Immunohistochemical and ultrastructural studies of stromal cells in hemangioblastoma.

In order to shed more light on the controversial tissue histogenesis of the stromal cells (SC), light microscopic, immunohistochemical and electron microscopic studies were performed on surgical specimens of hemangioblastomas (36, 26 and 7 cases, respectively). SC were immunoreactive for vimentin, S-100 protein, and neuron specific enolase (NSE) in all cases. Occasional SC were also positive for desmin, smooth muscle actin, Factor VIII, Ulex europaeus lectin receptors, GFAP, and Factor XIIIa. However, majority of these cells were negative with all the endothelial and smooth muscle cell markers used. Electron microscopy demonstrated several different types of SC that were reminiscent of pericytes, smooth muscle cells and abnormal endothelium as well as the intermediate forms between all the above cell types. Few SC were found lining the vascular lumina. Some SC formed small cavities reminiscent of early capillaries. However, typical Weibel-Palade bodies were not found in these SC. It is concluded that SC represents a heterogeneous population of lipidized cells, derived predominantly from the vasogenic mesenchyme. Although immunohistochemistry failed to reveal any consistent antigenic property of SC, ultrastructural findings strongly support the hypothesis that these cells are modified or abnormally differentiated endothelial cells and pericytes.

Diagnosis of Alzheimer's disease with commercially available anti-beta peptide (beta A4) antibodies following microwave oven pretreatment.

Alzheimer's disease (AD), the most common presenile dementia is underdiagnosed in Poland, thus every attempt to make the frequency of this diagnosis approaching standards of Western countries should be recommended. Deposits of beta A4 amyloid in a form of amyloid (senile) plaques, diffuse amyloid deposits and congophilic angiopathy is central to the pathogenesis of AD. These amyloid deposits are virtually invisible in routine pathological stainings like HE but may be visualized with Bielschowsky silver impregnation, other metallic impregnations, and following Thioflavine S or Congo red stainings. We report here that amyloid deposits are as easily immunolabeled with commercially available antibodies against beta A4 (DAKO) and such a staining was greatly enhanced by microwave oven pretreatment. In all cases of AD, the diagnosis could be easily made using either 4GM or commercial DAKO anti-beta A4 antibodies following pretreatment with formic acid or processing in microwave oven. Pretreatment in microwave oven even for only one second was already sufficient to visualize beta A4-immunopositive plaques while after 5 second the intensity of staining approached that obtained after formic acid pretreatment.

Adipohormones as prognostric markers in patients with nonalcoholic steatohepatitis (NASH).

Nonalcoholic steatohepatitis (NASH) represents one of the most common liver diseases. It is strongly associated with obesity and insulin resistance and is thought to be part of the metabolic syndrome. NASH can progress to cirrhosis and liver failure. Adipohormones, synthesized in adipose tissue, are involved in the pathophysiology of many acute and chronic liver diseases. The aim of this study was to evaluate the plasma concentrations of adiponectin, resistin, leptin, TNF-alpha and Il-6 in patients with NASH, as well as their correlation with the pathologic parameters. Serum concentration of leptin, adiponectin, resistin, insulin, TNF-alpha, IL-6 were measured with ELISA method. Liver biopsies were obtained from 18 (age 42.55+/-21 years) patients. NASH has been classified according to Dixon score. The control group was represented by 16 non-obese subjects. Mean serum concentration of adiponectin in patients with NASH was significantly lower than in healthy subjects (4.87+/-1.96 vs. 8.33+/-4.56 ng/ml; p<0.05). Mean serum levels of TNF-alpha in patients with NASH were significantly higher than in controls (34.2+/-19.7 vs. 20.7+/-15.5 ng/ml; p<0.05). In patients with more advanced inflammation (grade 2-3) and fibrosis (stage 2) in pathology, serum concentration of leptin was significantly higher than in patients with steatosis and less advanced inflammation (grade 1) and fibrosis (stage 1) (median 8.94 vs. 16.2 ng/ml; p<0.05). No significant differences of serum concentration of others adipohormones between these two groups of patients were stated. Moreover, we observed the correlation in serum levels (examined group vs controls) between: resistin and TNF-alpha (r = 0.62; p<0.05), adiponectin and IL-6 (r = -0.60; p<0.05) and leptin and insulin (r = -0.51; p<0.05). In conclusion, based on our study we speculate that changes of adipohormones levels may be markers of NASH and the serum level of leptin can be associated with more advanced form of NASH.

Expression of Bax, Bcl-xL and Bcl-2 proteins in relation to grade of inflammation and stage of fibrosis in chronic hepatitis C.

AIMS: To determine the expression of regulators of apoptosis in chronic hepatitis C. METHODS AND RESULTS: Expression of Bax, Bcl-xL and Bcl-2 proteins was assessed immunohistochemically in liver biopsy specimens obtained from 89 adults with chronic hepatitis C. Expression of Bax in hepatocytes correlated inversely with grade of inflammation (P < 0.001) and stage of fibrosis (P = 0.011), classified according to the Scheuer score; expression of Bcl-xL in hepatocytes did not correlate with grade of inflammation (P = 0.106) or stage of fibrosis (P = 0.078); maximum Bcl-xL expression was observed in grade 3 inflammation and stage 4 fibrosis. Expression of Bcl-2 protein in hepatocytes was present in only two cases (both with advanced disease); the expression of Bcl-2 protein in interlobular bile duct epithelial cells correlated with the grade of inflammation (P = 0.018), but not with stage of fibrosis (P = 0.154). The expression of Bcl-2 protein in lymphoid cells infiltrating portal zones and lobules did not correlate with grade of inflammation (P = 0.113) or stage of fibrosis (P = 0.815). CONCLUSION: Major differences in expression of studied proteins were observed in relation to grade of inflammation and stage of fibrosis in chronic hepatitis C.

Expression of tumour necrosis factor-related apoptosis-inducing ligand and caspase-3 in relation to grade of inflammation and stage of fibrosis in chronic hepatitis C.

AIM: To assess whether the distribution of the recently described proapoptotic ligand, tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), and the apoptosis effector, caspase-3 alters with the degree of inflammation and fibrosis present in liver biopsy specimens from patients with chronic hepatitis C virus infection. METHODS AND RESULTS: Expression of TRAIL and caspase-3 was assessed immunohistochemically in liver biopsy specimens obtained from 89 adults with chronic hepatitis C. Expression of TRAIL in hepatocytes correlated inversely with stage of fibrosis (P = 0.001), classified according to the Scheuer score; expression of caspase-3 in hepatocytes correlated with grade of inflammation (P = 0.012). Expression of TRAIL in hepatocytes was not correlated with grade of inflammation (P > 0.05); expression of caspase-3 was not correlated with stage of fibrosis (P > 0.05). Maximum expression of proapoptotic TRAIL protein was observed in cases with low grade inflammation (G0) and low stage fibrosis (S1). Maximum expression of caspase-3 in hepatocytes was observed in cases with high grade inflammation (G3-4) and high stage fibrosis (S3), but not with liver cirrhosis (S4). CONCLUSIONS: There is a significant decrease in TRAIL expression with increasing grade of inflammation, whereas caspase-3 expression is significantly increased with advanced fibrosis, short of cirrhosis.

Rabbit supraspinatus tendon detachment: effects of size and time after tenotomy on morphometric changes in the muscle.

We studied the effects of size and time after rabbit supraspinatus tenotomy (group A--small tenotomy, group B--large tenotomy) on muscle morphometric changes in 48 rabbits. Animals were killed 6 (subgroups A1 and B1), 12 (subgroups A2 and B2) and 24 weeks (subgroups A3 and B3) after tendon detachment. Statistically significantly greater increases in interstitium volume were noted in subgroups A1-A3 and B1-B3 than in controls. Reductions in type I and II fiber diameters were mainly due to the length of observation. However, statistically significant differences in comparison with controls appeared earlier after large tenotomy. The size of the tenotomy primarily affected muscle fiber composition. Our results suggest that these changes were caused by fiber transformation from type I to type II and vice versa. These findings indicate that the interdigitations between the supraspinatus and infraspinatus and between the supraspinatus and subscapularis tendons are important in dynamics and the degree of morphometric changes in the rabbit supraspinatus after tenotomy.

[Interferon alpha--treatment predictive response factors in group of adults patients with chronic hepatitis C]. / Czynniki predykcyjne odpowiedzi na leczenie interferonem--alpha 2b u doroslych chorych na przewlekle zapalenie watroby typu C.

INTRODUCTION: Chronic hepatitis C (CHC) is a serious clinical problem. Serious sequelae of the disease such as liver cirrhosis and hepatocellular cancer warrant the search for efficacious therapeutic methods. It is assumed that some initial clinical and laboratory data may predict the therapeutic response. However, before they can be used in clinical practice, objective evaluation of their value is to be performed. AIM OF THE STUDY: Evaluation of IFN-alpha efficacy in treatment of patients with chronic hepatitis C. Evaluation of factors predictive for results of treatment (therapeutic response) in patients with chronic hepatitis C treated with IFN alpha2b. Evaluation of cumulative prognostic value of the significant features. MATERIAL AND METHODS: Fifty four adult patients with chronic hepatitis C were treated with IFN-alpha for 6 months. Follow up was continued for 18 months after the end of treatment period. In the studied group, 19 clinical, biochemical and histologic features were the subjects of statistical analysis. RESULTS AND CONCLUSIONS: In 18.5% of patients the therapeutic response on IFN-alpha2b therapy was sustained, in 48.2% the response was transitory. In the remaining 33.3% of patients no therapeutic response were achieved. Discriminatory analysis revealed that duration of HCV infection, alcalic phosphatase activity in serum, histologic activity index, extensiveness of inflammation and biliary canaliculus proliferation in liver biopsy specimen before the IFN alpha2b treatment, are useful features predictive for the results of treatment. The cumulative prognostic value of these features in patients with chronic hepatitis C is 69%.

Clinical picture of chronic hepatitis C in children--Polish experience.

The aim of this study was to investigate long-term clinical, virologic and histologic outcome of hepatitis C virus infection in children. Sixty children (16 girls and 44 boys) have been followed for 1 to 5 years (mean 1.7 +/- 0.9 years). HCV RNA and anti-HCV were checked every six months. Biopsy specimens were evaluated for the grade of inflammation and stage of fibrosis (scores 0-4). ALT was measured every 3 months. Presumed duration of HCV infection was from 1 to 16 years (mean 7.4 +/- 3 years). Fifteen (25%) children could have been infected by blood transfusion, 5 (8%) during surgical procedures, 29 (50%) were multiply hospitalized. Twenty-five children infected as neonates had lower staging score than 24 infected later in life (p = 0.021). Two girls (aged 13 and 14) were diagnosed with acute hepatitis C, with maximum ALT of 1272 U/l and 1638 U/l respectively. In 11 children (18%) median ALT of more than 3 times the normal value (> 105 U/l) was noted. Six children (10%) had continuously normal ALT. Histopathology revealed mild to moderate inflammatory activity (0-2 points) in 52 children (87%). Seven specimens (11%) were scored for 3 to 4 staging points, 3 of them (5%) were diagnosed with liver cirrhosis. We have found statistically significant correlation between median ALT and grading (r = 0.36; p = 0.005) as well as staging scores (r = 0.32; p = 0.016), median AST and grading (r = 0.36; p = 0.006) as well as staging (r = 0.36; p = 0.007) scores but also median GGT and staging score (r = 0.39; p = 0.004).

Interferon alpha for eradication of HCV and remission of chronic hepatitis in children.

UNLABELLED: Thirty two children (8 girls and 32 boys), aged 4-14, with chronic hepatitis C were treated with interferon alpha at a dose of 3 MU or 5 MU, given times weekly for 6 months. Five children (16%) were complete responders (defined as disappearance of viremia), none of them relapsed, 6 (19%) were partial responders (normalization of ALT), 3 (9%) were transient responders (ALT initially decreased but then rebounded to pre-treatment levels) and 18 (56%) were non responders. During the treatment, continuous decrease of median ALT, AST and GGT was noted, statistically significant differences were found for comparisons of values measured before and after treatment (for ALT p = 0.023, for AST p = 0.021 and for GGT p = 0.012). Histological evaluation revealed one case of progression (to liver cirrhosis) out of three investigated complete responders, 1/5 such cases of partial responders, 1/2 of transient responders and 14/18 of non-responders. Remission of hepatitis was found in 2/18 (10%) non-responders. There was no statistically significant difference between 16 children previously infected with HBV (anti-HBc positive but HBsAg negative at the time of our study) and the others with respect to biochemical and histological severity of hepatitis or in the response to treatment rate. CONCLUSIONS: Chronic hepatitis C in children may cause severe hepatitis or even liver cirrhosis. Virological, biochemical and histological features should be considered to evaluate the severity of the disease and response to interferon treatment. Past infection with HBV does not affect interferon sensitivity in children with chronic hepatitis C.

Angiogenic histogenesis of stromal cells in hemangioblastoma: ultrastructural and immunohistochemical study.

Controversy regarding the origin of characteristic stromal cells (SC) is responsible for the placement of hemangioblastoma as a single entity in the category of "tumors of uncertain histogenesis" in the current WHO classification of brain tumors. This subclassification of hemangioblastoma is, to a large extent, a consequence of a remarkable antigenic heterogeneity of SC demonstrated in many, often contradictory immunohistochemical studies. In contrast, most of the electron microscopic studies demonstrated a number of features indicating angiogenic nature of SC and, therefore, hemangioblastoma. This study reevaluated the histogenesis of SC, applying immunohistochemistry as well as electron microscopy and immunoelectron microscopy. Immunohistochemical studies confirmed most of the previous results indicating a very frequent expression of vimentin, S-100 protein, neuron-specific enolase, and cytokeratins. SC were less commonly immunoreactive for desmin, factor XIIIa, and Ricinus communis lectin receptors, and only occasionally for factor VIII and Ulex europeus lectin. They were negative for other markers of endothelial, neuronal, glial, neuroendocrine, and smooth muscle differentiation. Approximately 1% of SC showed Ki67 immunoreactivity, indicating their slight proliferative activity, consistent with the benign nature of the tumor. In contrast to the inconclusive results of the immunohistochemistry, electron microscopy demonstrated a clear relationship of SC to endothelial cells, smooth muscle cells, and pericytes. Occasional SC were found within the vascular lumina. SC often showed intracellular caveolae consistent with the formation of early capillary lumina. Moreover, occasional SC contained small Weibel-Palade bodies positive for factor VIII in immunoelectron microscopy. SC represent a heterogeneous population of abnormally differentiating mesenchymal cells of angiogenic lineage, with some morphological features of endothelium, pericytes, and smooth muscle cells. Occurrence of SC in hemangioblastoma could be related to a limited ability of angioformative stromal cells to develop an architecture of capillary lumina integrated with the vascular network of the tumor. Hemangioblastoma should be reclassified and included together with other vascular tumors of the central nervous system.

Bilateral uveal melanoma presenting simultaneously.

A case of bilateral uveal melanoma in a 54-year-old man is described. On admission of the patient for a choroidal melanoma of the left eye, an asymptomatic ciliary-body tumour was detected in his right eye. A thorough general examination did not reveal any metastases. The left eye was enucleated and local excision of the tumour in the right eye was performed 2 months later. Histology confirmed the presence of malignant melanomas in both eyes. The tumours were of similar A/B spindle-cell type. The patient remained healthy, showing no sign of metastasis or local recurrence of melanoma 9 months after the date of diagnosis. The visual acuity in his remaining eye remained 6/6. Immunological assessment of the blood serum revealed abnormally high interleukin 1 beta (IL-1 beta) and IL-2 levels. Possible implications of these findings are discussed.