László Tiszlavicz

Hepatocellular carcinoma in ten children under five years of age with bile salt export pump deficiency

Hepatocellular carcinoma (HCC) is rare in young children. We attempted to see if immunohistochemical and mutational‐analysis studies could demonstrate that deficiency of the canalicular bile acid transporter bile salt export pump (BSEP) and mutation in ABCB11, encoding BSEP, underlay progressive familial intrahepatic cholestasis (PFIC)—or “neonatal hepatitis” suggesting PFIC—that was associated with HCC in young children. We studied 11 cases of pediatric HCC in the setting of PFIC or “neonatal hepatitis” suggesting PFIC. Archival liver were retrieved and immunostained for BSEP. Mutational analysis of ABCB11 was performed in leukocyte DNA from available patients and parents. Among the 11 nonrelated children studied aged 13‐52 months at diagnosis of HCC, 9 (and a full sibling, with neonatal hepatitis suggesting PFIC, of a tenth from whom liver was not available) had immunohistochemical evidence of BSEP deficiency; the eleventh child did not. Mutations in ABCB11 were demonstrated in all patients with BSEP deficiency in whom leukocyte DNA could be studied (n = 7). These mutations were confirmed in the parents (n = 14). With respect to the other 3 children with BSEP deficiency, mutations in ABCB11 were demonstrated in all 5 parents in whom leukocyte DNA could be studied. Thirteen different mutations were found. In conclusion, PFIC associated with BSEP deficiency represents a previously unrecognized risk for HCC in young children. Immunohistochemical evidence of BSEP deficiency correlates well with demonstrable mutation in ABCB11. (HEPATOLOGY 2006;44:478–486.)

DNAI2 Mutations Cause Primary Ciliary Dyskinesia with Defects in the Outer Dynein Arm

Primary ciliary dyskinesia (PCD) is a genetically heterogeneous disorder characterized by chronic destructive airway disease and randomization of left/right body asymmetry. Males often have reduced fertility due to impaired sperm tail function. The complex PCD phenotype results from dysfunction of cilia of the airways and the embryonic node and the structurally related motile sperm flagella. This is associated with underlying ultrastructural defects that frequently involve the outer dynein arm (ODA) complexes that generate cilia and flagella movement. Applying a positional and functional candidate-gene approach, we identified homozygous loss-of-function DNAI2 mutations (IVS11+1G > A) in four individuals from a family with PCD and ODA defects. Further mutational screening of 105 unrelated PCD families detected two distinct homozygous mutations, including a nonsense (c.787C > T) and a splicing mutation (IVS3-3T > G) resulting in out-of-frame transcripts. Analysis of protein expression of the ODA intermediate chain DNAI2 showed sublocalization throughout respiratory cilia. Electron microscopy showed that mutant respiratory cells from these patients lacked DNAI2 protein expression and exhibited ODA defects. High-resolution immunofluorescence imaging demonstrated absence of the ODA heavy chains DNAH5 and DNAH9 from all DNAI2 mutant ciliary axonemes. In addition, we demonstrated complete or distal absence of DNAI2 from ciliary axonemes in respiratory cells of patients with mutations in genes encoding the ODA chains DNAH5 and DNAI1, respectively. Thus, DNAI2 and DNAH5 mutations affect assembly of proximal and distal ODA complexes, whereas DNAI1 mutations mainly disrupt assembly of proximal ODA complexes.

Involvement of oxygen-derived free radicals in L-arginine-induced acute pancreatitis

This study was aimed at an assessment of the role of oxygen-derived free radicals in the pathogenesis of L-arginine (Arg)-induced acute pancreatitis in rat, by measuring the levels of malonyl dialdehyde (MDA), glutathione peroxidase (GPx), catalase, and superoxide dismutase (Mn- and Cu,Zn-SOD) in the pancreatic tissue, and evaluating the protective effect of the xanthine oxidase inhibitor allopurinol. Acute pancreatitis was induced in male Wistar rats by injecting 2 × 250 mg/100 g body weight of Arg intraperitoneally in a 1-hr interval, as a 20% solution in 0.15 M NaCl. Control rats received the same quantity of glycine. Allopurinol, 100 or 200 mg/kg, was administered subcutaneously 30 min before the first Arg injection. Rats were killed at 6, 12, 24, and 48 hr following Arg administration, and acute pancreatitis was confirmed by a serum amylase level elevation and typical inflammatory features observed microscopically. The serum level of amylase reached the peak level at 24 hr after the Arg injection (30,800 ± 3813 vs 6382 ± 184 units/liter in the control) and normalized at 48 hr. The tissue concentration of MDA was significantly elevated at 24 hr and reached the peak value at 48 hr (5.00 ± 1.75 vs 0.28 ± 0.05 nM/mg protein in the control). The catalase and Mn-SOD activities were significantly decreased throughout the study, while the GPx activity was significantly reduced at 6 and 12 hr, and the Cu,Zn-SOD activity was significantly lower at 12 hr after the Arg injection as compared with the controls. Allopurinol treatment markedly reduced the serum amylase elevation (12.631 ± 2.257 units/liter at 24 hr) and prevented the increase in tissue MDA concentration (0.55 ± 0.09 nM/mg protein at 48 hr). Both doses of allopurinol significantly ameliorated the pancreatic edema, necrosis, and inflammation at 48 hr after Arg administration. Oxygen-derived free radicals are generated at an early stage of Arg-induced acute pancreatitis. Prophylactic allopurinol treatment prevents the generation of reactive oxygen metabolites, reduces the serum amylase concentration, and exerts a beneficial effect on the development of histopathological changes.

Prognostic significance of endogenous adhesion/growth-regulatory lectins in lung cancer

Objective: To determine the expression of endogenous adhesion/growth-regulatory lectins and their binding sites using labeled tissue lectins as well as the binding profile of hyaluronic acid as an approach to define new prognostic markers. Methods: Sections of paraffin-embedded histological material of 481 lungs from lung tumor patients following radical lung excision processed by a routine immunohistochemical method (avidin-biotin labeling, DAB chromogen). Specific antibodies against galectins-1 and -3 and the heparin-binding lectin were tested. Staining by labeled galectins and hyaluronic acid was similarly visualized by a routine protocol. After semiquantitative assessment of staining, the results were compared with the pT and pN stages and the histological type. Survival was calculated by univariate and multivariate methods. Results: Binding of galectin-1 and its expression tended to increase, whereas the parameters for galectin-3 decreased in advanced pT and pN stages at a statistically significant level. The number of positive cases was considerably smaller among the cases with small cell lung cancer than in the group with non-small-cell lung cancer, among which adenocarcinomas figured prominently with the exception of galectin-1 expression. Kaplan-Meier computations revealed that the survival rate of patients with galectin-3-binding or galectin-1-expressing tumors was significantly poorer than that of the negative cases. In the multivariate calculations of survival lymph node metastases (p < 0.0001), histological type (p = 0.003), galectin-3-binding capacity (p = 0.01), galectin-3 expression (p = 0.03) and pT status (p = 0.003) proved to be independent prognostic factors, not correlated with the pN stage. Conclusion: The expression and the capacity to bind the adhesion/growth regulatory galectin-3 is defined as an unfavorable prognostic factor not correlated with the pTN stage.

Leiomyosarcoma of the larynx. Review of the literature and report of two cases.

Leiomyosarcoma of the larynx is an extremely rare malignancy. Until now, only 35 cases have been reported, and only 19 were reported in English. We present 2 new cases observed in the last 3 years at our department. Correct histologic diagnosis can only be made on immunohistochemical and electron microscopic grounds. A 65-year-old man previously treated for a premalignant vocal cord lesion and a 31-year-old woman who previously underwent operation on a benign vocal cord lesion are presented. Only surgical treatment (total laryngectomy and endolaryngeal laser resection) was performed. Thirty-six and 22 months after the initial surgical intervention, both patients are alive and well, with no evidence of local recurrence or distant metastasis.

Novel Role for Pendrin in Orchestrating Bicarbonate Secretion in Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)-expressing Airway Serous Cells

In most HCO3−-secreting epithelial tissues, SLC26 Cl−/HCO3− transporters work in concert with the cystic fibrosis transmembrane conductance regulator (CFTR) to regulate the magnitude and composition of the secreted fluid, a process that is vital for normal tissue function. By contrast, CFTR is regarded as the only exit pathway for HCO3− in the airways. Here we show that Cl−/HCO3− anion exchange makes a major contribution to transcellular HCO3− transport in airway serous cells. Real-time measurement of intracellular pH from polarized cultures of human Calu-3 cells demonstrated cAMP/PKA-activated Cl−-dependent HCO3− transport across the luminal membrane via CFTR-dependent coupled Cl−/HCO3− anion exchange. The pharmacological and functional profile of the luminal anion exchanger was consistent with SLC26A4 (pendrin), which was shown to be expressed by quantitative RT-PCR, Western blot, and immunofluorescence. Pendrin-mediated anion exchange activity was confirmed by shRNA pendrin knockdown (KD), which markedly reduced cAMP-activated Cl−/HCO3− exchange. To establish the relative roles of CFTR and pendrin in net HCO3− secretion, transepithelial liquid secretion rate and liquid pH were measured in wild type, pendrin KD, and CFTR KD cells. cAMP/PKA increased the rate and pH of the secreted fluid. Inhibiting CFTR reduced the rate of liquid secretion but not the pH, whereas decreasing pendrin activity lowered pH with little effect on volume. These results establish that CFTR predominately controls the rate of liquid secretion, whereas pendrin regulates the composition of the secreted fluid and identifies a critical role for this anion exchanger in transcellular HCO3− secretion in airway serous cells.

The role of nitric oxide in edema formation in L-arginine-induced acute pancreatitis.

Introduction Nitric oxide (NO) has been implicated in the regulation of the pancreatic circulation, the promotion of the capillary integrity, and the inhibition of leukocyte adhesion. Aims To investigate the rates of changes in the pancreatic constitutive NO synthase (cNOS) and inducible NOS (iNOS) activities and the role of NO in the vascular permeability changes during the development of L-arginine (Arg)–induced acute pancreatitis. Methodology Acute pancreatitis was induced in male Wistar rats by injecting 250 mg/100 g body weight of Arg i.p. twice at an interval of 1 hour, as a 20% solution in 0.15 M NaCl (group I). The control rats received the same quantity of glycine (group II). In group III, 30 mg/kg NG-nitro-L-arginine methyl ester (L-NAME) was injected i.p. 19 hours after the first Arg injection. The rats were killed at 6, 12, 24, or 48 hours following Arg administration, and the plasma amylase concentration and the pancreatic weight/body weight (pw/bw) ratios were evaluated. NOS activity was determined via the conversion of L-14C-Arg monohydrochloride to 14C-citrulline. The vascular permeability was examined by means of the extravasation of Evans blue dye (20 mg/kg bw) into the pancreatic tissue. Results The serum amylase level was already increased at 6 hours in group I animals, peaked at 12 hours after the Arg injection (11.800 ± 590 versus 6.618 ± 252 U/L in group II), and returned to the control level at 48 hours. The pw/bw ratio peaked at 24 hours in group I (6.63 ± 0.52 versus 4.02 ± 0.22 mg/g in group II) and returned to the control level at 48 hours. The cNOS activity was depleted at 6 hours in group I (0.02 ± 0.003 versus 0.23 ± 0.02 pmol/min/mg protein in group II); it then gradually increased to a level significantly higher than that in group II and decreased thereafter (0.45 ± 0.03 and 0.13 ± 0.01 pmol/min/mg protein at 24 and 48 hours). The iNOS activity was significantly increased at 24 and 48 hours versus that in group II (0.15 ± 0.05 and 0.07 ± 0.01 versus 0.04 ± 0.01 pmol/min/mg protein). The pancreatic concentration of Evans blue dye was significantly higher in group I than in group II (138.59 ± 11.04 versus 43.57 ± 2.67 (g/dry weight). Treatment with L-NAME significantly reduced the amylase activity, pw/bw, Evans blue concentration, and cNOS activity of the pancreas but did not exert any beneficial effect on the histologic score at 24 hours after the onset of pancreatitis, as compared with those values in group I (6.528 ± 673 U/L, 4.56 ± 0.65 mg/g, 86.84 ± 3.9 (g/dry weight, 0.14 ± 0.04 pmol/min/mg protein). Conclusion Endogenous NO is involved in the formation of pancreatic edema in Arg-induced acute pancreatitis by increasing the vascular permeability and protein extravasation. L-NAME treatment decreased the cNOS activity and edema formation but did not prevent the histologic damage in Arg-induced acute pancreatitis.

Genetic Polymorphisms of NOD1 and IL-8, but not Polymorphisms of TLR4 Genes, Are Associated with Helicobacter pylori-Induced Duodenal Ulcer and Gastritis

Background:  Intracellular pathogen receptor NOD1 is involved in the epithelial cell sensing Helicobacter pylori, which results in a considerable interleukin (IL)‐8 production. The aim of this study was to evaluate the relationship between NOD1 and IL‐8 genetic polymorphisms and the development of H. pylori‐induced gastritis and duodenal ulcer (DU), as compared with TLR4 polymorphisms.

Bronchioloalveolar lung cancer: Occurrence, surgical treatment and survival

OBJECTIVE The prevalence of pulmonary adenocarcinoma has risen worldwide. Bronchioloalveolar carcinoma (BAC) was studied with regard to whether it exhibits a similar tendency, and its typical features were analysed. METHODS Between 1992 and 2001, 278 lung resections were carried out for adenocarcinoma. Of these, 67 (24.1%) proved to involve BAC. Whereas BAC accounted for 6.9% of the cases in 1992, in 2001 the proportion was 46.9%. There were 37 men (55.2%) and 30 women (44.7%); the average age was 60.5 years. 58.2% of them had no complaints. Of the 26 non-smokers, 69.2% were women; of the 41 smokers, 29.2% were women. In consequence of the tumour, 49 lobectomies, three bilobectomies, six pneumonectomies and nine wedge resections were performed. RESULTS The surgical mortality was 1.6%. The pathology revealed that 26 (38.8%) tumours were in stage I/A. In 15 cases (22.4%), tuberculosis (TB) could be revealed besides the BAC: by skin tests in four cases, by CT in three cases, by case history in four cases, and by pathology in four cases. For the overall group of 67 patients, the 5-year survival rate was 61.9%, and the mean survival time was 75.7 months. The 5-year survival rate among the women (74%) was significantly better than that among the men (37%) (P=0.030). There was no significant difference in survival with regard to the multiple BAC (85%). The 5-year survival rate was significantly worse in the mixed BAC group (20%) than in the non-mucinous (62.7%) and in mucinous (59%) group. The overall 5-year survival rate among the smokers and TB patients was 61 and 79%, respectively, which is higher than that among the non-smokers (47%) and non-TB patients (56%). The survival rate for the wedge resection cases was 37%, which was lower than that for the cases involving major resections (60%) (P=0.939). CONCLUSION BAC has a favourable survival, particularly in women. In spite of this, resection smaller than lobectomy is recommended only as a compromise. A multiple appearance does not imply a worse survival. The best survival rate was found in the non-mucinous BAC among the histological groups. TB seems to be frequent among BAC patients.

Human dirofilariosis in Hungary: an emerging zoonosis in central Europe

ZusammenfassungEINLEITUNG: In den gemäßigten Regionen Europas, vorwiegend in Italien, Frankreich und Griechenland, und in den letzten Jahren auch in Ungarn, führte ein spezieller Wurm, die Dirofilaria (Nochtiella) repens, zu Infektionen beim Menschen und verursacht eine autochthone humane Dirofiariose. D. repens ist ein Parasit von Hunden, Katzen und einigen anderen Raubtieren, und wird durch Mücken übertragen. Wir berichten über einen weiteren Fall und fassen die bisher in Ungarn beobachteten Fälle zusammen. PATIENTEN UND METHODEN: Die Autoren präsentieren einen Fall einer humanen Dirofilariose bei einem 56-jährigen Patienten mit Jucken und brennendem Gefühl in seinem rechtem Auge. Ein sich langsam bewegender, weißlicher, Zwirn-ähnlicher Wurm wurde durch Spaltlampenuntersuchung entdeckt. Die parasitologische Untersuchung identifizierte ihn als junge männliche Dirofilaria repens. Eine aktualisierte Übersicht über die ungarischen Fälle von 1880–2002 wird vorgestellt und zusätzlich 16 neuen Fälle analysiert, die in der Periode von 2001 zu 2006 diagnostiziert wurden. Das Durchschnittsalter dieser 16 neuen Patienten war 60 Jahre, acht Patienten hatten eine okulare, sieben eine subkutane Lokalisation. Bei einem Patienten mit akuter lymphatischer Leukämie wurde histopatologisch aus einem axillärem Lymphknoten diagnostiziert. Keiner der Patienten hatte sich im Ausland aufgehalten. KONKLUSION: Die Mehrheit der in Urgarn beobachteten Fälle von Dirofilariasis muss als autochthon aquiriert angesehen werden. Die zunehmende Zahl an diagnostizierten Fällen spricht dafür, dass diese Zoonose auch in Zentraleuropa Verbreitung erlangt.SummaryINTRODUCTION: In the temperate regions of Europe, mainly Italy, France, Greece, and in recent years Hungary, a specific filarioid worm, Dirofilaria (Nochtiella) repens, a parasite of dogs, cats and some other carnivores, transmitted by mosquitoes, has occasionally been contracted, causing dirofilariosis in humans. We report a case of dirofilariosis and review all previously observed cases in Hungary. MATERIALS, METHODS AND RESULTS: The authors present a case of human subconjunctival dirofilariosis in a 56-year-old man with mild itching and a burning sensation in his right eye. A slow-moving, whitish, thread-like worm was detected by slit-lamp examination. The worm was surgically removed and parasitological examination identified it as an immature male Dirofilaria repens. A review of Hungarian cases from 1880 to 2000 is presented, with updates of earlier reports. In addition, 16 new cases (8 men; 8 women) diagnosed between 2001 and 2006 are reviewed. The mean age of these 16 patients was 60 years; eight cases had ocular localization, seven were subcutaneous and one was diagnosed in a histopathological section of removed axillary lymph node in a patient with lymphoid leukemia. Twelve of the 16 identified Dirofilariae were females, four were males. According to their recorded histories the patients had not traveled internationally. CONCLUSION: Most of the above cases of dirofilariosis are certainly autochthonous infections. The increasing number of diagnosed cases suggests that this zoonosis is becoming more important in central Europe and that the incidence may increase with the improvement of clinical diagnosis.