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Background Understanding the progression of prostate cancer to androgen-independence/castrate resistance and development of preclinical testing models are important for developing new prostate cancer therapies. This report describes studies performed 30 years ago, which demonstrate utility and shortfalls of xenografting to preclinical modeling. Methods We subcutaneously implanted male nude mice with small prostate cancer fragments from transurethral resection of the prostate (TURP) from 29 patients. Successful xenografts were passaged into new host mice. They were characterized using histology, immunohistochemistry for marker expression, flow cytometry for ploidy status, and in some cases by electron microscopy and response to testosterone. Two xenografts were karyotyped by G-banding. Results Tissues from 3/29 donors (10%) gave rise to xenografts that were successfully serially passaged in vivo. Two, (UCRU-PR-1, which subsequently was replaced by a mouse fibrosarcoma, and UCRU-PR-2, which combined epithelial and neuroendocrine features) have been described. UCRU-PR-4 line was a poorly differentiated prostatic adenocarcinoma derived from a patient who had undergone estrogen therapy and bilateral castration after his cancer relapsed. Histologically, this comprised diffusely infiltrating small acinar cell carcinoma with more solid aggregates of poorly differentiated adenocarcinoma. The xenografted line showed histology consistent with a poorly differentiated adenocarcinoma and stained positively for prostatic acid phosphatase (PAcP), epithelial membrane antigen (EMA) and the cytokeratin cocktail, CAM5.2, with weak staining for prostate specific antigen (PSA). The line failed to grow in female nude mice. Castration of three male nude mice after xenograft establishment resulted in cessation of growth in one, growth regression in another and transient growth in another, suggesting that some cells had retained androgen sensitivity. The karyotype (from passage 1) was 43–46, XY, dic(1;12)(p11;p11), der(3)t(3:?5)(q13;q13), -5, inv(7)(p15q35) x2, +add(7)(p13), add(8)(p22), add(11)(p14), add(13)(p11), add(20)(p12), -22, +r4[cp8]. Conclusions Xenografts provide a clinically relevant model of prostate cancer, although establishing serially transplantable prostate cancer patient derived xenografts is challenging and requires rigorous characterization and high quality starting material. Xenografting from advanced prostate cancer is more likely to succeed, as xenografting from well differentiated, localized disease has not been achieved in our experience. Strong translational correlations can be demonstrated between the clinical disease state and the xenograft model. Prostate 75: 628–636, 2015. © The Authors. The Prostate published by Wiley Periodicals, Inc.

Translocations or deletions involving the 11q23 region have been observed in acute lymphoblastic leukemia (ALL), acute myelocytic leukemia (AML), myelodysplastic syndrome (MDS), and chronic lymphocytic leukemia (CLL). BAC probes encompassing the D11S29 and D11S924 markers and flanking the MLL gene were used in dual color fluorescence in situ hybridization. Fifteen patients with hematologic malignancies and cytogenetic abnormalities of 11q23 were analyzed. The BAC and MLL probes demonstrated split signals in five of 7 ALL or AML cases with translocations of 11q23. Of the remaining 2 cases, one had normal signals for both probe sets and the other had a submicroscopic deletion of the MLL 3′ region. In one case of AML with del(11)(q23), deletion of the MLL 3′ region and the region telomeric to the MLL gene was seen. Three CLL cases with deletion of part or the entire 11q23 region showed deletion of one copy of MLL , but retention of the region telomeric to MLL . However, in four MDS cases with deletions involving the 11q23 region, deletions of both the MLL gene and the flanking regions of the MLL gene were observed. Hence, the deletions on 11q23 are different but overlapping for CLL and MDS, implicating different genes involved for these diseases.

Background Cytogenetic analysis performed at diagnosis is considered to be the most valuable prognostic factor in acute myeloid leukemia (AML). Large systematic studies of cytogenetic abnormalities in AML patients from Southeast Asia are not available. The karyotypic patterns in AML patients from a single center in Singapore were studied and compared with reports from other regions of the world to identify possible geographic heterogeneity. Methods Analysis was performed on 501 consecutive de novo AML patients diagnosed according to the FAB criteria in the Singapore General Hospital. The cytogenetic findings were analyzed for possible associations between karyotypic pattern and the age, gender, ethnicity as well as morphological (FAB) subtypes. Results A total of 454 patients were studied of which 275(61%) had abnormal cytogenetics(median age 48 years). The t(15;17) and trisomy 8 were the most frequent karyotypic abnormalities - seen in 52(11%) and 33(7.3%) cases, respectively. Inv(lf) and t(16;16) were uncommon, seen only in five (1.1%) cases. The abnormalities del 5/5q and del 7/7q were seen in 30(6.6%) and 32(7%) of the cases. Complex karyotypes were seen in 78(17%) of the cases.Recurrent cytogenetic abnormalities correlated with the FAB subtypes. In all, 21 novel cytogenetic abnormalities were observed. Conclusions Certain differences such as the age at presentation and frequency of recurrent balanced translocations were noted in comparison to previous reports. These point to the need for extensive epidemiological studies to clarify the role of genetic as well as geographic heterogeneity in the pathogenesis of AML.

Synovial sarcoma (SS) is an uncommon soft tissue tumor that occurs primarily in the extremities of young adults, especially in the periarticular region. In this report, we describe the rare occurrence of primary SS of the pleura in a 15-year-old boy. Histologically, the tumor demonstrated monophasic morphologic findings and showed positive staining with vimentin and Bcl-2 and focally for cytokeratin CK7. Fluorescent in situ hybridization identified t(X;18) translocation. The patient developed recurrences 20 months following resection of the tumor. The literature on this uncommon entity is reviewed, and its histogenesis, differential diagnoses, and cytologic features are also discussed.

This study investigated if formalin fixation duration affects HER2/neu gene amplification detection by fluorescence in situ hybridization (FISH) in breast cancer. Tumor tissues from 35 cases were divided into three groups and subjected to two formalin fixation protocols per group (12 hr, 27 hr in the first; 2 hr, 17.5 hr in the second; 28.5 hr, 541 hr in the third) before FISH analysis. There was no significant difference in FISH signal detection between the two different fixation protocols in the first two groups. In the third, no signal was detected in 4/6 cases fixed for an extended duration.

The first report of a fragile site (FS) on a human chromosome was published in 1965 by DEKABAN. Since then there have been many reports of FS on several different chromosomes (reviewed by MCKUSICK 1982). FS on chromosomes 2 , 9-12, 16, 20 and X are included in the review of MCKUSICK and in addition to these there exists another FS on chromosome 6 ( L E V E R S H A ~ ~ al. 1981). Furthermore, on each of chromosomes 10 and 16 there are two separate FS. The first report to demonstrate that FS can be heritable came in 1968 (LEJ E U N E et al.). Most FS have been regarded as normal variants, but the FS on the long arm of the X chromosome has been found t o be associated with one form of X-linked mental retardation in males (reviewed by BKQNDUM NIELSEN et al. 1981). An abnormal X chromosome in a family with such mentally retarded boys was first described in 1969 by LUBS. Recently psychotic-like symptoms have also been reported in this kind of patients ( N I E L S E N ~ ~ al. 1982). Also the FS on chromosome 2 seems t o be associated with various abnormalities though some subjects have been normal (reviewed by ANNEREN and GUSTAVSON 1981). It has been demonstrated that the expression of most FS is highly dependent on the culturing conditions (SUTHEKLANLI 1979). The most important factors are low serum content ( 5 9%) and a deficiency of folic acid and thymine in the culture medium. There are three studies of FS in normal subjects. In one of these studies a very low frequency (0.00098) of individuals having FS was found (SUTHERLAND I982a), whereas another study indicated that some FS might be much more common in the normal population than has been thought up to now (ZHOU et al. 1982). The third study of FS in normal subjects concerns the BrdU-requiring FS at 10q25 for which a gene frequency of

Chromosomal in situ hybridization (ISH) has extended the scope of cytogenetic analysis to nondividing cells by the use of chromosome-specific probes detected by nonisotopic techniques. This provides a rapid and sensitive method for identifying chromosomes in interphase cells, and is useful in gauging engraftment following bone marrow transplantation, particularly when the number of cells obtained is minimal. We have performed ISH using a Y-heterochromatin-specific probe to monitor patients with malignant hematological disease who have received a sex-mismatched transplant. The results have been compared with those obtained from concurrently performed standard cytogenetic analysis. Host cells were detected by interphase cytogenetics in all patients posttransplant, at times varying from 28-1,825 days, whereas routine analysis detected host cells in only 4 patients, 3 of whom were found to be in relapse. The significance of the persistence of host cells is unknown, but it does not appear to indicate impending relapse. © 1995 Wiley-Liss, Inc.

Parathyroid hormone-related protein (PTHRP) is expressed in a large number of tumors and is the mediator of parathyroid hormone-like effects seen in humoral hypercalcemia of malignancy. The gene coding for PTHRP has been localised to the short arm of chromosome 12. This is at the same region as the oncogene KRAS2, and amplification of KRAS2 has previously been found in human lung cancer. The BEN cell line which is known to express PTHRP was established from a patient who had squamous cell carcinoma of the lung with hypercalcemia. Cytogenetic analysis of the BEN cell line revealed a very complex karyotype with many marker chromosomes. Chromosomal in situ hybridization with biotinylated DNA probes visualized by a biotin-streptavidin-polyalkaline-phosphatase complex was used to analyse two dicentric marker chromosomes containing homogeneously staining regions (hsr) in BEN. The hsr were found to contain amplified PTHRP and KRAS2 at levels of 30-fold and 14-fold per cell, respectively. The higher level of amplification of the PTHRP gene would suggest that PTHRP is the target gene of amplification in the amplicon. This is the first report of gene amplification of PTHRP and in addition its co-amplification with KRAS2. © 1993 Wiley-Liss, Inc.

Disorders of eye development such as microphthalmia and anophthalmia (small and absent eyes respectively), anterior segment dysgenesis where there may be pupillary and iris anomalies, and associated cataract and glaucoma, often lead to visual impairment or blindness. Currently treatment options are limited, as much is unknown about the molecular pathways that control normal eye development and induce the aberrant processes that lead to ocular defects. Mutation detection rates in most of the known genes are generally low, emphasizing the genetic heterogeneity of developmental ocular defects. Identification of the disease genes in these conditions improves the clinical information available for affected individuals and families, and provides new insights into the underlying biological processes for facilitation of better treatment options. Investigation of chromosomal rearrangements associated with an ocular phenotype has been especially powerful for disease gene identification. Molecular characterization of such rearrangements, which pinpoints the region by physically disrupting the causative gene or its regulatory sequences, allows for rapid elucidation of underlying genetic factors that contribute to the phenotype. Genes includingPAX6, PITX2, FOXC1, MAF, TMEM114, SOX2, OTX2andBMP4have been identified in this way to be associated with developmental eye disorders. More recently, new methods in chromosomal analysis such as comparative genomic hybridization (CGH) microarray, have also enhanced our ability in disease gene identification.

In this study we present two novel anaplastic thyroid carcinoma (ATC) lines (HTh 104 and HTh 112) and further characterize six frequently used ATC lines (HTh 7, HTh 74, HTh 83, C 643, KAT-4, and SW 1736). Three of the lines carried a heterozygous BRAF mutation V600E, which is in line with reports of BRAF mutations in primary ATC and papillary thyroid cancer. Several nonrandom breakpoints were identified by spectral karyotyping (SKY) and G-banding in these lines including the novel 1p36 and 17q24-25 as well as 3p21-22 and 15q26 that are also implicated in well-differentiated thyroid cancers. Comparative genomic hybridization showed frequent gain of 20q, including the UBCH10 gene in 20q13.12, which was further confirmed by array-comparative genomic hybridization and fluorescence in situ hybridization analyses. Our results concur with previous studies in both primary tumors and cell lines, indicating that gain of chromosome 20 is important in the pathogenesis of ATC and/or progression of differentiated thyroid cancers to ATC.

Amplification of the chromosomal region 11q23 encompassing the MLL gene has been observed in patients with acute myeloid leukemia (AML). Patients with this abnormality often have a poor response to chemotherapy and short survival. We have studied the regions flanking the MLL gene using 8 bacterial artificial chromosome (BAC) probes and dual color fluorescence in situ hybridization (FISH) analysis. Two contigs of BAC probes flanking the MLL gene, were constructed by standard primer walking and BAC end sequencing. For comparison, metaphase chromosome preparations were also hybridized with a commercial MLL specific probe. Metaphase chromosomes were prepared from the bone marrow sample of an 80-year old female patient with AML-M1 and the cytogenetic aberration der(11)hsr(11) (q23). FISH analysis on metaphase chromosomes showed amplification on the homogeneously staining region (hsr) and marker chromosomes for both the MLL gene and the BAC probes. Using dual color FISH, probes proximal to MLL showed greater amplification than those distal to MLL, as represented by additional red signals on both metaphase and interphase chromosomes. Ratios of the copy numbers of the BAC probes relative to the chromosome 11 centromere copy number confirmed a higher copy number for probes proximal to MLL. These results suggest that other gene(s) proximal to MLL could be the target gene(s) of amplification in this case and not the MLL gene as previously assumed.

We evaluated HER2/neu gene amplification by fluorescence in situ hybridization (FISH) in archival paraffin-embedded breast cancer tissues. Tumors from 63 human invasive breast cancers were categorized into two groups depending on whether the paraffin-embedded tissue blocks had been stored for more or less than 12 months duration. These were subjected to routine and modified FISH protocols. As microwave oven formalin fixation of tissues was carried out in the majority of the older archived specimens, the effect of this fixation method was also analyzed. FISH signals were obtained in all 13 archival specimens stored for less than 12 months. However, in 50 specimens stored for more than 12 months duration, the procedure was successful in only 10 specimens (20%), for which the pretreatment procedure had to be individually optimized for each specimen. There was no significant difference in the detection of FISH signals between microwave oven and routinely fixed specimens.

Genetic alterations of thymomas are rarely described in the literature. In this study, a previously unreported instance of aberrant karyotypic change consisting of 45,XX,pseu dic(16;12) (q11;p11.2) [cp23]/87-90,idemx2[cp4] in a Masaoka Stage II mixed thymoma or type AB thymoma affecting a 56-year-old Chinese woman is detailed. Abnormalities involving 12p containing important tumor suppressor-like genes have been documented especially in hematological malignancies. Recently, recurrent losses involving 16q, a locus known to harbor several tumor suppressor genes, have been described in type C thymomas (squamous cell carcinoma), suggesting a possible relationship between type AB thymoma and type C thymoma. Whether these genes are involved in the pathogenesis of type AB thymoma remain to be clarified and it is currently unclear if cytogenetic studies may eventually play a role in the classification of thymic tumors.

We report a male newborn infant of Chinese descent with the Smith-Magenis (SMS) syndrome who presented with a severe cyanotic congenital heart disease. This report adds pulmonary atresia and ventricular septal defect to the spectrum of cardiac defects seen in SMS.

Among chromosomal abnormalities described in myeloma (MM), deletion of chromosome 13 (Δ13) has been linked to a poor outcome. However, prognosis appears to vary according to the detection method used: conventional cytogenetics (CC) or FISH. We have investigated this inconsistency in a large multi-center cytogenetic study. Bone marrow samples from 814 MM patients were provided from 95 UK hospitals. FISH on purified plasma cells (n=697) and/or CC (n=577) was performed centrally: 458 samples had both tests. Sample quality was often poor, with low cell counts (median 2x107 and low plasma cell (PC) content (median 9%), precluding both tests on all samples. FISH for Δ13, t(4;14), t(11;14), gain of chromosomes 5, 9, 15 and, where possible, 3, 7 &17 was carried out. Overall survival was used as the end point with median follow-up of 22mo (range 12–53mo). Abnormalities were detected by CC in 216 cases (37%). Non-hyperdiploidy (non-HRD) (≤47 chromosomes, n=82, 14% of total, 38% abnormal) was a stronger indicator of poor prognosis than hypodiploidy (≤45 chromosomes, n=55, 10% of total, 25% abnormal) (p96% and sensitivity >92%. There was complete agreement between CC and FISH for individual abnormalities. CC showed a highly significant difference in prognosis between abnormal karyotypes with (n=95, 44%) and without Δ13 (median 15 months vs not reached, p20% PC, n=322, 46%) to have the worst outcome among the individual abnormalities (median 30mo vs not reached, p50% or >80% PC, or by considering only Δ13 in non-HRD cases (median 29mo vs 49mo, p=0.001), consistent with our CC results. When patients with results from both CC and FISH were examined, the outcome for Δ13 by CC was significantly worse than by FISH only (median 15mo vs 33mo, p Fig 1. Survival curves for Δ13 by FISH (A) & by CC &/or FISH (B) Fig 1. Survival curves for Δ13 by FISH (A) & by CC &/or FISH (B)

Frequencies of HLA A, B, C, and DR antigens were studied in 100 schizophrenic patients and 919 controls from South Sweden. The patients were diagnosed according to the DSM III criteria and divided into four clinical subgroups (hebephrenic, paranoid, residual, and undifferentiated). In the schizophrenic patients as a whole significant increases were found for A2, A3, B17, B27, and Cw2 and decreases for A1, A11, and B8. A previous positive association with A9 from the same population was not confirmed. A significant heterogeneity between the four clinical subgroups was found for A3 and Bw35. Most of the associations between HLA antigens and schizophrenia reported in the literature appear to be fortuitous and dependent on the large number of trials made. However, confirmed increases have been found for A9 and B17, and confirmed decreases have been observed for A1 and B7. Some evidence for a heterogeneity between clinical subgroups was found in the present as well as in previous investigations.

The phenotype frequencies of properdin factor B (Bf) were studied in patients with (n = 47) and without (n = 66) a family history of schizophrenia and in controls. In patients with a family history of schizophrenia, a significant decrease of the FS type was found. No significant difference was found between patients without a family history of schizophrenia and controls.

Frequencies of HLA antigens (A, B, C and DR) were studied in patients with (n = 49) and without (n = 67) a family history of schizophrenia and in controls. Among the patients with a family history of schizophrenia significant increases were found in the A3 and B5 antigens while significant decreases were observed for the A1, A11 and B8 antigens. In a material of schizophrenic siblings the sharing of HLA haplotypes was consistent with normal segregation.

The proportion of erythroid cells in mitosis was determined in direct preparations and in synchronized and unsynchronized short-term cultures from normal human bone marrow. The stimulating effect on the proportion of erythroid mitoses of bovine serum albumin (BSA), erythropoietin (EP) and human leucocyte conditioned medium (HLC) derived from stimulated lymphocytes, was also analyzed in short-term marrow cultures. In synchronized and unsynchronized cultures, the proportion of erythroid mitoses was markedly lower than in direct preparations, with a mean of 75% in the direct compared to 11% in the unsynchronized cultures and 7% in synchronized. Statistical analysis showed that EP had a substantial influence on the erythroid cells while HCL had a minor effect.

Ten separate groups containing a total of 51 rat sarcomas induced by the polycyclic hydrocarbons were examined for inducing agent-related abnormalities in their DNAs. The levels of deoxyadenylate (dA) regions in normal and tumor rat DNAs were estimated from annealing [ 3 H ]-polyuridylic acid to DNA and treatment of the resultant hybrids with ribonuclease A. All the tumor material had significant reductions in the (dA) regions of their DNAs when the levels were compared to normal rat DNA. Although the (dA) reductions were rather heterogeneous from one tumor group to the other no matter the inducing agent, the greatest decreases tended to be found with tumors induced by 20 -methylcholanthrene (MC) and 7,12 -dimethylbenz(a)anthracene (DMBA). No relationship was found between (dA) alterations and the types of chromosomal aberrations in the tumor cell populations. However, normal and tumor DNAs, fractionated into A + T rich and G + C rich DNA by thermal elution from hydroxylapatite columns, showed specific neoplastic-associated (dA) disturbances in the A + T rich DNA of tumors induced by 3,4 -benzopyrene (BP), MC and DMBA. The base compositions of the tumor material were also analyzed and compared to normal rat DNA. Tumors induced by DMBA showed highly significant decreases in % A + T whereas BP and MC induced tumors tended to show only small increases in % A + T. Some tumors induced by BP, MC and DMBA were unaltered in base composition. Together our results are consistent with the hypothesis that DMBA is the strongest carcinogen in the series because it interacts preferentially with A + T Trich DNA, which is the type of DNA most important for polycyclic hydrocarbon-induced neoplasm in the rat.

Gc subtypes were studied by isoelectric focusing in schizophrenic patients and controls. No significant differences between patients and controls were found. The results so far on Gc groups and schizophrenia show no consistent pattern and the significant associations reported have not been confirmed.