Renal calcium phosphate and oxalate deposition in prolonged vitamin B6 deficiency: studies on a rat model of urolithiasis.

OBJECTIVE: To determine the effect on kidney oxalate-salt deposition of a prolonged diet which induced vitamin B6 deficiency in adult rats, as there are reports of the pathogenic involvement of vitamin B6 deficiency in the formation of renal calcium oxalate calculi. MATERIALS AND METHODS: The study comprised 24 6-month-old male albino Wistar rats; 12 were fed with a purified vitamin B6-deficient diet and the others provided with the same diet but supplemented with 6 mg/kg of vitamin B6. After 12 weeks, all rats were killed, and their kidneys fixed in formalin and routinely processed to paraffin for morphological examination; some fragments were fixed in glutaraldehyde and prepared for ultrastructural examination. From each rat consecutive sections of both kidneys were cut and stained with haematoxylin and eosin, periodic-acid Schiff, Sirius red and the Von Kossa method for calcium. Sections were examined in polarized light and by electron microscopy. RESULTS: The histopathological and ultrastructural features of the kidney of vitamin B6-deficient rats were those of tubular-interstitial nephritis, characterized by tubular atrophy, interstitial fibrosis and chronic inflammatory infiltration. Oxalate and phosphate crystals were present in the papillary and parenchymal connective tissue. Ultrastructural features confirmed severe tubular epithelial lesions and the presence of an interstitial and intraepithelial inflammatory infiltrate; there was mild interstitial fibrosis. None of these features were apparent in the kidney of control rats. CONCLUSIONS: Histopathological and ultrastructural data indicate that a prolonged vitamin B6-deficient diet may contribute to the formation and deposition of calcium phosphate and oxalate crystals, which lead to severe damage of the renal parenchyma. This phenomenon may occur not only in growing rats, which have more active protein metabolism and consequently higher vitamin B6 requirements, but also in adult rats.

[Intracranial location of a case of sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease) mimicking meningioma]. / Localizzazione intracranica di un caso di istiocitosi dei seni con linfoadenopatia massiva (malattia di Rosai-Dorfman) simulante un meningioma.

Sinus histiocytosis with massive lymphadenopathy was first described in 1969 in lymphnodes by Rosai and Dorfman ad subsequently in other organs including skin, bone ad soft tissue. Extranodal involvement has also been recognised, but central nervous system manifestations are extremely rare and clinical and radiological findings are suggestive of meningioma. We report a meningeal nodule in a young woman, clinically diagnosed as possible meningioma. A review of literature of previously reported intracranial lesions is presented.

Estimating a cardiac age by means of heart rate variability.

A data set of R-R intervals recorded for at least 15 min in 141 healthy individuals of different ages and under two different conditions ("resting" and "tilted" states) has been considered. The data have been subjected to spectral analysis by fast Fourier transform methods and considered in view of the possibility to work out a model in which the chronological and cardiac age could be compared. Understanding the results was greatly facilitated by 1) working out a number of derived variables from the original ones to highlight the presence of small but conceptually important variability factors; 2) extraction of the principal components from the original as well as from the derived variables to exclude redundancies and correlation effects; and 3) automatic clustering of the subjects in age classes, which allowed removal of individual variability within each class. The main conclusion is that, within the examined individuals, cardiac and chronological ages do not match for ages higher than approximately 50 years; this could reflect the presence of subtle (and difficult-to-envisage) biases in the data analysis or a real discrepancy. The latter hypothesis should be confirmed by similar observations in different systemic contexts. The use of a simple equation relating chronological and cardiac age, derived from a careful regression analysis on our data set and of general use for screening purposes, is demonstrated.

Progressive approach to the cytologic diagnosis of retroperitoneal spindle cell tumors.

OBJECTIVE: To assess the utility of fine needle aspiration biopsy (FNAB) in retroperitoneal spindle cell tumors with difficult tumor typing. STUDY DESIGN: Thirty-six cases of spindle cell tumors of the retroperitoneum were studied. Cytological diagnoses were set progressively: first exclusively by morphologic criteria on routinely stained slides, and then with the aid of immunocytochemistry (ICC), electron microscopy (EM) and clinical data. RESULTS: The morphologic diagnosis of benignity or malignancy was first made in double blind fashion by two researchers; it permitted an exact diagnosis in 31 cases (86%) by one examiner and 27 cases (75%) by the second. Using a progressive approach, benignity or malignancy was definitively determined in 35 cases (97%). As far as cytologic tumor typing is concerned, first it proved to be possible in 27 cases (75%) and then in 30 cases (83%) with the progressive approach. CONCLUSION: FNAB proved to be a useful tool in the diagnosis of retroperitoneal spindle cell tumors. A progressive approach to cytopathologic diagnosis, correlating morphology with ICC and EM results and matching them with clinical data, permitted a better differential diagnosis between benign and malignant spindle cell tumors and increased the possibility of correct tumor typing.

Histogenesis of primary liver carcinomas: strengths and weaknesses of cytokeratin profile and albumin mRNA detection.

To assess the utility of cytokeratin (CK) profile and albumin mRNA detection (as revealed by in situ hybridization) in the differential diagnosis of primary liver carcinomas (PLCs) we evaluated a series of surgically resected PLCs, comprising 20 "pure" hepatocellular carcinomas (HCCs) (10 well-differentiated, 10 poorly differentiated), 15 cholangiocarcinomas (CCs) (6 peripheral, 5 hilar, and 4 major duct ones) and 10 hepatocholangio-carcinomas (HCC-CCs). 11 of 20 (55%) of the pure HCCs expressed CKs of pure hepatocytic lineage (CK 8 and CK 18); 2 of 10 (20%) of the HCC-CCs displayed only hepatocytic profile, whereas 12 of 15 (80%) of the CCs evidenced mature bile duct cell phenotype (CK 8, CK 18, CK 7, CK 19). All HCCs expressed varying distributions of albumin mRNA, whereas 4 of 6 (67%) peripheral CCs showed cells with focal positivity for albumin mRNA. This suggests that the phenotypic expression of PLC cells are often not fixed, and in particular: (1) peripheral CCs have a different phenotype from hilar and large duct ones; (2) the CK profile and albumin mRNA expression in peripheral CCs show many similarities with those of some HCCs. Furthermore, the results show that a mixed biological phenotype (ie, CK 8, CK 18 and CK 7 and/or CK 19) can be found both among morphologically pure HCCs and peripheral CCs, suggesting that these two forms could share a common histogenesis. We think that special attention should be given to cases in which CK profile and albumin mRNA reveal mixed phenotype, as these tumors could have different biological behavior and respond differently to therapy.

A Markovian formalization of heart rate dynamics evinces a quantum-like hypothesis.

Most investigations into heart rate dynamics have emphasized continuous functions, whereas the heart beat itself is a discrete event. We present experimental evidence that by considering this quality, the dynamics may be appreciated as a result of singular dynamics arising out of non-Lipschitz formalisms. Markov process analysis demonstrates that heart beats may then be considered in terms of quantum-like constraints.

Micro- and macrostructure of learning in active avoidance: a quantitative approach.

The dynamics of learning in the Active Avoidance test was analyzed at the trials level as well as at the level of daily sessions, each comprising numerous trials. The two scales (large scale for the sessions and small scale for the trials) were demonstrated to be mutually independent. The intermediate derived scales (blocks of trials) were found consistent among themselves and with small scale but independent of the large one. Moreover, the two extreme scales were kinetically discriminable. These results point to the existence of two independent mechanisms for large and small scale learning together with the need to postulate a consolidation process during the rest period.

Multiple expression patterns of biopathological markers in primary invasive breast carcinoma: a useful tool for elucidating its biological behaviour.

BACKGROUND: Commonly used clinical and morphologic criteria have been reported to be of limited value in predicting the outcome of malignant tumours of the breast. Integrated information from the quantitative analysis in tumour tissue of biological parameters such as oestrogen and progesterone receptors (ER and PGR), proliferative activity, and proto-oncogene p53, c-erB2, and bcl-2 expression, may be useful for defining the biology of growth of breast carcinoma and to plan effective therapeutic strategies. PATIENTS AND METHODS: Immunohistochemistry with antibodies recognizing ER, PGR, Ki-67, and the p53, c-erbB2, and bcl-2 encoded proteins was performed on 291 primary breast carcinomas. Results were integrated with clinico-pathological indicators and examined with multivariate statistical procedures and modeling. RESULTS: P53, c-erbB2, and bcl-2 gene products were detected, respectively, in 30.6%, 31.6%, and 85.9% of the examined invasive breast carcinomas, revealing variable associations with cellular differentiation and proliferation as defined by ER/PGR status, Ki-67, tumour mass and histologic and nuclear grading. A multivariate graphical display on a subset of the most informative cases revealed that bcl-2 expression parallels ER/PGR status and is of importance in separating tumour clusters with different degrees of aggressiveness. CONCLUSIONS: The results of this study indicate that multivariate explorative analyses conducted on biological and clinico-pathological parameters might constitute an integrated approach to data analysis useful for distinguishing different biological behaviours and therapeutic groups in breast carcinoma. Our findings also suggest that bcl-2 expression may play a pivotal role in tumours lacking ER-mediated growth regulation.

A rare case of peritoneal mesothelioma in a male with ascites.

Mesothelial lesions can present significant diagnostic problems. An accurate identification of the lesion is important in deciding what course of action to follow, as well as what the required treatment will be. We report the case of a man with untreatable ascites. Four years after the first symptom appeared, the patient suddenly began to lose weight. The CT scan showed a thickening of the peritoneal profile; a diffuse lesion of the mesothelium was then observed at laparotomy. Histological examination led to a diagnosis of malignant mesothelioma in a well-differentiated mesothelial lesion with diffuse involvement of the serosa of the small intestine and the lesser omentum.

Immunocytochemical study of epidermal growth factor receptor, transforming growth factor alpha, and "squamous differentiation" in human endometrial carcinoma.

The expression of epidermal growth factor receptor (EGFr) and transforming growth factor alpha (TGF-alpha) was compared with the presence of "squamous differentiation" (SD) visualized in various histotypes of endometrial carcinoma by using a panel of monoclonal antibodies. The results of the current study demonstrate that EGFr and TGF-alpha are present in routinely processed endometrial carcinoma. The highest positive EGFr and TGF-alpha expression was seen in the group of adenocarcinomas with SD. The more intense EGFr and TGF-alpha immunoreactivity was observed in "squamous" foci both in adenoacanthomas (AA) and in adenosquamous carcinomas (AS). These EGFr- and TGF-alpha-positive squamous areas prevalently displayed a "stratification-related" cytokeratin (CK) immunoprofile characterized by the expression of CKs 1, 4, 5, 10, 13, 14, and 16. No correlation was found between EGFr- and TGF-alpha-positive status and depth of myometrial invasion or surgical stage. These results clearly demonstrate that EGFr and TGF-alpha expression is related remarkably to endometrial carcinoma with "squamous" areas both morphologically and immunophenotypically. This specific association leads us to suggest that EGFr and TGF-alpha expression in endometrial carcinoma may be prevalently involved in the equilibrium of cell differentiation of the "squamous" foci commonly observed in this group of neoplasias.

Gelatinase A (MMP-2) and its mRNA detected in both neoplastic and stromal cells of tumors with different invasive and metastatic properties.

Simultaneous presence of gelatinase A (MMP-2) and MMP-2 messenger RNA (mRNA) in 30 malignant tumors with various degrees of differentiation and biological behavior was evaluated by immunohistochemistry and in situ hybridization. The series consisted of 10 gastric carcinomas, 10 colorectal carcinomas, five squamous skin carcinomas, and five basal cell skin tumors. MMP-2 was detected in all cases. MMP-2 mRNA was expressed in the stromal cells in all cases and was more marked in the less-differentiated gastric and colonic carcinomas; it was also detected in the neoplastic cells of poorly differentiated tumors, particularly in those of the signet-ring cell type, both in the colon and stomach. The study confirmed that stromal cells have a specific role in tumor invasion and suggests a direct relationship between neoplastic epithelium and stromal cells in the most aggressive varieties.

Overexpression of p53 protein and Ki67 proliferative index in hepatocellular carcinoma: an immunohistochemical study on 109 Italian patients.

The overexpression of p53 protein and the Ki67 proliferative index was evaluated in 96 hepatocellular carcinomas (HCC), 67 in cirrhotic livers and 29 in non-cirrhotic ones, and in 13 non-carcinomatous lesions, all surgically resected from Italian patients. Overexpression of p53 was detected only in carcinomatous lesions, and was significantly related to the grade of HCC (P < 0.001). In fact, p53 was observed in 7/7 (100%) cases of grade IV, 13/43 (30.3%) of grade III, and 10/46 (21.7%) of grade II. The relationship between p53 and Ki67 scores was determined in serial sections from corresponding areas of both diffuse and patchy immunoreactivity. In the overall population, p53-positive tumors showed a significantly higher Ki67 score (15.9 +/- 5.5% vs 9.2 +/- 4.3% [P < 0.001]). This observation was evident in all grades of HCC.

Expression of intermediate filaments in normal and neoplastic exocrine pancreas.

The intermediate filament (IF) proteins present in the normal and pathological exocrine human pancreas were studied by immunolocalization using antibodies to cytokeratins (CKs) and vimentin. Acinar cells of normal pancreas showed a presence of simple CKs 8 and 18. Duct epithelium consistently expressed CKs 7, 8, 18 and 19 whereas centroacinar cells were rather low in CK 7. A subpopulation of CK 4 cells was detected in inter-intralobular ducts. In addition, some ducts contained individual cells or groups of cells that were positive for the stratification-related CKs (CKs 4, 5, 13, 15, 16). All pancreatic ductal adenocarcinomas regularly expressed CKs 7, 8, 18, 19 and were also positive for the 34 beta E12 antibody. Cytokeratin 4 was detected in a minor population of tumor cells. Pancreatic carcinoma also contained minor amounts of stratification-related CKs in variable combinations. Mucinous cystoadenocarcinoma showed the presence of CKs 7, 8, 18, 19 and was also positive for 34 beta E12, whereas the serous microcystic tumor presented CKs 8, 18, 19 and a variable amount of CKs 4 and 7. The duct-ductular alterations of the exocrine pancreas contained a different combination and distribution of CK isoforms similar to normal pancreatic ductal system. Mucinous hypertrophy and pyloric gland metaplasia reacted with antibodies to CKs 7, 8, 18 and 19. Vimentin was focally present both in normal and neoplastic tissue. Our results indicate that pancreatic ducts are characterized by an intrinsic "biliary-pancreatic duct type" immunoprofile (CKs 7, 8, 18 and 19), in contrast to acinar cells expressing exclusively CKs 8 and 18. We also detected a subpopulation of ducts regularly expressing CK 4. Surprisingly, several stratification-related CKs were detected both in normal and neoplastic exocrine pancreas. Moreover, the differentiation phenotypes of pancreatic tumors were reminiscent of normal cellular compartments.

Different in situ expression of insulin-like growth factor type II in hepatocellular carcinoma. An in situ hybridization and immunohistochemical study.

Reexpression of the insulin-like growth factor type II (IGF-II) gene has recently been described in hepatocellular carcinoma (HCC). In this study, we used a nonisotopic in situ hybridization method to analyze the expression of IGF-II mRNA in a series of 28 HCCs arising on cirrhotic and noncirrhotic livers. An immunohistochemical method was used to detect IGF-II peptide. Hepatitis B virus (HBV) status and the histological differentiation degree were also evaluated. Increased expression of IGF-II mRNA was found in 4 of 28 HCCs, and 7 of 17 cirrhotic patients showed IGF-II mRNA in the cirrhotic nodules surrounding the HCC. A slightly higher rate of positivity for IGF-II mRNA was found in the HBV-negative patients than in HBV-positive ones. Positive immunostaining for the IGF-II peptide in the HCC and/or in surrounding cirrhotic nodules was found in 10 of 28 cases. The normal hepatocytes of the noncirrhotic patients were always negative for IGF-II peptide and mRNA. The similarities between our results and those from experimental models in woodchucks seem to support the concept that heterogeneous phenotypic groups could exist in human HCCs.

Expression of insulin-like growth factor II (IGF-II) in human hepatocellular carcinomas: an immunohistochemical study.

Recent results obtained using molecular biology techniques have suggested a possible role for insulin-like growth factor II (IGF-II) in the pathogenesis of hepatocellular carcinoma (HCC). To investigate this phenomenon, a monoclonal antibody was used against IGF-II to study 54 patients with HCC. The presence of HBsAg was also tested both in serum and liver tissue. A positive immunoreaction was found in 9/15 (60%) of the HCC arising in cirrhotic livers of patients who had serum markers for HBV (HBV + positive patients). These results provide further evidence that HBV might play a role in the expression of IGF-II. In HCC of patients without any markers of HBV infection (HBV- negative patients), IGF-II was detected in 10/39 (25.6%) of the tumors, and in some benign neoplastic lesions. It was found not only in neoplastic cells but also in some dysplastic nodules. The speculation arises that IGF-II expression may play a role in some steps of hepato-carcinogenesis.

Prognosis of gastric carcinoma revealed by interactions between tumor cells and basement membrane.

The extra-cellular matrix (ECM) related antigens, type IV collagen, laminin, M(r) 68,000 laminin receptor (LR), M(r) 72,000 type IV collagenase (MMP-2), its inhibitor TIMP-2, and alpha 2-macroglobulin expression have been immunohistochemically investigated in 100 cases of human gastric carcinoma with a 5-yr follow up. Basement membranes were inversely related to tumoral differentiation. At the early intramucosal stage of both intestinal and diffuse histological types, TIMP-2 was expressed by the majority of tumor cells (60/63%), whereas MMP-2+ and LR+ cells were in the minority (24/19%, 23/0%, respectively). At the early submucosal stage, TIMP-2+ cells moderately decreased in both histological types (49/49%), whereas a consistently higher number of both MMP-2+ and LR+ cells were detected only in the diffuse carcinomas (72%). In the advanced stage, the expression of TIMP-2 further declined (22/24%), although the other two antigens increased or maintained high levels of expression. AMG+ cells never exceeded 10% in either histological type at any stage. In the liver metastases, both MMP-2+ and LR+ cells were more numerous than in the primary tumor (P < 0.002 and P < 0.01). Patients who died from their primary tumor had higher percentages of LR+, MMP-2+, and AMG+ cells and lower percentages of TIMP-2+ cells with respect to survivors. We believe evaluation of ECM-related antigens, and especially TIMP-2, may help determine a confident prognosis for gastric cancer.

Detection of anti-sense transcripts of the insulin-like growth factor-2 gene in Wilms' tumor.

The expression of the insulin-like growth factor-2 (IGF-2) gene was studied by means of both in situ hybridization and immunohistochemistry in eight cases of Wilms' tumor with different histological features. An anti-sense cRNA IGF-2 probe revealed the presence of abundant IGF-2 mRNAs in all eight tumors examined, localized in the most undifferentiated tumoral cells (blastemal cells); none of the tumors showed immunoreactivity for the anti-IGF-2 antibody. Using a sense cRNA IGF-2 probe, we also detected anti-sense transcripts of the IGF-2 gene in five of eight tumors. These transcripts were exclusively localized in the cells expressing the IGF-2 mRNAs. Although the function of these anti-sense transcripts is unknown, we think that their presence could explain the lack of IGF-2 peptides in Wilms' tumors despite increased expression of IGF-2 mRNAs.