bluuebunny/crossref_metadata_2025 huggingface dataset:
Historical review of malarial control in southern African with emphasis on the use of indoor residual house‐spraying
Authors: Mabaso Musawenkosi L. H. , Sharp Brian , Lengeler Christian
Published: August 2004

Indoor residual house‐spraying (IRS) mainly with dichlorodiphenyltrichloroethane (DDT) was the principal method by which malaria was eradicated or greatly reduced in many countries in the world between the 1940s and 1960s. In sub‐Saharan Africa early malarial eradication pilot projects also showed that malaria is highly responsive to vector control by IRS but transmission could not be interrupted in the endemic tropical and lowland areas. As a result IRS was not taken to scale in most endemic areas of the continent with the exception of southern Africa and some island countries such as Reunion, Mayotte, Zanzibar, Cape Verde and Sao Tome. In southern Africa large‐scale malarial control operations based on IRS with DDT and benzene hexachloride (BHC) were initiated in a number of countries to varying degrees. The objective of this review was to investigate the malarial situation before and after the introduction of indoor residual insecticide spraying in South Africa, Swaziland, Botswana, Namibia, Zimbabwe and Mozambique using historical malarial data and related information collected from National Malaria Control Programmes, national archives and libraries, as well as academic institutions in the respective countries. Immediately after the inception of IRS with insecticides, dramatic reductions in malaria and its vectors were recorded. Countries that developed National Malaria Control Programmes during this phase and had built up human and organizational resources made significant advances towards malarial control. Malaria was reduced from hyper‐ to meso‐endemicity and from meso‐ to hypo‐endemicity and in certain instances to complete eradication. Data are presented on the effectiveness of IRS as a malarial control tool in six southern African countries. Recent trends in and challenges to malarial control in the region are also discussed.
Human African trypanosomiasis amongst urban residents in Kinshasa: a case‐control study
Authors: Robays J. , Ebeja Kadima A. , Lutumba P. , Miaka mia Bilenge C. , Kande Betu Ku Mesu V. , De Deken R. , Makabuza J. , Deguerry M. , Van der Stuyft P. , Boelaert M.
Published: August 2004

Background  Increasing numbers of human African trypanosomiasis (HAT) cases have been reported in urban residents of Kinshasa, Democratic Republic Congo since 1996. We set up a case‐control study to identify risk factors for the disease.

Methods  All residents of the urban part of Kinshasa with parasitologically confirmed HAT and presenting for treatment to the city's specialized HAT clinics between 1 August, 2002 and 28 February, 2003 were included as cases. We defined the urban part as the area with contiguous habitation and a population density >5000 inhabitants per square kilometre. A digital map of the area was drawn based on a satellite image. For each case, two serologically negative controls were selected, matched on age, sex and neighbourhood. Logistic regression models were fitted to control for confounding.

Results  The following risk factors were independently associated with HAT: travel, commerce and cultivating fields in Bandundu, and commerce and cultivating fields in the rural part of Kinshasa. No association with activities in the city itself was found.

Discussion  In 2002, the emergence of HAT in urban residents of Kinshasa appears mainly linked to disease transmission in Bandundu and rural Kinshasa. We recommend to intensify control of these foci, to target HAT screening in urban residents to people with contact with these foci, to increase awareness of HAT amongst health workers in the urban health structures and to strengthen disease surveillance.
Migrants’ risky sexual behaviours in India and at home in far western Nepal
Authors: Poudel Krishna C. , Jimba Masamine , Okumura Junko , Joshi Anand B. , Wakai Susumu
Published: August 2004

Objective  To examine Nepali migrants’ vulnerability to HIV/sexually transmitted infections (STIs) and their possible role in causing the epidemic in far western Nepal.

Methods  From August to October 2000, we conducted six focus group discussions among 53 returned migrants from India, mainly from Mumbai. Data were analysed by interpretative thematic analysis.

Results  Migrants commonly had multiple sexual encounters, changed partners, and used condoms infrequently both in India and at home. Several factors influenced them to practice high‐risk sexual behaviours. In India, these included peer norms and pressures, cheaper sex, lack of family restraint, drinking alcohol, and low perceived vulnerability to HIV/STIs. In Nepal, these factors included the migrants’ new status, frequent local festivals, and low perceived vulnerability to HIV/STIs. Participants displayed substantial deficits in their knowledge of HIV/STIs.

Conclusions  This study revealed Nepali migrants’ high‐risk sexual behaviours both abroad and at home. Understanding these realities will assist in the development of culturally appropriate HIV/STI interventions necessary to halt the spread of HIV/STIs in Nepal.
Weekly iron supplements given by teachers sustain the haemoglobin concentration of schoolchildren in the Philippines
Authors: Roschnik Natalie , Parawan Amado , Baylon Melba Andrea B. , Chua Teresita , Hall Andrew
Published: August 2004

Objectives  To examine the effectiveness of weekly iron supplements given for 10 weeks by teachers to children in rural schools in the Philippines.

Methods  Forty‐nine rural primary schools took part in the study and were randomly assigned to two groups: children in 25 schools received a weekly tablet providing 108 mg iron while children in 24 schools acted as controls. All children were dewormed before the start of the iron supplementation. The haemoglobin concentration of a systematic sample of one in three children in two classes in each school was estimated before and 5–17 weeks after the end of the iron supplementation.

Results  A total of 1510 children aged 7–12 years were studied at both surveys. The mean haemoglobin concentration of children in the intervention group did not change significantly; in the untreated group it fell by 3.8 g/l and the prevalence of anaemia rose from 14.3% to 25.6%. The difference between study groups was significantly larger amongst the younger children (7–8 years), and was observed in both anaemic and non‐anaemic children.

Conclusion  Even where anaemia is only a mild public health problem, weekly iron supplements given by teachers may prevent a fall in the haemoglobin concentration, and can benefit both anaemic and non‐anaemic children.
Toxoplasma gondiispreading in an urban area evaluated by seroprevalence in free‐living cats and dogs
Authors: Meireles L. R. , Galisteo A. J. , Pompeu E. , Andrade H. F.
Published: August 2004

Infection by the protozoan parasite,Toxoplasma gondiiis widely prevalent in humans and animals throughout the world. Transmission takes place mainly by ingestion of raw or undercooked meat that contains parasite cysts or by ingestion of oocysts excreted in cat faeces, which can contaminate water and raw vegetables. The incidence of toxoplasmosis in urban areas can thus be also related to environmental contamination with oocysts. A direct measure of this environmental contamination by oocyst counting is unfeasible for technical reasons. An interesting alternative for measuringT. gondiiurban spreading is the seroprevalence in free‐living urban animals, used as sentinels, once they are exposed to similar risks ofToxoplasmainfection‐like humans. With this aim, we tested serum samples from stray cats and dogs for antibodies toT. gondiiby indirect haemagglutination assay (IHA) and enzyme‐linked immunosorbent assay (ELISA). Antibodies toT. gondiiwere found in 40% (40 of 100) of the cats, less than the 50.5% (101 of 200) found in dogs by ELISA (P < 0.05). Haemagglutination showed low resolution and concordance, precluding their use for diagnosis ofT. gondiiinfection compared with ELISA. The prevalence ofT. gondiiwas lower among stray cats probably due to their selective alimentary habits and lower water and food intake.Toxoplasma gondiiseroprevalence in stray dogs and cats could be an indirect indicator of the parasite spreading in urban areas.
Infant schistosomiasis in Ghana: a survey in an irrigation community
Authors: Bosompem K. M. , Bentum Irene A. , Otchere J. , Anyan W. K. , Brown C. A. , Osada Y. , Takeo S. , Kojima S. , Ohta N.
Published: August 2004

We used a rapid, visually read, field applicable monoclonal antibody (MoAb)‐dipstick assay for specific diagnosis of urinary schistosomiasis together with microscopy to determine the prevalence of infant schistosomiasis in a community in the Awutu‐Efutu Senya District in the Central Region of Ghana. The study group consisted of 97 infants (51 males and 46 females) aged 2 months to 5 years. A total of 75 of 97 (77.3%) subjects submitted stool samples; none had Schistosoma mansoni. Three individuals (3.1%) had hookworms but there were no other intestinal helminths. The urinary schistosomiasis prevalence by MoAb‐dipstick (30%) was higher (P < 0.05) than that estimated by microscopy (11.2%). However, three of nine (33.3%) microscopically confirmed cases tested MoAb‐dipstick positive after pre‐treatment of the urine specimen with heat. The youngest infant to be found infected with S. haematobium microscopically was 4 months old. Fifteen of 71 S. haematobium egg negative individuals tested dipstick positive, giving a dipstick specificity of 78.9% as compared with microscopy as gold standard test. The relative sensitivity of the dipstick was 100%.
Viewpoint: Why and how tuberculosis control should be included in health sector reviews
Authors: Schreuder Bert , Visschedijk Jan , Gondrie Peter , Van Cleeff Maarten
Published: August 2004

Sector‐wide approaches (SWAps) are currently implemented in several, predominantly highly donor‐dependent low‐income countries, particularly in Sub‐Saharan Africa. SWAp intends to enhance health sector performance, especially efficiency, effectiveness, coherence and sustainability. SWAps have been criticized for not being able to produce tangible results and the credibility of a SWAp would increase substantially if such results in terms of health services outputs could be demonstrated. We argue that the monitoring of tuberculosis within the SWAp reviews held to assess sector performance deserves a higher profile. Tuberculosis constitutes in the first place and one of the major public health problems in most of the countries, where SWAps have been introduced, but provided that the programme is integral part of the sector policy and financial management, the indicators used for monitoring TB control programmes can also be used as excellent proxies for monitoring overall health sector performance.
The PfCRT (K76T) point mutation favours clone multiplicity and disease severity in Plasmodium falciparum infection
Authors: Ranjit M. R. , Das A. , Chhotray G. P. , Das B. P. , Das B. N. , Acharya A. S.
Published: August 2004

In Orissa, a malaria‐hyperendemic area of India, we assessed the relationship between the PfCRT (K76T) point mutation of Plasmodium falciparum and the clinical severity of malaria. Forty uncomplicated and 36 severe malaria cases were selected, and parasite species, density and schizontaemia determined by examination of Giemsa‐stained thick or thin blood films. The PfCRT point mutation was analysed by PCR–RFLP and genotypes of the parasite isolates investigated by nested PCR using the polymorphic region of the merozoite surface protein‐2. We found that (i) the prevalence of the PfCRT point mutation was significantly higher (P < 0.01) in severe malaria cases and that (ii) heavy parasitaemia along with clone multiplicity was statistically more common (P < 0.01) in severe cases. These associations may be due to progression of uncomplicated to severe disease after chloroquine treatment failure and/or increased virulence of chloroquine‐resistant parasites. The implications for antimalarial treatment policy are discussed.
A prospective study of bloodstream infections as cause of fever in Malawi: clinical predictors and implications for management
Authors: Peters Remco P. H. , Zijlstra Ed E. , Schijffelen Maarten J. , Walsh Amanda L. , Joaki George , Kumwenda John J. , Kublin James G. , Molyneux Malcolm E. , Lewis David K.
Published: August 2004

Objective  To determine the contribution of a blood culture service to the diagnosis of fever in a resource‐poor setting and to identify clinical predictors of specific bloodstream infections (BSI).

Methods  In a descriptive, prospective study at the Medical Wards at Queen Elizabeth Central Hospital, Blantyre, Malawi, we tried to identify a specific cause of fever in febrile patients, comparing the use of routinely available diagnostic methods with the same methods plus blood culture. Clinical predictors of specific BSIs were sought.

Results  A total of 352 patients admitted with fever (axillary temperature ≥37.4 °C) or a history of fever within the last 4 days were enrolled. Tuberculosis (TB) was the diagnosis most commonly suspected initially on clinical grounds (28%), followed by lower respiratory tract infection (16%), malaria (12%) and gastroenteritis (5%). Blood cultures were positive in 128 patients (36%); Mycobacterium tuberculosis was the most commonly isolated organism (57 patients). In most cases the diagnosis of TB had already been made using routinely available diagnostic methods, including chest radiography. In all 16 cases of Streptococcus pneumoniae bacteraemia, infection with this agent was clinically suspected, usually on the basis of pulmonary symptoms and signs. In contrast, in 30 of 65 patients (65%) with non‐typhi salmonellae (NTS) bacteraemia, there were no symptoms or signs specifically suggestive of this diagnosis. Fever ≥39 °C and splenomegaly predicted NTS bacteraemia with an odds ratio of 8.4 (95% confidence interval 3.4–20.6, P < 0.001).

Conclusion  BSIs are common among patients admitted with fever. While BSI with mycobacteraemia and S. pneumoniae can usually be predicted on clinical grounds and with routinely available diagnostic methods, NTS bacteraemia often presents as a primary BSI without localizing symptoms and signs. Splenomegaly in this population indicates NTS bacteraemia rather than malaria.
Failure to deplete anti‐Galα1‐3Gal antibodies after pig‐to‐baboon organ xenotransplantation by immunoaffinity columns containing multiple Galα1‐3Gal oligosaccharides
Authors: Mañez Rafael , Domenech Nieves , Centeno Alberto , Lopez‐Pelaez Eduardo , Crespo Fabian , Juffe Alberto , Duthaler Rudolf O. , Katopodis Andreas G.
Published: August 2004

Abstract:  Background:  The impact of anti‐Galα1‐3Gal (αGal) antibodies on the acute humoral xenograft rejection (AHXR) of pig organs transplanted in baboons is unclear.

Methods:  Twenty‐three baboons underwent heterotopic pig heart transplantation (Tx). Groups A (n = 5) and B (n = 6) received non‐transgenic and human decay accelerating factor (hDAF) pig hearts, respectively, without any treatment. Groups C (n = 5) and D (n = 7) were transplanted with non‐transgenic and hDAF organs, respectively, and the exclusive treatment was repeated extracorporeal immunoadsorptions (EIA) before and after Tx with an αGal column containing disaccharide (DI), trisaccharide (TRI) 2 and pentasaccharide (PENTA) oligosaccharides.

Results:  In group A, 3 of 5 xenografts underwent hyperacute rejection (HAR). No xenograft from groups B, C and D experienced HAR, most of them failing from AHXR. Immediately after Tx and up to day 2, the level of immunoglobulin (Ig)M and IgG anti‐αGal DI, TRI2 and TRI6, and anti‐pig hemolytic antibody (APHA) antibodies decreased in all the groups by 80 to 96% compared with the concentration present before Tx. From day 3 to AHXR, a sustained increase of anti‐αGal IgM DI, TRI2 and TRI6, and APHA occurred in all groups. EIA depleted anti‐αGal IgM and APHA before Tx, but it did not modify the increase of these antibodies after Tx. Baboon serum samples before Tx, pre‐incubated in vitro with 1 mg/ml of DI, TRI2 and TRI6, had an average of 93% reduction of anti‐αGal IgM antibodies specific against each one of these αGal oligosaccharides. In contrast, at AHXR, the average reduction after in vitro pre‐incubation with either 1 or 5 mg/ml of DI, TRI2 and TRI6 was 40%.

Conclusions:  The EIA reduces anti‐αGal and APHA antibodies, preventing the HAR of non‐transgenic pig hearts transplanted in baboons, as does hDAF expression. However, EIA does not modify the level of anti‐αGal IgM and APHA antibodies after Tx nor the AHXR of either non‐transgenic or hDAF pig organs. The increase in anti‐αGal IgM after Tx was similar for the different antibodies of the anti‐αGal polymorphism, and was only partially neutralized in vitro with the specific αGal oligosaccharide.
Thrombotic microangiopathy and graft arteriopathy in pig hearts following transplantation into baboons
Authors: Houser Stuart L. , Kuwaki Kenji , Knosalla Christoph , Dor Frank J.M.F. , Gollackner Bernd , Cheng Jane , Shimizu Akira , Schuurman Henk‐Jan , Cooper David K.C.
Published: August 2004

Abstract:  Background:  Acute humoral xenograft rejection (AHXR) is an immunologic barrier in pig‐to‐baboon organ transplantation (Tx). We report microvascular thrombosis and myocardial necrosis in a series of cardiac xenografts.

Methods:  Ten baboons underwent heterotopic heart Tx from pigs transgenic for human decay‐accelerating factor. Recipients were treated with soluble Gal glycoconjugates and multiple immunosuppressive agents. Grafts were removed when palpable contractions stopped. Stained tissue sections from harvested grafts were analyzed by light and fluorescence microscopy.

Results:  Xenograft survival ranged from 4 to 139 (mean 37, median 27) days. Some histology was typical for AHXR (n = 4; median survival 22 days). Hemorrhage and edema were only focal in the longer‐surviving grafts (n = 4, median survival 54 days). All grafts had multiple platelet‐rich fibrin thrombi occluding myocardial vessels. Ischemic damage was manifested by contraction band necrosis in four grafts, myocytolysis in eight, coagulative necrosis in nine, and patchy myocyte dropout in all grafts. A notable paucity of interstitial mononuclear cells was observed in all grafts. Marked intimal thickening resembling that of allograft vasculopathy was observed in one graft. Immunofluorescence showed immunoglobulin (Ig)G and/or IgM deposition in five grafts. Multivessel C4d deposition appeared in seven grafts. Significant C3 deposition was absent.

Conclusions:  Cardiac xenograft survival in the pig‐to‐baboon model can be significantly prolonged by vigorous immunosuppressive treatment of recipient animals. Additional efforts to block humoral activation of graft endothelial cells and/or to overcome species‐specific molecular coagulation pathway incompatibilities may prevent the development of microvascular thrombosis and myocardial infarction. Cardiac xenograft vasculopathy (chronic rejection) can occur with prolonged graft survival.
Reversal of diabetes in non‐immunosuppressed rhesus macaques by intraportal porcine islet xenografts precedes acute cellular rejection
Authors: Kirchhof Nicole , Shibata Satoshi , Wijkstrom Martin , Kulick David M. , Salerno Christopher T. , Clemmings Sue M. , Heremans Yves , Galili Uri , Sutherland David E.R. , Dalmasso Agustin P. , Hering Bernhard J.
Published: August 2004

Abstract:  Background:  The functional response and immunobiology of primarily non‐vascularized islet cell xenografts remain poorly defined in non‐human primates.

Methods:  We transplanted 20 000 adult porcine islet equivalents/kg (purified and cultured for 48‐h) intraportally into six streptozotocin‐diabetic and two non‐diabetic rhesus macaques. Two recipients were killed at various intervals post‐transplant for histologic examination of livers bearing xenografts.

Results:  Plasma glucose levels in diabetic recipients averaged 94 mg/dl at 12 h, 92 mg/dl at 24 h, 147 mg/dl at 48 h, and 157 mg/dl at 72 h post‐transplant. Serum porcine C‐peptide was present in eight of eight recipients at 12 h, in five of six at 24 h, in four of four at 48 h, and in one of two at 72 h post‐transplant. C3a and SC5b‐9 plasma levels increased at 12 h post‐transplant and returned to pre‐transplant levels by 24 h. IgG, IgM anti‐pig and anti‐Gal IgG serum antibody levels did not increase post‐transplant. Rejection was initiated by IgM and complement deposition on islets. Neutrophils dominated the cellular infiltrate at 12 h; CD4+ and CD8+ T cells were the main infiltrating cells at 24, 48, and 72 h; and macrophages increasingly infiltrated xenografts starting at 24 h post‐transplant. Numerous xenoislets were present at all time points; their proportion without intraislet infiltrates decreased from 65% at 24 h to 17% at 72 h post‐transplant.

Conclusions:  Pig‐to‐primate intraportal islet xenografts reverse diabetes and the majority of intraportally transplanted xenogeneic islets are not subject to hyperacute rejection. They undergo acute cellular rejection mediated by CD4+‐ and CD8+ T cells and macrophages.
Expression of Fas but not Fas ligand on fetal pig β cells
Authors: Bai Lijun , Maedler Kathrin , Donath Marc , Tuch Bernard E.
Published: August 2004

Abstract:  Background:  The aim of this study was to determine whether fetal pig insulin‐producing cells, a potential source of transplantable tissue for the treatment of type 1 diabetes, are affected by the Fas–FasL interaction, one of the cytotoxic pathways involved in T‐cell‐mediated autoimmune destruction of pancreatic β cells.

Methods:  Expression of Fas/FasL on fetal pig β cells was assessed by immunohistochemistry, flow cytometry, Western blot, and RT‐PCR. Apoptosis of fetal pig β cells induced by soluble FasL (sFasL) or anti‐Fas antibody (APO‐1) was detected by flow cytometry using PI. Expression of FLIP on fetal pig pancreatic tissue was detected by immunofluorescent staining and Western blot.

Results:  Fas was expressed on fetal pig pancreatic cells, both β and non‐β cells, and the level of expression could be upregulated by exposure to human interleukin‐1β (IL1β) 2000 pg/ml for 24 h. In contrast, FasL was not detected on fetal pig pancreatic cells but could be induced on both β and non‐β cells when the cells were treated with IL1β. Fas persisted on fetal pig β cells transplanted as islet‐like cell clusters into severe combined immunodeficient mice, with expression of this antigen at all times examined, 1 day, 2, 3 and 4 weeks. FasL was absent. Despite the presence of Fas on fetal pig β cells, addition of sFasL or anti‐Fas antibody failed to induce apoptosis of the fetal pig β cells. In contrast, pig lymphocytes, which express Fas, were destroyed by addition of both sFasL and APO‐1. A possible reason for this is the expression on the fetal pig pancreatic cells of FLIP, an inhibitor of Fas‐induced apoptosis.

Conclusions:  Fetal pig β cells are resistant to Fas–FasL destruction. Our data imply that fetal pig β cells transplanted into humans with type 1 diabetes may not be destroyed by activated T cells through the Fas–FasL‐mediated pathway.
In vivo gene transfer of endo‐β‐galactosidase C removes αGal antigen on erythrocytes and endothelial cells of the organs
Authors: Miki Yusuke , Maruyama Shoichi , Liu DaGe , Kobayashi Takaaki , Sato Fumihiko , Shimizu Hideaki , Kato Sawako , Sato Waichi , Morita Yoshiki , Yuzawa Yukio , Muramatsu Takashi , Matsuo Seiichi
Published: August 2004

Abstract:  Background:  The presence of Galα1‐3Galβ1‐4GlcNAc (αGal) in pigs is a formidable barrier for pig‐to‐primate xenotransplantation. We have reported that administration of recombinant endo‐β‐galactosidase C (EndoGalC) removes αGal on porcine erythrocytes and kidneys. The present study examined the effects of EndoGalC gene therapy on αGal suppression.

Methods:  Naked plasmid DNA encoding Igκ‐EndoGalC was given to rats by rapid tail vein injection. The expression of αGal in the heart and kidney were studied by lectin staining followed by computer‐assisted quantitative analyses. αGal expression on erythrocytes was determined by flow cytometric analyses. Enzymatic activity of EndoGalC in the serum was also determined by evaluating the capacity of EndoGalC in removing αGal epitopes. Elimination of αGal was further studied by injecting antibodies against αGal to rats 2 days after the gene transfer.

Results:  Administration of 1 mg of Igκ‐EndoGalC/pCAGGS plasmid eliminated αGal from the vascular endothelium of the heart and kidney on day 1 and day 2. Between days 4 and 7, αGal started to reappear, but remained suppressed. No serious adverse effect was observed in rats treated with EndoGalC. Flow cytometric analyses showed that EndoGalC digested 97% of αGal on erythrocytes when measured 4 days after the gene transfer. Enzymatic activity of EndoGalC in the serum peaked on day 1, and significant levels were still observed on day 7. When antibodies against αGal were given, rats treated with EndoGalC showed no change, while all control rats died within 40 min.

Conclusions:  The present study demonstrates the potential of an EndoGalC gene transfer, using a hydrodynamics‐based delivery system, in eliminating αGal from endothelial cells in vivo. The results also ensured that EndoGalC is not harmful suggesting that the production of pigs overexpressing EndoGalC would be a reasonable alternative to pigs deficient in α1,3galactosyltransferase.
Biological activity of pig islet‐cell reactive IgG antibodies in xenotransplanted diabetic patients
Authors: Lindeborg Ellinor , Kumagai‐Braesch Makiko , Tibell Annika , Christensson Birger , Möller Erna
Published: August 2004

Abstract:  Background:  The IgG antibody response in type I diabetic patients, transplanted with fetal pig islet‐like cell‐clusters, was investigated using purified immunoglobulin G (IgG) fractions from sera collected 7 to 9 yr after transplantation. From our earlier studies, we knew that the immunological specificities of xenoreactive IgG1 and IgG2 antibodies are different, and that IgG1 antibodies, in contrast to the IgG2 population, are mainly directed against non‐Galα1,3Gal epitopes. In this study our goal was to establish whether xenoreactive IgG1 and IgG2 antibodies react with pig islet cells and, if so, to identify the target cell type, the biological function as well as the specificity of such antibodies for islet cell antigens. Sera from xenotransplanted patients were compared with those of patients with diabetes, selected for high titres of islet‐cell specific autoantibodies.

Methods:  IgG antibody fractions from patient sera were purified on a protein G column. Surface expression of target antigens was studied using flow cytometry as well as immunofluorescence microscopy. The biological function of islet‐cell reactive sera was tested using antibody dependent cellular cytotoxicity with both xenogeneic adult pig islet cells and allogeneic human islet cells as targets. Antibody specificity was assessed using 2D Western blots with both fetal and adult pig islet as well as human islet cell antigenic preparations.

Results:  Some of the diabetic patients, who have been transplanted with xenogeneic fetal pig islet cells, continue to produce xenospecific IgG1 and IgG2 antibodies for 7 to 9 yr post‐transplantation. A separate analysis of IgG1 and IgG2 antibodies showed that IgG1 antibodies react with pig islet β cells, whereas IgG2 antibodies mainly react with non‐endocrine pig cells. Such antibodies are xenospecific, as they were found to mediate antibody‐dependent cellular cytotoxicity of adult pig, but not human islet target cells. The reverse was true for antibodies from non‐transplanted diabetic patients with high titres of autoantibodies against β cells. Fluorescence analysis as well as 2D gel Western blots revealed that the reactivity was variable between patient samples, indicating that target antigens for non‐Galα1,3Gal‐specific antibodies are heterogeneous.

Conclusion:  Thus, xenotransplantation of diabetic patients induces islet‐β cell reactive xenospecific IgG1 antibodies, which are biologically active and can mediate antibody‐dependent cellular cytotoxicity of pig islet cells.
Curative resection of a small cell carcinoma of the pancreas: Report of a case of long survival without chemotherapy
Authors: KINOSHITA KAZUO , MINAMI TAKESHI , OHMORI YOSHIKAZU , KANAYAMA SHUJI , YOSHIKAWA KOICHI , TSUJIMURA TAKAHIRO
Published: August 2004

Abstract  A 41‐year‐old asymptomatic man was, by chance, diagnosed as having a mass in the left upper quadrant of the abdomen by ultrasound. Computed tomography and magnetic resonance imaging showed a slightly enhanced heterogeneous mass, measuring about 5 cm in diameter, adjacent to the pancreas tail and spleen. On abdominal angiography the tumor was found to be fed by the splenic artery, and no encasement was observed. At operation the tumor was connected to the pancreas tail and attached to the spleen, and no metastasis was evident. As a result of these observations, a curative resection was performed. On histological examination the tumor cells had no ductal or architectural organization and were continuous to the normal pancreatic tissues. The cells were negative for Grimelius argylphile and the periodic acid schiff stain. Immunohistochemistry indicated that the tumor cells were positive for keratin and epithelial membrane antigen, but negative for chromogranin A, vimentin, α1‐antitrypsin and α1‐antichymotrypsin. The tumor was diagnosed as a small cell carcinoma of the pancreas, which is a rare disease. In previously reported cases curative resection of the tumor had not been possible because of its rapid progression and metastasis, and the prognosis is generally very poor. In our case, however, curative resection of the tumor was done, and the patient is healthy with no signs of recurrence for 56 months after the operation and with no additional therapy.
Genetic polymorphisms of bilirubin uridine diphosphate‐glucuronosyltransferase gene in Japanese patients with Crigler–Najjar syndrome or Gilbert's syndrome as well as in healthy Japanese subjects
Authors: TAKEUCHI KEISUKE , KOBAYASHI YOSHINAO , TAMAKI SHIGENORI , ISHIHARA TOMOAKI , MARUO YOSHIHIRO , ARAKI JUN , MIFUJI RUMI , ITANI TOSHIO , KURODA MAKOTO , SATO HIROSHI , KAITO MASAHIKO , ADACHI YUKIHIKO
Published: August 2004

Background and Aim:  Numerous mutations of bilirubin uridine diphosphate‐glucuronosyltransferase gene (UGT1A1) have been reported in patients with familial unconjugated hyperbilirubinemia. The UGT1A1 mutation appears to be considerably different among ethnic groups. To clarify the incidence of this gene mutation in the Japanese population, the presence of UGT1A1 mutation was investigated in a group of Japanese patients with Crigler–Najjar syndrome type 2 (CNS2) and Gilbert's syndrome (GS), as well as in healthy anicteric subjects.

Methods:  Four patients with CNS2, 63 patients with GS, and 71 healthy subjects were enrolled in the study. The promoter and coding regions of UGT1A1 were amplified by polymerase chain reaction (PCR) from genomic DNA isolated from leukocytes. The PCR products were directly sequenced by a dye terminating method. The UGT1A1 enzyme activity was determined in COS7 cells transfected with wild or P364L (1091 C > T) mutant DNA.

Results:  Homozygous Y486D was observed in all four patients with CNS2. The GS patients had UGT1A1 mutations with 13 different genotypes in the promoter and coding region. Homozygous TA insertion in the TATA box (TA7) of the promoter region (TA7/7; 33%), homozygous G71R (9%), and combination of TA7/6 and heterozygous G71R (17%) were the most frequent findings in GS patients. Homozygous or heterozygous Y486D (8%) and P229Q (8%) were also observed in GS. A novel mutation, heterozygous P364L, was also identified in a GS patient. In addition to GS patients, homozygous or heterozygous TA7, G71R, and heterozygous Y486D were also observed in healthy subjects. The allele frequency of G71R and TA7 was 0.183 and 0.113 in healthy subjects, respectively. The P364L UGT1A1 enzyme activity was 64.4% lower than the wild‐type enzyme activity.

Conclusions:  Polymorphisms in the coding region of UGT1A1 were commonly observed in Japanese patients with GS and in healthy subjects. The genetic basis of hyperbilirubinemia appears to be different between the Japanese and Caucasian populations.
Clinical significance of variceal hemorrhage in recent years in patients with liver cirrhosis and esophageal varices
Authors: PARK DONG KYU , UM SOON HO , LEE JAE WON , LEE JUNG BOK , KIM YOUNG SUN , PARK CHUL HEE , JIN YOON TAE , CHUN HOON JAI , LEE HONG SIK , LEE SANG WOO , CHOI JAI HYUN , KIM CHANG DUCK , RYU HO SANG , HYUN JIN HAI
Published: August 2004

Background and Aims:  Recent progress in the treatment of variceal bleeding might have reduced the impact of variceal bleeding on survival in patients with esophageal varices. We conducted a retrospective cohort study in an attempt to re‐evaluate the clinical significance of variceal bleeding.

Methods:  A cohort of 304 patients with liver cirrhosis and esophageal varices, who had no previous history of variceal bleeding and no prophylactic therapy, was studied.

Results:  During a median follow‐up period of 32 months, 55 patients (18%) bled from varices and 111 (37%) died. Variceal hemorrhages accounted for 15% of total deaths. The mortality of first variceal bleeding was 25% in the whole group, but was remarkably different depending on liver function at the time of bleeding (0% in grade Child A vs 55% in grade C; P < 0.05). Among the survivors of first bleeding, 30% experienced rebleeding. Form of varix, red color sign and heavy drinking were the independent risk factors for first variceal bleeding. Multivariate analysis revealed that variceal bleeding still had a significant (P < 0.001) impact on death in the whole cohort, when other independent prognostic factors such as age, ascites, encephalopathy, platelet count, serum albumin level and hepatocellular carcinoma were adjusted. Furthermore, in subgroup analyses, variceal bleeding was more strongly (P < 0.001) linked to death in patients with alcoholic cirrhosis than in those with non‐alcoholic cirrhosis, and showed a significant association with survival only for the patients in Child grade B.

Conclusions:  Variceal bleeding has various prognostic impacts depending on the etiology of cirrhosis or on the degree of liver dysfunction, and this needs to be taken into account in the prophylaxis against first variceal bleeding.
Experimental bile‐duct ligation resulted in accumulation of oxidized low‐density lipoproteins in BALB/c mice liver
Authors: CÖMERT MUSTAFA , TEKİN İSHAK ÖZEL , AÇIKGÖZ ŞEREFDEN , ÜSTÜNDAǦ YÜCEL , UÇAN BÜLENT H , ACUN ZEKI , BARUT FIGEN , SÜMBÜLOǦLU VILDAN
Published: August 2004

Background and Aim:  Oxidized low‐density lipoproteins (LDL), which are produced during oxidative stress by the process of lipid peroxidation, have also been proposed to have complex roles in many other immuno‐inflammatory mechanisms. It has been shown that bile‐duct ligation results in oxidative stress in the liver of animals. The aim of this study was to investigate if oxidized LDL are produced in the liver tissues of bile‐duct‐ligated mice.

Methods:  Obstructive jaundice was induced in BALB/c mice by the ligation and division of the common bile duct. Liver concentrations of glutathione and malondialdehyde were measured in the sham‐operated (n = 10) and bile‐duct‐ligated (n = 10) mice on the 10th day of obstructive jaundice. The presence of oxidized LDL in the liver tissue sections was evaluated using a special, novel immunofluorescent staining method. The final step was to explore the existence of oxidized LDL under fluorescent microscopy.

Results:  Compared with sham‐operated mice, jaundiced mice showed significantly higher levels of malondialdehyde and lower concentrations of reduced glutathione in the liver. While there was no staining in the sham‐operated group, bile‐duct ligation resulted in positive oxidized LDL staining in the liver tissues of mice. The present study testifies that bile‐duct ligation results in oxidative stress and enhanced lipid peroxidation in the hepatic tissues of BALB/c mice and moreover, that oxidized LDL accumulate in the liver of mice with experimental obstructive jaundice.

Conclusion:  Oxidized LDL may be an important and direct indicator of ongoing oxidative stress and enhanced lipid peroxidation in obstructive jaundice. The potential roles of this finding were also discussed, briefly.

© 2004 Blackwell Publishing Asia Pty Ltd
Comparison of biochemical and cytotoxic functions of hepatocytes from goat, pig and human fetuses
Authors: VIJAYALAKSHMI V , NASEEM B , KHAN AA , CAPOOR AK , HABIBULLAH CM
Published: August 2004

Background and Aim:  To overcome the problem of shortage of donor organs, xenotransplantation of cells offers an alternative to orthotopic transplantation. Of the higher animals, the pig is considered as a suitable donor because of the similarity in size and function of pig organs to human organs. However, successful transplantation of pig organs/cells for human therapy is limited by hyperacute rejection, improper functioning of xenografts and the risk of transmission of endogenous retroviruses to the recipient. Thus, there is a pressing need to explore an alternate mammalian source to bridge the gap between the donor and the recipient waiting for transplantation. This has warranted us to explore the application of goat hepatocytes as a treatment modality in acute liver failure.

Methods:  In the present investigation, isolated goat hepatocytes were assessed for their viability, membrane integrity, synthetic and cytotoxic functions, and compared with the hepatocytes of pig and human fetuses (28–36 weeks).

Results:  The isolated hepatocytes from goat, pig and human fetuses were comparable in their viability, membrane integrity and synthetic functions. However, the cytotoxic functions assessed using 3‐(4,5‐dimethyl‐2 thiazoyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide assay demonstrated a significant reduction in the viability of the pig hepatocytes (38%) as compared with the goat and human fetal hepatocytes, which retained their viability (98%) on incubation with normal human serum.

Conclusion:  These observations are significant as they suggest that goat hepatocytes probably can be explored as a source for cell therapy in the treatment of acute liver failure.
Diverse p53 gene aberration in hepatocellular carcinoma detected by dual‐color fluorescence in situ hybridization
Authors: KONDO MASAHIKO , MARUSAWA HIROYUKI , UEDA YOSHIHIDE , KATSURADA AKIRA , KAWASOME CHIE , TAKAMI SATOSHI , KINOSHITA MORITOSHI , IKAI IWAO , YAMAOKA YOSHIO , CHIBA TSUTOMU
Published: August 2004

Background and Aim:  In order to evaluate loss of the p53 gene more precisely, we performed dual‐color fluorescence in situ hybridization (dual‐color FISH) for chromosome 17 and p53 gene together with DNA polymorphism analysis of the p53 gene in hepatocellular carcinoma (HCC).

Methods:  Dual‐color FISH using probes specific for the centromere of chromosome 17 and the p53 gene was performed for 41 HCC and DNA polymorphism analysis was also performed for them.

Results:  Of the 34 HCC tested by dual‐color FISH, 20 had loss of at least one p53 gene (58.8%). In contrast, of the 32 HCC tested by DNA polymorphism analysis, 23 gave informative results, among which only eight had loss of heterozygosity (LOH) of the p53 gene (34.8%). Notably, among 14 cases positive for loss of the p53 gene by dual‐color FISH, seven cases were negative for LOH of the p53 gene. Moreover, dual‐color FISH revealed that the percentage of cells that lost at least one p53 gene increased as the HCC became less differentiated (P < 0.01), whereas LOH did not reveal any such correlation.

Conclusions:  These data suggest that loss of the p53 gene was present in a considerable number of HCC, and diversity of the p53 gene aberration increases with progression of HCC. Dual‐color FISH is an effective method for detection of p53 gene aberration, and it can provide new insight into oncogenesis in HCC.
Liver iron deposits in hepatitis B patients: Association with severity of liver disease but not with hemochromatosis gene mutations
Authors: MARTINELLI ANA LC , FILHO ANTONIO B ARAUJO , FRANCO RENDRIK F , TAVELLA MARLI H , RAMALHO LEANDRA NZ , ZUCOLOTO SERGIO , RODRIGUES SANDRA S , ZAGO MARCOS A
Published: August 2004

Background and Aims:  Iron deposits in the liver and abnormalities in serum iron biochemistry are frequently observed in patients with chronic liver diseases, but data for patients with hepatitis B virus (HBV) infection are scarce. Moreover, the role of HFE mutations in iron deposits in this condition remains unknown. The aim of the present study was to determine the prevalence of serum iron biochemical abnormalities and iron deposits in the liver of chronic HBV patients, and to evaluate the consequences for the activity and severity of liver disease. Additionally, we studied the role of HFE gene mutations in iron deposits.

Methods:  Eighty‐one male non‐cirrhotic HBV patients were studied. Serum iron biochemistry, liver enzymes and C282Y/H63D mutations were investigated. Liver biopsies were scored for necroinflammatory activity (histological activity index [HAI]), fibrosis and iron deposits.

Results:  Elevated transferrin saturation (TS) was found in 27.1% of patients and liver iron deposits in 48.7%; these deposits were mild in 68.4% and moderate in 31.6%. Patients with liver iron deposits exhibited significantly higher scores for HAI and fibrosis than those without iron deposits. HFE mutations were identified in 23.4% of patients (14 H63D heterozygotes, four H63D homozygotes, one compound mutation). No difference in the prevalence of C282Y and H63D mutations was observed between HBV patients (1.2% and 23.4%, respectively) and the general population (4.1% and 27.8%, respectively). No association was detected between HFE mutations and elevated TS or liver iron deposits.

Conclusions:  Elevated TS and liver iron deposits were frequent in non‐cirrhotic HBV patients. Iron deposits were mainly mild and associated with higher activity and severity of liver disease, but not with HFE mutations.
Effects of colchicine on the maximum biliary excretion of cholephilic compounds in rats
Authors: TACHIZAWA HIDETAKA , SANO NAOYO , TAKIKAWA HAJIME
Published: August 2004

Background and Aim:  Colchicine, an inhibitor of intracellular vesicular transport, has been reported to inhibit the biliary excretion of bile acids and organic anions, but the previous findings are controversial. In order to systematically evaluate the effect of colchicine on the biliary excretion of cholephilic compounds, we studied the effect of colchicine on the biliary excretion of substrates of various canalicular transporters, which were administered at or above the excretory maximum in rats.

Methods:  Substrates of various canalicular adenosine triphosphate‐binding‐cassette transporters were infused at or above the rate of maximum excretion into rats, and the effect of colchicine (0.2 mg/100 g), which was intraperitoneally injected 3 h before, on the biliary excretion was studied. Furthermore, the effect of tauroursodeoxycholate (TUDC) co‐infusion on the biliary excretion of taurocholate (TC) after colchicine treatment was also studied.

Results:  The biliary excretion of TC and cholate administered at the rate of 1 µmol/min/100 g was markedly inhibited by colchicine, whereas that of TUDC was not inhibited even with the infusion rate of 2 µmol/min/100 g. TUDC co‐infusion at the rate of 1 µmol/min/100 g increased the biliary excretion of TC (1 µmol/min/100 g), which was decreased by the colchicine pretreatment. The biliary excretory maximum of taurolithocholate‐sulfate and sulfobromophthalein, substrates of the multidrug resistance protein 2, of erythromycin, a substrate of the P‐glycoprotein, and of indocyanine green were not affected by colchicine.

Conclusions:  The different excretory maximums of TC and TUDC and the different effect of colchicine on the excretion of these bile acids are considered to be a result of different regulatory mechanisms of vesicular targeting of the bile salt export pump to the canalicular membrane by these bile acid conjugates. The vesicular targeting of the multidrug resistance protein 2 and the P‐glycoprotein to the canalicular membrane is considered to be colchicine insensitive in the absence of bile acid coadministration.
Gastric epithelial cell CXC chemokine secretion following Helicobacter pylori infection in vitro
Authors: SIEVEKING DANIEL , MITCHELL HAZEL M , DAY ANDREW S
Published: August 2004

Background and Aims:  Helicobacter pylori infection of the stomach is commonly associated with infiltration of neutrophils. Gastric epithelial cells are recognized as central mediators of tissue responses to this organism. The aim of the present study was to ascertain patterns of production of three neutrophil chemoattractant chemokines following infection of gastric epithelial cells with H. pylori in vitro.

Methods:  Two gastric cancer‐derived epithelial cell lines were infected with H. pylori organisms of previously defined cagE and cagA status for periods of up to 24 h. The production of three chemokines (interleukin [IL]‐8, epithelial neutrophil activating protein [ENA]‐78 and growth‐related oncogene [GRO]‐α) over this time was measured in culture supernatants using immunoassays.

Results:  Both IL‐8 and GRO‐α were produced by both AGS and KATO‐III cells in response to H. pylori infection, and in a cag PAI‐dependent manner. ENA‐78, however, was not increased following H. pylori infection.

Conclusions:  GRO‐α is expressed by epithelial cells following H. pylori infection along with IL‐8. Both may contribute to neutrophilic infiltration present in gastric mucosa associated with H. pylori infection. In contrast, H. pylori infection does not lead to an increased synthesis of ENA‐78, suggesting that this may not be as important in vivo.