Diatom detection in the diagnosis of death by drowning.

A medicolegal/algological collaboration lasting several years aimed at developing methods for dealing with dead bodies found in water where the circumstances are not clear, has led to an interdisciplinary procedure. To enable algological analysis, sample preservation and preparation must be free from contamination or carry-over at the beginning of the autopsy, although it should be noted that the demands on the digestion method are very high. One or more water samples from the site of drowning (from surface and bed) should be taken. Microscopic-algological analysis should record quantitative (diatom density), qualitative (species) and morphological (description of diatom valves) details for every sample. Furthermore, the species index and dominance identity similarity indices are calculated during the analysis procedure. The algological conclusions are based primarily on the separation values of Kater as well as on pair-matching. The final report is the result of interdisciplinary collaboration.

Germ cell tumors of the testis overexpress wild-type p53.

Several recent studies have suggested that testicular germ cell tumors express high levels of wild-type p53 protein. To clarify and confirm this unexpected result, we have investigated seminomatous and nonseminomatous germ cell tumors at the genomic, mRNA, and protein levels. Thirty-five tumors were examined for p53 overexpression using antibodies directed against the p53 (PAb1801, PAb240, and CM1), mdm2 (IF2), and p21Waf1/Clp1 (EA10) proteins. Thirty-two tumors were screened for p53 mutations by single-strand conformation polymorphism analysis. Eighteen tumors were screened with a functional assay that tests the transcriptional competence of human p53 protein expressed in yeast. On frozen sections, 100, 65, 35, 73, and 0% of tumors reacted with the CM1, PAb240, PAb1801, IF2, and EA10 antibodies, respectively. No p53 mutations were detected by single-strand conformation polymorphism or by functional assay. The fact that many tumors overexpress wild-type p53 but not mdm2 rules out mdm2 overexpression as a general explanation for the presence of wild-type p53 in these tumors. The absence of p21 overexpression suggests that p53 may be unable to activate transcription of critical target genes, which may explain why the presence of wild-type p53 is tolerated in this tumor type, although the mechanism for this transcriptional inactivity remains to be established.

Comparison of prognostic markers detected by immunohistochemistry in male and female breast carcinomas.

Male infiltrating breast carcinomas are rare and seem to have different characteristics, prognosis and sensitivity to hormonal treatment than those of female breast carcinomas. Our aim was to determine whether markers which have an established role in women are also important in men. 66 male infiltrating-ductal breast carcinomas were compared with 190 female breast carcinomas of the same type. Various markers were studied using immunohistochemistry. Tumour size at diagnosis, grade, number of axillary metastases and prognosis were comparable in male and female breast carcinomas. However, male breast carcinomas were characterised by a higher percentage of oestrogen receptor (OR) reactivity, and weekly associated with markers that, in women, are under oestrogen control. Male breast carcinomas were positive for markers under androgen control. Male breast carcinomas also differed from female carcinomas by the low percentage of p53+ and the high percentage of bcl-2+ tumours. The phenotype of male breast carcinomas has characteristics that could have repercussions on prognosis and on the choice of hormonal treatment. Only a few male breast cancers are p53+. OR, which are frequently present in male tumours, are probably not functional. In contrast, androgen receptors seem efficient, as several markers under androgen control are expressed. Therefore, the selection of hormonal therapy should not be based on OR status only.

Neuropeptide Y secretion from a human insulinoma.

Neuropeptide Y (NPY) is a 36 aminoacid peptide known to inhibit glucose-stimulated insulin secretion. NPY has been shown to be synthesized and secreted by rat islets of Langerhans. More recently, we described the presence on NPY within human islets of Langerhans and in several pancreatic endocrine tumors. In this report, we describe the case of a patient presenting with an insulinoma who underwent the surgical resection of the tumor and was studied in vivo and in vitro for NPY production. Using a highly specific and sensitive two-site amplified enzyme-linked immunosorbent assay, we detected high plasma NPY levels in the patient prior to the surgical resection of the tumor which returned to normal after surgery. NPY was secreted from the tumor when kept in primary cell culture. Furthermore, immunohistochemistry of the insulinoma revealed the presence of NPY and its C-flanking peptide together with insulin, chromogranin and neuron specific enolase. It is concluded that elevated circulating NPY levels observed in this patient with an insulinoma reflected in vivo secretion by the tumor and it is hypothesized that NPY could potentially be used as an endocrine marker in patients with suspected insulinoma.

bcl-2 protein in invasive ductal breast carcinomas.

The bcl-2 gene encodes a protein which inhibits programmed cell death (apoptosis). This protein was detected by immunohistochemical techniques in 48% of invasive ductal carcinomas of the breast. It was present in well-differentiated carcinomas with hormonal receptors, and proteins synthesized under the control of oestrogens: pS2, cathepsin D and ERD5. In contrast, bcl-2+ carcinomas are less frequently positive for p53 and have a Ki67 score under the mean. bcl-2 protects cells against apoptosis. Accumulation of p53 protein, which is indicative of p53 mutation, would have the same effect; however, these two proteins seem inversely related, an inverse correlation observed by others in breast cancer cell lines and in lymphomas. Tumours positive for bcl-2 escape apoptosis and have worse prognosis but this is not what is found; survival at 5 years, and particularly the absence of recurrence during the first 5 years after surgery, seem to be associated with bcl-2 positivity. The bcl-2 protein seems only to be an important prognostic factor in women over 54 years of age. Moreover, p53-bcl-2+ tumours have a better response to hormonal therapy than p53-bcl-2-tumours.

Immunolocalization of neuropeptide Y in human pancreatic endocrine tumors.

Neuropeptide Y (NPY) is a 36 amino acid peptide known to inhibit glucose-stimulated insulin secretion. NPY has recently been shown to be synthetized within rat islets of Langerhans and to be secreted in a differentiated rat insulin-secreting cell line, and as to this date the localization of NPY in human endocrine pancreas has not been reported. As NPY shares high amino acid sequence homology with peptide YY (PYY) and pancreatic polypeptide (PP), the polyclonal antibodies raised against these peptides often cross-react with each other. To demonstrate the presence of NPY in the human endocrine pancreas, we used a highly specific monoclonal antibody raised against NPY and another against its C-flanking peptide (CPON). We studied three cases of hyperplasia of Langerhans islets and 11 cases of endocrine tumors of the pancreas. NPY and CPON were detected in all three cases of hyperplasia. For the 11 pancreatic tumors, five and nine of the tumors were positive for the antibodies NPY and CPON, respectively. The two negative tumors for CPON immunoreactivity were differentiated insulinomas, which showed no evidence of other hormonal secretion. In normal Langerhans islet, NPY and CPON immunoreactivities were colocalized in glucagon-producing cells (alpha-cells) and in a few insulin-secreting cell (beta-cells).(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of p53 protein in gastric carcinomas. Association with histologic type and prognosis.

We searched for p53 protein accumulation in 72 gastric carcinomas. Of the 38 cases of the diffuse type, only four were positive for p53. Of the 34 cases of intestinal type, 24 had p53 protein accumulation. This difference (p < 0.0001) between histological types was present regardless of whether the carcinoma was superficial or infiltrative. Normal epithelial cells and intestinal metaplasia were never positive. No correlation was found between p53 protein accumulation and tumor size, lymph node metastases, age or sex of the patients. Although tumor size, lymph node metastases, and infiltrative character all have prognostic value, p53, either alone or in association with these parameters, does not; p53 mutations seem to play a role in oncogenesis only in the intestinal type of gastric carcinomas; however, p53 protein accumulation has no prognostic value in gastric tumors.

Desmin and neural marker expression in mesothelial cells and mesotheliomas.

The distribution of keratin, vimentin, desmin, muscular actin, S100, specific neuron enolase, and chromogranin was studied by immunoperoxidase staining in mesothelium, malignant mesotheliomas, and pulmonary carcinomas. The mesothelial cells were positive for keratin and vimentin on all smears of pleural and ascitic effusions; most of them were also positive for desmin but rarely for enolase and S100. None was positive for muscular actin. Sixteen malignant mesotheliomas expressed vimentin and keratin; six were also positive for desmin, three for desmin and neural markers, and five for neural markers. In comparison, none of the pulmonary carcinomas was positive for these markers. Mesothelial cells are able to express markers of divergent differentiation. Mesotheliomas also have such markers that are present in other malignant tumors and, in particular, in intra-abdominal desmoplastic small cell tumors with divergent differentiation. This entity and mesotheliomas probably originate from the same cell. Moreover, desmin, found in 56% of malignant mesotheliomas but absent in pulmonary carcinomas, may be useful in the differential diagnosis of these tumors.

p53 Gene alterations and p53 protein accumulation in infiltrating ductal breast carcinomas: correlation between immunohistochemical and molecular biology techniques.

One hundred and eighty-eight infiltrating ductal carcinomas of the breast were examined immunohistochemically (IMM) for p53, and the results were compared to those of single strand conformation polymorphism (SSCP). Of the 65 IMM+ cases (35%), 32 showed a genetic alteration in exons 5 to 9. In some of the IMM+ SSCP- cases, the number of 53+ cells in the tumor was too low to be detected by SSCP. Cases with only a few p53+ cells must not necessarily be considered negative, because a genetic alteration has been found in nine such cases. However, in a few cases, the accumulation of p53 protein could be caused by a factor other than mutation. Of the 123 IMM- cases, six showed gene polymorphism. p53 phenotype, as established with three monoclonal antibodies, did not correlate with genetic alteration in a particular exon. p53 IMM+ or SSCP+ tumors were generally ER-, grade III tumors and were uncommon in women older than 69 yr of age. The two methods have almost the same prognostic value. The accumulation of p53 protein is a good indicator of p53 mutation and therefore, immunohistochemistry remains a good method for the detection of such mutations.

Expression of p53 protein in prostate cancers of different histologic types.

Mutations and overexpression of p53 gene in prostate carcinoma have been found but their significance in the development and progression of cancer is so far unknown. We investigated the prevalence of abnormalities of p53 protein in a heterogeneous group of prostate carcinoma to verify whether acinar and non acinar carcinomas have a different expression of p53 protein. Paraffin sections of 45 prostate carcinomas (39 acinar, 3 ductal papillary, 1 transitional cell, 1 mucinous and 1 pure small cell) were examined for the expression of p53 protein using a panel of antibodies (monoclonal antibodies Pab 1801, D07 and polyclonal antibody CM1). No p53 expression was observed in any acinar carcinomas independent of grade and stage. For non acinar carcinomas only small cell and transitional cell carcinomas exhibited detectable amounts of p53 protein in tumour cell nuclei. The prevalence of p53 overexpression in prostate carcinoma is relatively low compared with that found in many other tumours. In the present study, the overexpression of p53 in a small cell carcinoma and in a transitional cell carcinoma suggest that the loss of suppressing role of p53 gene may be an important mechanism in the genesis and in the development of these uncommon tumours.

[Ovarian carcinoid tumors: immunohistochemical and ultrastructural study of 8 cases]. / Carcinoïdes ovariens: étude immunohistochimique et ultrastructurale de 8 cas.

Eight ovarian carcinoids, 4 trabecular and 4 strumal have been studied immunohistochemically for polypeptide hormones; three of them were also examined with the electron microscope. Seven of eight cases were positive for at least one polypeptide hormone. The trabecular parts of the 4 cases of strumal carcinoid were positive for pancreatic polypeptide. As the vesicular regions of these 4 cases were positive for thyroglobulin and for pancreatic polypeptide, we suspect the existence of "hybrid" thyroid and neuroendocrine cells. The electron microscopic examination revealed: the cell in the insular carcinoid had no distinct polarization and contained electron-dense granules of varying shape; in the trabecular part of the strumal carcinoid the cells were polarized and contained round electron-dense granules.

Prognostic value of p53 protein expression in breast carcinomas.

A group of 196 ductal infiltrating carcinomas of the breast was examined immunohistochemically for p53. The purpose of this study was to show whether frozen and fixed tissues are equally adequate for detection of p53 and which antibodies should be used to have a prognostic value. Detection was superior on frozen to that on formalin-fixed tissues. It was not possible with any method to improve results on fixed tissues. Detection of p53 was different for each antibody: M 1801 detected 41 cases on frozen tissues, M-240 52 cases, M-421 28 cases. Using all the antisera, and the rabbit antiserum CM1, it was possible to detect 71 cases (36%). The percentage was the same in infiltrating lobular carcinomas but higher (94%) in medullary carcinomas. p53 was associated with high grade and ER-tumours. In formalin-fixed tissues, p53 had no prognostic value. In frozen tissues p53 was not an independent factor of prognosis. However, it was important in sorting out cases with bad prognosis in the ER-carcinomas and in the carcinomas without metastases. The prognostic value was different for each monoclonal antiserum. Positivity with M421 associated with negativity for M240, and positivity only for M1801 sorted out cases with a poor prognosis (67% and 50% deaths at 5 years).

Oestrogen receptor, progesterone receptor, pS2, ERD5, HSP27 and cathepsin D in invasive ductal breast carcinomas.

Hormonal receptors and markers for prognostic evaluation were detected immunohistochemically in 196 infiltrating ductal breast carcinomas. Immunohistochemical detection of progesterone and oestrogen receptor is a method giving results generally concordant with those of the binding assay. However, immunohistochemical detection seems better. It allows the detection of hormonal receptors on small carcinomas, it is not modified by the endogenous hormones, and it has a slightly better correlation with prognosis and with the response to hormone therapy. Immunohistochemical detection of progesterone receptor has a prognostic value, sorting a negative subgroup with a poor prognosis from the oestrogen receptor positive tumours. These results can be obtained without quantitative immunohistological methods. ERD5, pS2, HSP27 and cathepsin D are associated with oestrogen receptor positivity. pS2 and HSP27 are interesting markers. They characterize a subgroup of oestrogen receptor negative tumours with a good prognosis. Moreover, pS2 is a marker of response to hormone therapy. ERD5 and cathepsin D do not appear to be of value as markers of prognosis.

Natural history of diffuse uveal melanocytic proliferation. Case report.

BACKGROUND: Diffuse uveal melanocytic proliferation is a rare paraneoplastic syndrome resulting in rapid bilateral visual loss due to proliferation of benign melanocytes within the choroid and ciliary body. Most of the previously reported cases have been seen with bilateral involvement and typical ocular features. PATIENT: The authors report the case of a 61-year-old man who presented with uniocular posterior pole lesions at the level of the retinal pigment epithelium and subsequently developed the typical bilateral lesions of diffuse uveal melanocytic proliferation. His clinical course was typical, with visual disturbance preceding signs and symptoms of malignancy by 5 months. Rapid decline ensued, and he eventually died 10 months after the onset of visual symptoms. RESULTS: Results of ocular pathologic examination showed the typical choroidal thickening due to the proliferation of melanocytes and the primary tumor was found to be an undifferentiated adenocarcinoma originating in either the pancreas or the esophagus. CONCLUSIONS: The very early funduscopic and fluorescein angiographic findings of diffuse uveal melanocytic proliferation are presented as well as the evolution, ocular pathology, and possible mechanisms for its development.

Mammary origin of metastases. Immunohistochemical determination.

Zinc-alpha 2-glycoprotein, gross cystic disease fluid protein 15, and estrogen receptors are expressed in a great proportion of breast carcinomas. These markers were investigated by immunohistochemistry in 28 metastases from breast carcinomas and for comparison on 24 metastases from other carcinomas. A group of 83 primary nonmammary tumors was also studied. Most (> 96%) breast carcinoma metastases expressed one or several markers, while all metastases of other origins were negative. This sensitive and apparently specific immunostaining proved to be of great utility in cases in which the mammary origin of metastases was difficult to establish. In four axillary lymph node metastases, it even led to the discovery of an occult homolateral breast carcinoma that was not detectable by clinical and mammographic investigations. This study indicates that the combined use of zinc-alpha 2-glycoprotein, gross cystic disease fluid protein 15, and estrogenic receptors represents a useful immunostaining technique that can help the pathologist in determining the origin of breast carcinoma metastases.

Gastrointestinal stromal tumours: an immunohistochemical study of 165 cases.

The phenotype of 165 gastrointestinal stromal tumours was studied by immunohistochemistry. In each case the phenotype was compared to the histological diagnosis. The phenotype was muscle in 49 tumours (30%), neural in 18 (11%), histiocytic in 20 (12%) and mixed in five (3%); 68 tumours (41%) were positive for vimentin only, four tumours had no markers and one tumour was positive for keratin only. Histologically, the tumours were classified as smooth muscle, probably smooth muscle, probably nerve sheath tumours or tumours of undetermined differentiation. In 30 histologically unequivocal muscle tumours, the phenotype was muscle in 28. Half of them, all benign, arose in the oesophagus or gastric cardia. Apart from this group, there was no correlation between phenotype, site of tumour and histological differentiation. Actin was a more sensitive muscle marker than desmin. With the exception of oesophageal tumours, the histological appearances alone could not establish a diagnosis of malignancy and were inadequate in evaluating differentiation. Immunohistochemical examination determined differentiation in 54% of the tumours, but this finding should be interpreted with caution in terms of histogenesis. It allowed us, however, to specify the differential diagnosis in 57 tumours in which the histological diagnosis was uncertain.

Prognostic value of serum proteins synthesized by breast carcinoma cells.

Several breast carcinoma cell lines or explants of such tumors, as well as examples of lactating or dysplastic breast tissue, synthesized three serum proteins (alpha 2-Zn-glycoprotein, alpha 1-antichymotrypsin, and alpha-lipoprotein) in vitro. These proteins were detected by immunoperoxidase techniques in 126 breast carcinomas that had been evaluated clinically for more than six years. Alpha-2-Zn-glycoprotein was present in 58% of the carcinomas, whereas alpha 1-antichymotrypsin was seen in 55% and alpha-lipoprotein in 52%. These markers showed a relationship with clinical outcome. Alpha-1-antichymotrypsin and alpha-lipoprotein were unfavorable determinants, whereas alpha 2-Zn-glycoprotein was detected in lesions with a favorable evolution. Taken individually, these markers have similar but rather weak associations with five-year survival rates; roughly 20% of patients in the "favorable" group died, compared with 33% in the "unfavorable" group. Alpha-2-Zn-glycoprotein in grade 1 tumors was a marker with marginally favorable significance, but alpha 1-antichymotrypsin significantly worsened the prognosis of grade 2 and 3 tumors. Furthermore, stratification of patients according to the number of positive unfavorable markers yielded striking results. Eight percent of patients with none of the unfavorable markers were dead at five years, compared with 55% of those whose lesions expressed three unfavorable markers.

[Is papilloma of the esophagus a preneoplastic lesion? Study of 33 cases]. / Le papillome de l'oesophage est-il une lésion prénéoplasique? Etude de 33 cas.

The etiology of esophagus papilloma is much debated: some authors attach greater importance to irritation factors, while others give preference to the viral hypothesis and suggest that this disease could eventually lead to squamous cell carcinoma of the esophagus. To verify the viral hypothesis, we reviewed the histological slides of the 33 cases of esophageal papilloma diagnosed in our Institute of Pathology between 1973 and 1988. We evaluated the histological diagnosis using Winckler's criteria. HPV typing was done using probes of HPV DNA types 6-11, 16-18, 31-33-35 applied to paraffin sections according to the in-situ hybridization method. Clinical and endoscopic data of 15 cases (from the CHUV) are reviewed. Oncological data was provided by the Vaud Cantonal Tumor Register. No patient in our series fulfilled all the histological criteria set by Winckler to diagnose an HPV condition. Viral DNA 31-33-35 was found in a small minority of the papillomas. The clinical impact of esophageal papilloma on epidermoid carcinogenesis is nil.

Immunohistochemistry in the differential diagnosis of liver carcinomas.

Immunohistochemical techniques were used to study 177 hepatic tumors (hepatocarcinoma, cholangiocarcinoma, hepatocholangiocarcinoma, adenocarcinoma of unknown origin, and metastatic carcinoma). Phenotypes suggestive of hepatocarcinoma included keratins 8 and 18, factor XIII a, alpha-fetoprotein. C-reactive protein, carcinoembryonic antigen (CEA) cross-reacting antigen; those in effect that excluded hepatocarcinoma were keratins 1, 5, 10, 11, 19, true CEA. C-reactive protein, used for the first time, proved to be a fairly sensitive and specific marker. Factor XIII a, which was thought to be synthesized only by histiocytes, was also present in hepatocytes. Immunohistochemistry appears to be an important tool in the diagnosis of hepatic tumors. As a result of this study, 32 cases were reclassified; several were found to be intermediate between hepatocarcinoma and cholangiocarcinoma. Sixteen cases apparently were true hepatocholangiocarcinomas. In 12 cases of hepatocarcinoma, some tumor cells expressed keratins of bile duct type. It was impossible to differentiate immunohistochemically cholangiocarcinoma from metastatic carcinoma, except in two cases with breast tissue markers.