Non-deleterious inhibition of endogenous peroxidase activity (EPA) by cyclopropanone hydrate: a definitive approach.

Endogenous peroxidase activity (EPA) poses a serious problem in immunoperoxidase localization of antigens unable to withstand deleterious effects of aldehyde fixatives, alcohols, and various oxidative reagents. This has forced the development of more selective inhibition methods. Of these, phenylhydrazine or azide combined with small amounts of H2O2 have proved quite effective.

Immunocytochemical diagnosis of early myocardial ischaemic/hypoxic damage.

The sensitive and reliable dinitrophenyl (DNP) hapten sandwich staining (DHSS) procedure (B. Jasani et al., Virchows Arch (Pathol.

Immunological events leading to destructive thyroiditis in the AUG rat.

Single or infrequent observations in patients or animals with autoimmune thyroid disease (AITD) have failed to elucidate the exact sequence of pathogenetic events leading to thyroid cell destruction. A detailed serial morphological and functional study of experimental AITD (EAITD) in the female AUG rat was therefore undertaken. Following induction of EAITD with thyroglobulin (Tg) in adjuvant antibodies to Tg were detectable one week after the initial immunization, at which stage Ia positive vascular endothelium was observed within the thyroid.

Light microscopic immunocytochemical demonstration of thyroid-stimulating hormone (TSH) receptors on normal rat thyroid cells.

We present a method for immunohistochemical demonstration of surface-associated thyroid-stimulating hormone (TSH) receptors on direct imprints of rat thyroid follicular cells at the light microscopic level, by use of dinitrophenyl (DNP)-labeled bovine TSH as the primary probe in a DNP-hapten sandwich staining (DHSS) procedure. The light microscopically invisible diaminobenzidine (DAB) product of the enzyme reaction was amplified with a DAB enhancement technique (silver amplification), which yielded a reliable and distinct light microscopically visible cell surface staining. The consistently negative control results after inhibition with TSH receptor-specific reagents provide evidence in support of immunohistochemical localization of surface membrane-associated TSH receptors.

Antigen CA125 in tumor tissue and serum from patients with adenocarcinoma of the ovary.

The presence of cancer antigen CA125 has been assessed immunohistologically in formalin-fixed tumor tissue and immunoradiometrically in serum from 41 women with residual epithelial ovarian carcinoma after diagnostic laparotomy. CA125 was detected in tumor tissue from the majority of patients (34/41), including those with normal serum levels (15/18). The major determinant of serum CA125 concentration appears to be the size of the residual tumor.

Tamm-Horsfall protein is a marker of renal and extra-renal rhabdoid tumours.

A monoclonal antibody (MAb) to Tamm-Horsfall protein (THP) was used to stain 6 renal rhabdoid tumours (RRT) and 2 primary extra-renal rhabdoid tumours (E-RRT). One of the E-RRT was a tumour from the posterior fossa of a 3-year-old child and the other was a lump from the right side of the neck in an 18-month-old girl. Five of 6 RRT and both cases of E-RRT were positive for THP.

Immunohistologically definable light chain restriction in autoimmune disease.

Analysis of serum immunoglobulins from patients suffering from autoimmune disease has shown that pathogenically relevant autoantibodies directed at organ specific antigens are light chain restricted, i.e., they are entirely lambda or kappa type in a given patient.

Study of childhood renal tumours using a monoclonal antibody to Tamm-Horsfall protein.

A monoclonal antibody to Tamm-Horsfall glycoprotein was used for the immuno-localization of Tamm-Horsfall protein in formalin fixed, paraffin embedded tissue sections of childhood renal tumours, normal children's kidneys, and human fetal kidneys. The procedure was a dinitrophenyl hapten sandwich staining method. The antibody, diluted 1/100,000, gave a very strong and specific staining of the loop of Henle and distal tubules of normal and fetal kidneys.

Synchronous duodenal neuroendocrine tumours in von Recklinghausen's disease--a case report of co-existing gangliocytic paraganglioma and somatostatin-rich glandular carcinoid.

The co-existence of a gangliocytic paraganglioma and a glandular psammomatous carcinoid in the duodenum of a patient with von Recklinghausen's disease and bilateral phaeochromocytomas is reported. The two lesions were considered to be distinctive by light microscopy, electron microscopy and immunocytochemistry. The cells of the glandular carcinoid showed strong cytoplasmic immunoreactivity for somatostatin and contained only scanty intracytoplasmic microfilaments on electron microscopy.

An improved method for the inhibition of endogenous peroxidase non-deleterious to lymphocyte surface markers. Application to immunoperoxidase studies on eosinophil-rich tissue preparations.

The inhibitory effect of phenylhydrazine and azide combined with either pre-formed or nascent hydrogen peroxide H2O2 upon endogenous peroxidatic activity, expressed by tissue eosinophils in different disease states, was investigated. It was found that whilst endogenous peroxidatic activity due to eosinophils in a Hodgkin's disease and a histiocytosis X case were adequately inhibited by phenylhydrazine combines with pre-formed or nascent H2O2, the eosinophils in the Onchocerca volvulus nodule were either not at all or only partly inhibited by the two regimens. On the other hand, a combination of azide with nascent H2O2 proved consistently effective against this resistant form of endogenous peroxidatic activity.

Chromogranin A and B and secretogranin II in medullary carcinomas of the thyroid.

We have studied the presence of chromogranins A and B and secretogranin II in medullary carcinomas of the thyroid using the methods of immunoblotting and immunohistochemistry. All three antigens were identified by immunoblotting. Chromogranin A and secretogranin II behaved identically to the adrenal antigens in electrophoresis, whereas the molecular size of chromogranin B in the thyroid tissue appeared slightly smaller, possibly due to a higher degree of proteolytic processing.

Presence and possible significance of immunohistochemically demonstrable prolactin in breast apocrine metaplasia.

Paraffin wax embedded formalin-fixed benign breast disease tissue taken from 17 patients (15 with microcystic disease and 2 with fibroadenoma) was studied for the presence of tissue bound prolactin using a rabbit antiserum against human prolactin applied in conjunction with a highly sensitive modified version of the dinitrophenyl (DNP)-hapten sandwich staining (DHSS) procedure. Sections taken from 14 of the 15 cases showing apocrine cystic changes exhibited strong prolactin staining restricted to the cytoplasm of metaplastic apocrine cells lining the cysts. Normal lobules and ducts and blunt duct proliferations were all negative, as were also the two cases of fibroadenoma.

Lack of sampling site variation in chorion villus biopsy as assessed by DNA, enzymatic, morphological and cytogenetical analyses.

A study of villus samples from eight random sites on five electively aborted chorion sacs was performed to determine any significant differences in yield, quality and composition of DNA, iduronate sulphate sulphatase activity and karyotype status. The villi were also examined for their histological characteristics (e.g.

Histological demonstration of immunoreactive metallothionein in rat and human tissues.

Antibody to rat liver metallothionein prepared by the method of Brady and Kafka (1979) was used to localise immunoreactive metallothionein using a sensitive DNP hapten sandwich technique applied to formalin fixed wax embedded tissues. Rat tissues examined were liver, kidney and small intestine, taken from normal animals, from animals fasted after receiving either an oral dose of water, or 1 ml zinc acetate solution either orally or by intraperitoneal injection, (3-4 mg Zn++/Kg body weight). Human tissues examined were 6 histologically normal liver biopsies and small intestine including histologically normal jejunal biopsies and samples of ileum obtained at operation.

Immunohistochemical localization of cell surface receptors using a novel method permitting simple, rapid and reliable LM/EM correlation.

A method is presented which allows correlative serial section analysis by light and electron microscopy of cell surface antigens in monolayer cultures. Sites of antigenicity are shown by deposition of diaminobenzidine after pre-embedding, immunoperoxidase immunocytochemistry. Osmication is replaced by the use of gold chloride which specifically enhances the electron density of diaminobenzidine.

Immunohistochemical identification of prolactin and 24K protein in secretory endometrium.

Inadequate endometrial differentiation is a cause of infertility and recurrent pregnancy loss. Diagnostic histologic dating criteria may be supplemented by the immunohistochemical identification of protein markers in late secretory endometrium. Late secretory endometrium has been shown to contain prolactin.

Multiple hormone storage by 'polycrine' cells in the pancreas (from a case of nesidioblastosis).

Pancreatic tissue from a case of neonatal hypoglycaemia with nesidioblastosis has been studied by routine light and electron microscope techniques and by highly sensitive light and electron microscope immunolocalization methods. A hyperplastic nodule within the pancreas from this case contained enlarged distorted haemorrhagic islets, with a variable rim of exocrine tissue. Islet cells in these areas were shown to contain more than one hormone in separate granules.

Immunogold electron-microscopic localisation of calpain I in skeletal muscle of rats.

By using a double-affinity-purified first antibody and colloidal gold-conjugated second antibody, it is shown that calpain I (a cysteine proteinase activated by micromolar concentrations of Ca2+) has a predominant intracellular location in the I-band region of the extensor digitorum longus (EDL) muscle of the rat, but is not exclusively associated with the Z-line.