bluuebunny/crossref_metadata_2025 huggingface dataset:
Granzyme expression in fine-needle aspirates from liver allografts is increased during acute rejection
Authors: Kuijf Mark M.L. , Kwekkeboom Jaap , Kuijpers Marianne A. , Willems Marc , Zondervan Pieter E. , Niesters Hubert G.M. , Hop Wim C.J. , Hack Erik C. , Paavonen Timo , Höckerstedt Krister , Tilanus Hugo W. , Lautenschlager Irmeli , Metselaar Herold J.
Published: October 2002

We investigated whether determination in fine-needle aspiration biopsy (FNAB) specimens of cells expressing granzymes (Grs) and Fas ligand would provide a reliable, easy, and quantitative measure of rejection activity in the transplanted liver. Retrospectively, 13 FNAB specimens obtained during clinical acute rejection, 10 FNAB specimens obtained during subclinical rejection, 12 FNAB specimens obtained during cytomegalovirus (CMV) infection, and 26 FNAB specimens obtained in the absence of rejection or infection were included on the study. Cytospin preparations of FNAB and peripheral-blood specimens were immunocytochemically stained for Fas-ligand and Gr, and increments in the liver were calculated by subtracting frequencies of positive cells in blood from those in FNAB specimens. Only sporadically Fas ligand-expressing, but many Gr-expressing, cells were detected in FNAB specimens. Increments in Gr-positive (Gr+) cells were significantly greater in FNAB specimens obtained during clinical rejection (median, 70 Gr+ cells; range, 0 to 312 Gr+ cells; P = .006) and tended to be greater in FNAB specimens obtained during subclinical rejection (median, 62 Gr+ cells; range, 5 to 113 Gr+ cells; P = .09) compared with those obtained in the absence of rejection (median, 16 Gr+ cells; range, 0 to 103 Gr+ cells). Increments obtained during clinical or subclinical rejection did not differ from those obtained during CMV infection (median, 27 Gr+ cells; range, 6 to 212 Gr+ cells). With the exclusion of specimens obtained during CMV infection, the sensitivity of Gr determination in FNAB specimens for the diagnosis of acute rejection (either clinical or subclinical) was 70%, and specificity, 69%. In FNAB specimens obtained during clinical and subclinical acute rejection episodes after liver transplantation, increased numbers of Gr-expressing cells were present; in the absence of CMV infection, their quantification provides a measure for rejection activity with moderate accuracy.
Retrospective analysis of the results of liver transplantation for adults with severe hepatopulmonary syndrome
Authors: Collisson Eric A. , Nourmand Hamid , Fraiman Mark H. , Cooper Christopher B. , Bellamy Paul E. , Farmer Douglas G. , Vierling John M. , Ghobrial Rafik M. , Busuttil Ronald W.
Published: October 2002

The hepatopulmonary syndrome (HPS), consisting of elevated alveolar-arterial oxygen gradient and intrapulmonary vascular abnormalities in the presence of advanced liver disease, is associated with high mortality. Liver transplantation (LT) has been used for the treatment of HPS; however, the success of LT for the treatment of HPS is not uniformly documented. We reviewed our experience over a 5-year period and identified eight adult patients with incapacitating respiratory symptoms compatible with HPS. Inclusion criteria included hypoxemia, normal lung volumes, reduced oxygen diffusing capacity (DLCO), and the presence of intrapulmonary shunting. Underlying liver disease was caused by hepatitis C (2 patients), primary biliary cirrhosis (1 patient), cryptogenic cirrhosis (1 patient), alcohol (2 patients), and hepatitis C with alcohol (2 patients). Six out of eight patients required preoperative oxygen support. Severe hypoxemia was present in seven patients (PaO2 51.5 ± 8.2 mm Hg). Three patients had complicating pulmonary hypertension. All patients exhibited a severely reduced DLCO (44.6 ± 12.2% of predicted value). Six patients were transplanted, with five requiring oxygen support at the time of discharge. Resolution of oxygen dependency occurred in all patients but was delayed in the two patients exhibiting complicating pulmonary hypertension (288.5 ± 37.4 v 53.5 ± 35.7 days). All patients exhibited O2 saturations greater than 98% on room air. Currently, three patients are alive and off oxygen. The current report documents successful resolution of hypoxemia after LT in this pilot cohort. This supports the newly implemented United Network for Organ Sharing (UNOS) criteria, that LT for HPS may be extended to include patients with PaO
2 < 60 mm Hg.
An examination of factors predicting prioritization for liver transplantation
Authors: Muir Andrew J. , Sanders Linda L. , Heneghan Michael A. , Kuo Paul C. , Wilkinson William E. , Provenzale Dawn
Published: October 2002

With the recent transition of the liver transplant allocation system to the Model for End-Stage Liver Disease, a major change is its reliance entirely on objective criteria. In previous reports, potential donor families and members of the transplant community have questioned the fairness of the subjective nature of previous systems. Therefore, we examined the United Network for Organ Sharing database to determine if the previous allocation system benefited a particular group in prioritization for transplant. We included adult patients with chronic liver disease listed for transplant in the year 2000. Patients who had ever been listed as status 2A or 2B were analyzed. A multivariable analysis examined the patient characteristics that predicted being uplisted to status 2A. Of the 9244 patients, 2376 (25.7%) had received a liver transplant as a status 2A or had been listed as status 2A. In the multivariate analysis, the strongest patient characteristics that predicted status 2A were listing in the western United States and shorter duration of registration. Other predictors include blood type O, college education, unemployment, and coverage with private insurance or a health maintenance organization/preferred provider organization. In addition, patients with Laennec’s cirrhosis were less likely to be uplisted to status 2A. Age, gender, and race were not predictors of uplisting to status 2A. In conclusion, these data show the wide range of practice patterns with the use of status 2A, and these findings suggest that certain patient groups might have received preference in the previous liver transplant allocation system.
Albumin dialysis and molecular adsorbents recirculating system (MARS) for acute Wilson’s disease
Authors: Sen Sambit , Felldin Marie , Steiner Christian , Larsson Bo , Gillett Godfrey T. , Olausson Michael , Williams Roger , Jalan Rajiv
Published: October 2002

Wilson’s disease presenting as acute liver failure (ALF) is potentially fatal, and liver transplantation (LTx) is the only option. We report two patients with Wilson’s disease and ALF treated with the Molecular Adsorbents Recirculating System (MARS). Both patients fulfilled criteria for poor prognosis. Because LTx was not available immediately in either case, MARS was used as a bridge to LTx. In Case 1, serum bilirubin decreased from 803 to 425 μmol/L after 3 treatments, but increased to 656 μmol/L during a break, decreasing again to 457 μmol/L with further treatment. Serum copper decreased from 53.7 μmol/L, to 35.8 μmol/L after first treatment session, and 17.4 μmol/L at treatment completion. In Case 2, MARS treatment over 2 weeks reduced serum bilirubin from 1200 to 450 μmol/L and copper from 35 to 13 μmol/L with marked improvement in encephalopathy and reduction in ammonia (59 to 34 μmol/L). Both patients were successfully bridged to LTx (days 9 and 28, respectively). Analysis of albumin-dialysate from the MARS circuit suggested that copper removal occurred mostly in the first few hours of treatment, partly being adsorbed by albumin and partly by the MARSFlux membrane (Teraklin AG, Rostock, Germany). These data suggest that MARS removes copper efficiently and can be used to bridge patients with Wilson’s disease and ALF to LTx.
Donor and recipient outcomes in right lobe adult living donor liver transplantation
Authors: Ghobrial Rafik M. , Saab Sammy , Lassman Charles , Lu David S.K. , Raman Steven , Limanond Piyagorn , Kunder Greg , Marks Karyn , Amersi Farin , Anselmo Dean , Chen Pauline , Farmer Douglas , Han Steven , Durazo Francisco , Goldstein Leonard I. , Busuttil Ronald W.
Published: October 2002

Severe donor organ shortage has provided the impetus for adult living donor liver transplantation (ALDLT). Despite rapid implementation and expansion of the procedure, outcome analysis of ALDLT is still incomplete. This study analyzed both donor and recipient outcomes after ALDLT at a single center. ALDLT performed at UCLA between August 1999 and November 2001 were reviewed retrospectively. Twenty recipients (14 men and 6 women) with a mean age of 48.8 ± 9.7 (29 to 66) years underwent right lobe ALDLT. By computed tomograpy (CT), graft/recipient weight ratio (GRWR) was 1.3 ± 0.3 (1 to 2.2). Overall 1-year patient and graft survival rates were 95% and 85%, respectively. One recipient died of heart failure with normal liver function 5 months after transplantation. Three grafts (14%) were lost and all three patients underwent successful cadaveric retransplantation. Complications were classified according to the Clavien grading system with all but 3 recipients encountering at least one complication. Nine (45%) had grade 1 (minor), 10 (50%) had grade 2 (potentially life threatening without residual disease/disability), 3 (14%) had grade 4A (retransplantation) and one grade 4B (death). Right lobectomy for living donation was performed in 20 patients (12 men, 8 women). Residual left lobe volumes were 36 ± 5.3 (23.9 to 47.9)% of total donor liver volume. No donor required intensive care unit admission and median hospital stay was 7.5 (6 to 14) days. One donor was aborted after intraoperative biopsy showed > 50% macrovesicular steatosis. No donor mortality or long-term complications were encountered. Five grade 1 minor complications, by Clavien Classification, occurred in 4 of 20 (20%) donors. ALDLT using right lobe grafts is an effective procedure to expand a severely depleted donor, but is associated with a high complication rate despite good survival outcomes. Continuous standardized reporting of ALDLT outcomes is required to allow successful and safe implementation of the procedure.
Delayed neuropsychologic dysfunction after liver transplantation for acute liver failure: A matched, case-controlled study
Authors: Jackson Elizabeth W. , Zacks Steven , Zinn Sandra , Ryan John , Johnson Mark W. , Gerber David A. , Andreoni Kenneth , Fair Jeffrey H. , Shrestha Roshan , Fried Michael W.
Published: October 2002

Although several studies have identified posttransplant neurologic sequelae in patients with acute liver failure (ALF), the effects of these sequelae on neuropsychologic functioning after transplant is unknown. This study compared neuropsychologic functioning of ALF patients with chronic liver disease patients after liver transplantation. After liver transplantation, seven ALF patients were compared with a matched control group of patients who had been transplanted for chronic liver disease. The patients were matched by gender, age (within 5 years), and time since transplantation (within 2 years). Patients completed a 2-hour battery of tests, which included measures of attention, memory, motor performance, abstract conceptualization, and visuospatial perception. There were no significant differences between the groups on measures of socioeconomic status or education. Significant differences were found on three separate tests: WAIS-III Vocabulary, WAIS-III Similarities, and WMS-III Paired Associate Learning II. Although these tests measure distinct functions (vocabulary knowledge, abstract conceptualization, and delayed verbal recall), they may be influenced by broader verbal functions, such as verbal fluency, conceptualization, and the ability to articulate ideas. When patients were asked what functions had noticeably deteriorated since transplantation, nearly all complained of memory difficulties, and there was no difference between groups. However, more ALF than chronic liver disease (CLD) patients complained of concentration difficulties. The results of this study suggest that ALF patients may experience more neuropsychologic dysfunction after transplant. Further studies are required to expand on these initial observations with the potential to improve patient care and referral to appropriate rehabilitative services.
Risk factors associated with the development of skin cancer after liver transplantation
Authors: Mithoefer Abigail B. , Supran Stacey , Freeman Richard B.
Published: October 2002

Skin cancer is a well-recognized long-term complication of transplantation and immunosuppression. Although risk factors for the development of skin cancer in the general population are well defined, risk factors for the development of these lesions have not been identified clearly in the liver transplant population. We surveyed 151 liver transplant (LTx) recipients for risk factors associated with cutaneous malignancies in the general population. Variables included were: demographics, primary liver disease, severity of disease at LTx, immunosuppression history, complexion, hair color, eye color, tanning profile, number of moles, occupational history, sun exposure history, sunburn history, family history of skin cancer, and any history of removed skin lesions. All skin cancers were confirmed histologically. There were 86 documented skin cancers in 34 patients: 56 squamous cell, 23 basal cell and 7 melanomas. Median follow-up was 1490 days. In a univariate analysis, age, male gender, red hair, brown eyes, primary sclerosing cholangitis (PSC), primary biliary cirrhosis (protective), cyclosporine, number of second degree sunburns, and frequent lifetime sun exposure were associated with the development of new skin cancers. In a multivariate model, age, male gender, red hair, brown eyes, PSC, and cyclosporine remain the strongest predictors. The incidence of skin cancer after liver transplantation is underestimated. In particular, there is a higher incidence of squamous cell carcinoma compared with the general population. Recipients with identified risk factors may be candidates for prophylactic treatment and should be followed more intensively after liver transplantation.
Acute graft-versus-host disease of the lung after liver transplantation
Authors: Knox Kenneth S. , Behnia Mehrdad , Smith Lisa R. , Vance Gail H. , Busk Michael , Cummings Oscar W. , Kwo Paul Y. , Wilkes David S.
Published: October 2002

We describe the first case of a man who developed acute graft-versus-host disease (GVHD), isolated to the lung, after an orthotopic liver transplant from a female donor. Our patient experienced dyspnea, worsening hypoxemia, and a progressive obstructive ventilatory defect 12 days after liver transplantation. Open-lung biopsy revealed grade 2 lymphocytic bronchiolitis, the pathologic and immunologic correlate of acute pulmonary GVHD. Fluorescent in situ hybridization confirmed donor cells at sites of peribronchiolar inflammation. High-dose corticosteroids were given with a return to baseline pulmonary function. The current case should alert clinicians to investigate pulmonary GVHD as a potential cause of postoperative dyspnea in liver transplant recipients.
Clinical impact and efficacy of lamivudine therapy in de novo hepatitis B infection after liver transplantation
Authors: Castells Lluís , Vargas Víctor , Rodríguez Francisco , Allende Helena , Buti Maria , Sánchez-Avila José F. , Jardí Rosendo , Margarit Carlos , Pumarola Tomás , Esteban Rafael , Guardia Jaime
Published: October 2002

De novo hepatitis B virus (HBV) infection after orthotopic liver transplantation (OLT) in patients negative for hepatitis B surface antigen (HBsAg) is between 1.7% and 3.5% in areas with a low prevalence of HBV infection. The importance of this problem and the efficacy of lamivudine treatment has not been defined in areas with a high prevalence of positivity to antibody to hepatitis B core antigen (Anti-HBc). To define the characteristics and the clinical impact of de novo HBV infection in OLT recipients and to evaluate the efficacy of lamivudine treatment in this context, 229 HBsAg (−) donors (145 men, 84 women) were retrospectively evaluated between June 1994 and June 2000. Forty-eight recipients were excluded for various reasons. The final study population included 181 patients that were prospectively followed up for more than 6 months after OLT. When de novo HBV infection was detected, liver allograft biopsy was performed and treatment with lamivudine was indicated if patients were HBV-DNA-positive with elevated ALT levels. Survival time was defined as the interval between diagnosis of HBV infection and death or last follow-up visit. Thirty-one of 229 liver donors (13.5%) were anti-HBc(+). After a mean follow-up of 54.4±30 months, 9 of the 181 recipients (5%) developed de novo HBV infection; 8 of 27 recipients (29.6%) of livers from anti-HBc(+) donors as compared with only one of 154 recipients (0.6%) of livers from anti-HBc(−) donors P < 0.005). Liver biopsies performed in 8 of 9 cases showed chronic active hepatitis in 7 patients and acute hepatitis in one patient who cleared HBV spontaneously during the first 3 months. Seven patients were treated with lamivudine for a mean period of 24.5 months; HBV-DNA became negative in 5 of 7 (71.4%), and HBeAg became undetectable in 3 of 6 patients (50%). Patient actuarial survival rates at 1, 3, and 5 years were 100%, 94.7%, and 81.2% for recipients of anti-HBc (+) livers and 95.2%, 83%, and 77.3% for recipients of anti-HBc (−) livers P = ns). In our area, the appearance of de novo HBV infection after OLT is related to grafting livers from anti-HBc (+) donors is associated with a benign outcome, with no liver failure or graft loss, and treatment with lamivudine is highly effective in the control of HBV replication.
Safety of right lobectomy in living donor liver transplantation
Authors: Suh Kyung-Suk , Kim Seong Hoon , Kim Sang Beom , Lee Hyuk-Joon , Lee Kuhn Uk
Published: October 2002

The goal of this study was to examine the safety and effectiveness of right lobectomy in living donor liver transplantation (LDLT). From January 1999 to January 2002, 100 cases of LDLT were performed at Seoul National University Hospital; 45 involved right lobectomy (RL), 17 involved extended left lobectomy (ELL), 37 involved left lateral segmentectomy (LLS), and 1 involved right posterior segmentectomy. The outcome of RL was compared with those of other types of hepatectomy. An RL resulted in a longer operative time (minutes) than an LLS (349.0 ± 65.1 versus 286.7 ± 54.0, P < .01), but not an ELL (351.2 ± 84.3, P =.99). The hospital stay (days) in the RL group (14.4 ± 3.1) was longer than for those in the ELL group (11.7 ± 1.7, P < .01) and the LLS group (11.7 ± 1.9, P < .01). The drain amount (mL) of the postoperative third day in the RL group (194.4 ± 143.4) was larger than for those in the ELL group (56.8 ± 84.1, P < .01) and the LLS group (46.5 ± 39.6, P < .01). The postoperative peak serum level of total bilirubin (mg/dL) was 3.0 ± 1.5 in the RL group, 1.9 ± 0.7 in the ELL group, and 1.9 ± 0.9 in the LLS group (P < .01, RL versus LLS, ELL). There was no mortality or major morbidity and no reoperation of donors. Right lobectomy is a relatively safe and effective procedure in LDLT, but brings more potential risks and morbidity in donors.
Chronic renal dysfunction late after liver transplantation
Authors: Cohen Ari J. , Stegall Mark D. , Rosen Charles B. , Wiesner Russell H. , Leung Nelson , Kremers Walter K. , Zein Nizar N.
Published: October 2002

With the increasing success of liver transplantation, more patients are developing late complications such as renal dysfunction. The goal of the current study was to assess the prevalence of renal dysfunction years after liver transplantation and to identify patients at risk for the development of this complication. Of the 527 liver transplantations performed at our institution between April 1990 and October 1998, 353 had pretransplantation and posttransplantation glomerular filtration rate (GFR) determinations by iothalamate clearance. From this entire group, 198 patients had actual 4-year follow-up and 52 had actual 6-year follow-up. In addition, 191 of these patients had intensive follow-up with GFR measurements pretransplantation and at 1 and 3 years posttransplantation (complete follow-up group). All patients received either tacrolimus- or cyclosporine-based immunosuppression. The overall mean GFR levels in both of these groups was acceptable and was not different in cyclosporine- versus tacrolimus-based immunosuppressive regimens. Renal dysfunction was progressive, with 27.5% of patients in the intensive group having a GFR < 40 mL/min/body surface area 5 years after transplantation. GFR pretransplantation did not correlate well with late renal function; however, GFR at 1 year identified patients with subsequent renal dysfunction. The cumulative incidence of renal failure for the entire group was 6.25% at 7 years and 10% at 10 years. Renal dysfunction is a major late complication after liver transplantation. The GFR at 1 year correlates best with late renal function. Patients with a low GFR at 1 year (< 40 mL/min/BSA) are a high-risk group that might benefit from early therapeutic interventions aimed at preventing subsequent renal failure.
Can a continuum solvent model reproduce the free energy landscape of a β-hairpin folding in water?
Authors: Zhou Ruhong , Berne Bruce J.
Published: October 2002

The folding free energy landscape of the C-terminal β-hairpin of protein G is explored using the surface-generalized Born (SGB) implicit solvent model, and the results are compared with the landscape from an earlier study with explicit solvent model. The OPLSAA force field is used for the β-hairpin in both implicit and explicit solvent simulations, and the conformational space sampling is carried out with a highly parallel replica-exchange method. Surprisingly, we find from exhaustive conformation space sampling that the free energy landscape from the implicit solvent model is quite different from that of the explicit solvent model. In the implicit solvent model some nonnative states are heavily overweighted, and more importantly, the lowest free energy state is no longer the native β-strand structure. An overly strong salt-bridge effect between charged residues (E42, D46, D47, E56, and K50) is found to be responsible for this behavior in the implicit solvent model. Despite this, we find that the OPLSAA/SGB energies of all the nonnative structures are higher than that of the native structure; thus the OPLSAA/SGB energy is still a good scoring function for structure prediction for this β-hairpin. Furthermore, the β-hairpin population at 282 K is found to be less than 40% from the implicit solvent model, which is much smaller than the 72% from the explicit solvent model and ≈80% from experiment. On the other hand, both implicit and explicit solvent simulations with the OPLSAA force field exhibit no meaningful helical content during the folding process, which is in contrast to some very recent studies using other force fields.
Familial hemiplegic migraine mutations increase Ca
2+
influx through single human Ca
V
2.1 channels and decrease maximal Ca
V
2.1 current density in neurons
Authors: Tottene Angelita , Fellin Tommaso , Pagnutti Stefano , Luvisetto Siro , Striessnig Joerg , Fletcher Colin , Pietrobon Daniela
Published: October 2002

Insights into the pathogenesis of migraine with aura may be gained from a study of human Ca
V
2.1 channels containing mutations linked to familial hemiplegic migraine (FHM). Here, we extend the previous single-channel analysis to human Ca
V
2.1 channels containing mutation V1457L. This mutation increased the channel open probability by shifting its activation to more negative voltages and reduced both the unitary conductance and the density of functional channels in the membrane. To investigate the possibility of changes in Ca
V
2.1 function common to all FHM mutations, we calculated the product of single-channel current and open probability as a measure of Ca
2+
influx through single Ca
V
2.1 channels. All five FHM mutants analyzed showed a single-channel Ca
2+
influx larger than wild type in a broad voltage range around the threshold of activation. We also expressed the FHM mutants in cerebellar granule cells from Ca
V
2.1α
1
−/−
mice rather than HEK293 cells. The FHM mutations invariably led to a decrease of the maximal Ca
V
2.1 current density in neurons. Current densities were similar to wild type at lower voltages because of the negatively shifted activation of FHM mutants. Our data show that mutational changes of functional channel densities can be different in different cell types, and they uncover two functional effects common to all FHM mutations analyzed: increase of single-channel Ca
2+
influx and decrease of maximal Ca
V
2.1 current density in neurons. We discuss the relevance of these findings for the pathogenesis of migraine with aura.
Therapeutic neonatal hepatic gene therapy in mucopolysaccharidosis VII dogs
Authors: Ponder Katherine Parker , Melniczek John R. , Xu Lingfei , Weil Margaret A. , O'Malley Thomas M. , O'Donnell Patricia A. , Knox Van W. , Aguirre Gustavo D. , Mazrier Hamutal , Ellinwood N. Matthew , Sleeper Meg , Maguire Albert M. , Volk Susan W. , Mango Robert L. , Zweigle Jean , Wolfe John H. , Haskins Mark E.
Published: October 2002

Dogs with mucopolysaccharidosis VII (MPS VII) were injected intravenously at 2–3 days of age with a retroviral vector (RV) expressing canine β-glucuronidase (cGUSB). Five animals received RV alone, and two dogs received hepatocyte growth factor (HGF) before RV in an attempt to increase transduction efficiency. Transduced hepatocytes expanded clonally during normal liver growth and secreted enzyme with mannose 6-phosphate. Serum GUSB activity was stable for up to 14 months at normal levels for the RV-treated dogs, and for 17 months at 67-fold normal for the HGF/RV-treated dog. GUSB activity in other organs was 1.5–60% of normal at 6 months for two RV-treated dogs, which was likely because of uptake of enzyme from blood by the mannose 6-phosphate receptor. The body weights of untreated MPS VII dogs are 50% of normal at 6 months. MPS VII dogs cannot walk or stand after 6 months, and progressively develop eye and heart disease. RV- and HGF/RV-treated MPS VII dogs achieved 87% and 84% of normal body weight, respectively. Treated animals could run at all times of evaluation for 6–17 months because of improvements in bone and joint abnormalities, and had little or no corneal clouding and no mitral valve thickening. Despite higher GUSB expression, the clinical improvements in the HGF/RV-treated dog were similar to those in the RV-treated animals. This is the first successful application of gene therapy in preventing the clinical manifestations of a lysosomal storage disease in a large animal.
The membrane trafficking protein calpactin forms a complex with bluetongue virus protein NS3 and mediates virus release
Authors: Beaton Andrew R. , Rodriguez Javier , Reddy Y. Krishnamohan , Roy Polly
Published: October 2002

Bluetongue virus, an arbovirus of the
Orbivirus
genus, infects and replicates in both insect and mammalian cells. However, the cytopathic effect (cpe) on each host is very different. Mammalian cells show substantial cpe, most likely a result of the mechanism of virus release, whereas insect cells show little cpe and appear to release virus without cell lysis. Expression analysis of each infected cell type shows one protein, the nonstructural (NS) protein NS3, to be differentially expressed in the different cell types, suggesting it may act in the virus egress pathway. The molecular basis of such an interaction, however, has never been clear. Here, by using yeast two-hybrid analysis, we show that NS3 interacts with a cellular protein p11 (calpactin light chain), part of the annexin II complex that is involved in exocytosis. We map the NS3 region of interaction with p11 to a 13-residue peptide found at the N terminus of the protein and show it effectively competes with p36 (annexin II heavy chain) for p11 ligand binding. Further, we show that the C-terminal domain of NS3 interacts with VP2, the outermost protein of the fully assembled virus particle, suggesting that NS3 forms a bridging molecule that draws assembled virus into contact with the cellular export machinery. Our data describe the first host protein involvement in orbivirus egress and provide new insights into understanding arbovirus interactions with their hosts.
The hDcp2 protein is a mammalian mRNA decapping enzyme
Authors: Wang Zuoren , Jiao Xinfu , Carr-Schmid Anne , Kiledjian Megerditch
Published: October 2002

Decapping of mRNA is a critical step in eukaryotic mRNA turnover, yet the proteins involved in this activity remain elusive in mammals. We identified the human Dcp2 protein (hDcp2) as an enzyme containing intrinsic decapping activity. hDcp2 specifically hydrolyzed methylated capped RNA to release m
7
GDP; however, it did not function on the cap structure alone. hDcp2 is therefore functionally distinct from the recently identified mammalian scavenger decapping enzyme, DcpS. hDcp2-mediated decapping required a functional Nudix (nucleotide diphosphate linked to an X moiety) pyrophosphatase motif as mutations in conserved amino acids within this motif disrupted the decapping activity. hDcp2 is detected exclusively in the cytoplasm and predominantly cosediments with polysomes. Consistent with the localization of hDcp2, endogenous Dcp2-like decapping activity was detected in polysomal fractions prepared from mammalian cells. Similar to decapping in yeast, the presence of the poly(A) tail was inhibitory to the endogenous decapping activity, yet unlike yeast, competition of cap-binding proteins by cap analog did not influence the efficiency of decapping. Therefore the mammalian homologue of the yeast Dcp2 protein is an mRNA decapping enzyme demonstrated to contain intrinsic decapping activity.
A resident Golgi protein is excluded from peri-Golgi vesicles in NRK cells
Authors: Cosson Pierre , Amherdt Mylène , Rothman James E. , Orci Lelio
Published: October 2002

To understand the structure and the function of the Golgi apparatus, it is essential to establish how resident Golgi enzymes are localized in only a few Golgi cisternae. In particular it is crucial to establish whether Golgi enzymes are retained specifically in cisternae, or if they are continuously transported from cisterna to cisterna. Here we report that a resident Golgi enzyme is largely excluded from peri-Golgi transport vesicles in normal rat kidney cells, a cell type in which conflicting results have been reported. Analysis of the lateral distribution of two markers within Golgi cisternae led to the same conclusion: a protein incorporated in vesicles (KDEL receptor) is concentrated at the rims of cisternae where vesicles form, while mannosidase II is not. These results suggest that localization of resident Golgi enzymes is achieved primarily by selective retention within cisternae and exclusion from transport vesicles. These observations cannot easily be reconciled with the vision of rapidly maturing Golgi cisternae as the principal means of intra-Golgi transport.
Reelin, Disabled 1, and β
1
integrins are required for the formation of the radial glial scaffold in the hippocampus
Authors: Förster Eckart , Tielsch Albrecht , Saum Barbara , Weiss Karl Heinz , Johanssen Celine , Graus-Porta Diana , Müller Ulrich , Frotscher Michael
Published: October 2002

The extracellular matrix molecule Reelin is required for the correct positioning of neurons during the development of the forebrain. However, the mechanism of Reelin action on neuronal migration is poorly understood. Reelin is assumed to act on neurons directly, but it may also affect the differentiation of glial cells necessary for neuronal migration. Here we show that a regular glial scaffold fails to form
in vivo
in the dentate gyrus of mice deficient of Reelin or Disabled 1, a neuronal adaptor protein in the Reelin signaling pathway. A subset of these defects is observed in mice that lack β
1
-class integrins, known to bind Reelin. Moreover, recombinant Reelin induced branching of glial processes
in vitro
. Our data suggest that Reelin affects glial differentiation via Disabled 1 and β
1
-class integrin-dependent signaling pathways.
Escaping free-energy minima
Authors: Laio Alessandro , Parrinello Michele
Published: October 2002

We introduce a powerful method for exploring the properties of the multidimensional free energy surfaces (FESs) of complex many-body systems by means of coarse-grained non-Markovian dynamics in the space defined by a few collective coordinates. A characteristic feature of these dynamics is the presence of a history-dependent potential term that, in time, fills the minima in the FES, allowing the efficient exploration and accurate determination of the FES as a function of the collective coordinates. We demonstrate the usefulness of this approach in the case of the dissociation of a NaCl molecule in water and in the study of the conformational changes of a dialanine in solution.
Sequence-specific DNA binding by the vnd/NK-2 homeodomain of
Drosophila
Authors: Wang Lan-Hsiang , Chmelik Rebecca , Nirenberg Marshall
Published: October 2002

The ventral nervous system defective (vnd)/NK-2 homeodomain and some flanking amino acid residues were expressed in
Escherichia coli
, purified to homogeneity, and the protein was covalently coupled to Sepharose. Oligodeoxynucleotides that contained 16-bp random sequences were purified by vnd/NK-2 affinity column chromatography, cloned, and sequenced. The consensus nucleotide sequence of the vnd/NK-2 homeodomain binding site was shown to be T(T/C)AAGTG(G/C). The apparent equilibrium dissociation constant (
K
D
) of the vnd/NK-2 homeodomain for the consensus sequence is 1.9 × 10
−10
M. In addition, results of competition between oligodeoxynucleotides for binding to the vnd/NK-2 homeodomain and determination of the apparent
K
D
values of oligodeoxynucleotides that differ from the consensus sequence by only a single base pair demonstrate that the four central nucleotides, AAGT, in this sequence play a major role in determining the affinity of binding.
The TC10-interacting protein CIP4/2 is required for insulin-stimulated Glut4 translocation in 3T3L1 adipocytes
Authors: Chang Louise , Adams Rachael D. , Saltiel Alan R.
Published: October 2002

The GTPase TC10 plays a critical role in insulin-stimulated glucose transport. We report here the identification of the TC10-interacting protein CIP4/2 (Cdc42-interacting protein 4/2) as an effector in this pathway. CIP4/2 localizes to an intracellular compartment under basal conditions and translocates to the plasma membrane on insulin stimulation. Overexpression of constitutively active TC10 brings CIP4/2 to the plasma membrane, whereas overexpression of an inhibitory form of TC10 blocks the translocation of CIP4/2 produced by insulin. Overexpression of mutant forms of CIP4/2 containing an N-terminal deletion or with diminished TC10 binding inhibits insulin-stimulated Glut4 translocation. These data suggest that CIP4/2 may play an important role in insulin-stimulated glucose transport as a downstream effector of TC10.
Working Time Preferences in Dual-Earning Households
Authors: Väisänen Mia , Nätti Jouko
Published: October 2002

This study analyses actual and preferred household working times in paid employment and the preferred division of paid work between partners in dual-earning households in European Union countries and Norway, using representative Employment Options of the Future data (1998). The study focuses on working time preferences, which have so far received little attention in research. The results indicate that there is a mismatch between the actual and preferred household working time. Overall, at the European level the dual-earning households would like to reduce their working time by twelve hours a week. Actual working time practices impact on preferences, especially regarding the division of paid work between partners. There is a slight preference towards more equal sharing of paid work between partners, although the current gender division between partners within the households seems to be constant.
Towards a Model of Quality of Life for Older Ethnic Minority Migrants in Germany, Austria and the UK
Authors: Brockmann Michaela
Published: October 2002

This paper addresses the relationship between models of quality of life and Europe's increasingly diverse older population. European social policy on old age is responding only slowly to the presence of older ethnic minority migrants. People who migrated primarily for economic reasons, and who may never have intended to stay in the host country, are now approaching retirement age and faced with crucial issues of how to manage old age in a foreign country. In view of a growing policy interest in extending the quality of life of Europe's older population, the paper examines current approaches to measuring quality of life. It is argued that existing models are not suited to addressing the particular circumstances of older ethnic minority migrants, as they omit crucial aspects related to migration and their impact on key life domains. The outline of a model will be constructed which takes account of these issues and may thus serve as a basis for a more informed discussion.
Locating Europe1
Authors: Soysal Yasemin Nuhoğlu
Published: October 2002

This paper describes the emerging European public space and identity, as it is built and exercised in educational spheres. Discussions on European public space are overlaid with assumptions and requisites of a shared identity and culture. I argue that the presumed inextricable link between such constructs as identities, cultures and political communities is neither tenable nor empirically suitable in the European case. My substantive examples come from an empirical analysis of history and civics school books, as well as public debates and conflicting claims surrounding education. As projected in educational spheres, European identity is a loose collection of civic ideals and principles, such as democracy, progress, equality and human rights. This assemblage of principles and their enactment is what affords the ties that bind in the European public space. European identity as it happens is not on course to replace the nation but reinterprets it as another repository of the same ideals and principles for which Europe stands. The nation still comes across as a source of identification but is no longer unique or commanding, far from its familiar self. The emerging Europe is a space for participation but it does not imply the existence of a European demos or polity in the conventional sense - based on consensus and uniformity. European public space is open to conflict and also creates its own conflicts. It includes multiple spheres and subjects, and is created through the activities of a growing contingent of social and political actors, who engage in the discourse of Europe and deploy strategic action, with or without institutionalized contact with the EU.
Sustainable use of the Amazon River floodplain: Problems and possibilities
Authors: Junk Wolfgang J.
Published: October 2002

The Amazon River floodplain (várzea) belongs to the few areas in central Amazonia with relatively high natural fertility and productivity, because the annual flooding with sediment-rich water add nutrients to the system. Many development planners consider the area to be of high agricultural production potential and of great importance for regional and national economy and development. Inland fishery provides an important part of the animal protein supply of the local population. However, agricultural production is low and does not satisfy the demand of large cities like Manaus, because of the specific ecological conditions of the várzea. Politicians have two options directing development planning: towards a small scale diversified agriculture to satisfy local and regional market demand or towards large scale export-oriented agro-industrial development. Decision makers face the problem that, because of the flood pulse, floodplain ecosystems behave differently from other ecosystems. There is little useful information available from other regions, because most of the cost-intensive management systems of temperate zone floodplains have failed and there is little information on low-cost management systems of tropical floodplains.

Principle ecological parameters of the Amazon floodplain and a cost benefit analysis of the actual production systems are presented. Data analysis shows serious economic, social and ecological restrictions favoring small scale labor-intensive farming systems in comparison with high-technological agro-industrial solutions.
Wise use of wetlands, the Ramsar Convention on wetlands, and the need for an Asian Regional Wetlands Training Initiative
Authors: Mitchell Arthur H.
Published: October 2002

Proper management and protection of tropical aquatic ecosystems, or wetlands, are essential for a healthy environment and for the sustainability of resources required by a growing human population. The definition of wetlands, as accepted by the Ramsar Convention and Wetlands International is presented together with a summary of wetlands benefits. The Convention on Wetlands of International Importance is described, including the eight General Objectives of the Convention'sStrategic Plan (1997-2002), and their twenty-eight Operational Objectives, which in turn identify more than 125 specific actions. The focus of this paper is on General Objective 4 and its two Operational Objectives, which focus on training and capacity building for wetlands management by the Contracting Parties to the Convention. The Ramsar List of Wetlands of International Importance, its objectives and criteria, is discussed. A summary of wetlands training needs assessments in the Asian region is briefly covered, followed by a proposed development of a Wetlands Training Initiative for Asia. This includes the main elements of a training initiative, potential target groups of a multi-layered and multi-stakeholder training approach, and the need for a Coordination Centre for the Asian Training Initiative.
What defines a ‘healthy’ lake? Evidence from Lake Naivasha, Kenya
Authors: Hubble D.S. , Harper D.M.
Published: October 2002

Aquatic ecosystem health is often defined in terms of an earlier ‘preferred’ state described by earlier observations or palaeolimnology. Many tropical lakes are deteriorating, but few have a known history. Lake Naivasha (Kenya) however may be unique in having a 30000-year palaeoecological history during which it has undergone major fluctuations. It has a written history beginning in the 19th century, and has since experienced extreme changes through drought and human activity. It is now a Ramsar site, and so provides an ideal case study to examine the meaning of ‘aquatic health’. Although it has periodically dried, in its ‘natural’ state the lake supported dense submerged and fringing vegetation, much of which has now been lost. It is moderately eutrophic, has an unbalanced water budget and has experienced a number of exotic species introductions. The history of scientific work at Lake Naivasha is summarised from the earliest expedition in 1929, and with palaeolimnological evidence, is used to chart changes in the lake system including its chemistry, phytoplankton, macrophytes, macroinvertebrates and vertebrates. As the lake shows broad natural fluctuations, no single ‘natural’ state can be described. Despite anthropogenic impacts, the lake can still be considered reasonably ‘healthy’ although many of its components have been changed by human activity. Ongoing pressures on the system give cause for concern about its future condition and management.
Freshwater biodiversity: An outlook of objectives, achievements, research fields, and co-operation
Authors: Bíró Péter
Published: October 2002

This paper reviews the issue of freshwater biodiversity, from the time of its introduction on national and international bases. The project was initiated in co-operation by the International Union of Biological Sciences, the International Society of Limnology Working Group on Biodiversity in Freshwater Ecosystems and the Hydrobiological Committee of Hungarian Academy of Sciences. The first International Union of Biological Sciences International Workshop on Freshwater Biodiversity was convened at Balatonfüred, Hungary during 25-28 August 1996. A framework was developed and a wide scale project, BIODIVERSITAS, was worked out by Prof. Hiroya Kawanabe and Dr. Christian Lévesque. This project became one of the main issues of the International Union and has been incorporated into the activities of the United Nations Educational, Scientific and Cultural Organization. Several national and international meetings have been held dealing with particular fields in relation to species richness of aquatic ecosystems so that the topic has became a broad scientific, social, popular, political and economic issue. Scientific questions already cover nearly all aspects of biodiversity, for example, how to measure indices, and how to manage and preserve species richness in the most endangered aquatic habitats. Inventorying, monitoring and conservation of biodiversity in freshwater are of vital importance. All freshwater plant and animal species (invertebrates and vertebrates) are more or less threatened by human activities. Any decrease in diversity of aquatic species may indicate harmful human impacts, and whenever these activities increase, the living world of rivers, lakes and reservoirs is degraded. The rehabilitation of degraded aquatic systems has been a major limnological issue all over the world.

Freshwater biodiversity is studied in all types of habitats: lakes, reservoirs, rivers and wetlands and includes species from bacteria to aquatic vertebrates, and focuses on functional aspects and habitat differences (lake shores/ecotones, open waters and benthic habitats). Research on freshwater biodiversity can best be organized according to selected water bodies on different continents.

The ‘status quo’ of freshwater stocks is of crucial importance on local, regional and global scales. A crucial point is the high-level training of specialists and improving public-relations in order to make communities understand all aspects concerning ecological crises (e.g., habitat and biodiversity erosion and their consequences on nature and human beings). However, special interest must also be afforded to soil and sediment biodiversity, microbial biodiversity, human dimensions and genetic diversity in both land and aquatic systems.