Gastric paraganglioma: case report and review of the literature.

Very few cases of gastric paragangliomas have been reported in the literature to date. We report a rare case of parietal gastric paraganglioma fortuitously detected during intraoperative exploration. A 82-years-old woman presented to our emergency room for abdominal pain. On physical examination abdomen was painful on palpation and Blumberg's sign was present. The laboratory exams showed a neutrophilia in absence of leukocytosis. Acute appendicitis was suspected and a laparoscopy was performed. At exploration, the vermiform appendix was normal while a lumpy, hard-fibrous and white-pinkish extraluminal lesion of the anterior wall of the gastric body near the greater curvature of about 2 cm in diameter was present. Laparoscopic resection of the gastric lesion was performed. The patient was discharged in good condition in the fourth postoperative day. Pathologic examination revealed a gangliocitic paraganglioma. The patient is alive and well without evidence of relapse 6 months after surgery. Gastric paraganglioma is a very rare tumor and its diagnosis is very difficult. Surgical excision is the treatment of choice which can be performed successfully with laparoscopy.

Immunohistochemical markers of CYP3A4 and CYP3A7: a new tool towards personalized pharmacotherapy of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) represents a major global health problem, since more than 90% of primary liver cancers worldwide are HCC. Most cases of HCC are secondary to viral hepatitis infection (hepatitis B or C), alcoholism and cirrhosis. Sorafenib, an oral tyrosine kinase inhibitor that suppresses tumor proliferation and angiogenesis, emerged as the first effective systemic treatment for HCC after 30 years of research, and is currently the standard-of-care for patients with advanced HCC. Sorafenib is metabolized by cytochrome P450 (CYP450), particularly from the 3A4 isoform, producing two main metabolites: the N-oxide and the N-hydroxymethyl metabolite. We studied 11 HCC sample showing the presence of CYP3A4 and CYP3A7 in most of the samples analyzed. Specifically, the immunoreactivity of CYP3A4 was more strong and widespread than that of CYP3A7. The CYP3A4 immunoreactivity was observed in surrounding hepatocytes in 8 out of 11 cases; while the CYP3A7 immunostaining was found in normal liver cells, in 7 out of 11 cases. These results suggest the existence of a marked inter-individual variability regarding the presence of the isoforms of CYP3A. In addition, since sorafenib is metabolized by CYP3A4, but not by CYP3A7, an overexpression of CYP3A4 may lead to an increase in the degradation of the drug and then to clinical ineffectiveness. These results might implicate the necessity of an individualized approach in the treatment of HCC as positivity to CYP3A4 in HCC liver samples might predict a scarce response to sorafenib.

Immunohistochemical markers of neural progenitor cells in the early embryonic human cerebral cortex.

The development of the human central nervous system represents a delicate moment of embryogenesis. The purpose of this study was to analyze the expression of multiple immunohistochemical markers in the stem/progenitor cells in the human cerebral cortex during the early phases of development.  To this end, samples from cerebral cortex were obtained from 4 human embryos of 11 weeks of gestation. Each sample was formalin-fixed, paraffin embedded and immunostained with several markers including GFAP, WT1, Nestin, Vimentin, CD117, S100B, Sox2, PAX2, PAX5, Tß4, Neurofilament, CD44, CD133, Synaptophysin and Cyclin D1. Our study shows the ability of the different immunohistochemical markers to evidence different zones of the developing human cerebral cortex, allowing the identification of the multiple stages of differentiation of neuronal and glial precursors. Three important markers of radial glial cells are evidenced in this early gestational age: Vimentin, Nestin and WT1. Sox2 was expressed by the stem/progenitor cells of the ventricular zone, whereas the postmitotic neurons of the cortical plate were immunostained by PAX2 and NSE. Future studies are needed to test other important stem/progenitor cells markers and to better analyze differences in the immunohistochemical expression of these markers during gestation.

WT1 expression in the human fetus during development.

Wilms' Tumor 1 (WT1) is a transcription factor involved in the development of the urogenital system. The purpose of this study was to analyze the immunoreactivity for WT1 protein in different tissues and organs in human fetuses in early phases of gestation. To this end, samples from multiple organs were obtained from 4 human fetuses, ranging from 7 up to 12 weeks of gestation. Each sample was formalin-fixed, paraffin embedded and immunostained for WT1. Our data show that WT1 is involved in development of multiple human organs in a more vast series of cells types than previously reported. Immunostaining for WT1 was characterized by a predominant cytoplasmic reactivity in the vast majority of cell types. Mesenchimal progenitors in the fetal lung, ductal plate progenitors in fetal liver, cap mesenchimal cells in the developing kidney, fetal zone cells in adrenal glands, atrial and ventricular cardiomyocytes in the fetal heart, radial glial cells in the fetal cerebral cortex and skeletal muscle cell precursors showed the highest levels of WT1 immunoreactivity. Future studies will be needed to detect differences in the expression of WT1 in various organs at different gestational ages, in order to better evaluate the role of WT1 in cell proliferation and differentiation during intrauterine human development.

Perinatal heart programming: long-term consequences.

OBJECTIVE: evaluate the relationship between impaired growth during intrauterine life and adult risk of cardiovascular disease and death. MATERIALS: review of the most important contributions to the relationship between intrauterine fetal life and heart disease insurgence in childhood and adulthood, starting with a schematic representation of the principal steps in human heart development, discussion of the new theory on the relevance of the number of cardiomyocytes that every heart shows at birth. RESULTS: intrauterine environment defines the epigenetic profile of newborns, with implications for the risk of developing diseases later in adult life. This means that the programming of cardiovascular risk and other pathologies, such as obesity, in adulthood takes place starting from intrauterine life. CONCLUSIONS: it can be hypothesized that by preventing and eventually treating cardiovascular diseases in the pediatric age, if these are already present in their early and/or in light forms, the long-term management of complications could be approached differently and more effectively than by postponing the treatment to adulthood. The future challenge in this fascinating field of clinical research is the discovery of the molecular mechanisms underlying the association between intrauterine growth restriction and fetal onset of adult cardiac disease, so as to make a dream come true by applying primary prevention of adult heart disease in the womb.

Cytochrome P450 genetic polymorphism in neonatal drug metabolism: role and practical consequences towards a new drug culture in neonatology.

The cytochrome P450 superfamily (CYP450) in humans is formed by 57 functional monooxygenases critical for the metabolism of numerous endogenous and exogenous compounds. The superfamily is organized into 18 families and 44 subfamilies. CYP nomenclature is based on the identity of amino acids. The most important functions of the CYP450 are related to metabolism of endogenous compounds, detoxification of exogenous xenobiotics and decomposition of the vast majority of currently used drugs. The expression of CYP450 enzymes in the human body is characterized by a marked substrate and tissue specificity, the most important being localized in the liver, but also present in kidney, lung, brain, breast, prostate and in the small intestine. The human cytochrome P450 3A gene family (CYP3A) accounts for the largest portion of CYP450 proteins in human liver and includes 4 genes: CYP3A4, CYP3A5, CYP3A7, CYP3A43. Multiple and complex genetic variations, marked interindividual, interethnic and gender variability have been reported regarding CYP3A isoform expression and activity. Multiple factors may affect CYP3A expression and activity, such as inducers like rifampicin, phenobarbital, 3-methylcholantrene, beta-naphtoflavone, and dexamethasone. The maturation of organ systems, paralleled by ontogeny of drug-metabolizing enzymes during fetal life and in the first months of postnatal life, surely exerts profound effects on drug disposition, probably being the predominant factor accounting for age-associated changes in drug clearance. In fact, drug dosage in the perinatal period represents a continuous challenge for neonatologists. The purpose of this article is to provide a brief review of the pharmacokinetic differences between neonates and adults, showing the peculiarities of liver CYP450-related drug metabolism in the perinatal period and at birth, and to report the toxic mechanisms of liver injury in neonates, due to the most frequently utilized drugs in NICU centers.

First genetic analysis of lattice corneal dystrophy type I in a family from Bulgaria.

PURPOSE: To report a new family belonging to a previously non-investigated geographic are a with a rare form of lattice corneal dystrophy (LCD). METHODS: Detailed ophthalmologic analysis was carried out on a Bulgarian woman, enrolled for perforating keratoplasty. In order to obtain a final diagnosis both histology and genetic analysis were performed. RESULTS: Upon transplantation, histologic analysis of the dystrophic cornea revealed the typical staining pattern and amyloid deposits of lattice corneal dystrophies. Genetic analysis of the subject and her daughter confirmed the presence of an autosomal dominant R124C mutation within exon 4 of the BIGH3 gene, encoding for keratoepithelin, while showing no abnormalities in her son. CONCLUSIONS: The identification of this mutation allows the unambiguous classification of this corneal dystrophy as LCD type I. A first case of LCD I in a family from Eastern Europe could help to better clarify the molecular epidemiology of the disease.

[Late onset Wilson's disease]. / Morbo di Wilson ad esordio tardivo.

BACKGROUND: Wilson's disease (W.D.) is a metabolic disorder that occurs predominantly in children, adolescents, young adults and, rarely, in patients over 35 years. AIMS: In order to verify the prevalence of the clinical presentation of W.D. in adulthood, we analyzed a wide number of clinical presentation of W.D. with particular attention to the age of onset and to the evolutive stage of liver disease at presentation. PATIENTS: Our study is relative to 11 Sardinian adult subjects, aged 36-57 years, 6 males and 5 females, selected from a series of 120 patients affected by W.D. The only criterion utilized to select the patients was their age at presentation, with a cut off of 35 years. METHODS: Liver biopsies were routinely processed and stained with rhodanine, rubeanic acid, orcein and Timm's methods. On the basis of the histological picture, liver biopsies were subdivided into four evolutive stages: stage I = steatosis; stage II = interface hepatitis; stage III = bridging fibrosis; stage IV = cirrhosis. Molecular characterization of W.D. for gene mutations in the Sardinian population was performed in 7 out of 11 cases (-441/-427 del, 5' UTR and 3436 G > A Exon 16). RESULTS: 3 patients showed histological features of the first evolutive stage, 2 of the second, 1 of the third, and 5 of the fourth stage. Histochemistry for copper resulted positive in 9 of 11 cases at least with one of the four employed methods. In the seven patients in whom molecular characterization was perfomed, the gene mutation of W.D. was the same observed more frequently in the Sardinian population (-441/-427 del, 5'UTR). CONCLUSIONS: Our data show that: 1) W.D. with late onset is not rare in Sardinian population; 2) in spite of the late clinical presentation, W.D. may present in the first (3/10) and in the second evolutive stage (2/10) with mild to moderate changes of the liver architecture; 3) patients may show, at presentation, a severe liver disease, characterized by bridging fibrosis or cirrhosis; 4) from a practical point of view, we ask to consider the diagnosis the W.D. in all patients with chronic liver disease of unknown etiology, even if aged over 35 years.

Ultrasound real-time imaging in the differential diagnosis of periapical lesions.

AIM: To assess whether the use of real-time ultrasound imaging (echography), together with the application of 'Colour Power Doppler', can help the differential diagnosis of periapical lesions based on their contents. METHODOLOGY: Eleven patients diagnosed with periapical lesions using conventional clinical and radiographic findings and who had been scheduled for endodontic surgery were examined using ultrasound imaging and Colour Power Doppler at the site of the lesions. Lesions were described in each case by an expert echographist, together with an endodontist, according to a scheme described previously. Information on the size of each lesion, its content and its vascular supply was gained and recorded by means of the ultrasound images; then, a tentative differential diagnosis between a cyst and a granuloma was made. The lesions obtained during endodontic surgery were processed and examined with a light microscope. The results from the biopsies were compared with the descriptions made with the ultrasound images. RESULTS: The differential diagnoses between periapical granulomas and cystic lesions, which were based on the echograpic findings, were confirmed by the results of the histopathologic examination in all the 11 cases: four being granulomas and seven being cystic lesions. CONCLUSIONS: Ultrasound real-time imaging is a technique that may help make a differential diagnosis between cysts and granulomas by revealing the nature of the content of a bony lesion. This technique may have further applications in the study of other lesions of the jaws.

[Microcystic lateral parodontal cyst. Report of a case]. / Cisti parodontale laterale microcistica. Descrizione di un caso clinico.

The lateral periodontal cyst (LPC) is an odontogenetic lesion of the maxillary bone representing about 0.8% of maxillary cystic lesions. It starts in the lateral periodontal area of a vital erupted tooth. Clinical symptoms are generally absent. Its diagnosis usually can be made on the basis of an occasional radiographic examination which shows an osteolytic monoloculate lesion with defined outlines. The botryoid cyst is a variation of LPC and is radiographically different and more aggressive than the common LPC. A clinical case of LPC which allows to make histopathological and clinical considerations on these cystic lesions is described.

Uncommitted gastrointestinal stromal tumour. Case report.

Gastrointestinal Stromal Tumours (GIST) are mesenchymal tumours with uncertain prognosis. Malignant variety represents about 2.0% of malignant gastroenteric tumours. The Authors report a clinical case of malignant gastric and duodenojejunal GIST, in which the only surgical treatment seems to be definitive. R. S., a 69-year-old female, was admitted for asthenia and fever in January 1997. Endoscopic exploration, ultrasonography and CT-scan of the abdomen demonstrated an exophytic tumour in the greater gastric curvature and one tumour of 5.5 cm of diameter in the Treitz's angle. We performed a resection of the gastric tumour and the duodenojejunal angle. Postoperative course was uneventful and the patient was discharged after 14 postoperative hospital days. Histological analysis showed two spindle cells stromal tumours with mitotic rate > 20/10 HPF. The immunohistochemistry demonstrated the uncommitted origin of tumour cells. The patient refused the chemotherapy treatment. There was no local recurrence or metastasis at a follow up of 47 months, in spite the high malignancy degree. For this reason and because of the uncertain behaviour of benign GIST, the authors propose a lifelong follow up of the patients managed with potentially curative surgical resection.

Early surgical treatment of acute cholecystitis. A retrospective comparative study of the laparoscopic and open approaches.

The aim of this retrospective study was to compare the results of laparoscopic and open early cholecystectomy in patients with acute cholecystitis. From January 1997 to October 2000, 168 patients underwent cholecystectomy in our institution. Of the 35 patients (20.8%) with acute cholecystitis, 20 patients (57.1%) were operated on laparoscopically and the other 15 patients (42.9%) with the traditional open approach. The two groups were similar in terms of age, sex and onset of symptoms. The postoperative morbidity was 15.0% in the laparoscopic group versus 40.0% in the open group. The average postoperative hospital stay in the laparoscopic group was 5.1 days as compared to 10.5 days in the open group (P = 0.013). The conversion rate to laparotomy was 5.0% (1 case). At follow-up there has been one case of incisional hernia in the open group. Early laparoscopic cholecystectomy for acute cholecystitis was associated with a lower postoperative morbidity rate and significantly earlier patient discharge.

Brain copper, iron, magnesium, zinc, calcium, sulfur and phosphorus storage in Wilson's disease.

PROJECT: Wilson's disease (WD) is an inherited disorder of copper metabolism characterised by juvenile liver cirrhosis and by neurological symptoms. Copper levels in brain in WD have been reported to be 10 to 15 fold normal values, depending on the different brain regions. Being very few data on copper distribution in central nervous system in WD available, it seemed of interest to study the concentration of copper and of other trace elements (Zn, P, Mg, Ca, Fe and S) in the brain of a patient died for WD. PROCEDURE: a 56 year old woman affected by WD was admitted to our hospital with signs of hepatic failure and died few days later. At autopsy, a brain slice extending from the left to the right hemisphere was divided in 28 samples. On each sample Copper, Iron, Magnesium, Phosphorus, Sulphur, Zinc and Calcium were determined by Induced Coupled Plasma Atomic Emission Spectroscopy. RESULTS: the mean concentration of copper, ranging from 88 to 158 microg/g of dry tissue in all the brain specimens was higher than literature reference values, while that of the other tested elements was considerably lower. CONCLUSIONS: 1) In the brain of WD patient examined the status of trace elements was extensively altered. Further studies are necessary to correlate the concentration of trace elements with pathological lesions and with clinical pictures. 2) The elements considered in our study showed an uneven distribution in different brain areas.

[CK20 expression in the gastrointestinal tract of the embryo and fetus]. / Espressione della CK20 nel tratto gastroenterico nell'embrione e nel feto.

A novel type of cytokeratin, cytokeratin 20 (CK20), was added in 1990 to the classic catalog of human cytokeratins, a heterogeneous group of proteins present in almost all epithelia. In man, the expression of CK20 is almost entirely confined to the gastro-intestinal epithelium, to the urothelium and to Merkel cells. Since only few data are available regarding the expression of CK20 in the developing human intestinal mucosa, we studied CK20 immunoreactivity in fetal and neonatal human gut. Immunoreactivity for CK20 was tested in fetuses and newborns, from the twelfth up to the fortieth week of gestation. In each subject, a specimen from the oesophagus, stomach, small intestine, colon, appendix was studied. Tissue samples were routinely processed and paraffin sections were stained with the CK20-specific antibody IT-Ks 20.8. CK20 immunoreactivity was absent in the oesophageal epithelium and it was unevenly distributed in the gastrointestinal mucosa. Three main patterns of immunoreactivity were observed during normal development: the first, found in the stomach and in the small bowel, is characterized by a progressive increase in CK20 expression during gestation; the second pattern, found in the duodenum, shows a progressive decrease in CK20 expression during gestation; in colon and appendix (third pattern), we did not find significant changes in the degree of immunoreactivity for CK20 during gestation. CK20 is unevenly expressed in developing human intestinal mucosa. The degree of positivity for CK20 appears to be related to the epithelial maturation stage only in gastric and small bowel mucosa. Further studies are needed to verify if the uneven CK20 immunoreactivity in the gastrointestinal tract persists even in adulthood.

Scanning electron microscopy of the interior of cells in Hürthle cell tumors.

Four cases of Hürthle cell tumor were examined by scanning electron microscopy after being macerated to remove all soluble components. By all morphological criteria, Hürthle cells are oncocytes with their usual augmented complement of mitochondria. The Hürthle cell mitochondria either are ovate with central stacks of cristae or elliptical or rod-like with cristae that often are finger-like. As in salivary gland oncocytes, the shelf-like cristae are anchored to the inner boundary membrane by tubular necks. In some Hürthle cells, all of the mitochondria exhibit reticulate cristae. A few mitochondria harbor a globular inclusion in their inner compartment. The Golgi apparatuses are relatively simple, consisting of imbricated saccules that are edged by small, bud-like structures. The rare lumina in the midst of clusters of Hürthle cells are lined by numerous microvilli. Thus, scanning electron microscopy of macerated Hürthle cell tumors has revealed a number of features, especially of their mitochondria, that have escaped detection by transmission electron microscopy.

[Histological picture of liver disease in thalassemia intermedia]. / Quadro istologico dell'epatopatia nella Talassemia intermedia.

Thalassemia Intermedia (TI) is a clinical definition in use for a spectrum of clinical conditions ranging in severity from the asymptomatic carrier status to the transfusion-dependent status. The histological lesions of the liver in patients affected by TI has not seen well characterized yet. The aim of this study was to define the histological picture of liver disease in TI. To this end we studied our pool of 22 Thalassemia Intermedia patients who underwent liver biopsy; none of them had blood transfusion. We took into consideration fibrosis according to Ishak's staging and iron overload according to Sciot's grading. Moreover, we determined the hepatic iron content by atomic absorption spectroscopy. Our results have shown that, in all patients studied, the histological picture was characterized by the increase in liver iron stores. By Perls's stain haemosiderin appeared to be stored mainly in hepatocytes in the majority of cases. Inspite of the large amounts of liver iron in these patients, fibrosis was absent or mild. We also noted the presence of iron-free-foci, in part of the patients. Our data show that the histological and histochemical picture of liver disease in Thalassemia Intermedia is very similar to that found in Thalassemia Major. The finding of iron-free-foci could induce to consider a follow up of these patients, even if asymptomatic.