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J J Going

Publications and source records attributed to J J Going.

At least 55 records · Page 3Linked to original sources
PubMed1996

Allelic imbalance at NME1 in microdissected primary and metastatic human colorectal carcinomas is frequent but not associated with metastasis to lymph nodes or liver.

Allelic imbalance at the NME locus on chromosome 17q21 was analyzed in colorectal cancer patients using a highly polymorphic microsatellite repeat sequence within NME1 itself. Duplicate samples of carcinoma and adjacent normal tissue was obtained by microdissection from 6 to 7-microns paraffin sections of 94 primary carcinomas (treatment years 1979-1993) and available lymph node and liver secondaries. In 55 patients informative (heterozygous) at this locus, allelic imbalance was examined in primary and secondary carcinomas. Microsatellite instability prevented assessment of allelic balance in two cases, and there was no evidence of homozygous loss at NME1 in any case analyzed. Allelic imbalance at the NME locus in carcinomas was frequent (27/53; 51%), and concordant results were obtained between primary carcinoma and secondary deposits in 30 of 33 (91%) cases. Three discordant cases showed allelic imbalance in secondary deposits but not the primary lesion. Although frequent, allelic imbalance at NME1 had no relationship to Dukes' stage at presentation or with subsequent hepatic metastasis, nor with the primary carcinoma site (proximal versus distal), tumor size, or mitotic or apoptotic index. Moreover, neither disease-free nor overall survival differed between patients with carcinomas showing NME1 allelic imbalance and patients with carcinomas that did not. Our results show that although allelic imbalance is frequent at the NME locus in primary and secondary colorectal carcinomas, there is no evidence to link this with clinical or pathological features or with metastatic potential. Microsatellite PCR and microdissection of enriched populations of carcinoma cells allowed uniformly successful analysis of samples from archival formalin-fixed paraffin-embedded tissue up to 15 years old and clear assessment of allelic imbalance in tumor specimens. Target sequences (e.g., microsatellites and minisatellites) up to approximately 200-250 bp may be reliably analyzed for allelic balance, suggesting that this method is of general utility in the genetic analysis of primary and metastatic neoplasia.

Adenocarcinoma↗
PubMed1996

Practical histological microdissection for PCR analysis.

Recovery of cells by histological microdissection is increasingly used for analysis by polymerase chain reaction (PCR) or microchemical techniques. This paper describes techniques of histological microdissection. Sections of archival formalin-fixed, paraffin-embedded tissue up to 15 years old were mounted on plain glass slides. Sections 6-7 microns in thickness stained with toluidine blue were dissected under proteinase K buffer solution, using an electrolytically sharpened tungsten needle in a bacteriological loop-holder and a Leitz mechanical micromanipulator (model M). Detached cell groups were recovered in a silicone-coated pipette tip for PCR analysis after digestion in 25-50 microliters of proteinase K (500/ml) in TRIS-HCl buffer (pH 8.3). Consistent amplification and analysis of microsatellite loci were obtained from 2 microliters of crude lysate using 28-30 cycles of PCR incorporating a 32P 5'-end-labelled primer, electrophoresis under denaturing conditions on 6 per cent polyacrylamide gels, and autoradiographic detection.

Alleles↗
PubMed1996

Cell kinetics in vivo of human breast cancer.

Rates of cell proliferation within tumours may provide prognostic information and help in the rational administration of chemotherapy and radiotherapy. In vivo labelling with 5-bromo-2'-deoxyuridine (BrdU) was used in 89 women with breast cancer to determine the labelling index for BrdU (BLI), the length of S phase (TS) and the potential doubling time of the tumour (Tpot). Kinetic data were obtained in 84 patients. The median BLI was 3.2 per cent, TS 12 h, and Tpot 12.5 days. There was no systematic difference in the labelling index determined by flow cytometry and by manual counting on sections. Labelling indices were significantly higher in aneuploid tumours, and in tumours not expressing oestrogen receptors, but were not correlated with tumour size, nodal status, or expression of c-erbB2. In vivo measurement of tumour cell kinetics can be made rapidly and reliably in the majority of human breast cancers, and may have a role to play in planning therapy.

Breast Neoplasms↗
PubMed1996

Three dimensional anatomy of complete duct systems in human breast: pathological and developmental implications.

AIMS: To reconstruct the arrangement in space of all major ducts and their branches from nipple to periphery of a human breast obtained at necropsy. METHODS: Duct tracing through cleared haematoxylin stained 2 mm sub-gross coronal slices of a complete necropsy breast and computer modelling of duct territories. RESULTS: All branches were traced for 10 complete duct systems of a single breast from a 19 year old girl. Their complexity prevented comprehensive modelling of individual ducts and rami using available computer software, but the territories (catchments) drained by individual duct systems did not overlap and could be reconstructed. Catchment volume and length of the central unbranched duct draining each catchment varied greatly. Duct spacing showed non-random uniformity which is also seen in rodent mammary glands. CONCLUSIONS: These spatial relations are consistent with mutual growth inhibition between duct systems during mammary development. Although there is no clear morphological distinction between mammary duct end buds and lateral buds in women, the present study does suggest that processes of branching morphogenesis occurring during development of the breasts in women do show some analogies with the growth of end buds/lateral branches/alveoli during rodent mammary gland development. Rodent models of mammary development may usefully suggest hypotheses about human breast biology. Less laborious methods of three dimensional reconstruction of mammary ducts and their branches from sub-gross slices, allowing more specimens to be studied, would be valuable for the study of normal human breast development and mammary intraepithelial neoplasia. Increasing power and decreasing costs of high definition image processing hardware and software may make such endeavours practicable.

Adult↗
PubMed1996

Quantitative radioimmunohistochemical measurements of p185(erbB-2) in frozen tissue sections.

The relationship between expression of the c-erbB-2 proto-oncogene and the biology of breast cancer has been investigated widely, most studies using immunohistochemistry in formalin-fixed, paraffin-embedded tissues. This technique is at best semiquantitative and there is a high degree of interstudy variability because of its subjective nature and poor methodological standardization. The relationship between the levels of expression and biology can be examined thoroughly only with an accurately quantitative technique. We have developed a radioimmunohistochemical assay to measure p185(erbB-2) in tissue biopsy specimens. The method involves incubating frozen sections with 125I-labeled monoclonal antibody, microautoradiograpy, and grain counting with image analysis. Sections of cell pellets with known c-erbB-2 levels are processed with each batch of samples as internal calibration standards. We have quantified c-erbB-2 expression in 60 breast carcinomas and compared the results with conventional immunohistochemistry. Radioimmunohistochemistry measured receptor levels throughout the range of expression in breast carcinomas, whereas conventional immunohistochemistry detected the protein only in the highest expressing tumors. The quantitative, objective data produced by radioimmunohistochemistry allow a more thorough evaluation of the relationship between c-erbB-2 expression and tumor biology. This technique may have applications in other fields where quantitative data is required and relevant monoclonal antibodies are available.

Breast Neoplasms↗
PubMed1995

Characterization of extensive genetic alterations in ductal carcinoma in situ by fluorescence in situ hybridization and molecular analysis.

BACKGROUND: The molecular genetic analysis of invasive breast cancer has identified breast cancer as a genetically complex disease. Ductal carcinoma in situ (DCIS) is thought to represent a preinvasive step in breast cancer progression, yet we know little about its biologic behavior or the genetic alterations present. Because of the increasing diagnosis of DCIS by mammography screening and the debate over how DCIS should be managed, there is a clear need to define the molecular events underlying the development of DCIS. PURPOSE: Our purpose was to identify patterns of genetic alterations in DCIS. METHODS: A group of 30 formalin-fixed, paraffin-embedded blocks of tissue collected from 1987 through 1989 from 21 patients with DCIS was studied. Chromosomal imbalances were determined by interphase cytogenetic analysis using the fluorescence in situ hybridization (FISH) technique. DNA probes were used that recognize chromosome-specific repetitive sequence loci at the centromeres of chromosomes 1, 3, 4, 6, 7, 8, 9, 10, 11, 16, 17, and 18. FISH was also used to detect ERBB2 gene amplification in DCIS. To complement the FISH studies, microsatellite analysis of markers near the BRCA1 region of chromosome 17 was done on tissue microdissected from multiple areas of DCIS. Chromosomal imbalances were determined by comparisons of chromosomal indices (total number of hybridization spots per total number of nuclei counted) of normal and DCIS tissue, using the two-sided Mann-Whitney test. RESULTS: Using FISH, we have identified patterns of DNA loss and gain of certain chromosome-specific centromeric markers in DCIS. We observed frequent gains of markers on chromosomes 3, 10, and 17 as well as loss of chromosome 18-specific centromeric sequences. ERBB2 gene amplification was detected in tumors from four of 15 patients studied and was clearly limited to the tumor cells within the ducts. Because of the availability of topologically distinct regions of tumors from individuals, we were able to show that paired tumor specimens from individuals share genetic alterations and also have unique ones, suggesting clonal diversity within tumors. The combination of FISH and microsatellite analyses suggested that alterations in chromosome 17 may be quite complex; three of five patients whose samples were analyzed had allelic imbalance at markers on the long arm of chromosome 17. CONCLUSIONS: FISH and microsatellite analyses are useful in detecting extensive genetic alterations in DCIS. Examinations of DCIS tissue using these techniques have identified chromosomes 1, 3, 10, 16, 17, and 18 as candidate sites worthy of immediate study. IMPLICATIONS: This approach may give direction to future research aimed at precisely mapping loci altered in DCIS and help in understanding the biologic events associated with tumor progression or recurrence.

Breast Neoplasms↗
PubMed1995

Specimen slice radiography of cancer in breast conserving excisions.

AIM: To investigate histologically the power of specimen slice radiography to record invasive and in situ carcinoma in breast conserving cancer excisions. METHODS: Twenty six cancer excisions were converted into 171 complete tissue slices, which were examined radiographically. From these slices, 373 histological blocks were processed and histological and radiographic assessments were compared. RESULTS: Radiography and histology mapped excision margins and lesions in detail. Radiographic prediction of histology was imprecise. Six invasive carcinomas were either undetectable by radiology or reached a radiologically clear margin. Six small invasive carcinoma satellites were not recognised. Adjacent ductal carcinoma in situ was undetectable in nine of 15 cases. CONCLUSIONS: Slice radiography and histology are capable of precise lesion mapping in breast cancer excisions, and clinical utility of such mapping merits investigation. Radiology alone is imprecise and to infer complete excision of breast carcinoma by radiography (of excision specimens or residual breast) alone may be unsafe.

Breast Neoplasms↗
PubMed1994

Efficiently estimated histologic cell counts.

Three different estimates were compared with complete nuclear counts made over 200 fields (20 sets of 10) selected from histologic sections of 18 breast carcinomas. All counting used a 10 x 10 square grid eyepiece graticule at x 400. Two area sampling techniques agreed well with overall counts (59,210 cells counted in all 200 fields; estimates were 58,500 and 59,805 cells), but an estimate based on linear intersection of nuclei yielded a lower estimate (23,382 cells). The most efficient estimate was 10 times the number of carcinoma cell nuclei in a random sample of 10 small graticule squares; summation over 10 fields yielded a mean ratio of estimated to counted total of 98.2% with limits of agreement of 89.9% to 107% for individual-case 10-field estimates. To improve efficiency further a simplified version of this estimate was devised and validated on carcinoma cell nuclei in 200 fields (20 sets of 10) selected from 20 invasive cervical carcinomas. An invariant sampling set was randomized to one of four rotations (0 degree, 90 degrees, 180 degrees, or 270 degrees). A total of 102,799 cells was counted in all 200 fields; the estimate was 101,280 cells. Summation over 10 fields yielded a mean ratio of estimated to counted total of 98.3% with limits of agreement of 89.8% to 108%. This simplified procedure facilitated estimation of cell numbers without detriment to agreement with denumerated counts and could be implemented as a custom-made counting graticule.

Breast Neoplasms↗
PubMed1994

Angiocentric invasion by lentigo maligna melanoma.

A case of angiocentric invasion by a lentigo maligna melanoma is reported. Pericapillary cuffing and permeative intimal expansion of veins by melanoma without luminal tumour cell emboli, thrombosis, or vascular destruction were associated with a lentigo maligna melanoma on the face of a 93 year old woman. This unusual infiltrative pattern suggests a specific interaction between melanoma cells and perivascular connective tissue elements, and may imply expression by tumour cells of receptors for molecules in the intima of blood vessels and in pericapillary stroma.

Aged↗
PubMed1994

Limb ischaemia after intra-arterial injection of Temazepam gel: histology of nine cases.

AIMS: To record the histopathological findings associated with intra-arterial injection of Temazepam gel by nine drug misusers. METHODS: Standard histological examination and immunocytochemistry for endothelial markers (factor VIII related antigen, Ulex europaeus lectin) were carried out. RESULTS: Intra-arterial injection of Temazepam gel may cause severe vascular injury and lead to amputation of fingers or limbs. Histological changes include myocyte necrosis, interstitial oedema, extensive arterial, venous, and capillary thrombosis, and sometimes vasculitis, endothelial swelling, and denudation. CONCLUSIONS: Inadvertent injection of Temazepam gel into arteries may cause catastrophic ischaemic damage, possibly as a result of toxic effects on endothelial cells.

Adult↗
PubMed1994

Cytopathology in the post mortem room.

AIM: To demonstrate the role of cytopathology in examining tumours found at post mortem examination. METHODS: Tumour deposits were found in 25 hospital necropsies. Cytological diagnosis made at the time of necropsy was compared with subsequent paraffin wax embedded histological sections. RESULTS: In 19 out of 20 cases with no previous histological diagnosis, cytology at the time of necropsy provided rapid and accurate assessment of tumour type. Subsequent histological examination of formalin fixed material merely refined the diagnosis in some cases. In the remaining five cases in which tumour type was known, cytological examination of deposits found at necropsy provided extra information that was useful for compiling a provisional report. CONCLUSIONS: Rapid cytological examination of tumours found during post mortem examinations provides accurate relevant information which can be used to produce a more comprehensive provisional necropsy report. The technique has advantages over frozen section histology and can provide useful cytological experience for histopathology trainees.

Adenocarcinoma↗
PubMed1993

Relationship of endothelial cell proliferation to tumor vascularity in human breast cancer.

Current studies of tumor angiogenesis rely on the concept that endothelium proliferates 30-40 times faster in tumors than in normal tissues. This evidence is based on histological autoradiographic data largely from animal studies. To assess endothelial cell proliferation in human cancer we used the more sensitive and specific technique of immunohistochemistry. We measured the frequency and distribution of endothelial cell proliferation and examined their relationship to tumor cell proliferation. For the first time, we also correlated endothelial and tumor cell proliferation with tumor vascularity. Twenty breast carcinomas from patients exposed to bromodeoxyuridine 3-8 h prior to surgery were double immunostained using antibodies to CD31 (as a marker of endothelium) and bromodeoxyuridine (as a marker of proliferation). The labeling index (LI) for both tumor and endothelial cells was determined and tumor vascularity was assessed by counting the number of CD31 positive vessels. Endothelial cell proliferation was predominantly at the tumor periphery while tumor cell proliferation occurred throughout the lesion. The mean LIs for endothelium and tumor were 2.2% (range, 0.8-5.3) and 7.3% (range, 1.3-17.1), respectively. There was no correlation between tumor and endothelial cell LI (P = 0.414) or between the tumor LI or endothelial cell LI and tumor vascularity (P = 0.08 and P = 0.39, respectively). These findings suggest that previous studies in animal tumors have significantly overestimated endothelial cell proliferation and that its importance in tumor angiogenesis may be related more to continual remodeling and migration of vessels than to proliferation alone.

Adult↗
PubMed1993

Influences on measurement of cellular proliferation by histology and flow cytometry in mammary carcinomas labeled in vivo with bromodeoxyuridine.

Field-selection procedures strongly influence histologically determined bromodeoxyuridine (BrdU) labeling indices (HLIs): For a series of 52 breast carcinomas, "high-labeling" fields had mean 2,000-cell HLIs that were, on average, more than double those for random fields, but the ratio was extremely variable. Although there may be a priori reasons for preferring particular field-sampling techniques, nonstandardized kinetic data from different laboratories will be uninterpretable and therefore of limited clinical application. Also, if heterogeneity of tumor cell proliferation is neglected, it is possible that incomplete or even misleading data may be gained. Total flow cytometric BrdU labeling indices (TLIs) were not, on average, higher or lower than HLIs for the same cases, but individual discrepancies could be pronounced. The best correlation of flow cytometric LIs with HLIs was obtained for aneuploid carcinomas with a restricted labeling index derived for aneuploid (presumptively neoplastic) cells only. Although flow cytometric analysis avoids the statistical problems associated with accurately determining a small percentage from relatively few cells, it lacks precise definition of the character of the cells counted, but even histologic methods are not entirely free of this difficulty.

Breast Neoplasms↗
PubMed1992

Ras p21 in breast tissue: associations with pathology and cellular localisation.

Immunocytochemistry with monoclonal antibody Y13-259 demonstrated p21 ras in paraffin sections of breast tissue from 171 women: 85 with invasive breast carcinoma, 14 with non-invasive carcinoma and 72 with benign changes only. Many different tissue elements contributed to ras expression. Semiquantitative assessment showed that intensity of immunostaining in the normal epithelium of large ducts, small extralobular ducts and terminal duct lobular units (TDLU) was usually exceeded by that of myoepithelial cells. Vascular smooth muscle and apocrine epithelium also stained strongly, but the flat epithelial cells lining cysts did not express detectable p21 ras. There was a progressive increase from normal epithelium through epithelial hyperplasia of usual type and atypical hyperplasia to carcinoma in situ, without further increase in invasive carcinoma. Expression in carcinomas was inversely related to oestrogen receptor content but independent of the prognosis-associated variables of size, histological type, vascular invasion or lymph node metastasis.

Antibodies, Monoclonal↗