bluuebunny/crossref_metadata_2025_split huggingface dataset:
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.
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

AbstractBackground 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

AbstractBackground 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

AbstractBackground 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

AbstractBackground 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

AbstractBackground 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

AbstractBackground 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.