XbaI RFLP of ETS-1 oncogene in chronic B-cell leukaemia.

The influence of a polymorphic variant of the ETS-1 oncogene on the predisposition to develop chronic B-cell leukemia (CLL) was investigated. A total of 59 patients with CLL and 59 controls were examined for the frequency of an XbaI restriction fragment length polymorphism RFLP of the ETS-1 oncogene which has been reported to occur more frequently in patients with hematological malignancies than in normal controls. We found no significant difference in the allele frequency between the CLL patients and the normal controls. These data suggest that the presence of the XbaI RFLP is not associated with CLL.

Elevated frequency of the C2 allele of the ETS-I oncogene in elderly subjects.

We studied the frequency of an Sst I polymorphism of the ETS-I oncogene in 122 elderly subjects (mean age 78.14 years) and 115 teenagers (mean age 16.9 years). No difference in the frequency of the three genotypes (C1C1, C1C2, C2C2) was found between the two groups. However, the C2 allele occurred more frequently in the elderly subjects (chi 2 = 5.49, P < 0.02). These data suggest that the presence of the C2 allele may be associated with survival to old age.

Methylation status of the 3rd exon of the c-MYC oncogene in B-cell malignancies.

We examined the methylation status of the third exon of the MYC oncogene in 39 patients with B-cell malignancies. DNA was digested with MspI plus EcoRI or HpaII plus EcoRI and hybridised with a probe specific for the third exon of MYC. Thirty four patients showed complete methylation of the CCGG site. Four patients, one with chronic B-cell leukaemia and one with pro-lymphocytic leukaemia (PLL) and two with B-cell lymphoma showed partial hypomethylation of the CCGG site, while another patient with PLL showed complete hypomethylation of the CCGG site. These results suggest that hypomethylation of the MYC oncogene is infrequent in B-cell tumours but may be involved in the development of some cases of B-cell malignancies.

Lack of 5' BCL2 rearrangements in B-cell leukemia.

We found no rearrangements in the 5' region of the BCL2 gene in DNA samples from 60 patients with chronic B-cell leukemia (CLL). This compares with the presence of these rearrangements in up to 10% of patients in other reports, and suggests that the incidence of 5' BCL2 rearrangements in CLL is considerably less than 10%.

Elevated frequency of a SstI polymorphism of the Ets-1 oncogene in non-Hodgkin's lymphoma patients.

We studied the frequency of a SstI polymorphism of the Ets-1 oncogene in 100 patients with non-Hodgkin's lymphoma, 44 patients with Hodgkin's disease, 49 patients with chronic myeloid leukemia, and 100 controls. There was no difference in the genotype frequency between the controls and patients with either Hodgkin's disease or chronic myeloid leukemia. In contrast, there was a highly significant difference in the distribution of the three genotypes between the patients with non-Hodgkin's lymphoma and the controls (X2 = 10.76, 2df, p = 0.004) with the C2 allele being more frequent in the lymphoma patients. Molecular cloning indicated that the polymorphic SstI site lay 304 bp from exon 7. This is the second association of the SstI polymorphism of the Ets-1 oncogene with a lymphoid disorder and suggests that the presence of the C2 allele is associated with a predisposition to develop a lymphoid malignancy.

Translocation (2;14) associated with complex rearrangements of the Ig heavy chain in non-Hodgkin lymphoma.

Cytogenetic analysis of a patient with non-Hodgkin lymphoma revealed the following karyotype: 49,XXX,t(2;14)(q21;q32),+4,+8,del(13)(q14q21). Southern blot analysis with an Ig region probe showed non-productive rearrangements indicative of a translocation involving the Ig locus. However, molecular cloning of the abnormal rearrangements did not show novel sequences derived from chromosome 2 but showed that the BCL-6 gene was juxtaposed to the IgH enhancer. Three further clones with abnormal rearrangements involving the Ig locus, particularly Iggamma3, were isolated. This suggests that the mature lymphoid cells, in this patient, were capable of undergoing indiscriminate switch cleavage and religation.

Complex karyotypic evolution in B-cell chronic lymphocytic leukemia.

Cytogenetic analysis at diagnosis in a female patient with chronic B-cell leukemia showed a single abnormal clone with a 4p+ abnormality, 46,XX, -4, +der(4)t(4;?)(p16;?). Six additional clones evolved from this clone during the following 4 1/2 years and showed 3p+, 4p-, and 11q- chromosomes in addition to the 4p+ abnormality. Immunoglobulin heavy chain gene rearrangement studies showed two rearranged bands and a faint germline band. Following splenectomy, a strong germline and faint rearranged bands were seen, suggesting that the majority of cells were normal, whereas cytogenetic studies showed that the karyotypically abnormal cells were still present. The combination of cytogenetic and Ig gene rearrangement studies provides detailed information regarding the number of circulating normal and leukemic cells.

Identification of amplified genes in a patient with acute myeloid leukemia and double minute chromosomes.

A case of acute myeloid leukemia (M2) with double minute chromosomes and complex karyotypic abnormalities was analyzed cytogenetically and molecularly. Comparative genomic hybridization (CGH) showed that the 8q24 region that contains the MYC oncogene was not amplified. Instead, amplification of chromosomal regions 11q23-->qter and 9p11-->pter was identified. Southern blot analysis confirmed the CGH findings and showed that the ETS1, FLI1, SRPR, NFRKB, and KCNJ5 genes located at 11q23-->24 were amplified, whereas the MLL at 11q23 was not amplified. Additionally, the IFN beta 1 and CDKN2A genes at 9p were amplified, but to a lesser degree. This is the first example of a case of acute myeloid leukemia with double minute chromosomes that has not involved amplification of either the MYC or the MLL genes.

Characterization of the C-MYC amplicon in a case of acute myeloid leukemia with double minute chromosomes.

We have characterized the double minute chromosomes in a case of acute myeloid leukemia (AML). Southern blot analysis showed that the C-MYC was amplified. Further analysis with probes located both 3' and 5' of MYC indicated that the amplicon was at least 700 kb in size, extending from the papilloma virus integration site situated 500 kb 5' of MYC to the PVT gene located 280 kb 3' of MYC. This appears to be the largest MYC-containing amplicon in human leukemia.

Aberrant rearrangements of the immunoglobulin heavy chain switch region in chronic B-cell leukemia.

Analysis of the organisation of the Cmu-switch region of the immunoglobulin heavy chain locus in B-lymphocytes from 80 patients with chronic B-cell leukemia revealed 25 patients with abnormal rearrangements that could not be explained by the normal recombination events that take place in B-lymphocytes. Detailed analysis with probes spanning the Cmu -switch region and various restriction digests localised the rearrangements in two thirds of the patients to a 1300 bp region at the 5' end of the switch region while in the remaining patients the rearrangements occurred in the switch region. The consequences of these aberrant rearrangements remain to be determined, but their clustering to a defined region of the switch region suggests a "hot spot" that may be involved in the aetiology of the disease.

A process for implementation of the strategic plan: from strategic planning toward strategic management.

Although the principles and process for developing a strategic plan are well established, little information exists on how organizations can implement these plans. The authors developed a framework and methodology for The Ontario Cancer Institute/Princess Margaret Hospital that connects the implementation of the strategic plan with resource allocation and the budget; thus, the budget is a quantified expression of the strategic plan. To accomplish this, a specific cycle of events leading to the development of a corporate calendar was undertaken, which results in the establishment of the annual budget.

Planning for shared services: lessons from experiences of the Princess Margaret Hospital.

The plan to rebuild The Princess Margaret Hospital (PMH) on University Avenue in Toronto has provided several opportunities for institutions in the area to share programs and services. The PMH planning process has raised questions about the factors that influence the development of shared services plans, and the organizational and management challenges posed by such plans. This experience suggests 13 principles to develop effective shared services.

Selection of test locations for regional trials of barley.

Three sets of regional six-row barley (Hordeum vulgare L.) trial data, representing cultivar x location x year, were grouped for locations based on the similarity of genotype x environment (GE) interaction. Locations were selected from each group (cluster) so that the structure of the GE interaction generated by the subsets of the locations would be approximately similar to that of the whole set (all locations). The purpose of this paper is to determine the number of locations where the GE interaction structure generated by these selected locations would be fairly consistent over years. Two statistics were used to measure the success of the selected locations: (1) the ratio of GE mean square (MS) associated with the selected location set relative to that associated with the best set (which gives the highest GE interaction MS) and (2) the rank correlation between the cultivar means averaged over the selected locations and those based on the entire data set. The results show that, for eastern Canada, 10-13 locations based on the cluster method can achieve a fairly consistent GE interaction structure over years.

Hypermethylation of the M27beta (DXS255) locus in chronic B-cell leukaemia.

We have investigated the methylation status of the M27beta (DXS255) locus in 21 female patients with chronic B-cell leukaemia and in 20 normal controls. DNA was digested with Pst1 and then with the methylation sensitive enzyme HpaII and probed with the M27beta probe. Eight patients (38%) showed hypermethylation of the M27beta locus which was not seen in any of the normal controls. Hypermethylation of the M27beta locus has also been found in acute myeloid leukaemia, acute lymphocytic leukaemia and lymphoma, suggesting that hypermethylation of the M27beta locus is associated with the leukaemic process.

Elevated frequency of an ETS-1 restriction fragment polymorphism in chronic B-cell leukaemia.

We studied the frequency of an SstI polymorphism in 70 patients with chronic B-cell leukaemia (CLL) and 100 normal controls. There was a highly significant difference in the distribution of the three genotypes between the CLL patients and the normal controls (chi 2 = 13.46, 2 df, P < 0.001). The C2 allele was found more frequently in CLL patients and may be a marker for a predisposition to develop CLL.

Diabetic neuroarthropathy in the foot: patient characteristics and patterns of radiographic change.

The diagnosis of diabetic neuroarthropathy of the foot can be difficult. A series of 96 patients (116 extremities) who had diabetes and peripheral neuropathy with bone and joint changes was reviewed. Typically, the patients were middle-aged or older, were taking insulin, and had had diabetes for more than 10 years. Retinopathy, nephropathy, and peripheral vascular disease were often present. There were abnormalities of vibratory sensation (94%) and of the gastrosoleus reflex (88%). The finding of specific radiographic abnormalities assisted but did not reliably differentiate neuropathy from infection. Three patterns of radiographic changes were noted: (1) at the metatarsophalangeal and interphalangeal joints, usually with underlying ulceration; (2) at the tarsometatarsal joints; and (3) in the anterior pillar-medial column of the foot, with talus, talonavicular, navicular, or naviculocuneiform destruction. Ulceration and infection in patients with patterns (2) and (3) were rare. When correlated, the demographic features, mode of presentation, physical signs, specific radiographic abnormalities, and patterns of change were distinctive and allowed the diagnosis of this complication of diabetes to be readily made.

Inverse acyl phosph(on)ates: substrates or inhibitors of beta-lactam-recognizing enzymes?

Acyl phosph(on)ates represent a new class of inhibitors of beta-lactam-recognizing enzymes. Previously described members of this class were aroyl phosph(on)ates. These compounds have been shown to acylate and/or phosphylate the active site serine residue, leading to either transient or essentially irreversible inhibition [Li, N., and Pratt, R. F. (1998) J. Am. Chem. Soc.120, 4264-4268]. The present paper describes the synthesis and evaluation as inhibitors of an inverse pair of acyl phosph(on)ates that incorporate the amido side chain that represents a major substrate specificity determinant of these enzymes. Thus, N-(phenylacetyl)glycyl phenyl phosphate and benzoyl N-(benzyloxycarbonyl)aminomethyl phosphonate were prepared. The former of these compounds was found to be a substrate of typical class A and C beta-lactamases and of the DD-peptidase of Streptomyces R61; it thus acylates the active site serine. In contrast, the latter compound was an irreversible inhibitor of the above enzymes, probably by phosphonylation of the active site serine. With each of these enzymes therefore, the amido side chain rather than the acyl group dictates the orientation of the bound phosph(on)ate and thus the mode of reaction.

Increased frequency of the S allele of the L-myc oncogene in non-Hodgkin's lymphoma.

We studied 100 patients with non-Hodgkin's lymphoma, 44 patients with Hodgkin's disease and 100 controls for the prevalence of the EcoRI restriction fragment polymorphism of the L-myc oncogene. No difference in the frequency of the three genotypes (LL, LS, SS) was found between the patient and control groups. However, the S allele was found to occur more frequently in the non-Hodgkin's lymphoma patients (chi 2 = 4.57, P = 0.032). These data confirm an earlier report and suggest that the presence of the S allele is associated with susceptibility to non-Hodgkin's lymphoma.