bluuebunny/crossref_metadata_2025 huggingface dataset:
Verified predominance of slow acetylator phenotype N‐acetyltransferase 2 (NAT2) in a Hmong population residing in Minnesota
Authors: Straka Robert J. , Burkhardt R. Todd , Lang Nicholas P. , Vang Ter , Hadsall Kelly Z. , Tsai Michael Y.
Published: June 2006

Southeast Asians known as the Hmong have a high prevalence of tuberculosis and select cancers. The slow acetylation (SA) phenotype for N‐acetytransferase 2 (NAT2) has been associated with toxicity from the anti‐tuberculosis drug, isoniazid and in increased risk of select cancers. Previous research indicates a 74.5% prevalence of SA in Hmong which differs from other Asian populations including the Japanese and Thai (range: 7%–45%). Given this contrast, the purpose of this study was to confirm or refute this unexpected predominance of the SA phenotype in Hmong. Unrelated, Minnesota Hmong between 18 and 65 years of age consented and participated by ingesting caffeine as the probe for NAT2. A urinary caffeine metabolic ratio AFMU/1X (<0.6) was used to classify subjects as slow acetylators. Among 51 analysable samples provided by 61 enrollees (27 male, 33 female, 1 sex unknown, age 30±11 years [mean±SD]) there were 47 (92.2%) slow and 4 (7.8%) rapid acetylators. The prevalence of the SA phenotype (92.2%) from this study exceeds the 74.5% (p<0.02 by chi‐square test) previously noted in Minnesota Hmong (n=98). The predominance of the SA phenotype within Minnesota Hmong is confirmed. Further studies evaluating this unexpected prevalence, its genetic basis and potential clinical relevance to drug toxicity and disease are warranted. Copyright © 2006 John Wiley & Sons, Ltd.
Nutrition and management of enterocutaneous fistula
Authors: Lloyd D A J , Gabe S M , Windsor A C J
Published: June 2006

Background

The management of enterocutaneous fistula is challenging, with significant associated morbidity and mortality. This article reviews treatment, with emphasis on the provision and optimal route of nutritional support.

Methods

Relevant articles were identified using Medline searches. Secondary articles were identified from the reference lists of key papers.

Results and conclusion

Management of enterocutaneous fistula should initially concentrate on correction of fluid and electrolyte imbalances, drainage of collections, treatment of sepsis and control of fistula output. The routine use of somatostatin infusion and somatostatin analogues remains controversial; although there are data suggesting reduced time to fistula closure, there is little evidence of increased probability of spontaneous closure. Malnutrition is common and adequate nutritional provision is essential, enteral where possible, although supplemental parenteral nutrition is often required for high-output small bowel fistulas. The role of immunonutrition is unknown. Surgical repair should be attempted when spontaneous fistula closure does not occur, but it should be delayed for at least 3 months.
Redefining the R1 resection in pancreatic cancer
Authors: Verbeke C S , Leitch D , Menon K V , McMahon M J , Guillou P J , Anthoney A
Published: June 2006

Background

Resection margin (RM) status in pancreatic head adenocarcinoma is assessed histologically, but pathological examination is not standardized. The aim of this study was to assess the influence of standardized pathological examination on the reporting of RM status.

Methods

A standardized protocol (SP) for pancreaticoduodenectomy specimen examination, involving multicolour margin staining, axial slicing and extensive tissue sampling, was developed. R1 resection was defined as tumour within 1 mm of the RM. A prospective series reported according to this protocol (SP series, n = 54) was compared with a historical matched series in which a non-standardized protocol was used (NSP series, n = 48).

Results

Implementation of the SP resulted in a higher R1 rate overall, and for pancreatic (22 of 26 85 per cent) compared with ampullary (four of 15) and bile duct (six of 13) cancer. Sampling of the circumferential RM was more extensive in the SP series and correlated with RM status. RM involvement was often multifocal (14 of 32), affecting the posterior RM most frequently (21 of 32). Survival correlated with RM status for the entire SP series (P < 0·001), but not for the NSP series. There was a trend towards better median and actuarial 5-year survival after R0 resection in the SP pancreatic cancer subgroup.

Conclusion

Standardized examination influences the reporting of RM status.
Sustainability practices of SMEs: the case of NZ
Authors: Lawrence S. R. , Collins E. , Pavlovich K. , Arunachalam M.
Published: June 2006

While individually small and medium sized enterprises (SMEs) may have small social, environmental and financial impacts, cumulatively their impact is significant. One of the fundamental questions is how a single economic entity, especially a small‐scale enterprise, can be engaged in the uptake of sustainability practices. This question is particularly pertinent to New Zealand, where 98% of enterprises are SMEs. In this paper questions are raised about the conventional models of ‘business ethics’ and accountability and their relevance to SMEs. The paper reports on actual practices and discusses the possibility of small enterprises having accountability for their social and environmental impacts. Ways of linking individual firm activities to sustainability, such as a communitarian model of accountability, are discussed and illustrated. Copyright © 2006 John Wiley & Sons, Ltd and ERP Environment.
Radiotherapy alone, versus radiotherapy with amifostine 3 times weekly, versus radiotherapy with amifostine 5 times weekly
Authors: Jellema Anke Petra , Slotman Ben J. , Muller Martin J. , Leemans C. Rene , Smeele Ludi E. , Hoekman Klaas , Aaronson Neil K. , Langendijk Johannes A.
Published: June 2006

BACKGROUND.

The main objective of this study was to investigate whether nondaily intravenous administration of amifostine was as effective as daily intravenous administration with regard to the reduction of the incidence of Grade 2 or greater xerostomia in patients with head and neck cancer.

METHODS.

Ninety‐one patients who received bilateral irradiation for head and neck cancer were included. Thirty patients received no amifostine (AMI‐0), 31 patients received amifostine at a dose of 200 mg/m2 3 times weekly (AMI‐3), and 30 patients received amifostine at a dose of 200 mg/m2 daily (5 times weekly) (AMI‐5). Acute and late xerostomia and quality of life (QOL) were assessed at baseline, 6 weeks later, and at 6‐month intervals from 6 months to 24 months postradiotherapy.

RESULTS.

Grade 2 or greater late xerostomia differed significantly at 6 months (AMI‐0 74% vs. AMI‐3 67% vs. AMI‐5 52%; P = .03), but not thereafter. During follow‐up, patient‐rated xerostomia deteriorated more in AMI‐0 patients (mean difference score:, 52 for AMI‐0 compared with 25 for AMI‐3, and 29 for AMI‐5; P = .01). Nausea and emesis were reported most frequently as side effect, but Grade 2 or greater toxicity was observed in only 4 patients. However, 28% of patients discontinued amifostine before the end of radiotherapy.

CONCLUSIONS.

Long‐term, patient‐rated xerostomia was less for the AMI‐3 and AMI‐5 groups through 2‐year follow‐up, but no difference was noted between the AMI‐3 and AMI‐5 groups. For late xerostomia according to the Radiation Therapy Oncology Group criteria, the same effect was observed at 6 months, but not thereafter. Cancer 2006. © 2006 American Cancer Society.
Long‐term effects of the treatment of polycythemia vera with recombinant interferon‐α
Authors: Silver Richard T.
Published: June 2006

Patients with polycythemia vera (PV) are most often treated with phlebotomy‐only (PHL‐O) or phlebotomy plus hydroxyurea (PHL + HU). Such treatment is often unsatisfactory because of persistent susceptibility to thrombosis owing to inadequate control of abnormal erythropoiesis and thrombopoiesis. Recombinant interferon‐α (rIFNα) inhibits erythroid progenitors and affects megakaryocyte function and thus may be a more effective treatment, but reports of its use have been of relatively short duration. The long‐term use (median, 13 years) of rIFNα in 55 patients previously treated with PHL alone or with PHL + HU was studied. Data pertaining to the natural history of the disease were also examined. Patients achieved partial response of their disease by 6 months, and complete response by 1‐2 years (phlebotomy‐free, HCT ≤45%, platelets ≤600,000/μL); spleen size was reduced in 27 of 30 patients with prior splenomegaly. The initial dose of rIFNα was 1 mega unit 3 times a week (1 MU/tiw) for the majority of patients, with periodic dose increases as required and as tolerated. The maintenance dose, usually 3 MU/tiw, could be decreased after the second year of treatment in half the patients. Toxicity was acceptable. Disease‐free survival was marked by no thrombohemorrhagic complications reflecting both the effect of rIFNα and total patient care. Evidence is presented indicating that rIFNα effectively reduces PHL requirements, thrombocythemia, splenomegaly, and thrombohemorrhagic events. It is an effective drug for treating PV with acceptable toxicity. Cancer 2006. © 2006 American Cancer Society.
Impact of chemotherapy regimens prior to endocrine therapy
Authors: Buzdar Aman U. , Guastalla Jean‐Paul , Nabholtz Jean‐Marc , Cuzick Jack ,   on behalf of the ATAC Trialists' Group
Published: June 2006

BACKGROUND.

The 33‐month median follow‐up of the ATAC (anastrozole and tamoxifen, alone or in combination) trial showed a potential interaction between anastrozole and previous chemotherapy; however, this was much smaller at 47 months' median follow‐up. When the effects of different chemotherapy regimens were evaluated at that time, the apparent interaction was limited to patients who had received cyclophosphamide, methotrexate, and 5‐fluorouracil (CMF).

METHODS.

In this retrospective analysis of 68‐month data, we investigated the impact of prior chemotherapy, including different chemotherapy regimens, on time to recurrence. The chemotherapy regimens were 1) CMF only; 2) anthracycline‐containing regimens (anthracycline or anthracycline and CMF); and 3) other chemotherapy regimens, including taxane‐containing combinations.

RESULTS.

No evidence was found for an interaction between prior chemotherapy and anastrozole (hazard ratio [HR] 0.89 vs. 0.74 for those with or without prior chemotherapy, respectively; P = .21 for interaction). For those with prior chemotherapy, the HR of anastrozole when compared with that of tamoxifen shifted from 0.98 (95% confidence intervals [CI], 0.76–1.28) at 47 months' median follow‐up to 0.89 (95% CI, 0.71–1.12) at 68 months' median follow‐up and was closer to the overall treatment effect (HR, 0.79; 95% CI, 0.70–0.90). No differences according to type of chemotherapy were seen, and a benefit for anastrozole was also now apparent for patients receiving prior CMF (HR, 0.89; 95% CI, 0.63–1.24).

CONCLUSIONS.

On the basis of the 5‐year Completed Treatment Analysis, the ATAC trial does not indicate that the relative treatment benefits of anastrozole differ significantly between patients who received prior chemotherapy and those who did not. Cancer 2006. © 2006 American Cancer Society.
Role of chemotherapy for patients with recurrent platinum‐resistant advanced epithelial ovarian cancer
Authors: Rocconi Rodney P. , Case Ashley S. , Straughn J. Michael , Estes Jacob M. , Partridge Edward E.
Published: June 2006

BACKGROUND.

Current chemotherapy in platinum‐resistant ovarian cancer patients has demonstrated minimal to no improvements in survival. Despite the lack of benefit, significant resources are utilized with such therapies. Therefore, the objective in the current study was to assess the cost‐effectiveness of salvage chemotherapy for patients with platinum‐resistant epithelial ovarian cancer (EOC).

METHODS.

A decision analysis model evaluated a hypothetical cohort of 4000 platinum‐resistant patients with recurrent EOC. Several chemotherapy strategies were analyzed: 1) best supportive care (BSC); 2) second‐line chemotherapy‐monotherapy; 3) second‐line chemotherapy‐combination therapy; 4) third‐line chemotherapy after disease progression on second‐line monotherapy; and 5) third‐line chemotherapy after disease progression on second‐line combination therapy. Sensitivity analyses were performed on all pertinent uncertainties.

RESULTS.

Using costs alone, BSC was the only definitive cost‐effective treatment for platinum‐resistant recurrent ovarian cancer patients, and second‐line monotherapy was a reasonable cost‐effective strategy with an incremental cost‐effectiveness ratio (ICER) of $64,104. The cost‐effectiveness ranged from $4,065 per month of overall survival (OS) for BSC to $12,927 for third‐line previous combination therapy. Compared with BSC, second‐line monotherapy gained an additional 3 months of OS, with a cost‐effectiveness of $4,703 per month of OS. Second‐line combination therapy and third‐line therapies exhibited unfavorable ICER.

CONCLUSIONS.

The current decision analysis was intended to be thought‐provoking and bring awareness to the high costs of subsequent chemotherapy with limited effectiveness in patients with recurrent platinum‐resistant EOC. Although actual patients may receive multiple lines of chemotherapy, from the perspective of costs alone this model using a hypothetical cohort demonstrated that best supportive care was the only cost‐effective strategy, with second‐line monotherapy appearing to be a reasonable cost‐effective strategy given current chemotherapeutic options. Cancer 2006. © 2006 American Cancer Society.
Additional axillary metastases and stage migration in breast cancer patients with micrometastases or submicrometastases in sentinel lymph nodes
Authors: van Rijk Maartje C. , Peterse Johannes L. , Nieweg Omgo E. , Oldenburg Hester S. A. , Rutgers Emiel J. Th. , Kroon Bin B. R.
Published: June 2006

BACKGROUND.

In patients with breast cancer, micrometastases and submicrometastases are increasingly found in sentinel nodes when step sectioning and/or immunohistochemical staining are applied. The aims of the current study were to investigate the incidence of micro‐ and submicrometastases in the sentinel node, to estimate the risk of additional metastases in the remaining axillary lymph nodes, and to consider implications for staging and treatment.

METHODS.

A total of 2150 breast cancer patients who had undergone axillary sentinel node biopsy between 1999 and 2004 were retrospectively evaluated.

RESULTS.

In all, 649 patients (30%) had a tumor‐positive axillary sentinel node. Of these 649 patients, 148 had (23%) micrometastases and 105 (16%) submicrometastases. Of the 148 patients with micrometastases, 106 underwent axillary lymph node dissection (ALND) and additional metastases were found in 20 patients (19%). Sixteen (15%) had macrometastases and were upstaged. The other 4 patients had additional micrometastases. Seven of the 106 patients (7%) received additional systemic treatment based on the findings in the axillary lymph nodes. Fifty‐four of the 105 patients with submicrometastases underwent ALND. Two (4%) of them had additional macrometastases and were upstaged and 2 had additional micrometastases. None received additional treatment based on the ALND findings.

CONCLUSIONS.

Of the involved sentinel nodes, 23% contained micrometastases and 16% submicrometastases. Additional macrometastases were found in 15% and 4%, respectively, and treatment was altered in 7%. Based on these findings, offering additional treatment of the axilla is suggested in patients with micrometastases, but refraining from ALND in patients with submicrometastases in their sentinel node. Cancer 2006. © 2006 American Cancer Society.
Single‐Molecule Dynamic Triangulation
Authors: Plakhotnik Taras
Published: June 2006

A proposal for using single molecules as nanoprobes capable of detecting the trajectory of an elementary charge is discussed in detail. Presented numerical simulations prove that this single‐molecule technique allows determination of a three‐dimensional single‐electron displacement within a few seconds with an accuracy better than 0.006 nm. Surprisingly, this significantly exceeds the accuracy with which the probe molecule itself can be localized (given the same measuring time) by means of single‐molecule microscopy. It is also shown that the optimal concentration of probe molecules in the vicinity of the electron (i.e. the concentration which provides the best accuracy of the inferred electron displacement) is of the order of 10−5 m.
Subpopulations of motor and sensory neurons respond differently to brain‐derived neurotrophic factor depending on the presence of the skeletal muscle
Authors: Geddes Allison J. , Angka Heather E. , Davies Kimberley A. , Kablar Boris
Published: June 2006

The aim of our study was to assess the ability of brain‐derived neurotrophic factor (BDNF) to rescue motor and sensory neurons from programmed cell death. It is clearly demonstrated that the administration of a single injection of a putative neurotrophic factor to mouse embryos in utero on embryonic day (E) 14.5 is sufficient to significantly reduce the death of motor neurons when assessed on E18.5. However, the trophic requirements of somatic neurons have not been unequivocally determined in a mammalian species in vivo. Indeed, the unexpectedly high numbers of surviving neurons observed in neurotrophin and tyrosine kinase receptor knockout mice are probably the consequence of functional redundancy between the neurotrophins and their receptors. We studied spinal cord and facial motor nucleus neurons and proprioceptive neurons in the dorsal root ganglion and mesencephalic nucleus. The action of BDNF was assessed in wild‐type fetuses to gain insight into its ability to rescue neurons from naturally occurring programmed cell death. In addition, we used Myf5−/−:MyoD−/− embryos, which completely lack skeletal musculature, to assess the ability of BDNF to rescue neurons from excessively occurring programmed cell death. We found that BDNF differentially rescued neurons from naturally vs. excessively occurring cell death and that its ability to do so varied among neuronal subpopulations. Developmental Dynamics 235:2175–2184, 2006. © 2006 Wiley‐Liss, Inc.
Sema3D and Sema7A have distinct expression patterns in chick embryonic development
Authors: Bao Zheng‐Zheng , Jin Zhe
Published: June 2006

By RT‐PCR, we isolated a partial cDNA clone for the chick Semaphorin7A (Sema7A) gene. We further analyzed its expression patterns and compared them with those of the Sema3D gene, in chick embryonic development. Sema3D and Sema7A appeared to be expressed in distinct cell populations. In mesoderm‐derived structures, Sema7A expression was detected in the newly formed somites, whereas Sema3D expression was found in the notochord. In ectoderm‐derived tissues, Sema3D is expressed broadly in the surface ectoderm, lens and nasal placodes. Sema3D is also expressed in the developing nervous system including diencephalon, dorsal neural tube, optical and otic vesicles. In the limb bud, Sema3D expression was found throughout the ectoderm excluding the apical ectoderm ridge (AER), where Sema7A is concentrated. Although both genes appeared to be expressed in the migrating neural crest cells, Sema3D expression is limited to neural crest cells migrating out of the midbrain/hindbrain regions, while Sema7A expression is widespread in both cranial and trunk neural crest cells. Developmental Dynamics 235:2282–2289, 2006. © 2006 Wiley‐Liss, Inc.
Composition of the concrete, absolute, headspace and essential oil of the flowers of Michelia champaca Linn.
Authors: Rout Prasant K. , Naik S. N. , Rao Y. R.
Published: June 2006

Extraction of fresh flowers of Michelia champaca L. with pentane afforded the concrete in 1.5 ± 0.05% yield. Precipitating the waxes from the concrete with methanol gave absolute (80%). Distillation of the flowers in a Clevenger‐type apparatus furnished the essential oil in 0.03% yield. The chemical composition of the concrete, absolute and essential oil were determined by GC and GC–MS and compared. The headspace volatile composition of flowers while still attached to the branch of the tree and after plucking was determined by solid phase micro‐extraction on PDMS fibre and these results are also presented. Interestingly, the concrete and absolute contained a much smaller amount of sesquiterpene hydrocarbons, which constitute more than 60% in the headspace. Most of the esters present in the concrete and absolute are not present in the essential oil, which on the other hand contains several monoterpene hydrocarbons not isolated from the pentane extract. The sesquiterpenes detected in the headspace are present in the essential oil. Copyright © 2006 John Wiley & Sons, Ltd.
Contrasting multi‐site genotypic distributions among discordant quantitative phenotypes: the APOA1/C3/A4/A5 gene cluster and cardiovascular disease risk factors
Authors: Payseur Bret A. , Clark Andrew G. , Hixson James , Boerwinkle Eric , Sing Charles F.
Published: June 2006

Most tests of association between DNA sequence variation and quantitative phenotypes in samples of randomly chosen individuals rely on specification of genotypic strata followed by comparison of phenotypes across these strata. This strategy often succeeds when phenotypic differences are caused by one or two single nucleotide polymorphisms (SNPs) among the surveyed markers. However, when multiple‐SNP haplotypes account for observed phenotypic variation, identification of the best partitioning requires examination of an inordinate number of SNP combinations. An alternative approach is to rank individuals by their phenotypic measures and ask whether attributes of the genotypic variation show a non‐random distribution along this phenotypic ranking. One simple version of this strategy selects the top and bottom tails of the distribution, and then tests whether genotypes from these two samples are drawn from a single population. This framework does not require the recovery of phased haplotypes and allows contrasts between large numbers of sites at once. We use a method based on this approach to identify associations between plasma triglyceride level, a risk factor for cardiovascular disease, and multi‐site genotypes located in the APOA1/C3/A4/A5 cluster of apolipoprotein genes in unrelated individuals (1,071 African‐American females, 780 African‐American males, 1,036 European‐American females, and 930 European‐American males) sampled from four US cities as part of the Coronary Artery Risk Development in Young Adults (CARDIA) study. Method performance is investigated using simulations that model genealogical variation and different genetic architectures. Results indicate that this multi‐site test can identify genotype‐phenotype associations with reasonable power, including those generated by some simple epistatic models. Genet. Epidemiol. 2006. © 2006 Wiley‐Liss, Inc.
Stratified false discovery control for large‐scale hypothesis testing with application to genome‐wide association studies
Authors: Sun Lei , Craiu Radu V. , Paterson Andrew D. , Bull Shelley B.
Published: June 2006

The multiplicity problem has become increasingly important in genetic studies as the capacity for high‐throughput genotyping has increased. The control of False Discovery Rate (FDR) (Benjamini and Hochberg. [1995] J. R. Stat. Soc. Ser. B 57:289–300) has been adopted to address the problems of false positive control and low power inherent in high‐volume genome‐wide linkage and association studies. In many genetic studies, there is often a natural stratification of the m hypotheses to be tested. Given the FDR framework and the presence of such stratification, we investigate the performance of a stratified false discovery control approach (i.e. control or estimate FDR separately for each stratum) and compare it to the aggregated method (i.e. consider all hypotheses in a single stratum). Under the fixed rejection region framework (i.e. reject all hypotheses with unadjusted p‐values less than a pre‐specified level and then estimate FDR), we demonstrate that the aggregated FDR is a weighted average of the stratum‐specific FDRs. Under the fixed FDR framework (i.e. reject as many hypotheses as possible and meanwhile control FDR at a pre‐specified level), we specify a condition necessary for the expected total number of true positives under the stratified FDR method to be equal to or greater than that obtained from the aggregated FDR method. Application to a recent Genome‐Wide Association (GWA) study by Maraganore et al. ([2005] Am. J. Hum. Genet. 77:685–693) illustrates the potential advantages of control or estimation of FDR by stratum. Our analyses also show that controlling FDR at a low rate, e.g. 5% or 10%, may not be feasible for some GWA studies. Genet. Epidemiol. 2006. © 2006 Wiley‐Liss, Inc.
Particle beam radiotherapy for head and neck tumors: Radiobiological basis and clinical experience
Authors: Jereczek‐Fossa Barbara Alicja , Krengli Marco , Orecchia Roberto
Published: June 2006

Background.

Head and neck tumors are often located near critical organs, making it impossible to deliver a dose of conventional radiotherapy high enough to eradicate the disease. Our aim was to review the potential benefits and available clinical experience of particle beam therapy (hadrontherapy) in the treatment of these tumors.

Methods.

A review of the literature was carried out through a MEDLINE search (publications between 1980 and 2005).

Results.

A review of the available clinical data shows that particle beam therapy can offer several radiobiological and physical advantages over conventional photon radiotherapy: improved dose distribution permits dose escalation within the target and optimal sparing of normal tissue. Preclinical and clinical studies suggest that there may be benefits to using hadrontherapy for tumors characterized by poor radiosensitivity and critical location. At present, the most used hadrons are protons and, as yet on an experimental basis, carbon ions. It is now well accepted that there are certain indications for using proton therapy for skull base tumors (chordoma and chondrosarcoma), paranasal sinus carcinomas, selected nasopharyngeal tumors, and neutron/ion therapy for salivary gland carcinomas (in particular, adenoid cystic tumors). Its viability in other cases, such as locally advanced squamous cell carcinoma, melanoma, soft tissue sarcoma, and bone sarcoma, is still under investigation.

Conclusions.

Hadrontherapy can be beneficial in the treatment of tumors characterized by poor radiosensitivity and critical location. Further clinical and radiobiological studies are warranted for improved selection of patient population. © 2006 Wiley Periodicals, Inc. Head Neck, 2006
Atp8b1 deficiency in mice reduces resistance of the canalicular membrane to hydrophobic bile salts and impairs bile salt transport
Authors: Paulusma Coen C. , Groen Annemiek , Kunne Cindy , Ho-Mok Kam S. , Spijkerboer Astrid L. , de Waart Rudi D. , Hoek Frans J. , Vreeling Heleen , Hoeben Kees A. , van Marle Jan , Pawlikowska Ludmila , Bull Laura N. , Hofmann Alan F. , Knisely A. S. , Oude Elferink Ronald P. J.
Published: June 2006

Progressive familial intrahepatic cholestasis type 1 (PFIC1, Byler disease, OMIM 211600) is a severe inherited liver disease caused by mutations in

ATP8B1

. ATP8B1 is a member of the type 4 subfamily of P-type ATPases, which are phospholipid flippases. PFIC1 patients generally develop end-stage liver disease before the second decade of life. The disease is characterized by impaired biliary bile salt excretion, but the mechanism whereby impaired ATP8B1 function results in cholestasis is unclear. In a mouse model for PFIC1, we observed decreased resistance of the hepatocanalicular membrane to hydrophobic bile salts as evidenced by enhanced biliary recovery of phosphatidylserine, cholesterol, and ectoenzymes. In liver specimens from PFIC1 patients, but not in those from control subjects, ectoenzyme expression at the canalicular membrane was markedly deficient. In isolated mouse livers Atp8b1 deficiency impaired the transport of hydrophobic bile salts into bile.

In conclusion

, our study shows that Atp8b1 deficiency causes loss of canalicular phospholipid membrane asymmetry that in turn renders the canalicular membrane less resistant toward hydrophobic bile salts. The loss of phospholipid asymmetry may subsequently impair bile salt transport and cause cholestasis.

Supplementary material for this article can be found on the HEPATOLOGY website
(http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
Management of patients with biliary atresia in France: Results of a decentralized policy 1986-2002
Authors: Serinet Marie-Odile , Broué Pierre , Jacquemin Emmanuel , Lachaux Alain , Sarles Jacques , Gottrand Frédéric , Gauthier Frédéric , Chardot Christophe
Published: June 2006

This study analyzed the results of the decentralized management of biliary atresia (BA) in France, where an improved collaboration between centers has been promoted since 1997. Results were compared to those obtained in England and Wales, where BA patients have been centralized in three designated centers since 1999. According to their birth dates, BA patients were divided into two cohorts: cohort A, with patients born between 1986 and 1996, had 472 patients; and cohort B, with patients born between 1997 and 2002, had 271 patients. Survival rates were calculated according to the Kaplan-Meier method and compared by using the log rank test and the Cox model. Four-year overall BA patient survival was 73.6% (95% CI 69.5%-77.7%) and 87.1% (CI 82.6%-91.6%) in cohorts A and B, respectively (

P

< .001). Median age at time of the Kasai operation was 61 and 57 days in cohorts A and B, respectively (NS). Four-year survival with native liver after the Kasai operation was 40.1% and 42.7% in cohorts A and B, respectively (NS): 33.9% (cohort A) and 33.4% (cohort B) in the centers with two or fewer caseloads a year, 30.9% (cohort A) and 44.5% (cohort B) in the centers with 3-5 cases/year, 47.8% (cohort A) and 47.7% (cohort B) in the center with more than 20 caseloads a year. In cohorts A and B, 74 (15.7%) and 19 (7%) patients, respectively, died without liver transplantation (LT). Four-year survival after LT was 75.1% and 88.8% in cohorts A and B, respectively (

P

= .006).

In conclusion

, BA patients currently have the same chance of survival in France as in England and Wales. The early success rate of the Kasai operation remains inferior in the centers with limited caseloads in France, leading to a greater need for LTs in infancy and early childhood.
An MLCK-dependent window in late G1 controls S phase entry of proliferating rodent hepatocytes via ERK-p70S6K pathway
Authors: Bessard Anne , Coutant Alexandre , Rescan Claude , Ezan Frédéric , Frémin Christophe , Courselaud Brice , Ilyin Gennady , Baffet Georges
Published: June 2006

We show that MLCK (myosin light chain kinase) plays a key role in cell cycle progression of hepatocytes: either chemical inhibitor ML7 or RNA interference led to blockade of cyclin D1 expression and DNA replication, providing evidence that MLCK regulated S phase entry. Conversely, inhibition of RhoK by specific inhibitor Y27632 or RhoK dominant-negative vector did not influence progression in late G1 and S phase entry. Inhibition of either MLCK or RhoK did not block ERK1/2 phosphorylation, whereas MLCK regulated ERK2-dependent p70S6K activation. In addition, DNA synthesis was reduced in hepatocytes treated with p70S6K siRNA, demonstrating the key role played by the kinase in S phase entry. Interestingly, after the G1/S transition, DNA replication in S phase was no longer dependent on MLCK activity. We strengthened this result by

ex vivo

experiments and evidenced an MLCK-dependent window in late G1 phase of regenerating liver after two-thirds partial hepatectomy.

In conclusion

, our results underline an MLCK-dependent restriction point in G1/S transition, occurring downstream of ERK2 through the regulation of p70S6K activation, and highlighting a new signaling pathway critical for hepatocyte proliferation.
In vivo assessment of liver cell apoptosis as a novel biomarker of disease severity in nonalcoholic fatty liver disease
Authors: Wieckowska Anna , Zein Nizar N. , Yerian Lisa M. , Lopez Rocio A. , McCullough Arthur J. , Feldstein Ariel E.
Published: June 2006

In patients with nonalcoholic fatty liver disease (NAFLD), a liver biopsy remains the only reliable way to differentiate simple steatosis from nonalcoholic steatohepatitis (NASH). Noninvasive methods are urgently needed. Increasing evidence suggests hepatocyte apoptosis is a key mediator of liver injury in NAFLD. The aim of this study was to quantify hepatocyte apoptosis in plasma from patients with NAFLD and correlate it with histological severity. Plasma was obtained from 44 consecutive patients with suspected NAFLD at the time of liver biopsy. Histology was assessed blindly. Caspase-3–generated cytokeratin-18 fragments were measured

in situ

via immunohistochemistry and

in vivo

using a novel enzyme-linked immunosorbent assay. Plasma cytokeratin-18 fragments were markedly increased in patients with NASH compared with patients with simple steatosis or normal biopsies (median [interquartile range]: 765.7 U/L [479.6–991.1], 202.4 U/L [160.4–258.2], 215.5 U/L [150.2–296.2], respectively;

P

< .001). Cytokeratin-18 fragment levels independently predicted NASH (OR 1.95; 95% CI 1.18–3.22;

P

= .009 for every 50 U/L increase). A cutoff value of 395 U/L calculated using the receiver operating characteristic curve approach showed a specificity of 99.9%, a sensitivity of 85.7%, and positive and negative predictive values of 99.9% and 85.7%, respectively, for the diagnosis of NASH.

In conclusion

, these findings strongly suggest that determination of hepatocyte caspase activation in the blood is a strong and independent predictor of NASH in human subjects. These data highlight the potential usefulness of this test as a noninvasive diagnostic means of determining histological disease severity in patients with NAFLD.
A double-blind randomized trial of adefovir dipivoxil in Chinese subjects with HBeAg-positive chronic hepatitis B
Authors: Zeng MinDe , Mao YiMin , Yao GuangBi , Wang Hao , Hou JinLin , Wang YaoZong , Ji Beulah N. , Chang Chai-Ni P. , Barker Keith F.
Published: June 2006

Four hundred and eighty Chinese subjects with hepatitis B e antigen (HBeAg)–positive chronic hepatitis B (CHB) were enrolled in a multicenter, double-blind, randomized, placebo-controlled study of adefovir dipivoxil (ADV) 10 mg once daily. There was a significant difference in reduction of serum hepatitis B virus (HBV) DNA after 12 weeks between subjects who received ADV and those who received the placebo (3.4 and 0.1 log10 copies/mL, respectively,

P

< .001). Further reductions in serum HBV DNA and increases in the proportion of subjects with an HBV DNA level of at most 105 copies/mL, with HBV DNA undetectable, and with ALT normalization were observed in ADV-treated subjects at week 52 (median HBV DNA reduction of 4.5 log10 copies/mL, 67% with HBV DNA ≤ 105 copies/mL, 28% with HBV DNA undetectable, and 79% with ALT normalization). Subjects who initially received ADV lost some treatment benefit after being rerandomized to the placebo in week 40. Subjects with YMDD mutant HBV at baseline had virological, biochemical, and serological responses to treatment that were similar to those of subjects with wild-type HBV. The incidence of clinically adverse events was similar in nature and severity between the treatment groups, and there was no evidence of renal toxicity. No adefovir-related HBV mutations were identified.

In conclusion

, treatment with ADV 10 mg daily over 52 weeks was safe and effective in Chinese subjects with HBeAg-positive CHB and did not lead to the emergence of drug resistance. The study is continuing for an additional 4 years with all subjects on open-label ADV 10 mg daily.
Side population purified from hepatocellular carcinoma cells harbors cancer stem cell–like properties
Authors: Chiba Tetsuhiro , Kita Kaoru , Zheng Yun-Wen , Yokosuka Osamu , Saisho Hiromitsu , Iwama Atsushi , Nakauchi Hiromitsu , Taniguchi Hideki
Published: June 2006

Recent advances in stem cell biology enable us to identify cancer stem cells in solid tumors as well as putative stem cells in normal solid organs. In this study, we applied side population (SP) cell analysis and sorting to established hepatocellular carcinoma (HCC) cell lines to detect subpopulations that function as cancer stem cells and to elucidate their roles in tumorigenesis. Among four cell lines analyzed, SP cells were detected in Huh7 (0.25%) and PLC/PRF/5 cells (0.80%), but not in HepG2 and Huh6 cells. SP cells demonstrated high proliferative potential and anti-apoptotic properties compared with those of non-SP cells. Immunocytochemistry examination showed that SP fractions contain a large number of cells presenting characteristics of both hepatocyte and cholangiocyte lineages. Non-obese diabetic/severe combined immunodeficiency (NOD/SCID) xenograft transplant experiments showed that only 1 × 103 SP cells were sufficient for tumor formation, whereas an injection of 1 × 106 non-SP cells did not initiate tumors. Re-analysis of SP cell–derived tumors showed that SP cells generated both SP and non-SP cells and tumor-initiating potential was maintained only in SP cells in serial transplantation. Microarray analysis discriminated a differential gene expression profile between SP and non-SP cells, and several so-called “stemness genes” were upregulated in SP cells in HCC cells.

In conclusion

, we propose that a minority population, detected as SP cells in HCC cells, possess extreme tumorigenic potential and provide heterogeneity to the cancer stem cell system characterized by distinct hierarchy.

Supplementary material for this article can be found on the HEPATOLOGY website
(http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
Development of autoimmune hepatitis in patients with typical primary biliary cirrhosis
Authors: Poupon Raoul , Chazouilleres Olivier , Corpechot Christophe , Chrétien Yves
Published: June 2006

Primary biliary cirrhosis (PBC)–autoimmune hepatitis (AIH) overlap syndrome is a clinical entity characterized by the occurence of both conditions at the same time in the same patient. In addition to PBC-AIH overlap syndrome, transitions from one autoimmune disease to another have been reported, but no systematic series have been published. We report a series of 12 patients with consecutive occurrence of PBC and AIH (

i.e.

, PBC followed by AIH). Among 282 PBC patients, 39 were identified who fulfilled criteria for probable or definitive AIH. AIH developed in 12 patients (4.3%). The baseline characteristics of the patients were similar to those of patients with classical PBC. Time elapsed between the diagnosis of PBC and the diagnosis of AIH varied from 6 months to 13 years. Patients with multiple flares of hepatitis at the time of diagnosis of AIH had cirrhosis on liver biopsy. Ten patients were given prednisone ± azathioprine; short-term as well as sustained remissions were obtained in 8 of these, while two had multiple relapses and eventually died 8 and 7 years after diagnosis of AIH.

In conclusion

, the development of superimposed AIH could not be predicted from baseline characteristics and initial response to UDCA therapy. If not detected early, superimposed AIH can result in rapid progression toward cirrhosis and liver failure in PBC patients.
Fatigue in primary biliary cirrhosis is associated with excessive daytime somnolence
Authors: Newton Julia L. , Gibson John G. , Tomlinson Mark , Wilton Katharine , Jones David
Published: June 2006

A significant proportion of patients with primary biliary cirrhosis (PBC) suffer from severe fatigue. The aim of this study was to characterize patterns of daytime sleep in patients with PBC (using both objective and subjective assessment approaches) and to study the association between sleep abnormality and fatigue severity. Fatigue severity was assessed in 48 female subjects with PBC (using a disease-specific quality of life instrument (the PBC-40) and a generic fatigue measure (Fatigue Impact Scale [FIS]) as well as 48 case-matched normal controls. All participants also completed the Pittsburgh Sleep Quality Index (PSQI) and the Epworth Sleepiness Scale (ESS, which assesses daytime hypersomnolence). Objective sleep assessment was performed using accelerometry over 7 days. Global sleep quality assessed by the PSQI was significantly lower in the PBC group compared to controls (

P

< .0001). ESS scores were significantly higher in patients with PBC than controls (

P

= .0001), suggesting significantly greater daytime somnolence in the patients with PBC. Objective sleep assessment confirmed that subjects with PBC were sleeping on average almost twice as long as controls during the daytime. Both degree of daytime somnolence (ESS) and actual daytime sleep activity (accelerometry) correlated strongly with fatigue severity in the patient group (

r

2 = 0.5,

P

< .0001 and

r

2 = 0.2,

P

< .01, respectively).

In conclusion,

Sleep abnormality, in the form of excessive daytime somnolence, is present in a significant proportion of patients with PBC, with the degree of daytime somnolence correlating strongly with the degree of fatigue. Existing agents effective at reducing daytime somnolence (such as modafinil) hold potential for the treatment of fatigue in PBC.

Supplementary material for this article can be found on the HEPATOLOGY website
(http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).
Elevated levels of von Willebrand Factor in cirrhosis support platelet adhesion despite reduced functional capacity
Authors: Lisman Ton , Bongers Tamara N. , Adelmeijer Jelle , Janssen Harry L.A. , de Maat Moniek P.M. , de Groot Philip G. , Leebeek Frank W.G.
Published: June 2006

Cirrhosis of the liver is frequently accompanied by complex alterations in the hemostatic system, resulting in a bleeding tendency. Although many hemostatic changes in liver disease promote bleeding, compensatory mechanisms also are found, including high levels of the platelet adhesive protein von Willebrand Factor (VWF). However, conflicting reports on the functional properties of VWF in cirrhosis have appeared in literature. We have measured a panel of VWF parameters in plasma from patients with cirrhosis of varying severity and causes. Furthermore, we assessed the contribution of VWF to platelet adhesion, by measuring the ability of plasma from patients with cirrhosis to support adhesion of normal or patient platelets under flow conditions. VWF antigen levels were strongly increased in patients with cirrhosis. In contrast, the relative collagen binding activity, as well as the relative ristocetin cofactor activity, was significantly lower in patients as compared with controls, indicating loss of function. Accordingly, patients had a reduced fraction of high-molecular-weight VWF multimers. Both strongly elevated and reduced activity and antigen levels of the VWF cleaving protease ADAMTS13 were found in individual patients. Adhesion of either normal or patient platelets to a collagen surface was substantially increased when these platelets were resuspended in plasma of patients with cirrhosis, as compared with control plasma.

In conclusion

, highly elevated levels of VWF in patients with cirrhosis contribute to the induction of primary hemostasis despite reduced functional properties of the molecule. This phenomenon might compensate for defects in platelet number and function in patients with cirrhosis.
Different anti-HCV profiles of statins and their potential for combination therapy with interferon
Authors: Ikeda Masanori , Abe Ken-ichi , Yamada Masashi , Dansako Hiromichi , Naka Kazuhito , Kato Nobuyuki
Published: June 2006

We recently developed a genome-length hepatitis C virus (HCV) RNA replication system (OR6) with luciferase as a reporter. The OR6 assay system has enabled prompt and precise quantification of HCV RNA replication. Pegylated interferon (IFN) and ribavirin combination therapy is the world standard for chronic hepatitis C, but its effectiveness is limited to about 55% of patients. Newer therapeutic approaches are needed. In the present study, we used the OR6 assay system to evaluate the anti-HCV activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, called statins, and their effects in combination with IFN-α. Five types of statins (atorvastatin, fluvastatin, lovastatin, pravastatin, and simvastatin) were examined for their anti-HCV activities. Fluvastatin exhibited the strongest anti-HCV activity (IC50: 0.9 μmol/L), whereas atorvastatin and simvastatin showed moderate inhibitory effects. However, lovastatin, reported recently as an inhibitor of HCV replication, was shown to exhibit the weakest anti-HCV activity. The anti-HCV activities of statins were reversed by the addition of mevalonate or geranylgeraniol. Surprisingly, however, pravastatin exhibited no anti-HCV activity, although it worked as an inhibitor for HMG-CoA reductase. The combination of IFN and the statins (except for pravastatin) exhibited strong inhibitory effects on HCV RNA replication. In combination with IFN, fluvastatin also exhibited a synergistic inhibitory effect.

In conclusion

, statins, especially fluvastatin, could be potentially useful as new anti-HCV reagents in combination with IFN.
Induction of Gas6 protein in CCl4-induced rat liver injury and anti-apoptotic effect on hepatic stellate cells
Authors: Lafdil Fouad , Chobert Marie Noële , Couchie Dominique , Brouillet Arthur , Zafrani Elie Serge , Mavier Philippe , Laperche Yannick
Published: June 2006

The protein product of the growth arrest–specific gene 6 (Gas6) is a secreted ligand for tyrosine kinase receptors, among which Axl is the most widely distributed and displays the highest affinity for Gas6. The Gas6/Axl signaling pathway has been increasingly implicated in growth and survival processes occurring during development and tissue repair. In liver, after an acute or chronic injury, repair involves macrophages and hepatic stellate cells (HSC) activated into myofibroblastic cells (HSC/MFB), which produce cytokines and matrix proteins. We investigated the expression and the role of Gas6 and its receptor Axl in liver repair. Three days after CCl4-induced liver injury in the rat, we detected the expression of Gas6 in ED1-positive macrophages as well as in desmin-positive HSC, which accumulated in injured areas. Axl, the high-affinity receptor for Gas6, was detected in macrophages, HSC, and HSC/MFB.

In vitro

, expression of γ-carboxylated Gas6 was strongly induced in HSC along with their transformation into myofibroblasts, and it exerted an anti-apoptotic effect on both HSC and HSC/MFB mediated by the Axl/PI3-kinase/Akt pathway.

In conclusion

, Gas6 is a survival factor for these cells and we suggest that Gas6, secreted by macrophages and HSC/MFB

in vivo

after liver injury, promotes HSC and HSC/MFB survival and might support transient HSC/MFB accumulation during liver healing.

Supplementary material for this article can be found on the HEPATOLOGY website
(http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html).