Expression of the calcium receptor in human breast cancer--a potential new marker predicting the risk of bone metastases.

AIMS: This study investigates whether the calcium-sensing receptor (CaR) is commonly expressed in primary breast cancers. The CaR controls secretion of PTHrP in several breast cancer cell lines and PTHrP is known to stimulate osteolysis during metastatic bone resorption. Whether this could explain the propensity of breast cancers to develop bone metastases has not been explored. METHODS: With Ethical Committee approval, immunohistochemistry was performed using a commercially available antiCaR antibody (AffinityBioReagents, Cambridge, UK) on archived histological sections of primary tumours from patients who died with advanced breast cancer. Intensity of CaR expression was assessed by two independent observers on a 6-point scale. RESULTS: One hundred and eight patients with breast cancer were found to have positive bone scans, 42 patients had died. Of the patients with negative bone scans, 23 had liver or lung metastases. Most patients with strongly expressed CaR (score 4-5 on immunohistochemistry) had bone metastases (13/15 patients) compared with 2/23 patients with normal bone scans (p < 0.001, chi(2) test). Other clinical/pathological markers (ER, PR, c-erb B-2, LN status) were not significantly different between patients with CaR-positive or CaR-negative tumours. CONCLUSIONS: CaR expression is common in a selected group of patients with advanced primary breast cancers. A prospective study should investigate if patients with CaR-positive tumours are more likely to develop bone metastases.

Preoperative mammotome biopsy of ducts beneath the nipple areola complex.

AIM: To evaluate the role of ultrasound guided mammotome biopsy of the ducts beneath the nipple areola complex (NAC), as a new technique in detecting the occult involvement of the NAC in breast cancer patients prior to nipple preserving subcutaneous mastectomy. METHOD: A prospective study where 33 women requesting nipple preserving mastectomy for invasive or in situ disease were offered the procedure to determine if leaving the nipple was safe. A 5 mm skin incision was made after infiltration with local anaesthetic and the 11G mammotome needle was positioned beneath the nipple under ultrasound guidance which was turned through 360 degrees as the biopsies were taken. The procedures were performed by trained non-radiologists. RESULTS: Thirty-three women had 36 procedures. Seven out of the 36 had a positive mammotome biopsy. Twenty-three patients had 26 NAC preserving mastectomies with immediate reconstruction. Three had bilateral procedures. Ten patients had NAC sacrificed. The histopathology of the mastectomy specimen correlated 100% with the mammotome biopsy. CONCLUSION: Preoperative ultrasound guided mammotome biopsy of the ducts beneath the NAC is a safe, reliable and accurate technique and is evolving as an oncologically safe procedure. The large mammotome needle can be visualized easily under high resolution, near field high frequency scanners and this increases the accuracy of the biopsy. It can replace the traditional frozen section and be used as an alternate. It can be performed safely by an appropriately trained non-radiologist (surgeon/breast clinician).

Sonographically guided mammotome excision of ducts in the diagnosis and management of single duct nipple discharge.

AIM: To describe our experience in the use of ultrasound guided mammotome in the diagnosis and management of single duct nipple discharge. METHODS: Patients for whom surgical excision of the single duct had been advised for single duct nipple discharge were offered ultrasound guided mammotome excision of the duct as an alternative to surgical excision. The procedure was performed in the breast clinic by a surgeon or a breast clinician who had interventional ultrasound skills. RESULTS: Seventy-seven patients had 81 procedures. Follow-up at a mean time of 16 months revealed resolution of the presenting problematic discharge in 95% of patients. Nipple discharge recurred in four patients. Two patients had microdochectomy and two had a repeat mammotome for recurrence of symptoms. Complications were mild and infrequent. CONCLUSION: Ultrasound guided mammotome excision is a new tool in the work up and management of single duct nipple discharge. It can be performed under local anaesthetic by a surgeon/breast clinician or radiologist with interventional ultrasound skills. It has the potential to replace surgical excision (microdochectomy) as a treatment for nipple discharge.

Behavioral indicators to detect ovarian phase in the dromedary she-camel.

This pilot study was conducted to test the hypothesis that female camels behave differently in various ovarian phases in the presence of a restrained male camel. The aim was to identify behavioral patterns which could be used as indicators to detect ovulatory phase by visual observation in the presence of a restrained virile bull. Twenty-four healthy, nonpregnant, and nonlactating adult females were used. Transrectal ultrasonography was performed for each animal once a week over a 3-week period to determine the phase of the ovarian cycle. Females were considered to be in the ovulatory phase (O) when there was at least one preovulatory follicle (12<Ø<19 mm) protruding from the ovarian surface, and in the nonovulatory phase (NO), when growing follicles, regressing follicles, or corpora lutea were detected. Immediately after examination, each female was freely exposed to a restrained bull for 15 minutes, and her behaviors were filmed. The videos were analyzed through a focal animal-sampling ethogram (states: looking at the male; looking outside; standing close to the male; searching; and lying down; events: interaction with the male; urination; defecation; sound emission; and steps). A score for tail position (tail score: 1 = close to the vulva, 2 = horizontal, 3 = vertical) and for interest in the bull (male time score: from 1 to 5; 1 = <20% of observation period spent near the bull; 5 = more than 80%) were recorded. Ovulatory phase camels showed higher interest in the male than nonovulatory phases: they stood close to the male for longer periods (P = 0.0159), interacted with the male more frequently (P = 0.0004), and tended to lie down in front of him (P = 0.1202). Moreover, ovulatory phase had a significant effect on male time score (P < 0.01), mature follicular ovarian phase being associated with higher scores. Seeking the male has already been proposed as a behavioral indicator of estrus in camels, this has now been confirmed using a standardized ethogram. The present results clarify that camels behave differently in different ovarian phases and that monitoring their behavior in the presence of a restrained bull could help detect their ovulatory phase. This would have profound implications for enhancing fertility in dromedary camels by improving timing of mating or artificial insemination.

Segmental pulmonary veno-occlusive disease secondary to lung cancer.

This report describes two cases of segmental pulmonary vein occlusion secondary to lung malignancy in which lung biopsies showed histological features of veno-occlusive disease. These are the first cases to be reported in the literature in which such lung parenchymal histological changes are described in association with lung malignancy.

Perivascular nerves of the human basal cerebral arteries: I. Topographical distribution.

In the present study the topographical distribution of the intrinsic nerve plexuses of the basal cerebral arteries in humans was quantified and the relation between vessel diameter and nerve density was investigated. Whole-mount preparations of various segments of the basal cerebral arteries from middle-aged patients were stained for protein gene product (PGP) 9.5. The deep nerve plexuses, located at the adventitial-medial border, were quantified by image analysis. Confocal scanning laser microscopy was used to study nerve plexuses throughout the adventitia. Transverse cryostat sections were stained for PGP 9.5, tyrosine hydroxylase and neurofilament, and quantified. The results showed a three-layered configuration of the adventitial nerves. Measurements on whole-mounts demonstrated that nerve densities were highest in the posterior communicating artery (PCom), and next highest in the proximal parts of the posterior cerebral artery (PCA) and anterior choroidal artery. There appeared to be no clear relation between nerve density and vessel diameter. The measurements on sections confirmed the high nerve densities in the PCom and PCA. Tyrosine hydroxylase- and neurofilament-immunoreactivities appeared to demonstrate separate subpopulations of the overall nerve plexuses, representing sympathetic and, possibly, sensory fibers, respectively. Densities of both subgroups generally followed those of PGP 9.5-immunoreactive nerves. Transmission electron microscopy suggested motor function of the deep nerve plexuses. The results indicate a stronger neuronal influence on this part of the cerebral circulation than hitherto reported. It is concluded that human basal cerebral arteries display a topographical distribution of deep perivascular nerves, and that nerve density is determined by locality rather than by vascular diameter.

Epithelial-myoepithelial carcinomas of the bronchus.

Epithelial-myoepithelial carcinomas are very rare in the lung, and little is known about the relationship of their histologic features to prognosis. We describe five primary pulmonary epithelial-myoepithelial carcinomas with details on clinical presentation, histology, and immunohistochemical profiles. We also reviewed the literature to detail further their prognosis. The patients' ages ranged from 33 to 57 years (average 51 years). The tumors were all endobronchial and the patients presented with symptoms or imaging features of airway obstruction. The tumors were completely resected; none showed nodal involvement. All five patients are alive and free of disease 4 months to 8 years (average 4.2 years) after surgery. Four tumors showed a mixed pattern of glands lined by a dual layer of cells and solid sheets of either spindle cells or clear cells, the glandular and solid components being present in variable proportions. The fifth tumor comprised purely spindle cells. The mitotic rate was <1/20 high power fields in both the glandular and spindle/clear cell components. In one case there was focal nuclear pleomorphism. The inner layer of the glands stained for cytokeratins and epithelial membrane antigen, and the outer layer for S-100 and smooth muscle actin. In one case the spindle cells stained for CD34. A review of published cases shows the majority of tumors behave in an indolent fashion, the rare aggressive tumors being predominantly myoepitheliomatous. Nevertheless, the term epithelial-myoepithelial carcinoma is preferred because of their malignant potential. A high mitotic rate, tumoral necrosis, and nuclear pleomorphism appear to be adverse prognostic factors.

Substance P-, calcitonin gene-related peptide- and C-flanking peptide of neuropeptide Y-immunoreactive fibres are present in normal synovium but depleted in patients with rheumatoid arthritis.

By means of antisera to cytoplasmic components of nerve fibres and neuropeptides which are known to be present in sensory or sympathetic nerves we have examined the distribution of both total and different types of nerve fibres in normal and inflamed human synovial tissue. Samples of synovia were obtained at surgery from five normal and five rheumatoid patients (age range 10-77 years). In order to map the overall neural innervation of the synovium, antiserum to the general neuronal marker protein gene product 9.5 was employed. Substance P and calcitonin gene-related peptide antisera were employed to identify sensory fibres and antisera to the C-flanking peptide of neuropeptide Y to distinguish sympathetic nerves. In normal synovium protein gene product 9.5-immunoreactive fibres were numerous, in particular, the vasculature was densely innervated. Free protein gene product 9.5-immunoreactive fibres were less numerous but were present in all synovia examined, and in many cases these extended to the intimal layer. Neuropeptide immunostaining was predominantly found in perivascular networks. Fibres immunoreactive for the C-flanking peptide of neuropeptide Y were exclusively located around blood vessels whereas free fibres were immunoreactive for substance P or calcitonin gene-related peptide. As with free protein gene product 9.5-immunoreactive fibres, fibres expressing substance P or calcitonin gene-related peptide immunoreactivity were often seen in the intimal cell layer. In rheumatoid arthritis a similar innervation to that seen in normal synovium was apparent in the deep tissue but fibres immunoreactive for protein gene product 9.5, the C-flanking peptide of neuropeptide Y, substance P or calcitonin gene-related peptide were not visible in the more superficial tissues or the intimal cell layer. In addition, immunostaining of neuropeptides in the deep tissue was weaker in the diseased tissues than in normal controls. The data unequivocally demonstrate that synovial tissues are richly innervated and confirm the presence of both sensory and sympathetic nerves. The absence of nerves which innervate the superficial synovium in rheumatoid arthritis might suggest that there is increased release of substance P, calcitonin gene-related peptide and the C-flanking peptide of neuropeptide Y, reducing the stores in the nerves to levels below that detectable by immunocytochemistry. However, since protein gene product 9.5-immunoreactive nerves were not seen in the inflamed tissue it is probable that synovial growth outflanks neural growth and consequently as the disease progresses neural structures become restricted to deeper tissues.(ABSTRACT TRUNCATED AT 400 WORDS)

Observer variation in the histopathological classification of thymoma: correlation with prognosis.

AIMS: To assess the ability of three histopathologists, experienced in thoracic surgical reporting, consistently to classify thymomas as cortical, medullary, or mixed pattern tumours. METHODS: Three histopathologists classified 74 thymomas (none frank carcinomas) as of either cortical, medullary, or mixed pattern, on two separate occasions. Kappa statistics were used to assess inter- and intra-observer agreement. Tumour type was compared with surgical stage as a predictor of biological behaviour. RESULTS: Inter- and intra-observer agreement were only moderate (kappa 0.48 and 0.52, respectively). For only 26 of 74 tumours could a categorisation be consistently agreed on. Follow up information was obtained for 73 cases, with a mean follow up period of five years. The prognoses for those 26 of 74 cases appeared to be at variance from previously reported studies, and showed internal inconsistency, with the mixed pattern category showing a worse survival than the cortical category. For the group as a whole, however, stage at presentation was related to survival, with an overall five year survival of 78% (100% for stage I, 84% for stage II, 27% for stage III and 0% for stage IV). CONCLUSIONS: The classification of thymomas into cortical, medullary, or mixed pattern tumours is difficult to apply. Surgical stage remains a better guide to prognosis.

Practical use of a word processor in a histopathology laboratory.

Some of the facilities available with a commercially purchased word processing program, linked to a DEC PDP 11/23 computer are described, together with an account of the practical histopathological use. The system is based on a share of the computer with a Clinical Chemistry Department. Development was time-consuming and required the constant availability of the Department of Physics. However, once working, considerable saving in secretarial time has resulted and a number of projects have been started which would not have been contemplated without the use of the word processor and its linked computer.

Abnormal p53 expression in lung neuroendocrine tumors. Diagnostic and prognostic implications.

The diagnostic and prognostic implications of p53 immunostaining have been investigated in 59 pulmonary neuroendocrine tumors, including typical carcinoids (n = 15), so-called "atypical carcinoids" (n = 22), and small cell lung carcinomas (SCLCs; n = 22). Immunocytochemistry was performed on formalin-fixed, paraffin-embedded samples using the monoclonal antibody PAb1801, which has been shown to be suitable for staining fixed and embedded tissue sections. p53 immunoreactivity was restricted to atypical carcinoids (45% of the cases being immunoreactive) and to SCLCs (which were positively stained in 59% of the cases), whereas it was consistently lacking in typical carcinoid tumors. When the group of the so-called "atypical carcinoids" was further reclassified, p53 immunostaining was strictly confined to those cases belonging to the histologically more aggressive subsets (well differentiated neuroendocrine carcinoma subsets II and III). Within the same tumor type, however, p53 immunoreactivity did not correlate with the clinical outcome of the disease and was not predictive of the length of survival. The data indicate that abnormal p53 expression (which is strictly dependent on structural abnormalities of the p53 gene) is detectable in the majority of neuroendocrine carcinomas of the lung and might represent a useful adjunct in the differential diagnosis of pulmonary neuroendocrine neoplasms, particularly in routinely fixed and embedded small bronchoscopic biopsies.

Immunohistochemistry of nitric oxide synthase demonstrates immunoreactive neurons in spinal cord and dorsal root ganglia of man and rat.

The presence of nitric oxide synthase in spinal cord and dorsal root ganglia was investigated by immunohistochemistry using antibodies against the constitutive neuronal form of nitric oxide synthase (NOS). NOS immunoreactivity was present in both man and rat with similar distribution, being present in primary sensory neurons of dorsal root ganglia and their afferent terminals in dorsal horn of spinal cord. NOS immunoreactive interneurons were found in the superficial layer of the dorsal horn, around the central canal and in the intermediolateral cell column. NOS immunoreactivity was also present in numerous motoneurons in the ventral horn. The widespread distribution of NOS in both sensory and motor nervous system is indicative of the involvement of nitric oxide in different neural functions.

Correlation of pS2 expression of involved lymph nodes in relation to primary breast carcinoma.

Expression of the pS2 protein in breast carcinoma is a useful guide to prognosis and response to tamoxifen. We have investigated pS2 protein expression in both the primary tumour and lymph node metastases (LNM) using a computer-assisted image analysis system. In a consecutive series of 208 patients undergoing surgical excision of primary breast cancer with axillary clearance, 89 patients were found to have involved lymph nodes. We found a highly significant correlation between pS2 expression in primary tumours and their LNM when 5% was taken as the cut-off for positive staining (Fischer Exact, P < 0.0001). There was also a highly significant correlation between the proportion of positive staining between the local metastases and primary tumours (Spearman's rank order correlation = 0.87; P < 0.0001). We conclude that the pS2 status of LNM can be accurately predicted from the pS2 status of the primary tumour. As such, appropriate adjuvant therapy for primary breast cancer, or second line therapy for disseminated disease can be selected on the pS2 status of the primary tumour alone.

Solitary fibrous tumour of the diaphragm.

Solitary fibrous tumours are uncommon neoplasms that have been described as occurring in the pleura, peritoneum, lung, mediastinum, pericardium, nose and paranasal sinuses. This report describes the first known case of a solitary fibrous tumour of the diaphragm, occurring in a 60 year old woman with a two year history of respiratory symptoms. The lesion was initially misdiagnosed as an elevated left hemidiaphragm. The operative findings and histological appearance of the tumour are described. A review of the literature examines the various types and presenting features of solitary fibrous tumours as well as their clinical behaviour and postulated origin.

Lymphangiosarcoma of the hand arising in a pre-existing non-irradiated lymphangioma.

Lymphangioma is an uncommon lesion which is widely believed not to undergo malignant change. Lymphangiosarcoma is even rarer and most of the cases reported in the literature were encountered in the setting of chronic lymphoedema, although occasional cases have been reported in irradiated lymphangiomata. This brief communication describes a case of multifocal lymphangiosarcoma arising in a non-irradiated, long standing lymphangioma of the hand. To our knowledge no similar case has been reported previously in the English literature, which is briefly reviewed.

A Melanoma Registry based on a computer-run word and data processing system.

A Melanoma Register, previously organised on a filing card system, has been largely transferred to a computer which utilises both word and data processing programs. This transfer has revolutionised not only the analytical potential of the material, but the follow up of all patients, both old and new. The system is applicable to many other tumours and diseases.