κ-positive. Immunoglobulin heavy chain gene (IGH) sequence analysis showed no identity between the two clones. We conclude that Burkitt lymphoma can occur in patients with SMZL, although not necessarily of identical clonal origin.

Chronic myeloid leukemia (CML) is characterized by the presence of the Philadelphia chromosome (Ph), which results from a balanced translocation between chromosomes 9 and 22, the t(9;22)(q34;q11.2). In 5e10% of the cases, variants of the Ph (vPh) are detected, involving various breakpoints in addition to 9q34 and 22q11.2. Deletions on the der(9) and der(22) can be detected in approximately 10e15% of CML patients. The frequency of a deletion of the der(9) in vPh CML is variable. Most studies have shown high frequencies (30e45%) in this subgroup. We report the cytogenetic evaluation of 25 vPh cases, which represents 6.8% of the CML cases diagnosed at one institution in 20 years. The breakpoints of the partners of the vPh in our patients agree with those reported previously, except for a novel 18q23. We found a low incidence of deletions of the der(9) (10%) and der(22) (5%) in these patients, contrasting with several reports in the literature. This finding may reflect the extensive spectrum of aberrations in vPh, and the possibility that a considerable group of these aberrations may not affect the genetic stability of 5 0 ABL1 and 3 0 BCR. Epidemiologic differences may also exist and could explain our results. These differences would require further investigation.

In normal state of a cell, endogenous antioxidant enzyme system maintains the level of reactive oxygen species generated by mitochondrial respiratory chain. Mitochondrial superoxide dismutase [SOD; manganese SOD (MnSOD) or SOD2] neutralizes highly reactive superoxide radical (O.-2), the first member in the plethora of mitochondrial reactive oxygen species. A polymorphism in the target sequence of MnSOD enzyme, Val16Ala, is known to disrupt proper targeting of the enzyme from cytosol to mitochondrial matrix
where it acts on O.-2 to dismutate it to hydrogen peroxide (H2O2). A change in the level of O.-2 and of H2O2 in mitochondria modulates the molecular mechanisms of apoptosis, cellular adhesion, and cell proliferation and thus play key role in cancer development. Previous studies investigating the association between MnSOD Val16Ala polymorphism and cancer risk have revealed inconsistent results. We conducted a meta-analysis on these studies. Our meta-analysis on total of 7,366 cancer cases and 9,102 controls from 13 published case-control studies showed no overall association of this polymorphism either with breast cancer risk or for cancer risk
as such (for Ala homozygous odds ratio, 0.98; 95% confidence interval, 0.90-1.07 and odds ratio, 1.02; 95% confidence interval, 0.91-1.14, respectively). Also, there was no major effect in either recessive or dominant model for the MnSOD Val16Ala. However, a proper evaluation of this polymorphism with cancer link demands experiments involving large sample size, cross-tabulation of gene-gene, gene-environment interactions, and linkage studies, as cell biological experiments clearly correlate critical levels of mitochondrial O.-2 and H2O2 to carcinogenesis.

Background: Postpartum hemorrhage is the most common cause of mortality in women with vaginal or cesarean delivery. WHO statistics shows that about 500 thousands women have died of complications related to pregnancy or during childbirth in 2013. haemorrhage probability is the main reason to order blood requests in delivery and cesarean units. The purpose of this study is to evaluate the ratio of cross-matched to transfused blood in pregnant women during one year before and after the implementation of health reform program. Materials and Methods: In this retrospective descriptive study, the requests of blood reserves for pregnant patients with the gravid of 34±7 weeks and the age of 30±16 years old in two periods, before and after the implementation of health reform program were being collected and compared. Blood group antiserums and anti-human globulins with bovine albumins were purchased from LORN company and Baharafshan Company respectively. Results: The total number of requests ...

Genetic testing has been implemented in clinical practice. However, data on physician's practices and education related to cancer genetics, risk assessment and clinical management in Greece, is limited. In Greece, genetic counseling is performed by treating physicians in collaboration with clinical laboratory geneticists due to the absence of medical geneticists and genetic counsellors. We evaluated treating physicians' experience on genetic testing for hereditary cancer and counseling practices in Greece, thus providing critical areas for improvement of genetic counseling processes. A 28-question survey was used to assess physicians' experience with genetic testing practices, factors that affect their clinical management and decision making and limitations in their education. Of 250 physicians, 208 (83%) completed the survey; of whom 89 (42.8%) were medical oncologists, 88 general surgeons (42.3%), 26 gynecologists (12.5%) and 5 (2.4%) of other specialties. Overall, 91.8% of participants referred patients for genetic testing, with 51.8% recommending multigene panel testing. While most clinicians (84%) reported lack of a clinical genetics department at their institution, 75.7% referred patients for genetic counseling at available departments or healthcare professionals with expertise in genetic counseling. Overall, 68.8% of respondents reported no training or moderate training on cancer genetics. A higher proportion of medical oncologists reported sufficient/very satisfactory training (40.9%) compared to general surgeons (27.3%) or gynecologists (11.5%) (p = 0.012). Time spent on pre-and post-testing sessions varied significantly among respondents. Of 199 physicians, 70% would manage patients with BRCA1 VUS as patients with pathogenic variants, mainly surgeons (83.1%) and gynecologists (80%), compared to oncologists (52.3%) (p < 0.001). Additionally, 64% of physicians treating patients with breast and ovarian cancer would recommend an intervention based on the presence of a BRCA1 VUS. Most respondents (87%) were interested in receiving additional education on cancer risk assessment. Limited consensus was observed during physicians' genetic testing, counseling practices and clinical management of patients with increased predisposition to cancer. Our findings highlight the need for improvement in physician education on cancer risk assessment and increase of genetic counseling resources and services.

The t(12;21) translocation is the most common translocation in childhood B-precursor ALL. Because G-banding cannot detect the cryptic t(12;21), the full characterization of additional aberrations involving chromosome 12 is often difficult. Spectral karyotyping (SKY) can distinguish a der(12)t(12;21) chromosome from a nonrearranged chromosome 12. We therefore applied SKY on an unselected series of 14 patients after G-banding analysis for the detection of additional chromosomal rearrangements and especially for aberrations involving chromosome 12. Chromosome 12 was the most frequently affected by structural aberrations, with involvement of both homologs. Structural aberrations of der(12)t(12;21) in addition to the typical t(12;21) were found in five patients and structural aberrations of the nonrearranged chromosome 12 were found in four patients. This study points to the susceptibility of der(12)t(12;21) and the other chromosome 12 homolog to further rearrangement. Ó

We report a case of acute myeloid leukemia (AML) subtype M2, with t(5;11)(q35;q13), in a 30year-old man. Conventional cytogenetic, spectral karyotyping, and fluorescence in situ hybridization (FISH) studies on bone marrow sample obtained at diagnosis revealed an abnormal karyotype in all cells examined. FISH analysis demonstrated absence of translocations in the region of the cyclin D1 gene and real-time quantitative reverse transcriptaseepolymerase chain reaction revealed normal expression of this gene. Similar to the 11q23 region, 11q13 changes can be found in both myeloid and lymphoid neoplasias with different chromosomes serving as donors in translocations. Ó

It gives me great pleasure to warmly welcome you here to Dubai and the UAE to conduct your important scientific deliberations in the field of human genomics. Your focus on 'Genomics of Human Diversity and Heritable Disorders' along with the 4 th Pan Arab Human Genetics Conference is extremely relevant and of great consequence globally. It is of particular interest to us here in the Arab region, for as many of you are acutely aware, the prevalence of genetic disorders is disproportionately high in our communities. Many factors contribute to this situation and we are looking forward to working with you in the future to address those factors.

It gives me great pleasure to warmly welcome you here to Dubai and the UAE to conduct your important scientific deliberations in the field of human genomics. Your focus on 'Genomics of Human Diversity and Heritable Disorders' along with the 4 th Pan Arab Human Genetics Conference is extremely relevant and of great consequence globally. It is of particular interest to us here in the Arab region, for as many of you are acutely aware, the prevalence of genetic disorders is disproportionately high in our communities. Many factors contribute to this situation and we are looking forward to working with you in the future to address those factors.

Individual cancer cells carry a bewildering number of distinct genomic alterations (e.g., copy number variations and mutations), making it a challenge to uncover genomic-driven mechanisms governing tumorigenesis. Here, we performed exome sequencing on several breast cancer cell lines that represent two subtypes, luminal and basal. We integrated these sequencing data and functional RNAi screening data (for the identification of genes that are essential for cell proliferation and survival) onto a human signaling network. Two subtype-specific networks that potentially represent core-signaling mechanisms underlying tumorigenesis were identified. Within both networks, we found that genes were differentially affected in different cell lines; i.e., in some cell lines a gene was identified through RNAi screening, whereas in others it was genomically altered. Interestingly, we found that highly connected network genes could be used to correctly classify breast tumors into subtypes on the basis of genomic alterations. Further, the networks effectively predicted subtype-specific drug targets, which were experimentally validated.

Nuclear factor kappa B (NFkB) a pro-infl ammatory transcription factor, plays a signifi cant role in carcinogenesis. An A to G variation detected in the 3’UTR of the NFkB1A gene has been linked to crohn’s disease, which in turn is associated with increased susceptibility to Colorectal cancer. No data are available regarding the frequencies of NFkB1A genotype in Malaysian population and its association with CRC susceptibility. So a case control study involving 118 CRC patients and 117 normal controls was carried out to determine the frequencies of A to G variant NFkB1A genotype in Malaysian population and to investigate its infl uence on CRC susceptibility risk, employing PCR-RFLP technique. On analyzing the CRC susceptibility associated risk, the AG genotype showed signifi cant risk with OR 1.920 (p = 0.020) and GG genotype showed non signifi cant risk with OR 1.24 (p = 0.469). When the heterozygous and homozygous variant genotypes were combined, the associated risk was signifi cantly more pronounced with OR 2.124 (p = 0.005). The results suggest that, the A to G variant genotype of NFkB1A has signifi cant infl uence on CRC predisposition in Malaysian population.

Background
DNA repair genes have critical role in protecting the genome against endogenous and environmental agents. We hypothesize that, polymorphisms DNA repair genes might be predisposing factors for CRC susceptibility. In order to check this hypothesis, a case control study was designed to examine the frequencies of polymorphism Lys751Gln of XPD/ERCC2, Arg399Gln of XRCC1 and Thr241Met of XRCC3 gene in Malaysian CRC patients and healthy controls and investigate the associated CRC susceptibility.
Methods
Genotyping for the above DNA repair polymorphisms were performed in 118 colorectal cancer patients and 118 healthy normal controls by using PCR-RFLP.
Results/Conclusion
No association was found between the XPD/ ERCC2 Lys751Gln, XRCC1 Arg399Gln and XRCC3 Thr241Met polymorphisms and risk of colorectal cancer susceptibility when studied singly or in combinations. Our preliminary results, showing the lack association of XPD/ERCC2 Lys751Gln, XRCC1 Arg399Gln and XRCC3 Thr241Met polymorphisms with CRC susceptibility risk, could be attributed to the small sample size, interaction of other polymorphic DNA repair genes and also low frequency of variant alleles for the polymorphisms studied in this population.

Cytogenetic analysis of 25 breast fibroadenomas (FA) showed clonal chromosome alterations in three cases. Insertion (12;?) (q15;?) and deletion (2) (q14q31 or q32) were detected as a sole change in cases I and 3, respectively. Case 2 displayed the karyotype 45, XX,t(1;16)(q25;q23;,add (7)(p14), rea(15), -1Z The present findings are discussed together with the reports on FA in the literature.

Deletions of the long arm of chromosome 20 represent a common chromosomal abnormality associated with myeloid malignancies, in particular with myeloproliferative disorders (MPD), myelodysplastic syndromes (MDS), and acute myeloid leukemia (AML). Using G-banding cytogenetic techniques, we found clones with del(20q) in 36 patients with hematological malignancies examined in our laboratory during the years 2001-2003: in 23 patients as a sole cytogenetic aberration and in 13 patients together with other chromosomal changes. Fluorescence in situ hybridization (FISH) with a probe specific for the 20q12 region was used in all cases to confirm the presence of the clone with deletion. For patients with additional or complex chromosomal rearrangements, multicolor FISH (M-FISH) analysis was performed. Statistical evaluation of the prognostic impact of sex, age, diagnosis, and karyotype was performed. The survival time correlated with the type of chromosomal aberration; no significant differences in survival were found for sex, age, and diagnosis. Ć 2005 Elsevier Inc. All rights reserved.

Classic chemotherapy has little or no specificity for cancer cells, normally resulting in low accumulation at the tumor region (inefficacy), and in severe side effects (toxicity). This challenge has resulted in the development of several delivery strategies for chemotherapy agents to improve their concentration at the tumor site, simultaneously increasing their anticancer efficacy, while reducing the associated adverse systemic effects. In this work, the potential of drug delivery strategies involving the use of nanocarriers for controlling the biodistribution of antitumor drugs is deeply revised: passive targeting (through the enhanced permeability and retention effect, EPR effect) and active targeting (including stimuli-sensitive carriers and ligand-mediated delivery). Special attention will be also focussed on the recent approaches for overcoming multi-drug resistance. Finally, a general view of the problem of “nanotoxicity” in cancer treatment is also given.

We have performed a cytogenetic analysis of 23 myelodysplastic syndromes (MDS) with complex karyotypes (CK) using GTG-banding and spectral karyotyping techniques. Fifty-five percent of cases were hypodiploid, 34% were hyperdiploid, and 11% were pseudodiploid. The most recurrent alterations were monosomy of chromosomes 18, 5, and 7; trisomy of chromosome 8; and deletion of 5q, 11q, and 12p. Ninety-two structural alterations were mostly identified as unbalanced. The chromosomes and regions more frequently affected were 16q12, 17p11, and 20q11. Eight of 92 structural alterations were reciprocal translocations. Two translocations were recurrent, t(X;20)(p11.4;q11.2) and der(17)t(5;17)(?;p11.2); each one was present in about 10% of cases (2 cases, t[X:20] and 3 cases, t[5:17]). Mutations of TP53 were observed in five cases (22%), all with rearrangements affecting 17p. Total or partial inactivation of TP53 was detected in six cases (26%) as a result of loss of either both copies (four cases) or just one copy (two cases). Fluorescence in situ hybridization analysis showed amplification of genes previously identified in myeloid and/or hematological processes, such as HER2neu, MLL, and AML1, which could represent frequent events in MDS with CK. Ć

Cytogenetic analysis of an acute myeloid leukemia (AML-M1) showed the karyotype 53, XY,t(6;21)(q22;q22), + 10, + 13, + 19, + 21. Only one AML with a massively hyperdiploid karyotype ('>50 chromosomes) and t(6;21) has been published before. A comparison of the two cases reveals similarities both with regard to the morphologic subtype (M1) and to which chromosomes were trisomic (+ 6, + 8, + 13, + 19, and + 21 were found in both cases). We surmise that the t(6;21) was the primary chromosomal abnormality and that the set of multiple trisomies occurred secondarily; this pattern of clona! evolution may have favored a more immature leukemic phenotype in these two cases than is regularly seen in t(6;21)-associated leukemias.

Background:
Colorectal cancer (CRC) results from the interaction between environmental exposures and genetic predisposition factors. Aims: A case control study was designed and to investigate the genotype frequencies of P53Arg72Pro polymorphism in Malaysian CRC patients and healthy controls and to determine the associated risk of this polymorphism with CRC predisposition.
Methods:
In this case-control study, peripheral blood samples of 202 sporadic CRC patients and 201 normal controls were collected, DNA extracted and genotyped using the polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) technique.
Results: Genotype analysis showed the frequency of homozygous variant (Pro/Pro) genotype (21%) to be significantly higher in cases compared to controls (13%), (p=0.013). On examining the association between variant genotypes and CRC risk, the Pro/Pro homozygous variant genotype showed significantly higher risk association with CRC susceptibility (OR: 2.047, CI: 1.063-4.044, p=0.033). When stratified according to age, we observed that, individuals aged above 50 years and carriers of pro/pro genotype had significantly higher risk with OR: 3.642, CI: 1.166-11.378, p=0.026. Conclusions:
Our results suggest that the codon 72 SNP which results in amino acid substitution of Arginine to Proline in cell cycle regulatory gene P53, is associated with sporadic CRC risk and carriers of Pro/Pro genotype and more than 50 years old may have high susceptibility.

Esta investigación presenta la propuesta filosófica de Xavier Zubiri como nuevo paradigma desde el que definir el desarrollo orgánico y comprender el dinamismo del embrión preimplantatorio. Además, pone de manifiesto las ambigüedades terminológicas que acompañan a las explicaciones sobre el hecho biológico en las descripciones científicas y en las reflexiones de carácter interdisciplinar. En la investigación se expone la teoría de la sustantividad y de la esencia de Xavier Zubiri, y se describe minuciosamente el desarrollo del embrión preimplantatorio a la luz de los datos científicos actuales, prestando especial atención a la epigénesis. La investigación muestra que desde el planteamiento zubiriano se puede, por un lado, describir con precisión la imbricación entre lo genético y lo epigenético y, por otro lado, definir, sin perder la unidad del organismo, la noción de desarrollo. Además, se realiza un análisis crítico de las principales posturas sobre el estatus ontológico del embrión humano y se muestra las ventajas de su abordaje desde el prisma zubiriano.

Women with clinically node-negative breast cancer have a better prognosis than do those with axillary lymph node metastasis. Nonetheless,~20% of node-negative patients die within 15 years of diagnosis, and thus additional prognostic markers are greatly needed. To identify specific copy number alterations (CNAs) that differed in frequency between 10-year survivors and deceased patients with node-negative breast cancer, array comparative genomic hybridization (aCGH) was applied to 41 primary node-negative breast tumors. Fisher's exact test was used to identify significantly different CNAs between 10-year survivors and deceased patients. Losses at 8p21.2~p21.3, 8p23.1~p23.2, Xp21.3, and Xp22.31~p22.33 were significantly more common in tumors from deceased patients, suggesting that these alterations may contribute to tumor aggressiveness. Gains at 1q25.2~q25.3 and 1q31.3~q41 were more prevalent in tumors from survivors; specific gains at these genomic regions may inhibit further tumor progression, resulting in a less aggressive form of node-negative breast cancer. Evaluation of the identified CNAs in an independent external data set verified the prognostic potential of the 1q31.3~q41 region. Although further extensive validation is needed, the prognostic CNAs identified in this work may in time facilitate the clinical assessment of breast cancer. Ó

Partial trisomy 2p is typically associated with partial monosomy of another chromosomal segment and results from a balanced translocation in one of the parents. Inverted duplications with terminal deletions have been reported in an increasing number of chromosomes. Several cases initially interpreted as terminal duplications have instead been documented to represent inverted duplications with terminal deletions. Inv dup del(2p) has been reported in patients who manifest the clinical findings of trisomy 2p syndrome. Here we report on a 2-month-old girl with inv dup del(2p) and clinical manifestations that overlap those found commonly in partial 2p trisomy, as previously reported in the literature. Her clinical picture helps delineate the phenotype of 2p duplication disorders. © 2009 Wiley-Liss, Inc.

Cytogenetic analysis of a metastasis of a human testicular germ cell tumor (seminoma) revea/ed multiple numerical and structural anomalies, including an abnormally banding region (ABR) present on the short arm of one of the chromosome 12 homologs. Fluorescence in situ-and comparative genomic hybridization experiments revealed that the ABB results from the amplification of 12p11.2-p12.1 derived sequences. We speculate that this particular region may harbor gene(s) relevant for testicular germ cell tumor progression.

A novel three-way t(7;21;8)(q11.2;q22;q22) in a patient with acute myeloid leukemia Acute myeloid leukemia (AML) with t(8;21)(q22;q22) is well characterized by its unique morphology of AML-M2 subtype and favorable prognosis [1]. This recurrent translocation is often associated with additional chromosomal changes such as deletion of chromosome 9, loss of sex chromosomes, and other numerical abnormalities. Variants of t(8;21) with three-or four-way translocations are infrequently detected. The clinicopathologic features of AML with such variants are less well characterized . Here, we report a novel case of three-way t(7;21;8)(q11.2;q22;q22) translocation in a patient with AML.

Chromosome aberrations observed at diagnosis are considered to be the most valuable prognostic factors in acute myeloid leukemia (AML). Some specific aberrations vary in frequency among different geographical areas and ethnic groups. There are only limited studies on the role of such variability in AML patients. Here, we report the results of a cytogenetic study on 63 ethnic Omani patients with de novo AML: 18 children (<16 years) and 45 adults. By sex, 41 were male and 22 female; median age at diagnosis was 25 years. The morphological diagnosis was based on the FrencheAmericaneBritish (FAB) WHO criteria. Chromosome abnormalities were present in 39 of 63 patients (62% overall, or 44% for adults and 18% for children). Karyotypes with a sole abnormality accounted for 20 of 63 patients (32%). Chromosome abnormalities were more common in patients with the FAB-M2 subtype (15 of 22; 68%), which was also the most frequent subtype observed (22 of 63; 35%). Among the normal karyotypes (24 of 63; 38%), M2 subtype was the also most frequent (7 of 24; 29%), followed by M4 (4 of 24; 17%). Balanced translocations, t(8;21) and t(15;17) were observed in 7 of 63 (11%) and 6 of 63 (10%), respectively. Inv(16) was seen in 2 of 63 (3%). Trisomy 8 was the most frequent numerical anomaly, found in 7 of 63 (11%). Monosomy 7 was seen in 3 of 63 (5%). The patterns in our study were similar to those reported from Saudi Arabia and Kuwait, but the frequency of abnormalities varied. Our population differed morphologically, with the M2 subtype as most common, whereas M4 and M3 were more commonly in those reports. A comparison of our findings was made with other geographic and ethnic groups. This is the first systematic cytogenetic study of an ethnic Omani population. Ó

We describe a new family with a novel germline BAP1 nonsense mutation, c.723T>G, which leads to a predicted truncated protein, p.Y241*, or nonsense-mediated decay of the BAP1 mRNA. The proband had uveal melanoma (UM), and his paternal family has a remarkable history of multiple cancers. The proband's father had both pleural malignant mesothelioma (MM) and cutaneous melanoma (CM); a paternal uncle had lung cancer, CM, and UM; and a grandmother had CM. The findings in this family provide further support for the existence of a BAP1 cancer syndrome that predisposes to MM, various melanocytic neoplasms, and potentially other cancers. The fact that several members of the family manifested two or more different types of cancer suggests widespread BAP1-related tumor susceptibility targeting tissues of multiple organs. In addition, a review of BAP1 cancer syndrome families reported to date indicates that the location of the BAP1 mutation does not have any bearing on the spectrum of cancer types observed, either for mesothelial or melanocytic tumors.

Malignant peripheral nerve sheath tumors (MPNST) are rare soft-tissue malignancies. The genetic basis of these tumors is still poorly understood. Cytogenetic analyses predominantly revealed complex karyotypes, precluding the identification of recurrent chromosomal changes. We report loss of 1p material in a near-diploid karyotype with few or no additional structural chromosome changes in two sporadic cases of MPNST, indicating an important role of 1p loss in MPNST development. In one of these two tumors, a distal 1p deletion (1p31.2 ‫ف‬ pter) was detected suggesting involvement of a tumor suppressor gene located within this distal region of 1p. Further evidence for recurrent 1p loss in MPNST was obtained by interphase fluorescence in situ hybridization, which showed loss of 1p material in 3 out of 13 tumors. These findings together with data from the literature suggest that loss of a tumor suppressor gene located within distal 1p is implicated in the pathogenesis of MPNST.

Perlman syndrome is a congenital overgrowth syndrome inherited in an autosomal recessive manner that is associated with Wilms tumor susceptibility. We mapped a previously unknown susceptibility locus to 2q37.1 and identified germline mutations in DIS3L2, a homolog of the Schizosaccharomyces pombe dis3 gene, in individuals with Perlman syndrome. Yeast dis3 mutant strains have mitotic abnormalities. Yeast Dis3 and its human homologs, DIS3 and DIS3L1, have exoribonuclease activity and bind to the core RNA exosome complex. DIS3L2 has a different intracellular localization and lacks the PIN domain found in DIS3 and DIS3L1; nevertheless, we show that DIS3L2 has exonuclease activity. DIS3L2 inactivation was associated with mitotic abnormalities and altered expression of mitotic checkpoint proteins. DIS3L2 overexpression suppressed the growth of human cancer cell lines, and knockdown enhanced the growth of these cells. We also detected evidence of DIS3L2 mutations in sporadic Wilms tumor. These observations suggest that DIS3L2 has a critical role in RNA metabolism and is essential for the regulation of cell growth and division.

The mitochondrial DNA 4977-bp deletion (6mtDNA4977) has been explored in various cancers, but its predictive or prognostic role in esophageal cancer is poorly understood. The objective of the present study was to investigate a possible role of 6mtDNA4977 in susceptibility and prognosis of esophageal cancer in a northern Indian population. The study was performed in 39 histopathologically confirmed cases with esophageal cancer. Tumor, normal tissues, and intravenous blood samples were taken for detection of 6mtDNA4977 through a duplex polymerase chain reaction technique. 6mtDNA4977 was detected in two tumors and one adjacent normal tissue sample, but in none of the blood samples. All three patients with 6mtDNA4977 were male, with squamous cell carcinoma in the middle third of the esophagus. Survival analysis suggested a role of 6mtDNA4977 in prognosis of esophageal cancer patients. Despite the low frequency of 6mtDNA4977 in esophageal cancer patients of northern India, this feature may have a role in esophageal cancer progression and prediction of survival outcome. Ó

Long-standing gallstones are generally present in 65e80% patients of gallbladder cancer (GBC). It has also been suggested that inflammation caused by gallstones may be involved in the development of GBC. Interleukin-1 receptor antagonist (IL-1RN) and interleukin-1 beta (IL-1B) are proinflammatory cytokine genes at the interleukin-1 locus, and polymorphisms of these genes have been associated with various inflammatory diseases. The aim of this study was to investigate whether polymorphism in the IL-1RN and IL-1B genes are associated with GBC patients with and without gallstones. Polymorphisms within the IL-1RN 86ebase pair VNTR (variable number tandem repeat) and IL-1B (À511C / T) were genotyped using polymerase chain reaction (PCR) and PCR restriction fragment length polymorphism in 166 healthy subjects and 124 GBC patients. The frequency of the IL-1RN, VNTR 2/2 genotype was significantly higher in GBC patients [P 5 0.017; odds ratio (OR) 5 3.25; 95% confidence interval (CI) 5 1.23e8.58]. CC genotype and 'C' allele of the e511IL-1B C / T polymorphism also showed high risk for GBC (P 5 0.033; OR 5 3.36; 95%CI 5 1.52e7.43, P 5 0.047, OR 5 1.41; 95%CI 5 1.00e1.98, respectively). The higher cancer risk due to the IL-1RN, 2/2 genotype was observed in GBC patients with or without stones (P 5 0.038; OR 5 3.58; 95%CI 5 1.08e11.65, P 5 0.035; OR 5 3.33; 95%CI 5 1.08e10.61). Risk due to the CC genotype of IL-1B, however, was confined to GBC patients harboring gallstones (P 5 0.0003; OR 5 6.92; 95%CI 5 2.65e18.03). The haplotype 1/C of IL-1RN and IL-1B was found to confer a significantly enhanced risk of GBC in cancer patients with gallstones (P 5 0.022; OR 5 2.19; 95%CI 5 1.12e4.27), while higher risk resulting from 2/C haplotype was of borderline significance (P 5 0.061; OR 5 3.04; 95%CI 5 0.95e9.70). Individuals with 1/C and 2/C haplotypes of IL-1RN VNTR and e511IL-1B C / T polymorphisms were more susceptible to develop GBC with gallstones compared to healthy controls in north India. Ó

BCR/ABL1-negative chronic myeloproliferative neoplasms (CMPNs) are a heterogeneous group of clonal hematological malignancies. Over recent years, some genetic events in tyrosine kinase (TK) genes have been described as causal events of these diseases. To identify new genetic aberrations underlying these diseases, we used denaturing high performance liquid chromatography and fluorescence in situ hybridization (FISH) to analyze 17 genes from two receptor-TK families (III and IV) and from three cytoplasmic-TK families (Syk, Abl, and Jak) on samples from 44 BCR/ ABL1-negative and JAK2(V617F)-negative CMPN patients with different clinical phenotypes. Although screening by FISH did not reveal novel chromosomal aberrations, several sequence changes were detected. None of them were frequent events, but we identified a new potential activating mutation in the FERM domain of JAK2(R340Q). None of the germline JAK2(V617F) singlenucleotide polymorphisms detected differed in distribution between patients and control subjects. In summary, data presented here show that these genes are not frequently mutated or rearranged in CMPNs, suggesting that molecular events causing these disorders must be located in other genes. Ó

Fluorescence in situ hybridization (FISH), as a new clinical test, is not presently standardized. For practical reasons, each laboratory must build its own criteria. In this work, we present our standardization criteria for clinical practice, which include not only the methods for cell fixation, specimen preparation, and hybridization conditions, but mainly the definition of false-positive range and the scoring criteria of microscopic analysis. These include signal assessment, difference between individual microscopists, evaluation of specimen homogeneity, and the minimum number of scored nuclei required for a clinically reliable result. For this purpose, we analyzed by FISH 24 healthy volunteer donors, 31 patients affected by non-chronic myelogenous leukemia (CML) hematological malignancies, 47 CML patients at diagnosis, and 82 CML patients during treatment for the BCR/ABL fusion. In this article, we present several quality control and assurance methods that can be useful in providing standardization of the FISH technique.

In this study of ovarian carcinoma, we extended previous findings by performing FISH using chromosome 19 paint and microFISH probes and patient samples with and without abnormalities of chromosome 19 identified by G-banding. Karyotype interpretations of der(19) were confirmed, while additional 19 traJ~slocations were also detected by FISH with 19WCP in some cases. Similar FISH studies of ovarian carcinoma cell lines found chromosome 19 abnormalities even after extensive in vitro culture. MicroFISH probes were generated by chromosome microdissection from two cases with hsr(19) and mapped to :I9q13.2 and 19q13.1-.2, respectively. FISH with these microFISH probes alone or in combination with a 19 WCP probe to four patient samples and seven cell lines showed that 65% of chromosome 19 structural abnormalities contained 19q13.1-q13.2 sequences, sometimes as large hsrs. Ovarian cancer cell L!nes showed amplification and overexpression of the AKT2 putative oncogene, but not the ERCC-2 DNA repair gene in this chromosomal region. In addition to AKT2, amplification and overexpression of other yet-unidentified genes in the 19q13.1-q13.2 region may contribute to ovarian carcinoma pathogenesis or progression.

Synovial sarcomas are high-grade malignant mesenchymal tumors that account for 10% of all softtissue sarcomas. Almost 95% of these tumors are characterized by a nonrandom chromosomal abnormality, t(X;18)(p11.2;q11.2), that is observed in both biphasic and monophasic variants. In this article, we present the case of a 57-year-old woman diagnosed with high-grade biphasic synovial sarcoma in which conventional cytogenetic analysis revealed the constant presence of a unique t(18;22)(q12;q13), in addition to trisomy 8. The rearrangement was confirmed by fluorescence in situ hybridization. The use of the whole chromosome painting probes WCPX did not detect any rearrangements involving chromosome X, although reverse-transcriptase polymerase chain reaction (PCR) analysis demonstrated the conspicuous presence of a SYT/SXX1 fusion gene. Spectral karyotyping (SKY) was also performed and revealed an insertion of material from chromosome 18 into one of the X chromosomes at position Xp11.2. Thus, the karyotype was subsequently interpreted as 47,X,der(X)ins(X;18) (p11.2;q11.2q11.2),der(18)del(18)(q11.2q11.2)t(18;22)(q12;q13),der(22)t(18;22). Real-time PCR analysis of BCL2 expression in the tumor sample showed a 433-fold increase. This rare finding exemplifies that thorough molecularecytogenetic analyses are required to elucidate complex and/or cryptic tumor-specific translocations. Ó