bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunohistochemical Differentiation of Hepatitis D Virus Genotypes
Authors: Hsu Sheng-Chieh , Syu Wan-Jr , Ting Ling-Tan , Wu Jaw-Ching
Published: June 2003

Determination of hepatitis D virus (HDV) genotypes is epidemiologically and clinically important. Phylogenic analysis based on sequencing analysis of multiple HDV strains isolated from sera of patients is not convenient for mass screening in routine laboratories. This study was designed to develop genotype–specific antibodies against hepatitis delta antigen (HDAg) and to apply these antibodies for immunohistochemical differentiation of HDV genotypes in formalin–fixed, paraffin–embedded liver biopsies of patients. Divergence in the carboxyl–terminal 19 amino acids of the large HDAg between genotypes I and II is more than 70%. Peptides covering these residues were conjugated to keyhole limpet hemocyanin and were used for immunization. The generated antibodies were confirmed for their specificity by binding to type–specific HDAgs expressed in DNA–transfected Huh–7 hepatoma cells. Liver biopsies from 6 patients who had dominant genotype I HDV and 33 patients who had dominant genotype II HDV in sera were stained with these antibodies. The accuracy for these antibodies was 94.9%, and the agreement between dominant HDV genotypes in serum and dominant hepatic HDV genotypes based on HDAg staining was nearly perfect (κ = 0.83). In summary, the carboxyl–terminal 19 amino acids of the large HDAg can be used as immunogens to generate genotype–specific antibodies. These antibodies were proven to be useful in immunohistochemical differentiation of HDV genotypes in liver biopsies.
Different Costimulation Signals Used by Cd4+ and Cd8+ Cells That Independently Initiate Rejection of Allogenic Hepatocytes in Mice
Authors: Gao Donghong , Li Jiashun , Orosz Charles G. , Bumgardner Ginny L.
Published: June 2003

The current study evaluated the role of CD40/CD40 ligand (CD40L) and CD28/B7 costimulation signals during alloimmune responses independently mediated by CD4+ or CD8+ T cells. Allogeneic hepatocytes were transplanted into CD8 or CD4 knock out (KO) mice under cover of costimulatory blockade. Rejection of FVB/N (H–2q) hepatocytes occurred by day 10 posttransplant in untreated CD8 or CD4 KO (H–2b) mice. Treatment of CD8 or CD4 KO mice with anti–CD40L monoclonal antibody (mAb; MR1) resulted in significant prolongation of hepatocyte survival indicating that CD40/CD40L interactions were critical in both CD4+ and CD8+ T–cell initiated hepatocyte rejection. Anti–CD40L mAb also prolonged hepatocyte survival in B–cell KO (H–2b) mice, indicating that the efficacy of CD40/CD40L blockade in preventing hepatocyte rejection was B–cell (and antibody) independent. In contrast, treatment with CTLA4 fusion protein (CTLA4Ig), prolonged hepatocyte survival in CD8 KO but not CD4 KO mice, showing that CD28/B7 interactions were important in CD4+ but not CD8+ T–cell initiated hepatocyte rejection. Under selected circumstances, such as in CD40 KO mice, both CD4+ and CD8+ T cells mediate hepatocyte rejection in the absence of CD40/CD40L costimulation and without a significant contribution from CD28/B7 costimulation signals. These results highlight the disparate roles of CD40/CD40L and CD28/B7 costimulation signals in CD4+ versus CD8+ T–cell mediated immune responses to allogeneic hepatocytes. The CD4+ T–cell independent, CD40L–sensitive, CD28/B7–independent pathway of CD8+ T–cell activation in response to transplantation antigens is novel.
Genetic Instability and Aberrant Dna Methylation in Chronic Hepatitis and Cirrhosis—A Comprehensive Study of Loss of Heterozygosity and Microsatellite Instability At 39 Loci and Dna Hypermethylation on 8 Cpg Islands in Microdissected Specimens From Patients With Hepatocellular Carcinoma
Authors: Kondo Yutaka , Kanai Yae , Sakamoto Michiie , Mizokami Masashi , Ueda Ryuzo , Hirohashi Setsuo
Published: June 2003

A study was conducted to examine the significance of genetic instability and aberrant DNA methylation during hepatocarcinogenesis. Genomic DNA was extracted from 196 microdissected specimens of noncancerous liver tissue that showed no marked histologic findings or findings compatible with chronic hepatitis or cirrhosis, and 80 corresponding microdissected specimens of hepatocellular carcinoma (HCC) from 40 patients. Loss of heterozygosity (LOH) and microsatellite instability (MSI) were examined by polymerase chain reaction (PCR) using 39 microsatellite markers, and DNA methylation status on 8 CpG islands was examined by bisulfite–PCR. In noncancerous liver tissues, LOH, MSI, and DNA hypermethylation were found in 15 (38%), 6 (15%), and 33 (83%) of 40 cases, respectively. The incidence of DNA hypermethylation in histologically normal liver was similar to that in chronic hepatitis and cirrhosis, although neither LOH nor MSI was found in histologically normal liver. In cancerous tissues, LOH, MSI, and DNA hypermethylation were found in 39 (98%), 8 (20%), and 40 (100%) of 40 cases, respectively. CpG islands of the p16 gene and methylated in tumor 1, 2, 12, and 31 clones were frequently methylated in cancerous tissues, although neither the thrombospondin–1 nor the human Mut L homologue (hMLH1) gene was methylated. Absence of silencing of the hMLH1 gene by DNA hypermethylation is consistent with the low incidence of MSI in HCCs. The results of this study indicate that LOH and aberrant DNA methylation contribute to hepatocarcinogenesis; DNA hypermethylation in particular, which precedes or may even cause LOH, is as an early event during hepatocarcinogenesis.
Liver support by extracorporeal blood purification: A clinical observation
Authors: Stange Jan , Mitzner Steffen R. , Klammt Sebastian , Freytag Jens , Peszynski Piotr , Loock Jan , Hickstein Heiko , Korten Gero , Schmidt Reinhardt , Hentschel Jörg , Schulz Martin , Löhr Matthias , Liebe Stefan , Schareck Wolfgang , Hopt Ullrich T.
Published: June 2003

Liver failure associated with excretory insufficiency and jaundice results in an endogenous accumulation of toxins involved in the impairment of cardiovascular, kidney, and cerebral function. Moreover, these toxins have been shown to damage the liver itself by inducing hepatocellular apoptosis and necrosis, thus creating a vicious cycle of the disease. We report a retrospective cohort study of 26 patients with acute or chronic liver failure with intrahepatic cholestasis (bilirubin level > 20 mg/dL) who underwent a new extracorporeal blood purification treatment. A synthetic hydrophilic/hydrophobic domain-presenting semipermeable membrane (pore size < albumin size, 100-nm thick) was used for extracorporeal blood detoxification using dialysis equipment. The opposite side was rinsed with ligandin-like proteins as molecular adsorbents that were regenerated online using a chromatography-like recycling system (molecular adsorbent recirculating system [MARS]). Bile acid and bilirubin levels, representing the previously described toxins, were reduced by 16% to 53% and 10% to 90% of the initial concentration by a single treatment of 6 to 8 hours, respectively. Toxicity testing of patient plasma onto primary rat hepatocytes by live/dead fluorescence microscopy showed cell-damaging effects of jaundiced plasma that were not observed after treatment. Patients with a worsening of Child-Turcotte-Pugh (CTP) index before the treatments showed a significant improvement of this index during a period of 2 to 14 single treatments with an average of 14 days. After withdrawal of MARS treatment, this improvement was sustained in all long-term survivors. Ten patients represented a clinical status equivalent to the United Network for Organ Sharing (UNOS) status 2b (group A1), and all survived. Sixteen patients represented a clinical status equivalent to UNOS status 2a, and 7 of these patients survived (group A2), whereas 9 patients (group B) died. We conclude that in acute excretory failure caused by a chronic liver disease, this treatment provides a therapy option to remove toxins involved in multiorgan dysfunction secondary to liver failure.
Liver transplantation for hepatitis C: Recurrence and disease progression in 300 patients
Authors: Testa Giuliano , Crippin Jeffrey S. , Netto George J. , Goldstein Robert M. , Jennings Linda W. , Brkic Borisa S. , Brooks Barbara K. , Levy Marlon F. , Gonwa Thomas A. , Klintmalm Goran B.
Published: June 2003

The time progression of allograft damage in patients with recurrent hepatitis C after orthotopic liver transplantation (OLT) is not precisely determined. The aim of this analysis is to study the progression of disease recurrence and its impact on patient and graft survival. Data for 300 patients who underwent OLT for hepatitis C were analyzed regarding the incidence of histological recurrence, risk factors, immunosuppressive regimen, rejection episodes, and survival. For patients with histological recurrence, the timing and risks for disease progression were analyzed. Data for 30 patients who underwent retransplantation were studied. Histological recurrence occurred in 40.3% of patients, 27.2% of whom progressed to bridging fibrosis or cirrhosis. Eighty-seven percent of the patients experienced recurrence of disease within 24 months of OLT. Patients with histological recurrence within 6 months of OLT had an increased risk for progression to cirrhosis compared with patients with recurrence later than 6 months (risk ratio, 2.3). Recurrence within 1 year was associated with decreased patient and graft survival rates at 1 and 5 years (65.1% and 56.4% versus 80.6% and 78.4%; P = .004 and P = .0008, respectively). Patients with histological recurrence had a greater incidence of acute cellular rejection, as well as multiple episodes of rejection, steroid-resistant rejections, and greater cumulative doses of corticosteroids. Histological recurrence after OLT for hepatitis C is common and usually occurs within 2 years of OLT. Early recurrence negatively affects patient and graft survival. Host factors impacting on recurrence need further study. A relation between the hepatitis C virus, allograft rejection, and immunosuppression exists and needs investigation.
Quality of life and cognitive function of liver transplant patients: A prospective study
Authors: Moore Kathleen A. , McL. Jones Robert , Burrows Graham D.
Published: June 2003

The presence of cognitive impairment in end-stage liver disease is well recognized, as are patient reports of an impoverished quality of life. The aim of this study is to systematically evaluate the effect of orthotopic liver transplantation (OLT) on these factors. Thirty-two adult patients activated for OLT participated in the study. Assessments were made on activation and at 1, 3, and 9 months post-OLT, with 24 transplant recipients available for reassessment at 9 months. Two control groups (10 patients with nonalcoholic cirrhosis and 10 healthy volunteers) also completed the test protocol at four 2-month intervals. The test battery included the Austin Quality-of-Life Scale, Weschler Adult Intelligence Scale-Revised, Benton’s Controlled Oral Word Test, and the Complex Figure of Rey. The OLT group showed significant improvement in cognitive performance and their reported quality of life. These changes were evident by 3 month post-OLT and remained stable at subsequent testing. The control groups typically remained stable over test occasions. Clearly, OLT extends life and, most importantly, improves patients’ quality of life and their cognitive functioning.
Home‐based teleworking and the employment relationship
Authors: Harris Lynette
Published: June 2003

As home‐based teleworking grows in the UK, more evidence is needed of how working from home shapes the employment relationship and the implications this may have for those line managers responsible for a home‐based workforce. The reported experiences of a sales team and their line managers at one large international drinks manufacturing company of teleworking during its first year of operation revealed the importance of developing understanding of the complex interface between the domains of work and home life. The findings suggest individual circumstances require close attention before implementing home‐based working with line managers recalibrating perceptions of the boundaries between home and work for positive employee relationships to develop within a new paradigm of “home‐work” relations.
Regulation of Pulmonary and Systemic Bacterial Lipopolysaccharide Responses in Transgenic Mice Expressing Human Elafin
Authors: Sallenave J.-M. , Cunningham G. A. , James R. M. , McLachlan G. , Haslett C.
Published: June 2003

The control of lung inflammation is of paramount importance in a variety of acute pathologies, such as pneumonia, the acute respiratory distress syndrome, and sepsis. It is becoming increasingly apparent that local innate immune responses in the lung are negatively influenced by systemic inflammation. This is thought to be due to a local deficit in cytokine responses by alveolar macrophages and neutrophils following systemic bacterial infection and the development of a septic response. Recently, using an adenovirus-based strategy which overexpresses the human elastase inhibitor elafin locally in the lung, we showed that elafin is able to prime lung innate immune responses. In this study, we generated a novel transgenic mouse strain expressing human elafin and studied its response to bacterial lipopolysaccharide (LPS) when the LPS was administered locally in the lungs and systemically. When LPS was delivered to the lungs, we found that mice expressing elafin had lower serum-to-bronchoalveolar lavage ratios of proinflammatory cytokines, including tumor necrosis factor alpha (TNF-α), macrophage inflammatory protein 2, and monocyte chemoattractant protein 1, than wild-type mice. There was a concomitant increase in inflammatory cell influx, showing that there was potential priming of innate responses in the lungs. When LPS was given systemically, the mice expressing elafin had reduced levels of serum TNF-α compared to the levels in wild-type mice. These results indicate that elafin may have a dual function, promoting up-regulation of local lung innate immunity while simultaneously down-regulating potentially unwanted systemic inflammatory responses in the circulation.
Identification ofActinobacillus pleuropneumoniaeGenes Important for Survival during Infection in Its Natural Host
Authors: Sheehan Brian J. , Bossé Janine T. , Beddek Amanda J. , Rycroft Andrew N. , Kroll J. Simon , Langford Paul R.
Published: June 2003

Actinobacillus pleuropneumoniaeis a strict respiratory tract pathogen of swine and is the causative agent of porcine pleuropneumonia. We have used signature-tagged mutagenesis (STM) to identify genes required for survival of the organism within the pig. A total of 2,064 signature-tagged Tn10transposon mutants were assembled into pools of 48 each, and used to inoculate pigs by the endotracheal route. Out of 105 mutants that were consistently attenuated in vivo, only 11 mutants showed a >2-fold reduction in growth in vitro compared to the wild type, whereas 8 of 14 mutants tested showed significant levels of attenuation in pig as evidenced from competitive index experiments. Inverse PCR was used to generate DNA sequence of the chromosomal domains flanking each transposon insertion. Only one sibling pair of mutants was identified, but three apparent transposon insertion hot spots were found—an anticipated consequence of the use of a Tn10-based system. Transposon insertions were found within 55 different loci, and similarity (BLAST) searching identified possible analogues or homologues for all but four of these. Matches included proteins putatively involved in metabolism and transport of various nutrients or unknown substances, in stress responses, in gene regulation, and in the production of cell surface components. Ten of the sequences have homology with genes involved in lipopolysaccharide and capsule production. The results highlight the importance of genes involved in energy metabolism, nutrient uptake and stress responses for the survival ofA. pleuropneumoniaein its natural host: the pig.
Different Strains ofToxoplasma gondiiInduce Different Cytokine Responses in CBA/Ca Mice
Authors: Araujo Fausto G. , Slifer Teri
Published: June 2003

To investigate the role that cytokines may have in the development of toxoplasmic encephalitis (TE), the levels of gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), interleukin-12 (IL-12 ]p40[), IL-10, IL-6, IL-4, and IL-2 in serum were examined in CBA/Ca mice infected with a type II strain (ME49 or FORT) ofToxoplasma gondii. These strains caused severe (ME49) or mild (FORT) TE in CBA/Ca mice. From weeks 1 to 8 of infection, the levels of IL-6, IL-10, IL-12, IFN-γ, and TNF-α in serum were significantly higher in the ME49-infected mice than in the FORT-infected mice, suggesting a role for these cytokines in the severity of TE in CBA/Ca mice. Since the ME49 and FORT strains are of the same type, our results suggest a role for the parasite in the development of severe TE through the increased production of proinflammatory cytokines and indicate that not all type II strains cause TE.
Swing-leg retraction: a simple control model for stable running
Authors: Seyfarth André , Geyer Hartmut , Herr Hugh
Published: June 2003

In running, the spring-like axial behavior of stance limbs is a well-known and remarkably general feature. Here we consider how the rotational behavior of limbs affects running stability. It is commonly observed that running animals retract their limbs just prior to ground contact, moving each foot rearward towards the ground. In this study, we employ a conservative spring-mass model to test the effects of swing-leg retraction on running stability. A feed-forward control scheme is applied where the swing-leg is retracted at constant angular velocity throughout the second half of the swing phase. The control scheme allows the spring-mass system to automatically adapt the angle of attack in response to disturbances in forward speed and stance-limb stiffness. Using a return map to investigate system stability, we propose an optimal swing-leg retraction model for the stabilization of flight phase apex height. The results of this study indicate that swing-leg retraction significantly improves the stability of spring-mass running,suggesting that swing-phase limb dynamics may play an important role in the stabilization of running animals.
A biomechanical analysis of intra- and interspecific scaling of jumping and morphology in CaribbeanAnolislizards
Authors: Toro Esteban , Herrel Anthony , Vanhooydonck Bieke , Irschick Duncan J.
Published: June 2003

Scaling models predict how functional variables change as animals grow or increase in size evolutionarily. However, few experimental studies have found support for the predictions of these models. Here, we use a force plate to investigate the scaling of functional variables associated with jumping within(for three species) and across adults of 12 species of Anolislizards. Both ontogenetically (with the exception of Anolis carolinensis) and across the 12 species examined, limb dimensions increased geometrically, making Anolis lizards an ideal study system to test the predictions of geometric scaling models. However, both the ontogenetic and interspecific scaling of functional variables deviated in several aspects from model predictions. Unexpectedly, the scaling of functional variables such as acceleration differed for different species. Whereas acceleration capacity increases with hindlimb length for A. carolinensis, no relationship was detected for the other two species. Interspecifically, the inclusion of two large species in our analysis appears to drive the absence of a correlation between acceleration capacity and hindlimb length across species. These data suggest that selection for enhanced jumping performance is relaxed in larger anoles and support the notion that no scaling model seems to be able to comprehensively predict changes in function with size across species; rather, natural selection seems to drive changes in the scaling relationships of some key variables such as force output or acceleration capacity.
Recording Congenital Differences of the Upper Limb
Authors: LUIJSTERBURG A. J. M. , SONNEVELD G. J. , VERMEIJ-KEERS CHR. , HOVIUS S. E. R.
Published: June 2003

Consistent classification of congenital differences of the upper limb is of paramount importance for the study of the pathogenesis. To overcome the inconsistencies of present classifications, a non-classifying recording method has been developed. This method records individual aberrations, including bone and soft-tissue defects. Between 1996 and 1998, a prospective study was performed to validate the method. Two hundred and thirty-one patients with upper limb differences were assessed, and all individual aberrations were recorded. These data can be transferred to any classification. It is concluded that the presented method will allow consistent grouping of patients without losing details about simple and complex differences.
Perioperative Complications Associated with Brachial Plexus Repair in Infants
Authors: GROSSMAN J. A. I. , PRICE A. E. , SADEGHI P.
Published: June 2003

This report details the complications experienced during 100 consecutive cases of brachial plexus surgery in infants. There were eight perioperative complications. There was no mortality or permanent sequelae from any complication.
Trapeziometacarpal Joint Osteoarthritis and Carpal Tunnel Syndrome: A New Surgical Approach for Concomitant Treatment
Authors: STAHL S. , SHAPIRA D.
Published: June 2003

A new approach for the concomitant surgical treatment of trapeziometacarpal joint osteoarthritis and carpal tunnel syndrome through the same incision is described. The technique was used in 25 patients (20 women, five men; mean age, 56 years). At a mean follow-up of 27 months, there was complete disappearance of the symptoms of carpal tunnel syndrome in 20 of the 25 patients, incomplete but substantial relief in four patients, and no improvement in one patient. One patient had scar tenderness and another had a superficial wound infection. The surgical technique is simple, safe and cost-effective as it avoids separate operations for both pathologies. The procedure is not suitable for severe carpal tunnel syndrome or when direct visualization of the median nerve and the carpal tunnel is necessary.
A Modification in Technique of Abductor Digiti Minimi (Huber) Opponensplasty
Authors: CAWRSE N. H. , SAMMUT D.
Published: June 2003

Abductor digiti minimi opponensplasty is a well-established method of restoring opposition following median nerve trauma or in congenital deficiency of thenar intrinsic musculature. The abductor digiti minimi is detached distally and rerouted through a subcutaneous tunnel for insertion into the area of the thumb metacarpophalangeal joint. The pivot point for this rotation is the pisiform bone and it is at this site that the ulnar nerve is vulnerable to compression by the transferred muscle. We describe a modification of technique which involves complete islanding of the muscle without re-insertion of its proximal end. This eliminates the possibility of this complication without detracting from the efficacy of opposition produced by the transferred muscle.
Glenoid version and rotator cuff tears
Authors: Tétreault Patrice , Krueger Andreas , Zurakowski David , Gerber Christian
Published: June 2003

The purpose of this study was to determine the relationship between rotator cuff (RC) tear and the orientation of the glenoid. Ninety‐six shoulders (94 patients) that underwent open RC repair were grouped according to the type of tear. We measured on MRI the acromio–glenoid angle (AG) and the supraspinatus fossa glenoid angle on the anterior–posterior (SGAP) and axial (SGAX) views. RC patients had a smaller AG angle (76 ± 7° vs. 86 ± 10°) and a larger SGAP angle (112 ± 6° vs. 102 ± 7°) compared to controls (p < 0.001). We also found a highly significant difference (p < 0.001) in glenoid version measured by SGAX between anterior cuff tears (–5 ± 4°) and posterior cuff tears (3 ± 3°). Furthermore, we identified an association between RC tear and the orientation of the glenoid relative to the axis of the supraspinatus fossa. Greater retroversion is predictive of an anterior cuff injury and greater anteversion is predictive of a posterior cuff injury. © 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.
Immunologic Considerations for Therapeutic Strategies Utilizing Allogeneic Hepatocytes: Hepatocyte–Expressed Membrane–Bound Major Histocompatibility Complex Class I Antigen Sensitizes While Soluble Antigen Suppresses the Immune Response in Rats
Authors: Scherer Marcus N. , Graeb Christian , Tange Stefan , Dyson Chrissie , Jauch Karl-Walter , Geissler Edward K.
Published: June 2003

Understanding the immunologic effects of hepatocytes is critical because of the potential to use these cells for bioartificial livers, as a vehicle for gene transfer, and as a means to induce donor–specific immunosuppression in organ transplantation. However, this understanding is complicated by the fact that hepatocytes express membrane–bound and soluble forms of major histocompatibility complex (MHC) class I antigen, each with the potential to induce different immune responses. In the present study we first determined the immunologic effect of normal donor–derived hepatocytes in a rat heart transplant model. We then usedex vivohepatocyte gene transfer to examine the immunologic effects of different forms of hepatocyte–expressed MHC class I antigen. Results showed that intrasplenic injection of purified, donor–strain-specific hepatocytes into recipients primes alloimmunity, as evidenced by acceleration of heart allograft rejection. Interestingly, injection of autologous hepatocytes transfectedex vivowith DNA encoding only membrane–bound donor MHC class I antigen (RT1.Aa) also accelerated allograft rejection. However, hepatocytes transfected to express only secreted donor MHC antigen prolonged transplant survival. Limiting–dilution analysis of lymphocytes from animals treated with hepatocytes producing only secreted alloantigen showed an antigen–specific reduction in cytotoxic T lymphocyte (CTL) and helper T lymphocyte (HTL) precursors. Further analysis of CTL populations by flow cytometry revealed a relatively high percentage of nonviable cells, implying that soluble antigen promotes allospecific CTL death. In summary, this study suggests that hepatocyte–expressed MHC class I molecules have opposing immunologic effects, with the membrane–bound antigen inducing immunologic sensitization, and the soluble antigen promoting donor–specific immunosuppression.
Early Identification of Recipients With Progressive Histologic Recurrence of Hepatitis C After Liver Transplantation
Authors: Sreekumar Raghavakaimal , Gonzalez-Koch Alvaro , Maor-Kendler Yaakov , Batts Kenneth , Moreno-Luna Laura , Poterucha John , Burgart Lawrence , Wiesner Russell , Kremers Walter , Rosen Charles , Charlton Michael R.
Published: June 2003

Approximately half of patients undergoing liver transplantation (LT) for hepatitis C virus (HCV) develop histologic evidence of recurrence within the first postoperative year. Early identification of recipients at risk for more severe recurrence of HCV may be useful in selecting patients for antiviral therapy. We determined whether recipients at greatest risk for more severe recurrence of HCV can be identified by pre– and/or early post–LT HCV–RNA levels in serum or tissue. Serum and tissue samples were prospectively collected pre–LT and at 7 days, 4 months, 1 year, and at 3 years posttransplantation from patients undergoing LT for HCV. Hepatitis activity index (HAI) and fibrosis stage (FS) were assessed in all liver biopsies. Forty–seven patients (32 men) were studied. Higher HCV–RNA levels at 4 months post–LT (≥109copies/mL, n = 29) were associated with higher HAI at 1 year and at 3 years post–LT. The HAI seen on protocol biopsies at 4 months correlated significantly with fibrosis stage (FS) at 1 year (r= .56,P≤ .001) and 3 years (r= .53,P= .002). Higher HCV–RNA levels at 7 days and 4 months post–LT were sensitive (66% and 84%, respectively) and specific (92% and 63%, respectively) in identifying recipients with an HAI greater than 3 at 3 years. Higher pre– and early post–LT HCV–RNA levels are associated with more severe recurrence of HCV. The correlation of early HAI with subsequent FS suggests that higher mean HAI will eventually translate into more advanced stages of fibrosis. Patients at risk for more severe post–LT recurrence of HCV can be identified by early posttransplant HCV–RNA levels.
Targeted prophylaxis with amphotericin B lipid complex in liver transplantation
Authors: Singhal Saket , Ellis Richard W. , Jones Stephen G. , Miller Sarah J. , Fisher Neil C. , Hastings Mark J.G. , Mutimer David J.
Published: June 2003

The purpose of this study is to prospectively evaluate a strategy in which prophylaxis with amphotericin B lipid complex at 3 different dosages was targeted to liver transplant recipients at high risk for the development of invasive fungal infection (IFI). High risk was defined as a postoperative requirement for prolonged (≥5 days) intensive care unit (ICU) treatment. Consecutive high-risk patients were administered prophylaxis with amphotericin B lipid complex from day 5 after orthotopic liver transplantation (OLT) until ICU discharge or death. The first 10 eligible patients were administered 5 mg/kg/d, the next 10 patients were administered 2.5 mg/kg/d, and a final 10 patients were administered 1 mg/kg/d. Drug safety and efficacy were assessed before each dosage reduction. During the study period, 130 adult patients underwent 137 OLTs. Thirty patients fulfilled the entry criteria and were administered prophylaxis with amphotericin B lipid complex. No patient developed proven IFI during prophylaxis. Cultures from normally sterile sites (blood and abdominal drain fluid) always showed negative results. All fungal isolates were sensitive in vitro to amphotericin B. There was no significant difference in colonization scores among the groups of patients administered different dosages of amphotericin B lipid complex. No death, serious adverse reaction, or nephrotoxicity was attributed to amphotericin B lipid complex. We conclude that prophylaxis with amphotericin B lipid complex targeted to patients requiring prolonged ICU treatment after OLT appears to be well tolerated and may prevent IFI. Our current policy is to use amphotericin B lipid complex, 1 mg/kg/d, as antifungal prophylaxis in this high-risk group.
Increased risk for posttransplant lymphoproliferative disease in recipients of liver transplants with hepatitis C
Authors: McLaughlin Kevin , Wajstaub Sandra , Marotta Paul , Adams Paul , Grant David R. , Wall William J. , Jevnikar Anthony M. , Rizkalla Kamilla S.
Published: June 2003

Posttransplant lymphoproliferative disease (PTLD) is associated with immunosuppression and lymphotrophic viral infections. Hepatitis C virus (HCV) has been identified as a risk factor for non-Hodgkin’s lymphoma, but no association between HCV and PTLD has been shown. To investigate this possibility, we identified patients with HCV who received their first orthotopic liver transplant at our unit between January 1, 1992, and December 31, 1996, and compared them with contemporary liver transplant recipients without HCV for incidence and risk factors for PTLD and survival. Fifty-seven patients with HCV and 127 patients without HCV were compared. There was no age difference (52 v 53 years; P = .85), but there were more men in the HCV group (man-woman ratio, 2.1:1 v 0.9:1; P = .006). No difference was observed in the follow-up period, graft survival, rejection episodes, or use of different immunosuppressive regimes (P > .05 for all). Four patients with HCV (7%) developed PTLD compared with 1 patient without HCV (0.8%; P = .02). The relative odds for developing PTLD in patients with HCV were 9.5. All patients who developed PTLD were administered OKT3 induction therapy. These data suggest that PTLD may be more prevalent in patients undergoing liver transplantation for HCV-related liver disease who also receive OKT3, and that HCV infection may be a risk factor for developing PTLD.
Posttransplantation hypertension related to calcineurin inhibitors
Authors: Textor Stephen C. , Taler Sandra J. , Canzanello Vincent J. , Schwartz Lora , Augustine Jo Ellen
Published: June 2003

Calcineurin inhibitors are a mainstay of transplant immunosuppression and commonly induce hypertension. They are highly lipid soluble and penetrate vascular smooth muscle cell membranes readily. Changes in vascular tone are universally observed during administration of these agents, particularly within the kidney, leading to diminished glomerular filtration and enhanced sodium retention. Disturbances of endothelial function are prevalent in many tissues, including stimulation of endothelin and impaired nitric oxide synthesis. Multiple additional pathways produce increased vasoconstriction, leading to an increase in arterial pressure. Clinical manifestations include disturbances in circadian blood pressure patterns, left ventricular hypertrophy, and acceleration of atherosclerotic and renal injury. Rapid increases in pressure occasionally produce accelerated hypertension and microangiopathic tissue damage. Principles of therapy require recognition of hazards of changing arterial pressures during calcineurin use and preferential use of vasodilating drugs, particularly dihydropyridine calcium channel blocking agents. Attention must be paid to interactions between antihypertensive agents and calcineurin inhibitor blood levels.
Influenza infection in patients before and after liver transplantation
Authors: Duchini Andrea , Hendry Michael R. , Redfield David C. , Pockros Paul J.
Published: June 2003

Infection with influenza virus poses specific problems in pediatric and adult liver transplant recipients, both before and after liver transplantation. These include a higher rate of pulmonary and extrapulmonary complications, development of rejection with graft dysfunction, prolonged shedding of influenza virus, and increased drug-resistance. Hepatic decompensation may occur during influenza infection in patients with cirrhosis. Current prophylaxis includes yearly vaccination with trivalent inactivated vaccine. Appropriate diagnosis and prompt treatment of any upper respiratory infections are indicated in these patients. In this review, we describe a case of influenza viral pneumonia in an adult liver transplant recipient, review basic and clinical aspects of influenza infection in this patient population, and discuss current modes of prevention and treatment in detail.
Staphylococcus aureusSusceptibility to Innate Antimicrobial Peptides, β-Defensins and CAP18, Expressed by Human Keratinocytes
Authors: Midorikawa Kazushige , Ouhara Kazuhisa , Komatsuzawa Hitoshi , Kawai Toshihisa , Yamada Sakuo , Fujiwara Tamaki , Yamazaki Kenshi , Sayama Koji , Taubman Martin A. , Kurihara Hidemi , Hashimoto Koji , Sugai Motoyuki
Published: June 2003

The antimicrobial peptides human β-defensin-1 (hBD1), hBD2, hBD3, and CAP18 expressed by keratinocytes have been implicated in mediation of the innate defense against bacterial infection. To gain insight intoStaphylococcus aureusinfection, the susceptibility ofS. aureus, including methicillin-resistantS. aureus(MRSA), to these antimicrobial peptides was examined. Based on quantitative PCR, expression of hBD2 mRNA by human keratinocytes was significantly induced by contact withS. aureus, and expression of hBD3 and CAP18 mRNA was slightly induced, while hBD1 mRNA was constitutively expressed irrespective of the presence ofS. aureus. Ten clinicalS. aureusisolates, including five MRSA isolates, induced various levels of expression of hBD2, hBD3, and CAP18 mRNA by human kertinocytes. The activities of hBD3 and CAP18 againstS. aureuswere found to be greater than those of hBD1 and hBD2. A total of 44S. aureusclinical isolates, including 22 MRSA strains, were tested for susceptibility to hBD3 and CAP18. Twelve (55%) and 13 (59%) of the MRSA strains exhibited more than 20% survival in the presence of hBD3 (1 μg/ml) and CAP18 (0.5 μg/ml), respectively. However, only three (13%) and two (9%) of the methicillin-sensitiveS. aureusisolates exhibited more than 20% survival with hBD3 and CAP18, respectively, suggesting that MRSA is more resistant to these peptides. A synergistic antimicrobial effect between suboptimal doses of methicillin and either hBD3 or CAP18 was observed with 10 MRSA strains. Furthermore, of several genes associated with methicillin resistance, inactivation of thefmtCgene in MRSA strain COL increased susceptibility to the antimicrobial effect mediated by hBD3 or CAP18.
Helicobacter pyloriInduces RANTES through Activation of NF-κB
Authors: Mori Naoki , Krensky Alan M. , Geleziunas Romas , Wada Akihiro , Hirayama Toshiya , Sasakawa Chihiro , Yamamoto Naoki
Published: June 2003

Helicobacter pylori-infected gastric mucosa displays a conspicuous infiltration of mononuclear cells and neutrophils. RANTES (short for “regulated upon activation, normal T cell expressed and secreted”) is a chemoattractant cytokine (chemokine) important in the infiltration of T lymphocytes and monocytes. RANTES may therefore contribute to the cellular infiltrate in theH. pylori-infected gastric mucosa. The aim of this study was to analyze the molecular mechanism responsible forH. pylori-mediated RANTES expression. We observed that gastric epithelial cells produced RANTES upon coculture withH. pylori. In addition,H. pyloriinduced RANTES mRNA expression and an increase in luciferase activity in cells which were transfected with a luciferase reporter construct derived from the RANTES promoter, in gastric epithelial cells, indicating that the induction of RANTES production occurred at the transcriptional level. Induction of RANTES was dependent on an intactcagpathogenicity island. Activation of the RANTES promoter byH. pylorioccurred through the action of NF-κB. Transfection of kinase-deficient mutants of IκB kinase (IKK) and NF-κB-inducing kinase (NIK) inhibitedH. pylori-mediated RANTES activation. In contrast, tumor necrosis factor alpha- or interleukin-1/Toll-like receptor signaling molecules—such as mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1, MyD88, and interleukin-1 receptor-associated kinase—did not play a role in RANTES activation byH. pylori. Collectively,H. pyloriinduced NF-κB activation through an intracellular signaling pathway that involved IKK and NIK, leading to RANTES gene transcription. RANTES induction byH. pylorimay play an important role in gastric inflammation.
Various Types ofDirofilaria immitisPolyproteins Selectively Induce a Th2-Type Immune Response
Authors: Tezuka Hiroyuki , Imai Shinjiro , Hidano Shinya , Tsukidate Setsuko , Fujita Koichiro
Published: June 2003

Dirofilaria immitispolyproteins (DiAgs) are found as 15-kDa monomeric and 30-kDa dimeric forms in exceretory-secretory products of the adult worm. We evaluated the ability of various types of recombinant DiAg (rDiAg; V1 and V2 as monomers and V1V2, V2V1, V1V1, and V2V2 as dimers) to influence Th1/Th2 immune responses. V1-, V1Vx- and V2-, V2Vx-driven nonspecific immunoglobulin E (IgE) production peaked at 21 and 14 days after administration, respectively. Dimer-induced IgE response was an interesting biphasic pattern with the second peaks on days 35 (V2Vx) or 42 (V1Vx). Absolute amounts of nonspecific IgE production induced with monomers were larger than those observed with dimers at the first peak. The magnitude of cell expansion and interleukin-10 (IL-10) production in mesenteric lymph node (MLN) B-cell induced with rDiAgs was linked to the levels of the first IgE peak in vivo and IgE produced by rDiAg plus IL-4-stimulated B cells in vitro. All rDiAgs failed to augment IgG2c production. V2 and V2Vx elicited IL-4 production by MLN cells more rapidly than V1 and V1Vx. The inhibitory effect of rDiAg on gamma interferon (IFN-γ) production was stronger in monomers than in dimers. Neutralization of IL-10 restored IFN-γ production, whereas the expression of IL-4 and IgE was partly prevented by depletion of IL-10. These results indicate that monomer rather than dimer is an efficient form of DiAg and suggest that the difference of IgE-inducing capacity among these DiAgs is closely associated with the pattern of both B-cell activation and IL-4 production.
Antigenic Properties of HpaA and Omp18, Two Outer Membrane Proteins ofHelicobacter pylori
Authors: Voland Petra , Hafsi Nadia , Zeitner Marco , Laforsch Stephanie , Wagner Hermann , Prinz Christian
Published: June 2003

Outer membrane proteins (OMPs) are incorporated into the outer plasma membrane ofHelicobacter pyloriand are important for, e.g., ion transport, adherence, structural and osmotic stability, and bacterial virulence but may also be antigenic due to their surface exposure. Previous proteome-based approaches withH. pylorilysates determined a strong serological reaction towards twoH. pyloriOMPs, HpaA (TIGR HP0797) and Omp18 (TIGR HP1125). PCR was used to detect DNA encoding the two proteins, and a positive signal was found in allH. pyloristrains tested. Proteins were cloned and expressed in the human kidney cell line HK293 with the QiaExpressionist system with a C-terminal His tag. Only sera from infected persons showed a positive reaction with the recombinant proteins. Recombinant HpaA (rHpaA) and rOmp18 were incubated with human peripheral blood mononuclear cells and induced secretion of interleukin-12 (IL-12) and IL-10 from these cells. To determine the effect on antigen-presenting cells, human blood monocytic and dendritic cells (DCs) were isolated by magnetic cell separation. rOmp18 and rHpaA strongly stimulated major histocompatibility class II and CD83 expression 7- to 10-fold on isolated DCs. rHpaA and rOmp18 failed to stimulate IL-8 secretion from monocytes but increased secretion of IL-12 and IL-10 from DCs significantly. In summary, HpaA and Omp18 are recognized by human dendritic cells and induce their maturation as well as antigen presentation. HpaA and Omp18 ofH. pylorithereby appear to have a specific antigenic potential in humans.
Host-Adapted
Borrelia burgdorferi
in Mice Expresses OspA during Inflammation
Authors: Crowley Helena , Huber Brigitte T.
Published: June 2003

Antibody responses to outer surface protein A (OspA) of
Borrelia burgdorferi
may occur during periods of arthritis late in the clinical course of untreated Lyme disease. These antibody responses are paradoxical, given the conclusive evidence demonstrating that
B. burgdorferi
transmitted to the mammalian host expresses little or no OspA. The parallel occurrence of OspA antibodies and arthritic episodes suggests that OspA expression is upregulated during infection with
B. burgdorferi
. We hypothesized that this was due to the inflammatory environment caused by the immune response to the spirochete. To test our hypothesis, we adapted an in vivo model that mimics the host-pathogen interaction. Dialysis chambers containing
B. burgdorferi
were implanted into the peritoneal cavities of mice in the presence or absence of zymosan, a yeast cell wall extract that induces inflammation. Spirochetes were harvested 2 days later, and OspA expression was assessed at the protein and transcription level by Western blotting and real-time reverse transcription-PCR, respectively. Flow cytometry was also utilized to evaluate OspA protein expression on individual spirochetes.
B. burgdorferi
maintained in an inflammatory in vivo environment show an increased OspA expression relative to
B. burgdorferi
kept under normal in vivo conditions. Furthermore, host-adapted
B. burgdorferi
with a low OspA phenotype upregulates OspA expression when transferred to an inflammatory in vivo environment. The results obtained by these techniques uniformly identify inflammation as a mediator of in vivo OspA expression in host-adapted
B. burgdorferi
, providing insights into the behavior of live spirochetes in the mammalian host.
Toll-Like Receptor 4-Defective C3H/HeJ Mice Are Not More Susceptible than Other C3H Substrains to Infection withMycobacterium tuberculosis
Authors: Kamath Arati B. , Alt Jennifer , Debbabi Hajer , Behar Samuel M.
Published: June 2003

Mycobacterium tuberculosisproduces a variety of molecules capable of activating Toll-like receptors, a family of pattern recognition receptors expressed by macrophages and a variety of other cells. To determine whether Toll-like receptor 4 (TLR4) was critical in resistance toM.tuberculosisinfection, we compared the morbidity and mortality of TLR4-defective C3H/HeJ mice to those of TLR4-sufficient C3H mouse substrains. TLR4-defective C3H/HeJ mice and TLR4-sufficient C3H/HeSnJ, C3HeB/FeJ, and C3H/HeOuJ mice were infected by the aerosol route withM.tuberculosis. TLR4-defective C3H/HeJ mice had levels of cytokines in their bronchoalveolar lavage fluids and in vitro mycobacterial antigen-specific recall responses similar to those of other C3H mouse substrains. In addition, bacterial replication and long-term survival of mice following infection appeared to be independent of TLR4. Interestingly, C3HeB/FeJ mice were significantly more susceptible toM.tuberculosisinfection, indicating that genetic heterogeneity among inbred C3H mouse substrains modifies resistance to infection. Therefore, cautious interpretation is required when the C3H/HeJ strain is used as a model of a TLR4-defective mouse strain, as there are significant allelic differences between C3H/HeJ and other C3H mouse substrains in response toM.tuberculosisinfection. With this caveat, our data indicate that TLR4 may not be required for optimal immunity of mice toM.tuberculosis.
Mis-Takes Across Paradigms
Authors: Pilkington F. Beryl , Mitchell Gail J.
Published: June 2003

In this column, the authors reflect on mis-takes across paradigms, misconceived or misunderstood meanings given and taken among scholars with differing worldviews. Mis-takes of paradigmatic perspectives, human science, distinctiveness, ways of knowing, and rationality are explored. It is proposed that mis-taken views of opposing perspectives are often deliberately advanced by scholars as ways to discredit the opposition, not to further dialogue. The authors call for an appreciation of difference in the context of the intellectual sparring and debating with regard to fundamental issues that characterize evolving disciplines.
Mis-Takes: Mistakes in the Nurse-Person Process
Authors: Bournes Debra A. , Flint Frances
Published: June 2003

This column supports the global mantra to develop mechanisms that address the worldwide crisis in patient safety and demonstrates, using nursing as an example, that the mechanisms currently being developed are too limited. Patient safety initiatives must be expanded to acknowledge and make ways to minimize mis-takes, that is, misconceptions of meaning that emerge when healthcare professionals dismiss, misconstrue, guess, or undervalue patients’ perspectives. Commitment to the human becoming school of thought as a guide for practice is suggested as one way to address mis-takes in the nurse-person process.
A Middle-Range Theory of Caregiver Stress
Authors: Tsai Pao-Feng
Published: June 2003

The Roy adaptation model is a widely used nursing theory, but only a few studies have used it in the context of caregiving for the chronically ill. Many caregivers with chronically ill relatives suffer from stress and its consequences, and in an aging society, more and more people will be affected by stress. In this article, the author presents a middle-range theory of caregiver stress based on the Roy adaptation model as the first step in understanding the applicability of the Roy adaptation model in the context of stress in caregivers.