Identification and intraspecific genetic diversity of Sarcocystis rileyi from ducks, Anas spp., in Lithuania and Finland.

Macroscopic Sarcocystis cysts were detected in the muscles of 28 Mallards ( Anas platyrhynchos ), 1 Eurasian Wigeon ( Anas penelope ), and 1 Common Teal ( Anas crecca ) hunted in Lithuania and Finland. According to the sequences of the 18S rRNA gene, 28S rRNA gene, and ITS-1 region, the macrocysts examined from all 30 ducks belonged to Sarcocystis rileyi. This parasite was found in the Eurasian Wigeon and the Common Teal for the first time. All S. rileyi isolates examined were identical to each other and differed from 2 S. rileyi isolates previously reported from 2 Mallards from the United States only by 1 nucleotide substitution within the ITS-1 region.

Liver transplantation for familial amyloidotic polyneuropathy (FAP): a single-center experience over 16 years.

Orthotopic liver transplantation (LTx) is currently the only available treatment that has been proven to halt the progress of familial amyloidotic polyneuropathy (FAP). The aim of this study was to assess mortality and symptomatic response to LTx for FAP. All 86 FAP patients transplanted at our hospital between April 1990 and November 2005 were included in the study. Five patients underwent retransplantation. The 1-, 3- and 5-year patient survival rates in patients transplanted during 1996-2005 were 94.6%, 92.3% and 92.3%, respectively, a significant difference from the rates of 76.7%, 66.7% and 66.7%, respectively, during 1990-1995 (p = 0.0003). Multivariate analysis revealed that the age at the time of LTx (>or=40 years), duration of the disease (>or=7 years) and modified body mass index (mBMI) (<600) were independent prognostic factors for patient survival. A halt in the progress of symptoms was noted in most patients, but only a minority experienced an improvement after LTx. To optimize the posttransplant prognosis, LTx should be performed in the early stages of the disease, and close post-LTx monitoring of heart function by echocardiography and of heart arrhythmia by Holter ECG is mandatory.

Outbreak of parasitic peritonitis in reindeer in Finland.

In 2003, there was an outbreak of peritonitis in reindeer in the southern and middle part of the Finnish reindeer herding area caused by the filarioid nematode Setaria species. In the province of Oulu, the proportion of reindeer viscera condemned owing to parasitic lesions increased from 4.9 per cent in 2001 to 40.1 per cent in 2003. In 2004, the focus of the outbreak moved approximately 100 km north. A total of 260 adult and pre-adult Setaria species nematodes were collected for morphological and molecular studies. The parasite was indistinguishable in terms of morphology and molecular biology from Setaria tundra. Samples of parasites were also collected from wild cervids. In elk, only a few cases of pre-adult encapsulated S tundra nematodes were detected on the surface of the liver but there was no peritonitis. Two roe deer had S tundra nematodes in their abdomen but no peritonitis. Of 34 wild forest reindeer, 21 had changes associated with S tundra. At meat inspection of the affected reindeer carcases, the changes observed included ascites, green fibrinous deposits, adhesions, and live and dead S tundra nematodes. There were histological lesions of granulomatous peritonitis with lymphoplasmacytic and eosinophilic infiltration. No specific bacterial growth was found. The parasitic infection had no significant effects on the pH or the organoleptic quality of the meat. There was a significant positive correlation between the worm count and the degree of peritonitis (P<0.001) and a negative correlation between the degree of peritonitis and the thickness of the back fat layer (P=0.015).

Liver transplantation for HCV cirrhosis at Karolinska University Hospital Huddinge, Stockholm.

Hepatitis C virus (HCV)-induced cirrhosis is the major indication for liver transplantation globally, and an increasing indication for liver transplantation in Sweden. We have retrospectively examined the 120 patients transplanted for HCV cirrhosis from 1987 through 2005, including 11 who received more than one graft. The 1-, 3-, and 5-year postoperative survivals for all patients transplanted for HCV with or without hepatocellular cancer (HCC) were 77%, 66%, and 53%, respectively. HCV patients without HCC had a 1-, 3-, and 5-year survivals of 78%, 73%, and 61%, compared with 84%, 79% and 74%, respectively, for patients transplanted with chronic liver diseases without cancer or HCV. The number of patients with HCV cirrhosis transplanted in our center is increasing. Compared with patients transplanted for other chronic liver diseases, we experienced inferior results among patients with HCV cirrhosis.

Molecular characterization of Echinococcus isolates of cervid origin from Finland and Sweden.

The species Echinococcus granulosus is made up of several genotypic strain groups, whose taxonomical classification is still undetermined. Genotypes in the cervid-wolf life-cycle are poorly known, especially in Europe. In this study, 33 Echinococcus isolates from cervids from Finland and Sweden were characterized using mitochondrial ND1 gene sequencing. In addition, phylogenetic analysis of E. granulosus strains using the mitochondrial ATP6, ND1, ND3 and CO1 genes was performed using maximum likelihood, neighbour-joining and maximum parsimony methods. The Finnish and Swedish cervid isolates were found to represent the genotype G10. In the phylogenetic analyses, the camel (G6), pig (G7), cervid (G8) and Fennoscandian cervid (G10) strains clustered in a well-supported monophyletic group. This group differed clearly from the common sheep (G1) and horse (G4, 'E. equinus') strains, but was closely related to the cattle strain (G5, 'E. ortleppi'). Our results support the previous studies suggesting that the genotypes G6-10 should be separated from the species E. granulosus sensu stricto. However, additional morphological studies are needed, and the relationship to the cattle strain ('E. ortleppi') should be thoroughly evaluated before a final decision of the taxonomical status of the G6-10 group can be made.

Inflammation and cytokeratin 7/20 staining of cardiac mucosa in young patients with and without Helicobacter pylori infection.

BACKGROUND: Both Helicobacter pylori and gastro-oesophageal reflux disease (GORD) may cause inflammation in cardiac mucosa. Intestinal metaplasia (IM) is found more often in GORD associated inflammation than in inflammation caused by H pylori, especially in young individuals. AIM: To examine morphological differences in chronic inflammation in these two conditions by immunohistochemistry. PATIENTS/METHODS: Tissue blocks from cardiac mucosa of patients <45 years were available as follows: 10 patients with chronic inflammation of cardiac mucosa (carditis) and H pylori gastritis (group 1); 10 patients with (possibly GORD related) carditis, but normal antrum and corpus (group 2); and 10 patients with non-inflamed cardiac mucosa and normal antrum and corpus (group 3). Haematoxylin and eosin staining and immunohistochemical staining for various inflammatory cells were performed for patients in groups 1 and 2 as follows: CD20 (B cells), CD3 (T cells), CD4 (T helper cells), CD8 (T suppressor cells), CD163 (macrophages), CD138 (plasma cells), and CD117 (mast cells). For all patients, cytokeratin 7/20 (CK7/20) staining was performed. RESULTS: No clear differences were seen in the morphology of chronic inflammation between groups 1 and 2. In both, plasma cells were most abundant. CK7/20 staining showed no differences between these groups. CONCLUSION: Helicobacter pylori negative (possibly GORD associated) and H pylori related carditis cannot be distinguished on a morphological basis. The stronger tendency towards IM in the first entity cannot be explained by differences in the type of inflammation. Barrett-type CK7/20 staining seems typical for cardiac mucosa, irrespective of the type of inflammation or presence of IM.

Redescription of Besnoitia tarandi (Protozoa: Apicomplexa) from the reindeer (Rangifer tarandus).

Besnoitia tarandi tissue cysts were found in naturally-infected reindeer (Rangifer tarandus) from Finland. Infectivity of its tissue cysts, bradyzoites, and tachyzoites to animals and cell culture was studied. The bradyzoites and tissue cysts were not infectious to out-bred mice, rabbits or gerbils. When fed tissue cysts, neither cats nor dogs excreted oocysts. However, the parasite was lethal to interferon-gamma gene knock out mice irrespective of the route of inoculation. The parasite was grown successfully in African Green Monkey cells from tissues of two reindeer for the first time. Non-dividing, uninucleate tachyzoites from smears from cell cultures were 5.6 x 1.4 microm (4.5-7.4 x 1.0-1.9, n=50) in size. Longitudinally-cut bradyzoites in tissue sections measured 7.4 x 1.3 microm (6.5-7.8 x 1.0-1.6, n=30). Ultrastructurally, tachyzoites and bradyzoites were similar to those in other Besnoitia species, and in particular to parasites described from cattle (Besnoitia besnoiti) and equids (Besnoitia bennetti) in that their bradyzoites lacked enigmatic bodies. Based on comparative analysis of three portions of nuclear ribosomal DNA (the small and large subunits and the first internal transcribed spacer) B. tarandi was found to be more closely related to the other congeners described from ungulates. The parasite was formally redescribed and specimens deposited in the US National Parasite Collection.

Outcome following liver transplantation for primary sclerosing cholangitis in the Nordic countries.

BACKGROUND: Primary sclerosing cholangitis (PSC) is the most common indication for liver transplantation in the Nordic countries. Because these patients are difficult to evaluate with regard to timing of liver transplantation, it is important to establish predictors of post-transplant survival. METHODS: Data from two groups of patients receiving liver allografts during 1982-2001 were recorded: (a) PSC patients and (b) comparison patients. Outcome following transplantation has been recorded for all patients. Regression analyses have been performed for PSC patients to analyse predictors of patient and graft survival. RESULTS: A total of 245 PSC and 618 comparison patients received a first liver allograft in the period 1982 until the end of the study. The overall 1-, 3- and 5-year patient survival rates were 82%, 77% and 75%, and 80%, 77% and 74% in the PSC group and comparison group, respectively. Survival following transplantation has increased with time in both the PSC and the comparison group. Recent year of transplantation, no previous hepatobiliary surgery and a lower MELD score were predictors of survival following transplantation for PSC patients. PSC patients had a higher rate of re-transplantations (13% versus 8%, P = 0.01). Predictors of re-transplantation in PSC patients were an episode of early rejection and vascular thrombosis. CONCLUSION: In PSC patients, year of transplantation, previous hepatobiliary surgery and MELD score are predictors of survival following transplantation and these patients are more frequently in need of re-transplantation compared to the comparison group.

Transcriptional regulation of the lactase-phlorizin hydrolase gene by polymorphisms associated with adult-type hypolactasia.

BACKGROUND AND AIMS: The mechanism of the developmental downregulation of the lactase-phlorizin hydrolase (LPH) gene underlying adult-type hypolactasia is unknown. We have determined the functional significance of the recently identified two single nucleotide polymorphisms (SNPs), C/T(-13910) and G/A(-22018), associated with adult-type hypolactasia by studying LPH mRNA levels in intestinal biopsy samples with different genotypes. METHODS: Intestinal biopsy samples were taken from 52 patients with abdominal complaints. Hypolactasia was diagnosed by determining lactase and sucrase activities and calculating their ratio (L/S ratio). The functional effect of the C/T(-13910) and G/A(-22018) genotype on expression of LPH mRNA was demonstrated in patients heterozygous for the C/T(-13910) and G/A(-22018) polymorphism and an informative expressed SNP located in the coding region of the LPH mRNA. Reverse transcription-polymerase chain reaction followed by solid phase minisequencing was used for accessing the relative expression levels of the LPH alleles using informative SNPs located in exons 1, 2, 6, 10, 13, or 17 as markers. RESULTS: Statistically significant differences between the three different genotypes CC(-13910) GG(-22018), CT(-13910) GA(-22018), and TT(-13910) AA(-22018) and their respective L/S ratios were observed. Relative quantitation of the expressed LPH alleles showed that the persistent allele represented 92 (6)% (mean (SEM), range 78-99%; n=14) of the expressed LPH mRNA. The patient with the homozygous persistent TT(-13910) AA(-22018), as well as hypolactasic patients with CC(-13910) GG(-22018), showed equal expression of both alleles (47 (1)%; n=7). CONCLUSIONS: Expression of LPH mRNA in the intestinal mucosa in individuals with T(-13910) A(-22018) alleles is several times higher than that found in individuals with C(-13910), G(-22018) alleles. These findings suggest that the two SNPs, C/T(-13910) and G/A(-22018), associated with adult-type hypolactasia, are associated with the transcriptional regulation of the LPH gene. The presence of the T(-13910) A(-22018) allele also shows significant elevation of the L/S ratio.

Inflammation and intestinal metaplasia at the squamocolumnar junction in young patients with or without Helicobacter pylori infection.

BACKGROUND: Intestinal metaplasia (IM) in the oesophagus is a known risk factor for adenocarcinoma of the oesophagus. The incidence of adenocarcinoma of the cardia and oesophagus has increased in Western countries simultaneously with a decrease in Helicobacter pylori prevalence. AIMS: To determine the association of H pylori infection with inflammation and IM at the squamocolumnar junction (SCJ) in young individuals. PATIENTS: A total of 168 (121 women; 72%) consecutive outpatients,

Serologic diagnosis of Helicobacter pylori infection in outpatients aged 45 years or less.

The diagnostic accuracy of serological tests for Helicobacter pylori was studied in 145 consecutive outpatients aged 45 years or less referred for gastroscopy. Helicobacter pylori infection can be detected by serological tests, including rapid whole-blood tests. The low prevalence of the disease in young people may have a negative effect on the positive predictive value of a test. In this study, the presence of Helicobacter pylori was assessed by a biopsy urease test and histological examination, and by several serological tests: a rapid whole-blood test on fingerstick blood, a latex agglutination serum test, a commercial enzyme immunoassay (EIA) test, and an in-house EIA for detection of antibodies of both the IgG and IgA classes. Helicobacter pylori infection was diagnosed with invasive tests in 21 (14.5%) patients. The sensitivity, specificity, and positive and negative predictive values of the EIA-based tests, compared to histological examination, were 100%, 96-97%, 81-84%, and 100%, respectively. The positive predictive value of the latex agglutination test was 78%, whereas it was only 47% for the whole-blood rapid test used. Although the results of the whole-blood rapid test were unsatisfactory, the quantitative EIA-based tests could reliably detect Helicobacter pylori among young patients, in whom the prevalence of the infection is low.

Haemogregarines of the genus Hepatozoon (Apicomplexa: Adeleina) in rodents from northern Europe.

We studied the prevalence and distribution of Hepatozoon infections in small rodents from Finland and other areas in northern Europe. Hepatozoon infections were more common in voles (Arvicolinae) than mice (Murinae) and more prevalent in voles of the genus Clethrionomys than in voles of the genus Microtus. Transmission electron microscopical examination of Hepatozoon erhardovae Krampitz, 1964 from bank voles Clethrionomys glareolus (Schreber) showed that intracellular lung meronts were located in alveolar septa. Meronts consisted of varying numbers of merozoites packed with amylopectin vacuoles inside electron-lucent parasitophorous vacuole. The size of the meronts was approximately 19 x 14 microm. Monozoic or dizoic cysts were frequent findings in the lung alveoles; the size of cysts was approximately 10 x 6 microm. Gametocytes were found inside eosinophilic granulocytes in the capillaries of lung tissue. Ultrastructurally, micronemes, microtubules, mitochondria, nuclei and lipid droplets were visible.

Evaluation of blood tests to predict normal gastric mucosa.

BACKGROUND: To determine the accuracy of blood tests in predicting normal gastric mucosa confirmed by histological examination of gastric biopsy specimens. METHODS: In total, 207 consecutive patients referred for upper endoscopy were included. Two biopsy specimens each from the antrum and corpus were assessed histologically for the presence of Helicobacter pylori, gastritis, and atrophy. Serum samples were studied for H. pylori antibodies by enzyme immunoassay (Pyloriset EIA-G and EIA-A) and by a rapid latex agglutination test (Pyloriset Dry); pepsinogen I was measured by an immunoenzymometric assay (Gastroset PGI), gastrin by radioimmunoassay, and parietal cell antibodies by indirect immunofluorescence. RESULTS: In 101 (49%) of 207 patients, the gastric mucosa on histologic examination was normal. In the 63 patients aged 45 years or less, H. pylori IgG serology was negative in all 47 patients with normal gastric mucosa and none had low serum pepsinogen I levels. Among 144 patients over age 45 years, 72 had negative H. pylori IgG serology. Combining the serum pepsinogen I assay with the results of H. pylori IgG serology, 12 patients with normal serology but low serum pepsinogen I were found. Thus, 60 patients, 52 of whom showed normal gastric histology, had normal IgG serology and serum pepsinogen 1. In the remaining eight patients with normal blood tests, the histologic changes were very mild. CONCLUSIONS: Although negative H. pylori IgG serology alone in younger patients, and in combination with normal serum pepsinogen I levels in older patients, reliably predicted the presence of normal gastric mucosa, gastroscopy is still recommended for patients over 45 years.

Atrophic gastritis and Helicobacter pylori infection in outpatients referred for gastroscopy.

BACKGROUND: Atrophic gastritis has been shown to be one of the long term sequelae of Helicobacter pylori infection. AIMS: To determine the prevalence of atrophic gastritis in outpatients, to study the accuracy of serological methods for revealing atrophy, and to define the association of H pylori infection with atrophic gastritis in these patients. PATIENTS/METHODS: A total of 207 consecutive outpatients referred for gastroscopy were included. Biopsy specimens from the antrum and corpus were assessed histologically according to the Sydney system. Serum samples were studied for H pylori IgG and IgA antibodies by enzyme immunoassay, CagA antibodies by immunoblot, pepsinogen I by an immunoenzymometric assay, gastrin by radioimmunoassay, and parietal cell antibodies by indirect immunofluorescence. RESULTS: Histological examination revealed atrophic gastritis in 52 (25%) of 207 patients. H pylori and CagA antibodies were strongly associated with atrophic antral gastritis but poorly associated with atrophic corpus gastritis. Low serum pepsinogen I was the most sensitive and specific indicator of moderate and severe atrophic corpus gastritis. All six patients with moderate atrophic corpus gastritis had H pylori infection but eight of 10 patients with severe atrophic corpus had increased parietal cell antibodies and nine had no signs of H pylori infection. CONCLUSIONS: Atrophic antral gastritis was strongly associated with CagA positive H pylori infection. Severe atrophic corpus gastritis was not determined by H pylori tests but low serum pepsinogen I, high gastrin, and parietal cell antibodies may be valuable in detecting these changes.

Association of CagA-positive infection with Helicobacter pylori antibodies of IgA class.

cagA gene, the best known virulence factor of Helicobacter pylori, codes for an immunodominant CagA protein. In this study, CagA antibodies of the IgG class were measured by immunoblot or enzyme immunoassay in subjects with positive H. pylori serology, and the presence of CagA antibodies was compared with that of H. pylori antibodies of IgA and IgG classes. Serum samples were available for a total of 1,481 subjects, including gastroscopied patients with biopsy-verified H. pylori infection, smoking men with a normal or low serum pepsinogen I level indicating atrophic corpus gastritis, and subjects who later developed gastric cancer and their matched controls. CagA antibodies were significantly more prevalent among individuals with elevated H. pylori antibody titres of the IgA class than in those with IgG antibodies only, with the exception of a small subgroup of individuals who later developed gastric cancer. CagA-positive H. pylori strains seem to induce an immune response with a markedly higher frequency of IgA than what is found in inflammation caused by CagA-negative strains. The presence of serum IgA antibodies to H. pylori seems to indicate a higher risk for CagA-positive H. pylori infection and possibly more severe late sequelae of the disease.

Liver transplantation in the Nordic countries, 1982-1998: changes of indications and improving results.

BACKGROUND: Liver transplantation has become an established therapeutic option for patients with life-threatening liver disease. The aim of the present study was to analyse the results of and developments in liver transplantation in the Nordic countries during a 15-year period. METHODS: Data on all patients receiving a liver allograft in the Nordic countries during 1982-98 and waiting list data for all patients listed for a liver transplantation after 1989 were obtained from the Nordic Liver Transplantation Registry. RESULTS: A total of 1485 first liver transplantations were performed during 1982-98. The annual number of first liver transplantations increased steadily up to 1993, thereafter remaining around 150-170 per year. There are major differences between countries both in the number of transplants adjusted to populations performed per year, with more than twice as many performed in Sweden as in Norway, and in the relative distribution of patients in accordance with diagnosis. The number of patients more than 60 years old increased and comprised 13%-14% of the total patient population during 1996-98. Primary biliary cirrhosis, primary sclerosing cholangitis, acute hepatic failure, malignant liver disease, and alcoholic cirrhosis are the five most frequent diagnoses. The over-all 1-year patient survival probability has increased from 66% among patients receiving a transplant in 1982-89 to 83% in 1995-1998. The waiting time remains stable, with a median waiting time of 35 days during 1990-98. The mortality of patients while on the waiting list is 7.4% and is not increasing. CONCLUSION: Results of liver transplantation in the Nordic countries are very similar to those obtained in other countries. Waiting time and mortality remain low. There are, however, major differences between the countries both as to the number of transplantations performed and as to distribution of diagnoses.

Expression of autoantibodies to specific cytochromes P450 in a case of disulfiram hepatitis.

BACKGROUND/AIMS: Immunological mechanisms are involved in many adverse drug reactions. In certain forms of drug-induced hepatitis, patients have been reported to express specific autoantibodies to hepatic drug-metabolising enzymes. The alcohol deterrent disulfiram is associated with a low frequency of severe liver toxicity, including hepatitis, but the mechanism of the toxicity is unknown. We investigated whether autoantibodies to cytochrome P450 enzymes were expressed in the serum of a 28-year-old male patient, who developed hepatitis after 7 weeks of disulfiram treatment and in whom possible causes of hepatitis other than disulfiram had been ruled out. METHODS: Patient serum IgG reactivity was analysed by immunoblotting or ELISA against test antigens consisting of recombinant/purified human or rat liver P450 enzymes, or isolated rat liver microsomes. RESULTS: A significant serum reactivity was found in immunoblotting against human cytochromes P450 1A2 and rat P450 3A1, using serum dilutions of up to 1:900 and 1:2400, respectively. In contrast, the reactivity against cytochromes P450 2E1, 2C9, 2D6, 3A4, and rat liver P450 reductase was either very low or undetectable. ELISA reactivity was low in general, indicating that the P450 epitopes were not surface exposed. Immunoblotting of rat liver microsomes revealed that autoantibodies recognised one major polypeptide corresponding to P450 3A. Autoantibody titres remained stable for at least 6 months after acute hepatitis. A similar reactivity was not found in any of ten control sera. CONCLUSIONS: The expression of autoantibodies directed against specific cytochromes P450 in a case of disulfiram hepatitis suggests that immunological mechanisms are involved in this adverse drug reaction, and that these P450 proteins should be evaluated as possible diagnostic test antigens in disulfiram hepatotoxicity.

Serosurvey of Toxoplasma gondii in North Atlantic marine mammals by the use of agglutination test employing whole tachyzoites and dithiothreitol.

Serum samples from North Atlantic populations of harp seal, Phoca groenlandica (n = 316), ringed seal, Phoca hispida (n = 48), hooded seal, Cystophora cristata (n = 78), and minke whale, Balaenoptera acutorostrata (n = 202), were tested for Toxoplasma gondii-specific IgG. The modified agglutination test (MAT) was slightly modified to be more user-friendly by replacing the 0.2 M 2-mercaptoethanol with 10 mM dithiothreitol. No positive samples were found at 1:40 dilution.

Evaluation of Pyloriset Screen, a rapid whole-blood diagnostic test for Helicobacter pylori infection.

Helicobacter pylori infection can be detected by several invasive tests based on gastroscopy and by noninvasive methods such as serologic assays. Noninvasive tests can be used not only in addition to invasive tests but also by themselves to screen for H. pylori infection in patients who are not in urgent need of endoscopy. Lately, rapid qualitative serologic tests have been developed. In the present study, the accuracy of a novel rapid whole-blood test, Pyloriset Screen, detecting immunoglobulin G (IgG) and IgA antibodies against H. pylori was evaluated. A total of 207 consecutive adult outpatients referred for upper endoscopy were enrolled. Gastric biopsy specimens were taken from the antrum and corpus for histologic examination and rapid urease testing. Cultures were available for 113 patients. Serum samples collected from all patients were tested for H. pylori antibodies by two enzyme immunoassays (EIAs) (Pyloriset EIA and an in-house EIA), a rapid latex agglutination test (Pyloriset Dry), and Pyloriset Screen. Patients were considered H. pylori positive if helicobacters were seen on histologic examination (77 patients) or, if in combination with histologically verified (although helicobacter-negative) gastritis, their IgG antibody titers were elevated in the two EIAs (five patients). The Pyloriset Screen test had a sensitivity of 95%, a specificity of 94%, a positive predictive value of 91%, and a negative predictive value of 97%. Among 63 patients under the age of 45 years, the Pyloriset Screen test did not miss a single H. pylori diagnosis, and only 1 patient had a false-positive result. Pyloriset Screen could be used reliably to screen for H. pylori infection.