Demonstration of gene-amplification by PCR in archival paraffin-embedded breast cancer tissue

Summary The polymerase chain reaction (PCR) is a powerful tool to examine genetic alterations in tumor samples. We describe a simple, rapid, nonisotopic PCR method to semi-quantitatively determine the number of gene copies in human formalin-fixed paraffin-embedded tissue. The procedure is exemplified by analysis of 15 years old breast cancer samples to determine the presence of amplification of c-erb-B2. The samples were obtained from routine specimens kept in pathological archives. Patients with amplified samples showed a poor prognosis, both for recurrences and death in breast cancer.     

Ultrasonic localization and cytologic identification of parathyroid tumors

Ultrasonic examination of the parathyroid glands was performed in 55 consecutive patients with subsequent surgically verified hyperparathyroidism. Ultrasound located 26 of 37 parathyroid adenomas in the neck. Eleven of 48 hyperplastic glands in the neck were visualized by ultrasound in 16 patients with primary or uremic hyperplasia. A parathyroid adenoma was revealed in all 3 patients with hypercalcemic crisis and in all 5 patients with an adenoma and a previously unsuccessful exploration of the neck. Two of 3 glands were visualized by ultrasound prior to secondary explorations in 3 patients with primary hyperplasia associated with the multiple endocrine neoplasia syndrome type I. Undetected parathyroid glands were generally small and located in regions of the neck difficult to detect by ultrasound. It was often difficult to unequivocally establish that identified lesions represented a parathyroid gland. Nodules and cysts of the thyroid and lymph nodes were misinterpreted for parathyroid lesions. Ultrasonically guided fine-needle biopsies were obtained from 8 parathyroid lesions, 7 thyroid tumors, and 2 lymph nodes. By cytologic examination these tissues could be discriminated after a differential staining of the aspirates.

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Resumen El examen ultrasónico de las glándulas paratiroides ha atraído considerable interés en los últimos años gracias a su capacidad para localizar glándulas aumentadas de tarnano antes de emprender el tratamiento quirúrgico, pero la identidad de tumores detectados por el ultrasonido puede ser difícil de establecer en forma inequívoca en el ecograma. Con el fín de lograr una forma de discriminación citológica, hemos utilizado un método de biopsia con aguja fine guiada por el ultrasonido.El examen ultrasónico de las glándulas paratiroides fue realizado en 55 pacientes consecutivos con hiperparatiroidismo verificado ulteriormente por cirugía. El ultrasonido pudo localizar 26 de 37 adenomas paratiroideos en el cuello. Once de 48 glándulas hiperplásicas fueron visualizadas por ultrasonido en 16 pacientes con hiperplasia urémica o de origen urémico. Se demostró la presencia de adenoma paratiroideo en todos los pacientes (3) con crisis hipercalcémicas y en todos los pacientes (5) con adenoma que habían sido sometidos a exploraciones previas y no exitosas del cuello. Dos de tres glándulas fueron visualizadas por ultrasonido antes de exploración secundaria en 3 pacientes con hiperplasia primaria asociada con el síndrome de neoplasia endocrina múltiple tipo I. Aquellas glándulas paratiroides que no pudieron ser detectadas generalmente fueron de tamaño pequeño y estuvieron ubicadas en regiones del cuello de difícil acceso ultrasónico. Con frecuencia fue difícil establecer en forma inequívoca si las lesiones visualizadas representaban una glándula paratiroides; nódulos y quistes tiroideos, así como ganglios linfáticos, fueron interpretados equivocadamente como lesiones paratiroideas. Biopsias con aguja fina guiadas por ultrasonido fueron obtenidas en 8 lesiones paratiroideas, 7 tumores tiroideos y 2 ganglios linfáticos. Estos tejidos pudieron ser discriminados mediante examen citológico y la coloración diferencial del material aspirado.

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Résumé L'échographie parathyroïdienne a été pratiquée consécutivement chez 55 malades dont l'hyperparathyroïdisme a été vérifié par l'intervention chirurgicale. L'échographie a permis de localiser 26 des 37 adénomes parathyroïdiens cervicaux. Onze parmi 48 glandes hyperplasiques cervicales ont été mises en évidence chez 16 malades qui présentaient un hyperparathyroïdisme primitif ou secondaire à un état urémique. Un adénome parathyroïdien fut découvert chez chacun des trois malades atteints d'hypercalcémie aiguë et chacun des 5 malades porteurs d'un adénome resté méconnu au cours d'une opération antérieure. Deux sur trois adénomes furent décelés avant une réintervention chez 3 malades qui étaient porteurs d'une hyperplasie primitive associée avec un syndrome MEN type I. Les parathyroïdes qui n'avaient pas été localisées étaient généralement de volume réduit et situées dans des régions difficilement accessibles à l'échographie cervicale. Souvent il fut délicat d'affirmer sans réticence que les lésions identifiées représentaient une glande thyroïde. Des nodules et des kystes thyroïdiens ainsi que des adénopathies furent confondus avec des adénomes parathyroïdiens. La biopsie à l'aiguille fine sous échographie fut pratiquée en présence de 8 lésions parathyroïdiennes, de 7 tumeurs thyroïdiennes et de 2 adénopathies. L'examen histologique après coloration des échantillons prélevés permit de définir leur nature avec exactitude.

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Presented at the International Association of Endocrine Surgeons at Hamburg, September 1983.

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Supported by the Swedish Medical Resarch Council (project no. 06264).     

Organization of a breast tumour clinic and aspects of data analysis of a clinical material.

With the aim of making it easier for patients with tumours or suspected tumours of the breast to obtain adequate medical advice and treatment, a special breast tumour clinic was established at the Department of Surgery of the University Hospital in Uppsala for women resident in the county of Uppsala. The waiting time was less than one week. In addition to palpation, fine-needle biopsy with cytological examination, mammography and xeroradiography were performed. Close co-operation between a few doctors responsible for the various diagnostic units made it possible to give the patients quick service and contributed to an efficient use of the diagnostic resources. The clinic functioned mainly within the framework of the existing medical care facilities and did not act as a screening project. Over a period of 15 months, 1244 women were examined. 116 had breast cancer, and 5 of these tumours were occult. A comparatively low mean tumour size and a low frequency of lymph node metastases would seem to indicate that the patients came under medical care at a relatively early stage. For recording of the case histories and examination findings, special medical records permitting data processing were used. This has facilitated follow-up of the patients, special checks on certain patient groups, statistical analyses and prospective investigations. We consider that, for the patient, a clinic of this kind offers considerable psychological and financial benefits, and that for the health service as a whole it means increased efficiency and a lower cost per patient. It seems probable that both the patient's and the doctor's delay are reduced.     

The diagnostic accuracy of palpation and fine-needle biopsy and an evaluation of their combined use in the diagnosis of breast lesions: report on a prospective study in 1244 women with symptoms.

In 1244 women with breast symptoms an evaluation by means of palpation was made with respect to diagnosis of malignancy according to a four-grade scale ranging from "definite cancer" to "no cancer." Aspiration biopsy and cytologic examination were then performed in 984 breast lesions. The diagnosis from the cytologic evaluation ranged from benign, through three grades of atypia (slight, moderate, grave) to cancer. A histologic diagnosis was made in 411 cases and in 28% it was cancer. Cancer was found in 92.5% of the patients with a palpatory diagnosis of "definite cancer," and in 50% of those with a palpatory diagnosis of "strong suspicion of cancer." In all patients in whom cancer was diagnosed cytologically, the same diagnosis was made at histology, while 87.5% of those with grave atypia at the cytologic examination were diagnosed histologically as having cancer. A false negative cytologic diagnosis was made in 4% of the cancer cases. With a combination of palpation and cytology, 91% of the cancer cases fell within the groups "definite cancer," "strong suspicion of cancer" (palpation)/"cancer," "grave atypia" (cytology). No patients with cancer were evaluated as "no cancer" (palpation)/"no atypia" (cytology). In this group of 697 patients, however, one cancer was discovered after 7 months. The investigation showed that a thorough palpatory evaluation is a prerequisite for a good result of aspiration biopsy, in particular to meet the risk of a false negative cytologic diagnosis. The cytologic examination revealed cancer in 6 and 12 cases, respectively, when palpation gave no or some suspicion of cancer, and in many cases it was able to eliminate malignancy suspected on palpation. Cytologic atypia indicated cancer in a relatively high per cent, but was also noted in many cases found to be benign histologically. The possibilities of reducing the number of "unnecessary" surgical biopsies by using a combination of palpation and cytology is discussed.     

Analysis of grey-level histograms as a method of classifying cells from the uterine cervix

An accurate single-cell classifier is an essential part of any system that uses image-processing and pattern-recognition techniques for prescreening cervical smears. The cell classifier, however, is heavily dependent on the feature extraction, which must be done automatically. This paper describes an attempt to use the grey-level histogram from a cell as a feature vector, in order to simplify part of the automatic feature-extraction process.     

Histopathological systems of breast cancer classification: reproducibility and clinical significance.

The inter- and intraobserver reproducibilities of the histopathological systems of breast cancer classification suggested by the World Health Organisation (WHO), the Armed Forces Institute of Pathology (AFIP) and Ackerman have been analysed. The reproducibilities of the three classification systems were only "fair" to "moderate" and no correlation with the five-year recurrence rate was found. Our results indicate that these classification systems are without biological significance and are useless for prognosis in the individual patient. When the tumours were classified according to degree of differentiation (high, moderate, low) or graded according to WHO (which includes both differentiation and nuclear atypia), however, there was a significant correlation with the five-year recurrence rate. Yet even such "reduced" subdivisions are of no value in judging prognosis for the individual patient at the time of diagnosis; rather, they are useful only in the follow-up analysis of groups of patients.