Imprint cytology of axillary lymph nodes as an intraoperative diagnostic tool.

AIMS: Frozen section histology of lymph nodes intraoperatively is associated with a false negative rate of twenty-five per cent. Imprint cytology is reported to have greater accuracy than frozen section. Accurate intraoperative examination of lymph nodes could help to prevent some patients from having a second axillary operation in breast cancer. This study assesses the sensitivity of imprint cytology to lymph node metastases using two different staining techniques. METHODS: Imprint cytology of 238 freshly excised axillary lymph nodes in 53 patients with stage T1-2, N0, M0 breast cancer was performed. Imprints were stained using toluidine blue dye and anti-pancytokeratin immunoglobulin to compare the two staining methods. A consultant histopathologist blinded to the routine histology results examined each set of imprints. A non-pathologist also examined each set of imprints to determine whether a technician could be used to screen slides in order to decrease the workload of the pathologist. RESULTS: Sensitivity was 82% with toluidine blue and 36.4% with anti-pancytokeratin immunoglobulin. Positive predictive values were 100% and 57% respectively for toluidine blue and anti-pancytokeratin. The false-negative rate was 18% for toluidine blue and 63.6% for anti-pancytokeratin antibody when examined by a consultant histopathologist. CONCLUSIONS: Imprint cytology using toluidine blue is as sensitive as frozen section in the detection of axillary lymph node metastases. A positive result may be acted upon with acceptable safety. Immunohistochemistry using antibody to pancytokeratin did not improve imprint sensitivity in this study. Imprint cytology could help to prevent a second operation in 80% of breast cancer patients with positive lymph nodes.

Outcome after extended follow-up in a prospective study of operable breast cancer: key factors and a prognostic index.

In 1990, 215 patients with operable breast cancer were entered into a prospective study of the prognostic significance of five biochemical markers and 15 other factors (pathological/chronological/patient). After a median follow-up of 6.6 years, there were 77 recurrences and 77 deaths (59 breast cancer-related). By univariate analysis, patient outcome related significantly to 13 factors. By multivariate analysis, the most important of nine independent factors were: number of nodes involved, steroid receptors (for oestrogen or progestogen), age, clinical or pathological tumour size and grade. Receptors and grade exerted their influence only in the first 3 years. Progestogen receptors (immunohistochemical) and oestrogen receptors (biochemical) were of similar prognostic significance. The two receptors were correlated (r=+0.50, P=0.001) and displaced each other from the analytical model but some evidence for the additivity of their prognostic values was seen when their levels were discordant.

Follistatin and activin A production by the male reproductive tract.

Follistatin is a binding protein for the activin and inhibin family of hormones, regulating their biological activity. In the male reproductive tract, the interaction of these factors is likely to be involved in the regulation of the proliferation of several cell types. We have investigated the presence of follistatin and activin A in seminal plasma using specific immunoassays and have localized follistatin and activin/inhibin subunits in the adult human testis, prostate and seminal vesicle to establish their likely sources. High concentrations of immunoreactive follistatin were present in seminal plasma in normal men (mean 97.9 ng/ml; 1.43 ng/ml in peripheral plasma) and were similar in men with oligo/azoospermia and following vasectomy. Follistatin immunoreactivity was localized to both Leydig and Sertoli cells of the testis, and to epithelial cells of the prostate gland and seminal vesicle, which are likely to be the predominant sources of the hormone in seminal plasma. Activin A was also present in seminal plasma in normal men but was undetectable following vasectomy, thus deriving from the testis. Consistent with this finding, the betaA-subunit was immunolocalized in Sertoli and Leydig cells but was not present in seminal vesicle or prostate gland. The functional significance of the high concentrations of follistatin secreted into seminal plasma by the prostate gland and/or seminal vesicle is uncertain, but they may regulate the biological activity of testis-derived activin A and inhibin B.

Prospective evaluation of prognostic factors in operable breast cancer.

In 215 patients with operable breast cancer (T1-T3, N0-1, M0) and no other or previous cancer, presenting to a single breast unit, sufficient tumour was available for the prospective determination of four putative biochemical markers of prognosis: oestrogen receptor (ER) activity, cathepsin D (cath D), epidermal growth factor receptor (EGFR) activity and cyclic AMP-binding proteins (c-AMP-b). There were significant inter-relationships between ER and EGFR (r = -0.26), c-AMP-b and cath D (r = +0.32) and ER and c-AMP-b (r = +0.14). After follow-up (median 36.2 months), a total of 55 recurrences (18 locoregional only) and 35 deaths were recorded. By univariate analysis, up to 10 of 18 biochemical, clinical and histopathological variables of potential prognostic value were significantly related to disease-free interval or death, but by multivariate analysis only oestrogen receptor concentration and node status contributed significantly to risk of both distant recurrence/death; in addition, tumour size made a small contribution to the risk for a distant recurrence only. Only two parameters, tumour grade and ER concentration, were significantly related to risk of locoregional recurrence by univariate analysis, but by multivariate analysis, only tumour grade was important. It is concluded that tumour ER concentration, axillary nodal status and tumour grade remain as the most important prognostic factors in the early years after presentation of operable breast cancer, with a minor influence of tumour size. At this time, the prognostic significance of quantitative measurements of ER concentration, carefully controlled for the quality of both assay and tumour specimen, is probably greater than is generally appreciated. We have yet to identify other factors, which add significantly to the short-term prognostic value of these key features.

N omega-nitro-L-arginine methyl ester reduces the incidence of IDDM in BB/E rats.

Evidence that nitric oxide (NO) is involved in cytokine-mediated islet beta-cell dysfunction and destruction in vitro has led to the hypothesis that increased production of NO may contribute to the pathogenesis of insulin-dependent diabetes mellitus (IDDM). This study demonstrates that oral administration of N omega-nitro-L-arginine methyl ester (an inhibitor of NO synthase) from 30 to 150 days of age significantly reduced (P < 0.05) the incidence of IDDM in diabetes-prone BB/E rats. This supports the idea that NO plays a significant role in the pathogenesis of IDDM in this animal model.

Analysis of disease distribution, activity and complications in the patient with inflammatory bowel disease.

Management strategies in Crohn's disease and ulcerative colitis should be based on up-to-date information on disease distribution, extent, activity and complications. A system of structured analysis is suggested, with separate consideration of destructive ulceration, inflammatory activity and other factors. Direct investigation of gut immunity by using whole gut lavage fluid (WGLF) is a valuable new technique of clinical investigation in IBD and related disorders. Recent studies have shown that the concentrations of plasma-derived proteins in WGLF provide objective measures of disease activity; and that this activity is a separate phenomenon from destructive ulceration and fibrosis. Neutrophils in the lumen can be in- investigated by cytology, or by assay of neutrophil elastase in WGLF. Cytokines and other immuno-regulatory mediators can also be detected. These new techniques can provide a description of intestinal immunity and inflammation, based on a non-invasive test of 2-4 h duration. Work in progress shows that patients who respond clinically to elemental diet treatment have unusually high concentrations of soluble IL2 receptor in WGLF; cytokine profiles may facilitate the selection of patients suitable for other new treatment modalities.

Negative testicular biopsy and asynchronous bilateral testicular germ cell tumour.

A case is described of a patient who was recently found to have a second primary testicular cancer 3 years after his first orchidectomy and negative contralateral testis biopsy.

Audit and its impact in the management of early prostatic cancer.

An analysis was made of a prospective database of 614 men with newly diagnosed carcinoma of the prostate who presented between January 1978 and December 1990; 3-monthly updates were available on their clinical, haematological and biochemical parameters and 6-monthly updates on chest X-rays, bone scans and skeletal X-rays. It was found that 107 men (mean age 73.5 years) had early disease at presentation. Their management was based on regular surveillance and the treatment deferred until disease progression or development of symptoms. The audit of outcome measured various clinical events. Four patients (3.7%) developed local failure, 11 (10.3%) developed bone metastases, 3 (2.8%) died of cancer with a median survival of 6.3 years, and 34 (31.8%) died of intercurrent disease with a median survival of 2.6 years. The observed survival for 12 years of the whole group was similar to the expected survival for an age-matched population in Scotland. The standardised mortality ratio was 81 (95% confidence limits 57-112).

Seminal fluid analysis and fine-needle aspiration cytology in the diagnosis of carcinoma in situ of the testis.

Surgical biopsies and fine-needle aspirates of peri-tumoural seminiferous tubules were taken from freshly-excised orchidectomy specimens. In addition, patients with suspected germ cell tumour provided a peri-operative sample of seminal fluid. All three tissue preparations were investigated using flow cytometry, immunochemistry for placental-like alkaline phosphatase and enzymochemistry for alkaline phosphatase. Biopsy and fine-needle aspiration cytology provide the greatest diagnostic accuracy for carcinoma-in-situ using these techniques. Seminal fluid analysis did not provide a satisfactory diagnostic yield in the series of patients presented. A seminal plasma placental-like alkaline phosphatase immunoassay failed to discriminate CIS because of the high level of background germ cell alkaline phosphatase.

Measurement of placental alkaline phosphatase activity in benign and malignant pleural effusions.

The usefulness of placental alkaline phosphatase (PLAP) as a diagnostic marker of malignancy was assessed in pleural fluid from 60 patients with effusions. Pleural fluid PLAP activities were measured by an enzyme linked immunoassay (ELISA) using the two monoclonal antibodies H17E2 and H317. Similar values were found in groups of patients with primary bronchial tumours (n = 12), secondary malignancies (n = 23), and "benign" conditions (n = 25). The highest values were found in a small subgroup of patients with metastatic ovarian carcinoma. However, the production of this enzyme by normal lung makes the measurement of PLAP in pleural fluid unhelpful as a diagnostic aid to distinguish "benign" from malignant effusions.

Post-menopausal atrophy in elderly women: is a vaginal smear necessary for diagnosis?

The diagnosis of post-menopausal atrophy should preferably be confirmed before any trial of oestrogen treatment for genito-urinary symptoms. The ideal method, cytological analysis of a vaginal wall smear, is not always feasible in elderly women. Might physical examination alone be sufficient? This study aimed to determine which clinical features might best predict atrophy on the smear. Of 120 consecutive women admitted to an acute geriatric unit (mean age 82 years), satisfactory smears were obtainable from 70 (58%). Sixty per cent of these showed severe atrophy but almost one-fifth showed no atrophic changes. Degree of atrophy correlated with physical thinness (p less than 0.01), low parity (p less than 0.01) and dryness on vaginal examination (p less than 0.001). However, none of the other physical findings traditionally thought to indicate atrophy were found to be associated, nor were age or medication. In contrast, severe atrophy was present in only one of nine women with breast cancer (p less than 0.01). This merits further study.

Prospective study of quantitative aspects of audit in a large general histopathology laboratory.

A non-computerised method of accurate prospective analysis of the quantitative aspects of the work of a general histopathology laboratory was devised. The method entails monitoring the progress of all biopsy and cytology specimens through the laboratory during selected monthly periods of study. The data collected include details of specimen type, block/section/stain details for each specimen, and information about the timed progress through technical, medical, and secretarial stages of specimen/report handling. The results give a detailed breakdown of the biopsy and cytology workload of this department. They give information about the rate of passage of specimens through the system and identify reasons for delay in the reporting of some cases. The method is easy to operate and will allow for the analysis of specific effects--for example, staff changes--on the efficiency of the department. The data will also provide some of the information required for the costing of histopathology laboratory services, and the type of information obtained will probably become a necessary requirement for laboratory accreditation.

Potentiation of hydrocortisone activity in skin by glycyrrhetinic acid.

The enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta-OHSD), which catalyses the conversion of cortisol to the inactive steroid cortisone in man (and corticosterone to 11-dehydrocorticosterone in rodents), was demonstrated by immunohistochemistry in skin biopsy samples from healthy volunteers and from patients with psoriasis and eczema. In-vitro studies confirmed the presence of the enzyme in skin from nude mice and showed that it is inhibited by glycyrrhetinic acid, the major active component of liquorice. By means of the skin vasoconstrictor assay, glycyrrhetinic acid was shown to potentiate the action of hydrocortisone. This work suggests a novel means of targeting glucocorticoid therapy.

De-differentiation with time in prostate cancer and the influence of treatment on the course of the disease.

There is little information on histological changes in prostate cancer during the course of the disease. We have studied 74 patients with carcinoma of the prostate who required 2 transurethral resections of the prostate (mean interval between resections 2.4 years). They constituted 18.4% of all patients with carcinoma of the prostate presenting to our clinic between January 1978 and April 1988. All tumours were staged by conventional methods and graded using the Gleason system. The Gleason sum score in those patients with tumour in both specimens increased in 49, remained constant in 12 and decreased in 7. Within this group were 34 patients who were treated expectantly. The mean Gleason sum scores in this group increased, with a concomitant increase in local tumour stage and development of metastases. Although this was not a randomised trial, there was no significant difference in survival between patients having "deferred" management and those treated immediately, either from time of diagnosis or from time of second resection. There was, however, a significant difference in the time to second resection, with the "deferred" group requiring repeat resection on average 1 year earlier. This study confirmed the concept of tumour de-differentiation with time and showed that this phenomenon occurs in both treated and untreated tumours. Although overall survival was not influenced by the type of initial therapy or its timing, local progression, as assessed by the need for further TURP, occurred earlier in those not receiving immediate therapy.

Surveillance of an at risk population: assessment of postal urine cytology.

The urine postal cytology kit Urotel was assessed against our standard cytological method; 184 patients attending for routine surveillance cystourethroscopy provided free flow urine specimens. The urine cytology was reported blind. Comparison of the 2 tests showed similar specificity but significantly higher sensitivity with the Urotel kit. These kits may have a place in the surveillance of patients at high risk of recurrent tumour or those with carcinoma in situ, reducing the frequency of hospital attendance.

The endocrinopathy of POEMS syndrome.

We describe the clinical features and post mortem findings in a patient with POEMS syndrome (P--polyneuropathy, O--organomegaly, E--endocrinopathy, M--M protein, S--skin changes), with particular emphasis on the evolution of the endocrinopathy over a three year period. In addition to gynaecomastia this case exhibited primary hypothyroidism and primary and secondary adrenal and gonadal failure. The widespread nature of the endocrine changes suggests that a circulating substance interferes with the action of trophic hormones, possibly by inhibiting second messenger production.

Carcinoma in-situ of the testis diagnosed on semen cytology.

The case is reported of a patient who presented to an infertility clinic following extensive treatment for testicular teratoma. An incidental finding on semen analysis was of abnormal cells suggestive of carcinoma in-situ. This diagnosis was later confirmed by biopsy of the remaining testicle. Radiotherapy will be given to the testis to ablate the germinal epithelium while sparing Leydig cell function.

Intravesical Evans Bacille Calmette-Guérin in the treatment of carcinoma in situ.

We report our experience in the treatment of carcinoma in situ of the urinary bladder using Evans strain Bacille Calmette-Guérin (BCG). The mean period of follow-up was 30 months; 18 patients received intravesical therapy and 16 of these were evaluable. They received 8 weekly instillations of 60 mg Evans strain BCG in 50 ml of saline. No intradermal BCG was given. The response rate was 93.75% at 3 months; this declined to 57% at 2 years. The complications of therapy were frequent but mild. Seven patients failed to complete the course. These results were comparable to those obtained with other strains of Bacille Calmette-Guérin.