Meningeal SWI/SNF related, matrix-associated, actin-dependent regulator of chromatin, subfamily B member 1 (SMARCB1)-deficient tumours: an emerging group of meningeal tumours.

AIMS: Bi-allelic inactivation of SWI/SNF related, matrix-associated, actin-dependent regulator of chromatin, subfamily B member 1 (SMARCB1; also known as INI1) and loss of immunohistochemical expression of SMARCB1 define the group of SMARCB1-deficient tumours. Initially highlighted in malignant rhabdoid tumours, this inactivation has subsequently been observed in several intra and extracranial tumours. To date, primary meningeal SMARCB1-deficient tumours have not been described. We report two cases of meningeal SMARCB1-deficient tumours occurring in adults. METHODS: We performed immunohistochemical analyses, comparative genomic hybridization, fluorescence in situ hybridization and targeted next-generation sequencing. RESULTS: The first meningeal tumour was a solitary mass, composed of rhabdoid, adenoid, chordoid and sarcomatoid areas. The second case presented as multiple, bilateral, supra and infratentorial nodules, was composed of fusiform and ovoid cells embedded in a myxoid stroma. Tumour cells were positive for epithelial membrane antigen (EMA), vimentin and CD34 and negative for SMARCB1 and meningothelial, melanocytic, muscular, glial markers. In the first case, one allele of SMARCB1 was completely deleted, whereas in the second case, loss of expression of SMARCB1 was observed as a consequence of a homozygous deletion of SMARCB1. CONCLUSIONS: The phenotype and genotype of these two cases did not fit diagnostically with entities already known to be SMARCB1-deficient tumours. As both tumours shared common features, they are regarded as belonging to an emerging group of primary meningeal SMARCB1-deficient tumours, not described to date. To facilitate the identification and characterization of these tumours, we recommend SMARCB1 immunohistochemistry for primary meningeal tumours which are difficult to classify, especially if immunopositive for EMA and CD34.

Well differentiated endometrioid adenocarcinoma of the uterus: a cancer unit or centre case?

OBJECTIVE: The purpose of this study was to investigate what proportion of cases showing a well differentiated endometrioid endometrial adenocarcinoma in the hysterectomy specimen removed at two UK cancer centres had adverse pathological features or advanced stage disease at the time of presentation. STUDY DESIGN: Ninety-eight patients who were operated on at either the South East London Cancer Centre, London or the Kent Oncology Centre, Maidstone had a histological diagnosis of well differentiated (grade 1) endometrioid adenocarcinoma in their hysterectomy specimen. These were identified using the multidisciplinary meeting database as well as the respective pathology department databases. The histology reports for these patients were examined and analysed for the purpose of this study. RESULTS: Of the initial 98 cases, 65 patients (66.3%) were referred with a preoperative curettage showing a well differentiated endometrioid adenocarcinoma, 25 cases (25.5%) were referred with atypical endometrial hyperplasia, seven patients (7.1%) were referred with a moderately differentiated endometrioid adenocarcinoma, and one case (1.0%) was referred with a possible malignant mixed Mullerian tumour. Subsequent histological examination of the hysterectomy specimens revealed that all of these cases had a well differentiated endometrioid adenocarcinoma. In 20 of the 98 cases (20.4%) there was no myometrial invasion, 56 cases (57.1%) showed invasion of the inner half of the myometrium and 22 cases (22.4%) showed outer half involvement. There was no cervical involvement in 78 cases (79.6%), endocervical gland involvement in eight patients (8.2%) and cervical stromal involvement in 12 patients (12.2%). The total percentage of cases with cervical involvement was 20.4%. Thirty-eight cases (out of the 98) underwent a bilateral pelvic lymphadenectomy. Of these 38 cases, four cases had locoregional nodal metastases (10.5% of the patients who underwent lymphadenectomy). There were ovarian metastases in one case and metastasis to one fallopian tube in another. From our study, 33.6% of cases with a well differentiated endometrioid adenocarcinoma of the uterus were Stage Ic or more at the time of presentation; 12.2% were at least FIGO Stage Ic, eight patients (8.2%) were FIGO Stage IIa, seven patients (7.1%) were Stage IIb and six patients (6.1%) were Stage III. In these patients a full surgical staging operation with a pelvic lymphadenectomy was indicated according to FIGO recommendation. CONCLUSION: A significant proportion (33.6%) of well differentiated tumours in a hysterectomy were found to have Stage Ic disease or more at the time of presentation, and thus full surgical staging including a lymphadenectomy should have been carried out in these cases. Cases with a preoperative biopsy showing atypical hyperplasia or well differentiated adenocarcinoma should have a preoperative MRI scan or preferably an intraoperative frozen section examination to identify those cases with adverse pathological features which need to be fully staged with pelvic and paraaortic lymphadenectomy.

Accuracy of frozen section diagnosis at surgery in pre- malignant and malignant lesions of the endometrium.

OBJECTIVE: The purpose of this study was to correlate the histological diagnosis made during intraoperative frozen section examination of hysterectomies with atypical hyperplasia or carcinoma, with the definitive paraffin section histology. STUDY DESIGN: Frozen section pathology results of patients with a preoperative biopsy showing atypical hyperplasia or endometrial carcinoma (87 patients) were compared retrospectively with paraffin section pathology findings. Those patients with curettage specimens showing atypical hyperplasia or curettings suspicious of endometrioid carcinoma had intraoperative frozen section to determine whether an invasive lesion was present and whether they required pelvic lymphadenectomy. The purpose of frozen section assessment in those patients who had a preoperative curettage specimen showing endometrial carcinoma was to identify poor prognostic pathological factors related to histological subtype, grade, depth of myometrial invasion and cervical involvement. RESULTS: The correlation between frozen sections and paraffin histology in patients with endometrial carcinoma was 98.6% (69/70) for histological sub-type and 84.3% (59/70) for grade of differentiation. Depth of myometrial invasion was accurately diagnosed in 94.3% (66/70) while cervical involvement was accurately assessed in 86.7% (52/60). Of the 37 patients with atypical hyperplasia or suspicious curettings on preoperative curettage who had intraoperative frozen section, 23 patients had invasive malignancy, which was confirmed in subsequent paraffin sections. Of the remaining 14 patients with a non-malignant frozen section diagnosis, 11 were confirmed with paraffin sections while three had a small well differentiated invasive lesion, two were FIGO Stage 1a and one had microscopic invasion into the myometrium. CONCLUSION: Intraoperative frozen section is a useful procedure to identify poor prognostic pathological factors as well as to diagnose endometrial cancer in patients undergoing hysterectomy for a preoperative biopsy diagnosis of atypical hyperplasia.

Evaluation of the Dade Behring Syva EMIT 2000 tacrolimus assay on the Bayer Advia 1650.

BACKGROUND: The Dade Behring Syva(R) EMIT (enzyme multiplied immunoassay technique) method for the measurement of tacrolimus in whole blood was evaluated against the Abbott IMx(R) microparticle enzyme immunoassay (MEIA) method. EMIT measures tacrolimus colorimetrically, whereas MEIA measures the analyte using fluorimetry. Both methods incorporate a protein precipitation step prior to measurement. METHOD: Whole blood specimens were treated by two types of precipitation technique followed by analysis for tacrolimus by either MEIA or EMIT on the Bayer Advia 1650. Linearity and precision were assessed and correlation analysis performed to evaluate the EMIT assay on the Bayer Advia 1650. RESULTS: The EMIT tacrolimus assay was linear over the concentration range 0.0-22.0 micro g/L; the limit of detection was 1.2 micro g/L. Correlation between the Syva EMIT and IMx tacrolimus assays was excellent (r = 0.959) and no significant bias existed between the two methods (mean difference, delta = 0.221 micro g/L). Calibration data for the EMIT assay was stable for a period of 24-48 h on the Advia between runs. CONCLUSION: The Syva EMIT assay for the measurement of tacrolimus in whole blood is suited for daily routine use on the Bayer Advia 1650.

Pigmented liver cell adenoma in two male patients.

We report two cases of hepatocyte neoplasia with extensive deposition of Dubin-Johnson-like pigment in men without Dubin-Johnson syndrome. This pigment has previously been described in hepatocellular carcinoma but not in liver cell adenoma. The tumors of both patients showed some atypical cytologic features, but no frank histologic evidence of malignancy. Long-term follow up for several years showed no evidence of recurrence after limited surgical excision. We conclude that tumors with this structure may be cured by limited surgical excision and should be considered as pigmented liver cell adenomas.

Results of a pilot program for training bar staff in preventing aggression.

One hundred and twenty-one staff from eight bars in Ontario, Canada participated in a three-hour training program that used a peer learning model to teach problem-solving skills regarding the prevention and management of aggressive behavior in bars. Participants showed significant positive changes in knowledge and attitudes regarding effective approaches to preventing aggression. The majority of participants reported that the training made them think about ways they handed problem situations and that they would change the way they handle problems in the future. Participants rated most aspects of the training as very useful, especially the group discussion. The program illustrates the potential for increasing skills and knowledge of bar staff in preventing aggression and associated injury.

The prevalence and severity of intestinal disaccharidase deficiency in human immunodeficiency virus-infected subjects.

BACKGROUND: Gastrointestinal symptoms are distressing features of human immunodeficiency virus (HIV) infection, and management is often empirical, including withdrawal of dietary lactose. We assessed the prevalence and severity of intestinal disaccharidase deficiency in vitro and in vivo. METHODS: Fifty-four HIV-seropositive patients (19 HIV well +/- mild diarrhoea, 7 acquired immunodeficiency syndrome (AIDS) well, and 28 AIDS with diarrhoea) were studied with a combined non-invasive absorption-permeability-disaccharidase test that enables quantitative assessment of the rate of intestinal hydrolysis of lactose, sucrose, and palatinose. Thirty patients had jejunal biopsy specimens suitable for histomorphometric assessment, and 36 had in vitro disaccharidase activity measurement. RESULTS: Patients with HIV (with mild diarrhoea) and AIDS (with and without severe diarrhoea) had frequent but mild histomorphometric changes in jejunal specimens. This was associated with frequent (21%-100%) and often severe in vitro jejunal disaccharidase deficiency. In vivo hydrolysis of lactose, sucrose, and palatinose was impaired in 25%-75% of patients, apart from HIV well patients, who were normal. The prevalence of the in vivo lactase and sucrase deficiency was significantly (P < 0.006) lower than in vitro and severe in about 30%. CONCLUSIONS: Intestinal disaccharidase deficiency is common both in vitro and in vivo in HIV-seropositive patients but sufficiently severe to consider lactose withdrawal only in about a quarter of the patients with AIDS and diarrhoea.

Community action research: who does what to whom and why? Lessons learned from local prevention efforts (international experiences).

This paper describes lessons learned about community action research, drawing upon papers written and presented at a recent international conference on community action research and the prevention of alcohol and other drug problems. Projects reflected both action and evaluation research traditions and focused on a variety of issues from moderation of drinking to alcohol-related violence, and on range of target populations from youth to specific ethnic groups. The interventions described ranged from policy-based prevention to education and training and to secondary prevention and treatment. Lessons identified in the papers are discussed within three broad areas: the community targeted for change; the implementation of community projects; and community action research projects generally. The common lessons emerging from these diverse projects provide useful lessons on which to base future progress in community action research.

Novel interaction between the transcription factor CHOP (GADD153) and the ribosomal protein FTE/S3a modulates erythropoiesis.

The transcription factor CHOP (GADD153) heterodimerizes with other C/EBP family members, especially C/EBPbeta, thus preventing their homodimerization and binding to DNA sequences specific for the homodimers. Some CHOP-C/EBP heterodimers apparently bind to alternative DNA sequence and thereby regulate the transcription of other genes. Recently, we demonstrated that CHOP is up-regulated during certain stages of erythroid differentiation and that ectopic overexpression of CHOP enhances this process (Coutts, M., Cui, K., Davis, K. L., Keutzer, J. C., and Sytkowski, A. J. (1999) Blood 93, 3369-3378). In the present study, we report that CHOP also interacts with another non-C/EBP protein designated v-fos transformation effector (FTE) (Kho, C. J., and Zarbl, H. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 2200-2204), which is identical to ribosomal protein S3a (Metspalu, A., Rebane, A., Hoth, S., Pooga, M., Stahl, J. , and Kruppa, J. (1992) Gene (Amst.) 119, 313-316). Bacterially expressed His-CHOP and in vitro translated (35)S-labeled FTE/S3a-Gal4 fusion protein co-immunoprecipitated using anti-CHOP antibodies, and both anti-CHOP and anti-FTE/S3a antibodies co-immunoprecipitated CHOP and FTE/S3a from lysates of Rauscher murine erythroleukemia cells overexpressing both proteins. The in vivo interaction of CHOP and FTE/S3a was also demonstrated in cells overexpressing FTE/S3a but with endogenous expression levels of CHOP. Western blot analysis demonstrated co-localization of CHOP and FTE/S3a in both the cytosol and the nuclei of non-transfected cells. Overexpression of FTE/S3a inhibited differentiation of Rauscher cells induced either by erythropoietin or by dimethyl sulfoxide. This inhibition was reversed partially by simultaneous overexpression of CHOP or of antisense fte/S3a. FTE/S3a appears to be a bifunctional ribosomal protein that regulates CHOP and, hence, C/EBP function during erythropoiesis.

v-Abl utilizes multiple mechanisms to drive G1/S progression in fibroblasts.

Transformation of 3T3 fibroblasts by the v-Abl tyrosine kinase replaces mitogenic and adhesion signals normally required for cell cycle progression. A 3T3 cell line conditionally transformed with v-Abl has been used to study v-Abl's effects on cell cycle in the context of either serum depletion or absence of adhesion signals. We show that E2F-dependent mRNAs, encoding proteins required for cell cycle progression, are induced by v-Abl. In addition, we identify two previously unknown targets of v-Abl signaling: (1) cyclin D1 and D2 mRNAs are induced upon v-Abl activation; and (2) the CDK inhibitor p27 is decreased upon v-Abl activation.

p53 deficiency increases transformation by v-Abl and rescues the ability of a C-terminally truncated v-Abl mutant to induce pre-B lymphoma in vivo.

Abelson murine leukemia virus (A-MuLV) is an acute transforming retrovirus that preferentially transforms early B-lineage cells both in vivo and in vitro. Its transforming protein, v-Abl, is a tyrosine kinase related to v-Src but containing an extended C-terminal domain. Many mutations affecting the C-terminal portion of the molecule block the pre-B-transforming activity of v-Abl without affecting the fibroblast-transforming ability. In this study we have determined the abilities of both wild-type and C-terminally truncated (p90) forms of v-Abl to transform cells from p53(-/-) mice. Lack of p53 increases the susceptibility of bone marrow cells to transformation by v-Abl by a factor of more than 7 but does not alter v-Abl's preference for B220(+) IgM(-) pre-B cells. p53-deficient mice have earlier tumor onset, more rapid tumor progression, and decreased survival time following A-MuLV infection, but all of the tumors are pre-B lymphomas. Thus, p53-dependent pathways inhibit v-Abl transformation but play no role in conferring preferential transformation of pre-B cells. Surprisingly, the C-terminally truncated form of v-Abl (p90) transforms pre-B cells very efficiently in mice lacking p53, thus demonstrating that the C terminus of v-Abl does not determine preB tropism but is necessary to overcome p53-dependent inhibition of transformation.

An unusual case of ileocaecal tuberculosis in an 80-year-old Caucasian male.

An 80-year-old Caucasian male presented with a seven-week history of diarrhoea and weight loss. Distal duodenal biopsies showed partial villous atrophy but he failed to respond to a gluten-free diet. Subsequently he developed a right iliac fossa mass associated with radiological evidence of ileocaecal ulceration. Colonoscopic biopsies from the caecum showed non-caseating granulomata and Ziehl-Neelsen (ZN) staining and culture for acid-fast bacilli (AFB) were negative. Crohn's disease was diagnosed and he was started on steroids. Although he showed an initial response, his condition then deteriorated and he died after six weeks. ZN staining of tissue at postmortem showed AFBs. Although a rare diagnosis in the UK, a high index of suspicion should be maintained for ileocaecal TB in patients with appropriate clinical features, even if classical risk factors for TB are absent.

Regulated expression and functional role of the transcription factor CHOP (GADD153) in erythroid growth and differentiation.

The hematopoietic growth factor erythropoietin (Epo) triggers changes in the expression of genes that encode important regulators of erythroid cell growth and differentiation. We now report that Epo markedly upregulates chop (gadd153) expression and that this transcription factor plays a role in erythropoiesis. Using a differential hybridization assay, we isolated a full-length cDNA of chop as an Epo upregulated gene in Rauscher murine erythroleukemia cells. RNase protection assays demonstrated that Epo or dimethyl sulfoxide induction increased steady-state mRNA levels 10- to 20-fold after 24 to 48 hours. Western blot analysis confirmed a marked increase in CHOP protein. Among the other c/ebp family members, only c/ebp beta was also upregulated during erythroid differentiation. Among normal hematopoietic cells examined, steady-state mRNA levels were highest in erythroid cells, with levels peaking during terminal differentiation. Transient overexpression of chop in Rauscher cells resulted in a significant increase in Epo- or dimethyl sulfoxide (DMSO)-induced hemoglobinization, further linking chop upregulation to erythroid differentiation. Artificial downregulation of chop in normal murine bone marrow cells with antisense oligodeoxynucleotides inhibited colony-forming unit-erythroid (CFU-E)-derived colony growth in a concentration-dependent manner. Burst-forming unit-erythroid (BFU-E)-derived colony growth was not affected. Using a Far Western type of analysis, we detected several potential CHOP binding partners among the nuclear proteins of Rauscher cells. Importantly, the number and relative abundance of these proteins changed with differentiation. The results strongly suggest that CHOP plays a role in erythropoiesis, possibly through interactions with both C/EBP and non-C/EBP family members.