bluuebunny/crossref_metadata_2025 huggingface dataset:
Histochemical staining and quantification of dihydrolipoamide dehydrogenase diaphorase activity using blue native PAGE
Authors: Yan Liang‐Jun , Yang Shao‐Hua , Shu Hongjun , Prokai Laszlo , Forster Michael J.
Published: February 2007

Mammalian mitochondrial dihydrolipoamide dehydrogenase (DLDH, EC 1.8.1.4) catalyzes NAD+‐dependent oxidation of dihydrolipoamide in vivo and can also act as a diaphorase catalyzing in vitro nicotinamide adenine dinucleotide (reduced form) (NADH)‐dependent reduction of electron‐accepting molecules such as ubiquinone and nitroblue tetrazolium (NBT). In this paper, we report a gel‐based method for histochemical staining and quantification of DLDH diaphorase activity using blue native PAGE (BN‐PAGE). Rat brain mitochondrial extracts, used as the source of DLDH, were resolved by nongradient BN‐PAGE (9%), which was followed by diaphorase activity staining using NADH as the electron donor and NBT as the electron acceptor. It was shown that activity staining of DLDH diaphorase was both protein amount‐ and time‐dependent. Moreover, this in‐gel activity‐staining method was demonstrated to be in good agreement with the conventional spectrophotometric method that measures DLDH dehydrogenase activity using dihydrolipoamide as the substrate. The method was applied to determine levels of DLDH diaphorase activity in several rat tissues other than the brain, and the results indicated a similar level of DLDH diaphorase activity for all the tissues examined. Finally, the effects of thiol‐reactive reagents such as N‐ethylmaleimide (NEM) and nitric oxide donors on DLDH diaphorase activity were evaluated, demonstrating that, with this method, DLDH diaphorase activity can be determined without having to remove these thiol‐reactive reagents that may otherwise interfere with spectrophotometric measurement of DLDH dehydrogenase activity. The gel‐based method can also be used as a means to isolate mitochondrial DLDH that is to be analyzed by mass spectral techniques in studying DLDH post‐translational modifications.
EDTA alkaline elution characteristics and measurement of DNA damage in unlabeled DNA using hoechst 33258 fluorescence
Authors: Ducore Jonathan M.
Published: February 2007

Hoechst 33258 fluorescence of single stranded DNA has been used to perform alkaline elution with unlabeled DNA. The high background fluorescence of “standard” elution solutions has prompted others to use EDTA but the elution characteristics of DNA in EDTA‐containing solutions and the comparability of results with those using “standard” tetrapropyl ammonium hydroxide solutions have not previously been examined. We report here the elution characteristics of DNA in EDTA and the relevant parameters for the successful use of EDTA as an elution solution. An increase in elution pH to 12.4 is required but elution solutions of higher pH cause alkaline hydrolysis of undamaged DNA. Drug‐treated DNA from which DNA‐protein crosslinks have been removed can be completely removed from the filters at the end of the elution by a Pronase filter digestion. The simplest and most efficient removal of DNA‐protein crosslinks is through the inclusion of proteinase‐K in an SDS containing lysis solution. EDTA elution can measure interstrand crosslinks and single strand breaks as easily as is performed using radiolabeled DNA under “standard” elution conditions and requires only 1.5–2 × 106 cells per elution filter. DNA‐protein crosslinking measurements were unsatisfactory, however, since even the Pronase digestion failed to completely remove protein‐crosslinked DNA from the elution filters.

Portions of this work were presented at the 77th Meeting of the American Association for Cancer Research, May 1986.
Measurement of unscheduled DNA synthesis and S‐phase synthesis in rodent hepatocytes following in vivo treatment: Testing of 24 compounds
Authors: Mirsalis Jon C. , Tyson C. Kim , Steinmetz Karen L. , Loh Erica K. , Hamilton Carol M. , Bakke James P. , Spalding Judson W.
Published: February 2007

The in vivo‐in vitro hepatocyte DNA repair assay has been shown to be useful for studying genotoxic hepatocarcinogens. In addition, measurement of S‐phase synthesis (SPS) provides an indirect indicator of hepatocellular proliferation, which may be an important mechanism in rodent carcinogenesis. This assay was used to examine 24 chemicals for their ability to induce unscheduled DNA synthesis (UDS) or SPS in Fischer‐344 rats or B6C3F1 mice following in vivo treatment. Hepatocytes were isolated by liver perfusion and incubated with 3H‐thymidine following in vivo treatment by gavage. UDS was measured by quantitative autoradiography as net grains/ nucleus (NG). Controls from both sexes of both species yielded <0.0 NG. Chemicals chosen for testing were from the National Toxicology Program (NTP) genetic toxicology testing program and most were also evaluated in long‐term animal studies conducted by the NTP. 11‐Aminoun‐decanoic acid, benzyl acetate, bis(2‐chloro‐1‐methylethyl)ether (BCMEE), C.I. Solvent Yellow 14, cinnamaldehyde, cinnamyl anthranilate, dichloromethane, dichlorvos, glutaraldehyde, 4,4′‐methylenedianiline (MDA), 4‐nitrotoluene, 4,4′‐oxydianiline, a polybrominated biphenyl mixture (PBB), reserpine, 1,1,2,2‐tetrachloroethane, 1,1,2‐trichloroethane, trichloroethylene, and 2,6‐xylidine all failed to induce UDS in rats and/or mice. Dinitrotoluene and Michler's Ketone induced positive UDS response in rat, while N‐nitrosodiethanolamine and selenium sulfide induced equivocal UDS results in mouse and rat, respectively. BCMEE, bromoform, chloroform, PBB, 1,1,2‐trichloroethane, and trichloroethylene were all potent inducers of SPS in mouse liver, while C.I. Solvent Yellow 14, and 1,1,2,2‐tetrachloroethane yielded equivocal SPS results in rat and mouse, respectively. These results indicate that most of the test compounds do not induce UDS in the liver; however, the significant S‐phase response induced by many of these compounds, especially the halogenated solvents, may be an important mechanism in their hepatocarcinogenicity.
Comparative study of the mutagenic and genotoxic activity associated with inhalable particulate matter in Rio de Janeiro air
Authors: Miguel Ann G. , Daisey Joan M. , Sousa Judy A.
Published: February 2007

We have determined the genotoxic and mutagenic activities associated with inhalable particulate matter (IPM) collected in Rio de Janeiro, Brazil, Camden, NJ, and Caldecott Tunnel, CA, and used these results to compare three different bioassays. Samples collected every 12 hr (Rio) or every 24 hr (Camden) were extracted sequentially with cyclohexane (CX), dichloromethane (DCM), and acetone (ACE), for a rough fractionation by polarity, and composites of the extracts were tested for mutagenicity using the Salmonella frame shift (TA98) and base substitution (TA100) tester strains, as well as for genotoxicity using the Rossman Microscreen bioassay based on the induction of γ‐prophage in a lysogenic Escherichia coli strain. All samples were tested without and with S9 metabolic activation. Maximum mutagenic and genotoxic activities were in the nonpolar (CX) and polar (ACE) fractions, respectively, indicating that these two assays detect different classes of compounds with different efficiencies. Oxidative aging of the Rio aerosol is indicated by a shift in activities in both tests from the less polar fractions in the day to the polar (ACE) fraction at night. The Rio TA98 mutagenic (18 rev/m3) and genotoxic (1.4 × 105 PFU/m3) activities were higher than those fcr Camden, an Eastern U.S. city, by factors of 1.4 and 2.8, respectively.
Sister chromatid exchange and chromosome aberrations in mice after in vivo exposure of green S‐A food colorant
Authors: Giri A. K. , Sivam S. Sai , Khan K. A. , Sethi N.
Published: February 2007

Sister chromatic exchanges (SCE) and chromosome aberrations (CA) in mice after in vivo exposure of Green S were carried out following single acute treatment. Except for the lowest dose (25 mg/kg body weight) a significant increase in the SCEs were observed in all the other doses (50, 100, and 200 mg/kg) tested. In CA study two higher doses (200 and 400 mg/kg) showed a significant increase in CA when compared with control. The minimum effective dose which induced SCE and CA was 50 and 200 mg/kg of body weight, respectively. The trend tests for the evidence of dose response effects were also significant for both SCE and CA. No significant differences were observed in cell replication kinetic (Rl) analysis. A significant increase in the mitotic index (Ml) was also observed in the highest dose (400 mg/kg) tested when compared with control. Thus the present study indicates that Green S can induce both SCE and CA in vivo in bone marrow cells of mice.
Genetic method for pre‐classification of genotoxins into monofunctional or cross‐linking agents
Authors: Vogel Ekkehart W. , Zijlstra Jaap A. , Nivard Madeleine J. M.
Published: February 2007

To characterize environmental carcinogens, there is a need to distinguish monofunctional genotoxic agents from those having cross‐linking potential, because chemicals which can cross‐link DNA are among the most potent carcinogens in rodents [Barbin and Bartsch, 1989] and humans [Allen et al., 1988; Kaldor et al., 1988]. Here we provide a genetic method for a pre‐classification of genotoxins with respect to their functionality—monofunctional versus cross‐linking. The procedure is based on the determination of relative clastogenic efficiency by a two‐endpoint comparison in Drosophila: (i) induction of chromosome loss (CL), (ii) incidence of recessive lethal mutations (RL). Analysis of CL/RL ratios of 53 genotoxins, all mutagens in Drosophila, permitted distinction of 45 into two major categories: (i) 21 monofunctional agents with CL/RL indices generally ≤1; (ii) 24 agents with ratios >2 exhibiting DNA cross‐linking properties. Within the group of monofunctional agents, CL/RL ratios tend to be low for SSN1 agents, i.e., for N‐ethyl‐N‐nitrosourea, N‐ethyl‐N′‐nitro‐N‐nitrosoguanidine, and for N‐nitroso‐diethylamine. With cross‐linking agents, the number of reactive groups appeared of minor importance as bi‐, tri‐, and tetrafunctional agents showed no significant differences in their CL/RL indices. Among 8 chemicals which could not be grouped into one of the two categories are two (adriamycin, daunomycin) regarded as intercalating agents. It is concluded that this two‐endpoint analysis in Drosophila has prognostic value and can assist in the characterization of genotoxic agents with unknown mode of action. © 1993 Wiley‐Liss, Inc.
Interchange and intra‐nuclear architecture
Authors: Savage John R. K.
Published: February 2007

For chromatin lesions to interact to form exchanges of any sort, it is obvious that contact between them must be made. However, the probability of such interaction is conditioned by other factors like time, initial separation, metabolic activity, and, in the case of chemically induced lesions, scheduled DNA synthesis.

The irradiated nucleus was, for a long time, regarded as a “bag of broken chromosomes” with the severed ends free to move around and find partners with which to form illegitimate reunions. Many of these would be seen at following metaphase as intra‐ and interchanges.

Evidence is rapidly accumulating which indicates that this picture of the nucleus is false. We know now that chromosomes occupy highly localised domains with limited movement, and that there is no massive intermingling; that much of the chromatin is compacted and splinted with proteins and so precluded from exchange‐type contact; that most of the chromatin is looped and “fixed” into an intra‐nuclear protein scaffold or skeleton; that some chromatin is spun‐out and associated with the nuclear envelope in the vicinity of the pore‐complexes.

Thus it would appear that movement, in the sense envisaged by early workers, is curtailed, and that only a proportion (probably a small proportion) of the chromatin is actually “at‐risk” with respect to interchange formation.

Where then does interchange take place? Are the “sites” pre‐existent, or can proximity requirements be realised after radiation exposure? In what ways will the intra‐nuclear architecture influence exchange? These are some of the questions which are considered in this paper. © 1993 Wiley‐Liss, Inc.
Genotoxic effects of the carbamate insecticide, methomyl. II. In vivo studies with pure compound and the technical formulation, “lannate 25”
Authors: Bolognesi Claudia , Peluso Marco , Degan Paolo , Rabboni Roberta , Munnia Armelle , Abbondandolo Angelo
Published: February 2007

The carbamate insecticide, methomyl, and the methomyl‐containing technical formulation, “Lannate 25”, were tested for the induction of DNA damage in vivo. Swiss CD1 mice were treated intraperitoneally with test substances and the following tests were performed: alkaline elution of liver and kidney DNA, 8‐hydroxy‐guanosine detection in liver DNA, and 32P‐postlabelling analysis of DNA adducts in liver DNA. The clastogenic activity of the two pesticide preparations was also evaluated as micronucleus frequency in bone marrow. No DNA adducts were detected in liver DNA of mice treated with pure methomyl, while a dose‐related increase in DNA adducts was found in Lannate 25‐treated animals. All other tests were positive with both methomyl and Lannate 25. A summary of genotoxic activity of methomyl is also presented. The hypothesis that the observed genotoxic effects of methomyl are induced indirectly, through formation of active oxygen species, is discussed. © 1994 Wiley‐Liss, Inc.
Preferential effects of the chemotherapeutic DNA crosslinking agent mitomycin C on inducible gene expression in vivo
Authors: Caron Rosemary M. , Hamilton Joshua W.
Published: February 2007

The immediate effects of a single dose of the chemotherapeutic DNA crosslinking agent, mitomycin C (MMC), on the expression of several constitutive and drug‐inducible genes were examined in a simple in vivo system, the 14 day chick embryo. We observed no effect of MMC on the steady‐state mRNA expression of the constitutively expressed β‐action, transferrin, or albumin genes. In contrast, MMC treatment significantly altered both the basal and drug‐inducible mRNA expression of two glutethimide‐inducible genes, 5‐aminolevulinic acid (ALA) synthase and cytochrome P450 CYP2H1. The basal expression of these genes was transiently but significantly increased over a 24 hr period following a single dose of MMC. Conversely, MMC significantly suppressed the glutethimide‐inducible expression of these genes when administered 1 to 24 hr prior to the inducing drug. The effects of MMC on both basal and drug‐inducible ALA synthase and CYP2H1 mRNA expression were principally a result of changes in the transcription rates of these genes. In contrast, MMC treatment had little or no effect on glutethimide‐induced expression of ALA synthase or CYP2H1. when administered 1 hr after the inducing drug, suggesting that a very early event in the induction process represents the target for these MMC effects. Covalent binding studies demonstrated that the effects of MMC on gene expression were closely correlated temporally with formation of [3H]‐porfiromycin‐DNA adducts. These results support the hypothesis that genotoxic chemicals specifically target their effects to inducible genes in vivo. © 1995 Wiley‐Liss, Inc.
Software package for the management of sequencing projects using lacl transgenic animals
Authors: De Boer Johan G.
Published: February 2007

The bacterial lacl gene has been used for many years as a mutational target for the study of the mechanisms of mutation. A wealth of information has been collected for many mutagenic treatments and in strains with diverse DNA repair backgrounds. Recently this gene has been used in the construction of a transgenic mouse, named Big Blue®, and a transgenic rat, as well as a rat cell line. The lacl gene in these animals and cells can conveniently be recovered and analyzed in bacteria. This makes it possible to study mutagenic potential of chemical compounds in vivo using a mammal. Tissue, strain, and gender specificity can be addressed. In addition, mutations recovered from tumour tissues or from animals with specific genetic backgrounds can be analyzed conveniently. The mammalian systems can produce large numbers of mutants that require computer assistance to manage the samples and the resulting DNA sequence data. Accordingly, a computer software system was develped. The system maintains an inventory of bacteriophage lambda lacl mutants and allows entry of mutant sequences while performing accuracy checks on the data. The software features several options for displaying lists of mutants. The system can perform several analyses, including mutant class compilations, mutational spectra comparisons, and clonal expansions analysis. An extensive database obtained from the bacterial lacl system is included with the software and can be analyzed along with mutants derived from transgenic animals. © 1995 Wiley‐Liss, Inc.
Proto‐oncogene amplification and homogeneously staining regions in human breast carcinomas
Authors: Saint‐Ruf Claude , Gerbault‐Seureau Michéle , Viegas‐Péquignot Evani , Zafrani Brigitte , Cassingena Roland , Dutrillaux Bernard
Published: February 2007

Cytogenetic studies on fresh human breast cancers revealed that homogeneously staining regions (HSRs), which are assumed to represent DNA amplification, are observed in almost half of the cases. To search for a possible relationship between HSRs and proto‐oncogene amplification, 16 proto‐oncogenes, including ERBB2, were studied by Southern blot analysis in four tumors with two or three HSRs, and in three tumors without HSRs. Only four proto‐oncogenes were found to be amplified in at least one tumor each: HST and INT2 (x3), MYC (x2–3), and FES (x > 10). The large sizes of the HSRs, which each corresponded to several percent of the haploid genome, were hardly compatible with the low rate of amplification, except for FES and then only if a large adjacent segment was co‐amplified. This incomplete correlation was demonstrated by in situ hybridization, using biotinylated probes, which showed fluorescent spots on only one HSR for FES in one tumor and for INT2 in another one. Our results indicate that most of the large amplifications corresponding to HSRs do not involve the proto‐oncogenes usually studied in breast cancer. The large amplification of FES, detected in one tumor, may be coincidental.
Cytogenetic findings in six posterior uveal melanomas: Involvement of chromosomes 3, 6, and 8
Authors: Sisley Karen , Rennie Ian G. , Cottam David W. , Potter Anthony M. , Potter Christopher W. , Rees Robert C.
Published: February 2007

Six posterior uveal melanomas were karyotyped after short‐term culture. One had a normal chromosome complement; the remaining five had limited chromosome changes. Involvement of chromosomes 1 and 6 was noted in two and four cases, respectively, and three ciliary body tumours demonstrated both monosomy 3 and i(8q).
The 12q13‐q15 translocation breakpoints in pleomorphic adenoma and clear‐cell sarcoma of tendons and aponeuroses are different from that in myxoid liposarcoma
Authors: Stenman Göran , Sahlin Pelle , Mark Joachim , Chaganti R. S. K. , Kindblom Lars‐Gunnar , Åman Pierre
Published: February 2007

The transcription factor gene CHOP was recently shown to be rearranged in myxoid liposarcoma with t(12;16)(q13;p11). We have analyzed whether the CHOP gene is the target of rearrangements in pleomorphic adenoma and clear‐cell sarcoma of tendons and aponeuroses with chromosome abnormalities of 12q13‐q15. Restriction fragment analysis showed that the CHOP gene and its flanking sequences were not rearranged in any of these tumor types, indicating that the 12q translocation breakpoints in pleomorphic adenoma and clear‐cell sarcoma are different from that in myxoid liposarcoma. © 1993 Wiley‐Liss, Inc.
Analysis of deletions of the long arm of chromosome 11 in hematologic malignancies with fluorescence in situ hybridization
Authors: Kobayashi Hirofumi , Espinosa Rafael , Fernald Anthony A. , Begy Catherine , Diaz Manuel O. , Beau Michelle M. Le , Rowley Janet D.
Published: February 2007

We studied samples containing deletions of the long arm of chromosome 11 (11 q) from patients with hematologic malignancies by using cytogenetic and fluorescence in situ hybridization (FISH) techniques. Cytogenetic analysis of 28 patients and of a cell line showed that all deletions included band 11q23. FISH analysis demonstrated that the proximal part of 11q23, including NCAM, was deleted in 13 of 15 patients and the cell line. Recurring chromosomal losses in human tumors have been regarded as evidence that the affected regions contain tumor‐suppressor genes. These results suggest that the putative tumor‐suppressor gene is proximal to the MLL gene which is also located in 11q23. © 1993 Wiley‐Liss, Inc.
Testicular germ cell tumors of adults show deletions of chromosomal bands 11 p13 and 11p15.5, but no abnormalities within the zinc‐finger regions and exons 2 and 6 of the wilms' Tumor 1 Gene
Authors: Looijenga Leendert H. J. , Gillis Ad J. M. , Oosterhuis J. Wolter , Abraham Mary , Sunders Grady F.
Published: February 2007

We have studied the involvement of chromosomal bands 11 p13 and 11p15.5 in 15 testicular seminomas (SE) and 18 testicular nonseminomatous germ cell tumors (NS). NO allelic imbalances were found in 40% of the SE and 44% of the NS. LOSS of heterozygosity (LOH) at 11 p 15.5 was seen in 21 % of the SE and 47% of the NS; the corresponding frequencies for 11 p 13 were 47% and 44%. Both regions were deleted in 13% of the SE and 44% of the NS, indicating that all NS with a complete LOH of 11 p 13 also lost the 11 p 15.5 region. In one (out of two) SE and in five (out of eight) NS, this was due to at least two separate deletions. Loss of the whole p‐arm was likely in one SE and two NS. No gross genomic changes of the Wilms' tumor 1 (WT1) tumor suppressor gene were found using a cDNA probe (WT33). Nor were aberrations found in the zinc‐finger regions and exons 2 and 6 of this gene, using polymerase chain reaction amplification, single stranded DNA polymorphism analysis, and sequencing. We suggest that loss of genetic information from the short arm of chromosome 11, without affecting the WT1 gene in the regions studied, is relatively frequent but not crucial in the pathogenesis of testicular germ cell tumors of adults. Genes Chrom Cancer 9:153‐160 (1994). © 1994 Wiley‐Liss, Inc.
Polysomy of chromosome 12 in 60 patients with non‐hodgkin's lymphoma assessed by fluorescence in situ hybridization: Differences between follicular and diffuse large cell lymphoma
Authors: Younes Anas , Goodacre Angela , Andreeff Michael , Pugh William , Katz Ruth , Rodriguez Maria A. , Hill Debbie , Cabanillas Fernando
Published: February 2007

Sixty consecutive evaluable specimens from patients with non‐Hodgkin's lymphoma (NHL) were studied for the incidence of polysomy of chromosome 12 by fluorescence in situ hybridization (FISH) with probes for the repetitive DNA sequence in the centromeric region of chromosome 12. Thirty‐six samples were from follicular lymphomas (FLs), and twenty‐four were from diffuse large cell lymphomas (DLCLs). Fifty‐two specimens (86%) were obtained by fine‐needle aspiration of a diseased node, seven (11.6%) were from involved bone marrows, and one specimen was from a pleural effusion. Twelve of the thirty‐six (33%) cases with FL had trisomy 12 in 3‐41% of the cells (median, 10%) (normal controls had three signals in 1.4 +M 0.7% of cells). Trisomy 12 was found in 62% of the patients who had had FL for more than 5 years. Nine of the twenty‐four cases (37%) with DLCL had more than two copies of chromosome 12 in 4‐92% of the cells (median, 78%), and all nine cases were of B‐cell phenotype. Unlike FL cells, some DLCL cells had 4‐6 copies of chromosome 12. In previously untreated patients, 54% of DLCLs and 26% of FLs had subpopulations of cells containing more than two copies of chromosome 12 (P = 0.04). Only 2/7 cases of DLCL with polysomy 12 had rearrangement of the BCL2 gene, indicating that the majority of DLCL cases with polysomy 12 did not result from histologic transformation of low grade follicular lymphomas. These data demonstrate that FISH of interphase cells is a sensitive method for detecting numerical abnormalities of chromosome 12 in NHL. The differences between FL and DLCL in the number of chromosomes 12 and the number of involved cells may be related to differences in the biology of these tumors. Genes Chrom Cancer 9:161‐167 (1994). © 1994 Wiley‐Liss, Inc.
Numerical chromosomal aberrations in thyroid tumors detected by double fluorescence in situ hybridization
Authors: Taruscio Domenica , Ried Thomas , Ward David C. , Carcangiu Maria Luisa
Published: February 2007

Double fluorescence in situ hybridization with DNA probes specific for the (peri)centromeric regions of chromosomes 3, 7, 9, 11, 12, 18, and X was performed on fresh isolated nuclei and frozen tissue sections prepared from 2 nodular hyperplasias, 2 adenomas, and 7 papillary carcinomas of the thyroid in order to detect numerical chromosomal changes. Numerical chromosomal aberrations were found in all malignant specimens examined. A consistent presence of at least two trisomies was detected in most cases, especially in the follicular variant specimens; the highest degree of trisomy was observed for chromosome 12. Isolated monosomies of moderate degree for different chromosomes were found in 1 adenoma and 2 papillary carcinomas. Severe monosomy of chromosome 9 was the only significant feature observed in the single metastatic papillary carcinoma. Genes Chrom Cancer 9:180‐185 (1994). © 1994 Wiley‐Liss, Inc.
Cytogenetic aberrations in 188 benign and borderline adipose tissue tumors
Authors: Mandahl Nils , Höglund Mattias , Mertens Fredrik , Mitelman Felix , Rydholm Anders , Willén Helena , Brosjö Otte
Published: February 2007

Chromosome studies of lipomas have revealed an extensive cytogenetic heterogeneity. To investigate the frequencies of previously recognized cytogenetic subgroups and to find out if more recurrent rearrangements can be identified, we have analyzed cytogenetically short‐term tissue cultures of 237 samples from 188 adipose tissue tumors obtained from 142 patients. Only one of 58 tumors from 18 patients with multiple lipomas (more than two tumors) had karyotypic changes. Among the sporadic lipomas, 20 tumors had supernumerary ring chromosomes of unknown origin, 55 had different aberrations involving chromosome segment 12q13‐15, 11 had changes of 6p or chromosome 13, but no rings or 12q13‐15 changes, and 14 had various other aberrations. Ring chromosomes were found in all cytogenetically abnormal lipomas histologically classified as atypical and in nine tumors classified as typical lipoma or spindle cell lipoma. Recombinations between 12q 13‐15 and a few other bands or segments were seen more than once: 3q27‐28 (15 tumors), 2p22‐24 and 2q35 (four tumors), 1 p32‐34 and 13q 12‐14 (three tumors), and 5q33 (two tumors). Recombinations of 12q 13‐15 with 2q35 and 13q 12‐14 have not been described before. Of eight tumors with chromosome 13 aberrations, five had loss of 13q material. Aberrations of 12q 13‐15, 6p, and/or chromosome 13 were found simultaneously in nine tumors. Two to four samples from the same tumor were investigated in 29 tumors with clonal aberrations. Thirteen of these tumors displayed clonal evolution, also noted in another 17 tumors in which only one sample had been investigated. Thus clonal evolution occurred in 30% of the tumors and was particularly frequent in atypical lipomas. Genes Chrom Cancer 9:207‐215 (1994). © 1994 Wiley‐Liss, Inc.
Deletion of chromosome arm 1p in a merkel cell carcinoma (MCC)
Authors: Gibas Zenon , Weil Susan , Chen Sing‐Tsung , McCue Peter A.
Published: February 2007

A case of neuroendocrine skin carcinoma (Merkel cell carcinoma) with a deletion of the short arm of chromosome 1 (1p) as the sole chromosomal abnormality was examined. The tumor originated in the skin of the left knee of a 67‐year‐old man. Histopathologic study showed an undifferentiated small cell tumor which expressed neuron‐specific enolase, chromogranin, and cytokeratin (CAM 5.2). Cytogenetic analysis of a lymph node metastasis from the groin showed a pseudodiploid cell population with a deletion of the short arm of chromosome 1 as the only abnormality: 46,XY,del( 1 )(p36.1). In situ hybridization with the DI22 probe specific for the terminal band of 1 p confirmed the terminal deletion. This is the first case of Merkel cell carcinoma in which only one chromosomal abnormality has been observed. Loss of the terminal portion of 1p suggests that a tumor supressor gene on 1p plays a role in the pathogenesis of Merkel cell carcinoma. Genes Chrom Cancer 9:216‐220 (1994). © 1994 Wiley‐Liss, Inc.
MDM2 gene amplification correlates with ring chromosomes in soft tissue tumors
Authors: Nilbert Mef , Mitelman Felix , Mandahl Nils , Rydholm Anders , Willén Helena
Published: February 2007

The human homolog of the murine double minute type 2 gene (MDM2) has been cloned and mapped to 12q 13‐14. The gene presumably functions as a cellular regulator and mediator of TP53 function. Amplification of the MDM2 gene has recently been observed in soft tissue sarcoma and in osteosarcoma. We studied MDM2 amplification in a series of 94 mesenchymal tumors and found 3‐20‐fold amplification in 20 tumors: in 10 of 49 malignant fibrous histiocytomas (MFH), in 1 of 2 pleomorphic liposarcomas, in 6 of 7 atypical lipomas, and in 3 of 12 typical lipomas. Normal hybridization patterns were detected in all 16 myxoid liposarcomas, in all 3 leiomyosarcomas, and in all 5 leiomyomas studied. The MDM2 amplification correlated with the presence of marker ring chromosomes; of the 10 MFH with MDM2 amplification, 5 had ring chromosomes, compared to 4 of 39 without MDM2 amplification, and all 9 lipomas with MDM2 amplification had ring chromosomes, in 5 of the tumors as the sole karyotypic anomaly. The correlation between ring chromosomes and MDM2 gene amplification indicates that the marker rings of MFH and of atypical lipoma often harbor genetic material derived from chromosome 12. Genes Chrom Cancer 9:26 1‐265 (1994). © 1994 Wiley‐Liss, Inc.
Evidence that wild‐type TP53, and not genes on either chromosome 1 or 11, controls the tumorigenic phenotype of the human fibrosarcoma HT1080
Authors: Anderson Michael J. , Fasching Clare L. , Stanbridge Eric J. , Casey Graham
Published: February 2007

The specific transfer of normal chromosomes via microcell fusion has been instrumental in identifying putative tumor suppressor gene loci in a variety of human cancers. Using this same technique it has been proposed that the tumorigenicity of the human fibrosarcoma cell line HT1080 is controlled by functionally distinct tumor suppressor genes on human chromosomes 1 and 11. To address these results and perhaps further localize the suppressive effect to particular regions on these two chromosomes, we transferred into HTI080 seven different fibroblast‐derived human chromosomes containing either intact or discrete portions of chromosome I or II. Interestingly, we found no evidence of genes on these chromosomes that could alter the growth of HT1080 either in vitro or in vivo. Based on these results we were left with the possibility that a gene, or genes, residing on an entirely different chromosome(s) was involved in the tumorigenesis of HTI080. Since TP53 mutation has been documented in a variety of human tumor types, and we found both copies of TP53 to be mutated in HTI080, we were prompted to examine its role by both cDNA transfection and chromosome transfer. Although by cDNA transfection we found that expression of exogenous wild‐type TP53 was incompatible with continued proliferation of HTI080 cells in vitro, chromosome 17 transfer studies revealed that a more physiologic expression of exogenous wild‐type TP53 could be tolerated in vitro while being completely incompatible with growth in vivo. These studies demonstrate a differential effect of TP53 growth inhibition and clearly show that TP53 tumor suppressing function can be independent from its potent growth suppressing effect in vitro. Genes Chrom Cancer 9:266‐281 (1994). © 1994 Wiley‐Liss, Inc.
Malignant rhabdoid tumor of the kidney: Involvement of chromosome 22
Authors: Shashi Vandana , Lovell Mark A. , Von Kap‐Herr Christopher , Waldron Peter , Golden Wendy L.
Published: February 2007

Cytogenetic and molecular studies have demonstrated that involvement of 22q is a non‐random finding in malignant rhabdoid tumors (MRTs) of the brain. We present an MRT of the kidney with the karyotype 47,XY, +i(I)(q 10), der(8)t(8;22)(q12;q11.2),der(22)t(8;22)(q23 or q24.l;q11.2). This unbalanced reciprocal translocation was confirmed by fluorescence in situ hybridization (FISH) with chromosome‐specific paints for chromosomes 8 and 22. Molecular analysis demonstrated a partial deletion of 22q in the BCR region at q 11.2, strengthening the suspicion that this is a critical region for the initiation or progression of these highly malignant neoplasms. Establishing non‐random cytogenetic changes in MRTs arising from the kidney may be of value in distinguishing these rare, but often fatal tumors from other renal neoplasms that mimick them histologically. The similarity in cytogenetic and molecular abnormalities between renal and extra‐renal MRTs argues against the concept that extra‐renal MRTs are only representative of a rhabdoid phenotype, rather than being true rhabdoid tumors. Genes Chrom Cancer 10:49–54 (1994). © 1994 Wiley‐Liss, Inc.
Detailed deletion mapping of chromosome segment 17q12‐21 in sporadic breast tumours
Authors: Nagai Maria A. , Yamamoto Lidia , Salaorni Sibeli , Pacheco Mércia M. , Brentani M. Mitzi , Barbosa Edson M. , Brentani Ricardo R. , Mazoyer Sylvie , Smith Simon A. , Ponder Bruce A. J. , Mulligan Lois M.
Published: February 2007

Linkage studies have indicated that a gene on chromosome arm 17q, designated BRCAI, confers susceptibility to familial breast and ovarian cancer. To investigate the possible involvement of the BRCAI gene in sporadic breast cancer we have analysed loss of heterozygosity (LOH) in a panel of 100 sporadic primary breast tumours using 10 PCR‐based polymorphic markers from 17q12–21. Allele losses were detected in 40 of 100 tumours informative for at least one of the markers analysed. Of these 40 deleted tumours, 27 showed partial or interstitial loss on 17q. The pattern of LOH in the tumours with partial or interstitial LOH revealed three putative distinct deleted regions on 17q12–21. The first lies on the proximal long arm between D17S250 and THRAI; the second one lies between D17S776 and D17S579, the region containing the BRCAI gene; and the third is telomeric to D17S733. The most frequently deleted region overlaps with the minimal region containing the BRCAI gene, suggesting that this gene might also be associated with the development or progression of a proportion of sporadic breast tumours.
AMLI fusion transcripts in t(3;21) positive leukemia: Evidence of molecular heterogeneity and usage of splicing sites frequently involved in the generation of normal AMLI transcripts
Authors: Sacchi Nicoletta , Nisson Paul E. , Watkins Paul C. , Faustinella Fabrizia , Wijsman Jacqueline , Hagemeijer Anne
Published: February 2007

The t(3;21)(q26;q22) is associated with chronic myelogenous leukemia in blast crisis (CML‐BC), leukemia evolving from (therapy‐related) myelodysplasia, and with leukemia following other hematopoietic proliferate diseases. Molecular cytogenetic analysis and cloning of a few t(3;21) cases indicate that the breakpoints are quite heterogeneous even within a specific clinical phenotype. Interestingly some of the (3;21) breakpoints involve the AMLI gene previously found rearranged in the t(8;21) associated with acute myelogenous leukemia. AMLI is related to the Drosophila gene runt and is the human counterpart of the gene for the α subunit of the nuclear polyoma enhancer binding protein (PEBP2) also known as the core binding factor (CBF). In the t(3;21) AMLI was found rearranged with EAP, a gene on chromosome 3 encoding a small ribosomal protein, as well as with EVII, another gene on chromosome 3. Here we report our study of six cases of t(3;21). By using fluorescence in situ hybridization (FISH) analysis and AMLI probes we could conclude that at least in two CML‐BC cases the breakpoint occurred in the AMLI intron that is disrupted by the t(8;21). An AMLI/EAP fusion transcript, different from the one described in a therapy‐related myelodysplasia, was detected in both CML‐BC cases. This transcript is expected to result in a predicted protein containing the AMLI nuclear binding domain with an attached stretch of 17 amino acids unrelated to the EAP small ribosomal protein. In the other t(3;21) patients we could not detect an AMLI/EAP transcript or an AMLI/EVII transcript. This result suggests heterogeneity of the t(3;21) at the molecular level. The AMLI chimeric transcripts identified so far, both in the t(3;21) and in the t(8;21), diverge from the normal transcripts either after exon 5 or exon 6. Here we show that in normal AMLI transcripts different splicing events are seen to occur after AMLI exon 5 as well as exon 6.
A constitutional bws‐related t(11;16) chromosome translocation occurring in the same region of chromosome 16 implicated in wilms' tumors
Authors: Newsham Irene , Kindler‐Röhrborn Andrea , Daub Douglas , Cavenee Webster
Published: February 2007

Beckwith‐Wiedemann syndrome (BWS) is a congenital overgrowth disorder with a varying spectrum of clinical manifestations including macroglossia, omphalocele, hemihypertrophy, and a predisposition to a subset of embryonal tumors, most frequently Wilms' tumor (WT). A variety of cytogenetic, genetic linkage, and molecular mapping data implicate a gene or genes on chromosome band 11p15.5 in BWS and its related tumors. However, some families with BWS do not show linkage to 11p15, and other alterations have been found in Wilms' tumors as well. One such alteration is loss of heterozygosity (LOH) for chromosome arm 16q. Here we have analyzed a balanced t(11;16)(p15;q13) chromosomal translocation associated with the BWS phenotype and mapped the breakpoint positions for both chromosomes 11 and 16 by using somatic cell hybrids and polymorphic markers. The chromosome 11 breakpoint was found to lie distal to the D11S12 locus, but proximal to TH on 11p15.5, a region shown previously to contain other BWS‐related chromosomal events. The chromosome 16 breakpoint was distal to D16S290 in 16q13, but proximal to loci D16S265, D16S267, and D16S164 in band 16q21. This area encompasses the region of LOH occurring through mitotic recombination in sporadic WT. This raises interesting possibilities for the genetic and epigenetic involvement of both chromosomal regions (11p15 and 16q13) in the pathogenesis of BWS and Wilms' tumor.
Allelic imbalance on chromosome I in human breast cancer. II. Microsatellite repeat analysis
Authors: Hoggard Nigel , Brintnell Bill , Varley Jennifer , Howell Anthony , Weissenbach Jean
Published: February 2007

We have determined regions of allelic imbalance in human breast cancer cells using highly polymorphic microsatellite markers, which can be rapidly typed by the polymerase chain reaction (PCR) using very small amounts of DNA. It appears that there are several regions of chromosome I which may be the targets of allelic imbalance, including some regions which have been identified previously by different groups. The detail with which we have mapped these regions of imbalance is, however, much greater than has been previously reported, and we have been able to localise these regions to small intervals of the genome. In addition we have identified previously uncharacterised regions of allelic imbalance on chromosome arm 1p, one of which (at 1p22–31) is lost in a high proportion of malignant lesions. We are currently attempting to analyse this latter region in detail in order to identify and characterise the sequence(s) involved. Study of such regions should help us understand some of the mechanisms underlying the development and progression of breast cancer.
Analysis of the joining sequences of the t(15;17) translocation in human acute promyelocytic leukemia: Sequence non‐specific recombination between the pml and rara genes within identical short stretches
Authors: Yoshida Hitoshi , Naoe Tomoki , Fukutani Hisashi , Kiyoi Hitoshi , Kubo Kazuaki , Ohno Ryuzo
Published: February 2007

Molecular analysis of the t(15;17) translocation in 70 patients with acute promyelocytic leukemia (APL) confirmed that the breakpoints of chromosome 15 were located in two regions of the promyelocytic leukemia (PML) gene, mainly introns 3 and 6, whereas the breakpoints of chromosome 17 were consistently in intron 2 of the retinoic acid receptor alpha (RARA) gene. To study the reason for the clustering of the breakpoints and the underlying mechanism of the chromosomal translocation, we characterized the joining sequences of der(15) and der(17) by polymerase chain reaction in samples from eight patients with APL. There was no cluster of the breakpoints within the introns, and no consensus sequence‐motif was found around them. One or nine extra nucleotides were inserted into two joining sites. There were identical stretches of one to seven nucleotides between the PML and RARA genes in the majority of the joining sequences. These data provide a potential model of the t(15;17) translocation: random DNA double strand cleavage, modification of DNA ends by enzymes including terminal deoxynucleotidyl transferase, and single strand base‐pairing within identical short stretches. Furthermore, APL develops only when the PML and RARA genes are rearranged within restricted genomic regions and a functional PML‐RARA chimeric product is produced, and this might lead to a clustering of the breakpoints.
Changes in mean telomere length in basal cell carcinomas of the skin
Authors: Wainwright Linda J. , Rees Jonathan L. , Middleton Peter G.
Published: February 2007

Human telomeres consist of arrays of the sequence TTAGGG up to 15–20 kb in length, which are essential for the maintenance of normal chromosomal stability. It has been suggested that genomic instability observed in tumours may be due to loss of telomere sequences. Somatic cells that are dividing continuously appear to progressively lose telomere sequences, and it would therefore be anticipated that cell type specific differences in mean telomere length may exist within an individual. Previous reports have suggested that mean telomere length may be different in human neoplasia when compared to control. Basal cell carcinomas are epidermal derived tumours and in order therefore to make valid cell type specific comparisons we have measured mean telomere length in 20 basal cell carcinomas as well as in both adjacent epidermis and dermis. Mean telomere length was significantly reduced in epidermis in comparison with dermis, from clinically normal skin immediately adjacent to the tumours (mean difference 2.5 kb). This result is not related to the presence of the tumour as similar results were obtained from skin samples of healthy volunteers. Basal cell carcinomas showed increased mean telomere length in 13/20 samples in comparison with matched epidermis (mean difference 3.1 kb), whereas in 7/20 mean telomere length was reduced (mean difference 2.2 kb). These results showing that mean telomere length varies from cell type to cell type underpin the importance of performing cell type specific controls when assessing changes in tumour telomeres.
Amplification of the met gene in glioma
Authors: Fischer Ulrike , Müller Hans‐W. , Sattler Hans‐P. , Zang Klaus D. , Meese Eckart , Feiden Klaus
Published: February 2007

We have previously reported the finding of MET amplification linked to double minutes (dmins) in a human glioblastoma (TX3095). Because dmins are found in approximately 50% of glioblastomas, 18 gliomas were analyzed for MET amplification. Three grade IV glioblastomas and one grade II astrocytoma showed amplification. We could also localize the MET amplicon to dmins in glioblastoma TX3095 by fluorescence in situ hybridization.
Identification of cytogenetically undetected 12p13 translocations and associated deletions with fluorescence in situ hybridization
Authors: Kobayashi Hirofumi , Rowley Janet D.
Published: February 2007

We performed fluorescence in situ hybridization (FISH) on bone marrow or peripheral blood cells thought to contain a del(12p) or an unbalanced 12p11–12 translocation from 17 patients who had various hematologic malignant diseases. We used 11 cosmid, phage, and plasmid probes which we had previously ordered on 12p. Cells from three patients with myeloid disorders were shown to have 12p13 translocations that involved chromosome 2 in two of them. Moreover, in all patients, FISH showed that the translocations were associated with proximal interstitial deletions which contributed to the difficulty in identifying these translocations. Our data suggest that some rearrangements of 12p which have been described previously as deletions or unbalanced translocations may, in fact, represent 12p13 translocations accompanied by an interstitial deletion.
Detection of hidden structural rearrangements by fish in pleomorphic adenomas
Authors: Sahlin Pelle , Mark Joachim , Stenman Göran
Published: February 2007

Previous cytogenetic analysis of pleomorphic adenomas of the salivary glands has revealed a subgroup of tumors with interstitial deletions of 8q with breakpoints at q12 and q21–23. In this paper we have re‐examined four adenomas with confirmed or suspected deletions of 8q by FISH. Painting of interphase nuclei and metaphase chromosomes with whole chromosome libraries revealed that in each of the four cases the deleted chromosome 8 material was found inserted or translocated onto other chromosomes. In two of the cases we were also able to resolve several other complex rearrangements. Our FISH data thus suggest that gross deletions of 8q do not occur in pleomorphic adenomas. These observations are of crucial importance for the molecular interpretations of the 8q12 rearrangements. The fact that all rearrangements with breakpoints at 8q12 seem to be translocations or insertions strongly indicates that they involve a similar molecular mechanism. Our findings illustrate that chromosome painting is a valuable and useful adjunct to conventional cytogenetic analysis of solid tumors.