Identification and comparative profiling of gonadal microRNAs in the adult pigeon (Columba livia).

1. MicroRNAs are small noncoding RNA molecules that play crucial roles in gene expression. However, the comparative profiling of testicular and ovarian microRNAs in birds are rarely reported, particularly in pigeon.2. In this study, Illumina next-generation sequencing technology was used to sequence miRNA libraries of the gonads from six healthy adult utility pigeons. A total of 344 conserved known miRNAs and 32 novel putative miRNAs candidates were detected. Compared with those of ovaries, 130 differentially expressed (DE) miRNAs were identified in the testes. Among them, 70 miRNAs showed down-regulation in the ovaries, while another 60 miRNAs were up-regulated.3. Combining the results of the expression of target gene measurements and pathway enrichment analyses, it was revealed that some DEmiRNAs from the gonad samples involved in sexual differentiation and development (such as cli-miR-210-3p and cli-miR-214-3p) could down-regulate AR (androgen receptor). Cli-miR-181b-5p, cli-miR-9622-3p and cli-miR-145-5p were highly expressed in both the ovaries and testes, which could co-target HOXC9, and were related to regulation of primary metabolic processes. KEGG enrichment analysis showed that DEmiRNAs may play biological and sex-related roles in pigeon gonads.4. The expression profiles of testicular and ovarian miRNA in adult pigeon gonads are presented for the first time, and the findings may contribute to a better understanding of gonadal expression in poultry.

Enterococcus faecium HDRsEf1 elevates the intestinal barrier defense against enterotoxigenic Escherichia coli and regulates occludin expression via activation of TLR-2 and PI3K signalling pathways.

Previously, we isolated a novel probiotic strain, designated HDRsEf1. In this study, we investigated the effects of this probiotic strain on intestinal barrier function and how it regulates the tight junction protein occludin in vitro. We used an ETEC-infected mouse model for the in vivo experiment. Briefly, 40 ICR mice were randomly divided into four groups: control group, assigned to saline gavage; prevention group, given HDRsEf1 before and saline after infection with ETEC; infection group, given saline both before and after infection with ETEC; treatment group, given saline before and HDRsEf1 after infection with ETEC. The weight loss was alleviated both in the prevention and treatment groups. The ETEC-induced intestinal inflammation was alleviated and the occludin mRNA expression levels in the jejuna of infected mice were increased in the prevention group. We explored the mechanism by which HDRsEf1 regulates occludin expression in vitro and found that HDRsEf1 prevented the downregulation of occludin expression in the prevention group. Simultaneously, we found that toll-like receptor-2 (TLR-2) and phosphoinositide 3-kinase (PI3K) play an important role in maintaining occludin expression. Therefore, we concluded that HDRsEf1 can prevent ETEC-induced infection by enhancing the intestinal barrier function and increasing the expression levels of occludin. SIGNIFICANCE AND IMPACT OF THE STUDY: Enterotoxigenic Escherichia coli (ETEC) is a major cause of infectious diarrhoea in children, and porcine ETEC has been the leading cause of post-weaning diarrhoea (PWD) in pigs. In our present study, we demonstrated for the first time that HDRsEf1 protects occludin from ETEC-induced suppression. Moreover, HDRsEf1 was found to regulate occludin expression via TLR-2 activation and the PI3K pathway. The results provide insights into the mechanism by which HDRsEf1 protects cells against ETEC infection and a rationale for the use of HDRsEf1 as a therapeutic and preventative agent.

Association of COL4A1 gene polymorphisms with cerebral palsy in a Chinese Han population.

The basement membrane (BM) is an extracellular matrix associated with overlying cells and is important for proper tissue development, stability, and physiology. COL4A1 is the most abundant component of type IV collagen in the BM, and COL4A1 variants can present with variable phenotypes that might be related to cerebral palsy (CP). We postulated, therefore, that variations in the COL4A1 gene might play an important role in the etiology of CP. In this study, six single nucleotide polymorphisms (SNPs) in the COL4A1 gene were genotyped among 351 CP patients and 220 healthy controls from the Chinese Han population. Significant association was found for an association between CP and rs1961495 (allele: p = 0.008, odds ratio (OR) = 1.387, 95% confidence interval (CI) = 1.088-1.767) and rs1411040 (allele: p = 0.009, OR = 1.746, 95% CI = 1.148-2.656) SNPs of the COL4A1 gene. Multifactor dimensionality reduction analysis suggested that these SNPs had interactive effects on the risk of CP. This study is the first attempt to investigate the contribution of polymorphisms in the COL4A1 gene to the susceptibility of CP in a Chinese Han population. This study shows an association of the COL4A1 gene with CP and suggests a potential role of COL4A1 in the pathogenesis of CP.

Complete mitochondrial genome of the Chinese Hwamei Garrulax canorus (Aves: Passeriformes): the first representative of the Leiothrichidae family with a duplicated control region.

The Chinese Hwamei Garrulax canorus, a member of the family Leiothrichidae, is commonly found in central and southern China, northern Indochina, and on Hainan Island. In this study, we sequenced the complete mitochondrial genome of G. canorus. The circular mitochondrial genome is 17,785 bp in length and includes 13 protein-coding genes, 22 transfer RNA (tRNA) genes, and two ribosomal RNA genes. In addition, two copies of highly similar putative control regions were observed in the mitochondrial genome. As found in other vertebrates, most of the genes are coded on the H-strand, except for one protein-coding gene (nad6; NADH dehydrogenase subunit 6) and eight tRNA genes (tRNA(Gln), tRNA(Ala), tRNA(Asn), tRNA(Cys), tRNA(Tyr), tRNA(Ser(UCN)), tRNA(Pro), and tRNA(Glu)). All the protein-coding genes start with ATG, with the exception of cox1 (cytochrome oxidase subunit 1), which starts with GTG. All tRNA genes have the potential to fold into the typical clover-leaf structure. Conserved sequences in three domains were observed in the two putative control regions. These results provide basic information for future phylogenetic analyses among species of the order Passeriformes.

Sapphire-based graphene saturable absorber for long-time working femtosecond lasers.

We report a long-time working femtosecond laser using metal-free sapphire-based graphene as a saturable absorber (SA). The sapphire-based graphene yielded excellent nonlinear saturable absorption properties and was demonstrated to be suitable as an SA for an ultrafast solid-state laser. Stable mode-locked pulses of 325 fs were obtained at a central wavelength of 1032 nm with a repetition rate of 66.3 MHz. At pump power of 8.23 W the average output power was 1.78 W and the highest pulse energy reached 26.8 nJ with a peak power of 72.6 kW. Our work opens up a facile route for making reliable graphene SA in the mode-locking technique and also displays an exciting prospect in making low-cost and ultrafast lasers.

Immunogenicity and Safety of the AS04-adjuvanted Human Papillomavirus-16/18 Cervical Cancer Vaccine in Malaysian Women Aged 18-35 years: A Randomized Controlled Trial.

INTRODUCTION: Cervical cancer is the third most common cancer in women worldwide. The HPV-16/18 AS04- adjuvanted vaccine (Cervarix©) has previously been shown to be highly immunogenic with a clinically acceptable safety profile. This phase IIIb, double-blind, randomized (1:1) and placebo controlled trial (NCT00345878) was designed to evaluate the vaccine immunogenicity against HPV-16 and HPV-18 as well as its safety and reactogenicity in Malaysian women. METHODS: Healthy women aged 18-35 years received intramuscularly three doses of either the vaccine (HPV group) or aluminium hydroxide (ALU group) at 0, 1, and 6 months. Antibody titers were measured by an enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 271 eligible subjects were enrolled and 266 subjects completed the study. Initially seronegative subjects in the HPV group showed 100% seroconversion one month post-dose-3 for anti HPV-16 and anti-HPV-18 antibodies with geometric mean titers of 11107.5 (95% CI: 9727.3-12683.4) EL.U/mL and 4273.5 (95% CI: 3771.8-4841.9) EL.U/mL, respectively. Over 96% of subjects in both groups received all three vaccine doses. Solicited local (pain) and general symptoms (myalgia, fatigue, arthralgia and headache) were commonly reported in both HPV and ALU groups. Eight serious adverse events were reported throughout the study (five in the HPV group; three in the ALU group), all considered by investigators to be unrelated to vaccination. CONCLUSION: The HPV-16/18 AS04-adjuvanted vaccine was immunogenic and generally well tolerated in Malaysian women aged 18-35 years.

Cytokine gene expression in the livers of ducklings infected with duck hepatitis virus-1 JX strain.

Duck hepatitis virus type 1 (DHV-1) causes a highly contagious disease in ducklings and is often associated with liver necrosis, hemorrhages, and high mortality. In the current study, the expression levels of gene transcripts encoding proinflammatory cytokines and the virus were measured by quantitative reverse-transcription PCR in duck livers after infection with a DHV-1 JX isolate obtained from natural cases in Hubei Province, China. In addition, sera IL-1ß, IL-6, and alanine aminotransferase levels were quantified. Liver histopathology was examined following DHV-1 infection. The ducklings died within 1 to 2 d postinfection (d.p.i.) because of typical liver degeneration, hemorrhage, necrosis, and bile-duct epithelial cell proliferation. Transcripts of the cytokines IFN-α, IL-6, TNF-α, and IL-10 decreased by 0.5 d.p.i. and then gradually increased at 1 d.p.i. Similarly, DHV-1 JX 3D gene levels in the liver sharply increased at 1 d.p.i. and then maintained a high level. In contrast, liver TNF-α and IL-1ß transcripts showed no increased expression of the cytokine gene postinfection and significantly decreased compared with the expression at 0.25 d.p.i., only the expression of IFN-α transcripts increased 128-fold by 1 d.p.i. Changes in the serum IL-6 level remained relatively stable postinfection and not significantly different compared with that of the control (P > 0.05), whereas serum levels of IL-1ß significantly decreased at 0.5 d.p.i. and increased from 1 d.p.i. onwards (P < 0.05). Serum alanine aminotransferase levels significantly increased 2 d.p.i. compared with that of the control group (P < 0.01), which seemed to keep with the number of dead ducks. The cytokines exhibited a biphasic pattern following DHV-1 JX infection. Taken together, the data indicated that duckling liver inflammatory responses were produced following experimental DHV-1 JX infection involving multiple cytokines.

Clinical Research Investigating Alzheimer's Disease in China: Current Status and Future Perspectives Toward Prevention.

Based on the background of research investigating brain aging and neurodegenerative diseases in China, the present review addresses Alzheimer's disease (AD), one of the most common types of neurodegenerative diseases, clinical research progress, and prospects for future development in China.

Total inferior border ostectomy versus T-shape genioplasty for chin narrowing combined with mandibular contouring.

The objective of this study was to compare the indications and outcomes of the total inferior border ostectomy and T-shape genioplasty. A retrospective study was conducted using the clinical notes and records of patients who underwent total inferior border ostectomy (group 1, n = 42) and T-shape genioplasty (group 2, n = 60). The outcomes were evaluated by assessment of computed tomography images combined with medical records and photographs. Lower facial height, chin width, chin symmetry, and facial proportions, as well as patient satisfaction and complications were investigated. The data were collected preoperatively and 6-24 months postoperatively. All 102 patients showed an improved lower facial contour. No severe complications were observed during the follow-up period. Although the postoperative lower to midfacial height ratios were similar in the two groups (P = 0.080), both the preoperative and postoperative chin width to lower facial height ratios were lower in group 1 (both P < 0.001). A larger amount of chin narrowing, as well as better chin symmetry were observed in group 1 (P < 0.001). In conclusion, compared to the T-shape genioplasty, the total inferior border ostectomy is well suited for a longer, wider, and more asymmetrical chin. The surgical options should be considered and chosen quantitatively to achieve aesthetically pleasing results.

Reduced placental size and increased apoptosis are associated with prenatal nicotine exposure in rats.

OBJECTIVE: Smoking during pregnancy has been linked to a variety of negative embryonic and neonatal consequences. Nicotine is the major constituent of tobacco smoke, which was associated with adverse impacts on histological and functional features of the placenta. This study aims to investigate the potential influence of nicotine exposure on the rat placenta and fetus. MATERIALS AND METHODS: Nicotine was administrated through the drinking water of female pregnant rats. The placental size, as well as the fetal body weight and size, were measured at E20. The mRNA expression of the Bax gene (pro-apoptotic), the Bcl-2 gene (anti-apoptotic) and the angiogenic genes VEGF, Flt-1, and HIF1 were measured in placental tissue. Furthermore, Immunohistochemistry (IHC) using Bax, caspase 9 and VEGF antibodies were performed on placental sections. RESULTS: The results of the current study showed a significant reduction in the size of the placenta along with fetal body weight in nicotine treated group compared to the control group. Apoptosis was observed to be boosted in the placentas of the nicotine-treated group. This was associated with upregulation of Bax expression combined with no change in the expression of Bcl-2 in the treated group. On the other hand, there was no difference in the expression of angiogenic factors VEGF, Flt-1, or HIF1 between tested groups. CONCLUSIONS: In utero nicotine exposure in pregnant rats showed deleterious impacts on fetus growth and weight, as well as placental size. These were accompanied by increased apoptosis within the placenta, as revealed by Bax gene upregulation.

Human papillomavirus-16/18 AS04-adjuvanted cervical cancer vaccine: immunogenicity and safety in healthy Chinese women from Hong Kong.

OBJECTIVE: To assess the immunogenicity and safety of human papillomavirus-16/18 AS04-adjuvanted cervical cancer vaccine in Chinese women aged 18 to 35 years enrolled from Hong Kong. DESIGN: Double-blind, randomised controlled trial with vaccine and placebo groups. SETTING: Single-centre study in Hong Kong. PARTICIPANTS: Three hundred women enrolled (150 per group) between March 2006 and June 2007. INTERVENTIONS: Subjects received three doses of human papillomavirus-16/18 vaccine or placebo (aluminium hydroxide), administered intramuscularly at 0, 1, and 6 months. MAIN OUTCOME MEASURES: Human papillomavirus-16/18 seroconversion rates and geometric mean titres at month 7 (in human papillomavirus-16/18 recipients); reactogenicity and safety (in all subjects). RESULTS: A total of 294 women completed the study (148 in the vaccine group, 146 in placebo group). All initially seronegative subjects in the vaccine group had seroconverted for human papillomavirus-16/18 antibodies by month 7. Anti-human papillomavirus-16 and anti-human papillomavirus-18 antibody geometric mean titres were 10 422 (95% confidence interval, 8730-12 442) EL.U/mL and 4649 (3975-5437) EL.U/mL, respectively. High compliance (99% in both groups) was observed for the three-vaccination course. The frequencies of local injection site reactions were higher in the vaccine than placebo group; pain being the most common symptom in both groups. Regarding solicited symptoms, fatigue and myalgia were the most frequent in both groups. Five serious adverse events (four in vaccine group, one in placebo group) were reported, but all were considered unrelated to the vaccinations. CONCLUSION: The human papillomavirus-16/18 AS04-adjuvanted vaccine was highly immunogenic, safe, and generally well tolerated in Chinese women from Hong Kong.

Identification of differentially expressed genes in the growth plate of broiler chickens with thiram-induced tibial dyschondroplasia.

Tibial dyschondroplasia (TD) is characterized by expansion of the proximal growth plates of the tibiotarsus that fail to form bone, lack blood vessels, and contain non-viable cells. Thiram (a carbamate pesticide), when fed to young broiler chicks, induces TD with high regularity and precision. We used this experimental model to understand the cause of the defects associated with TD by selecting and identifying the genes differentially expressed in the TD growth plate of broiler chickens. Broiler chicks at 7 days of age were randomly divided into two groups. After fasting overnight, they were fed with regular diet (control) or the same diet containing 100 mg/kg thiram for 96 h to induce TD (thiram-fed). mRNA was purified from the growth plates of control and thiram-fed broilers. Forward and reverse-subtracted cDNA libraries were generated by suppression subtractive hybridization technology. Ten selected genes from cDNA libraries were identified by real-time quantitative polymerase chain reaction. All were differentially expressed in TD growth plates (P<0.05 or P<0.01). The levels of collagen type X (Col X), pro-alpha-1 collagen type I (Col I alpha1), collagen type IX (Col IX), NADH dehydrogenase (NADH DH), cytochrome C oxidase subunit III (COX III), enolase 1, alpha (ENO1), carbonic anhydrase II (CA2) and heat shock protein 90 (Hsp90) mRNA transcripts were up-regulated, while the expression levels of Matrilin 3 (MATN3) and chondromodulin-I (ChM-I) were down-regulated. Col I and Hsp90 were detected by immunohistochemistry at different stages. Given that these genes are involved in matrix formation, endochondral ossification, developmental regulation, electron transport in the mitochondrial respiratory chain and vascularization, our findings may provide new insights into understanding the pathogenesis of TD.

Assessing pharmacokinetic variability directly induced by drug intake behaviour through development of a feeding behaviour-pharmacokinetic model.

Variability in drug intake is increasingly recognized as a major source of variability in drug response. The non-uniform access to medicated feed, influenced by swine individual feeding behaviour, is a determinant of antibiotic exposure, recalling the intrinsic similarity with human compliance to drug regimens. In this paper, we developed a feeding behaviour-pharmacokinetic (FBPK) model of in-feed chlortetracycline (CTC) and established, in a definite way, the effect of feeding behaviour and its induced pharmacokinetic (PK) variability. Based on reported animal behaviour, we mathematically formulated swine feeding behaviour by incorporating its main characteristics: intense feeding periods that repeat on a daily basis and random feeding periods of free access to feed, along with growth stage factors. This behaviour model was then integrated into a PK model of CTC. Moreover, we analysed the effect of each feeding behaviour component and assessed the corresponding PK variability. We have been able to delineate the impact of different feeding behaviour components and characterize the induced PK variability. We have compared different therapeutic assumptions to our model and shown that random features underlying the feeding behaviour have dramatic influence on the PK variability. A practical tool to adopt the dosing regimen in terms of dose and age has been proposed. The method developed here can be generalized to other therapeutic contexts and incorporated into medical practice, particularly to make long-term projections of drug-intake behaviour, to explain possible treatment failure and guide practitioners in adjusting the dosing regimen.

Support vector networks in adaptive friction compensation.

This paper presents our research on how support vector regression (SVR) and parametric adaptive learning, which are normally used independently, can be exploited together to benefit adaptive neural control. In the context of friction compensation for servo-motion control systems, we present the notion of support vector networks which play an essential role in combining SVR and adaptive neural network (NN) in cooperation for friction estimation. The analysis shows that the proposed support vector network contributes not only to the performance improvement but also to the practical usefulness in adaptive friction compensation. Experimental results are reported to demonstrate the effectiveness of the proposed approach.

Effect of diet with thiram on liver antioxidant capacity and tibial dyschondroplasia in broilers.

1. An experiment was conducted to study the effect of thiram on liver antioxidant capacity and incidence of tibial dyschondroplasia in broilers. 2. One hundred and twenty Avian commercial broilers were allotted at random to three treatments: control group, low thiram group (50 mg/kg) and high thiram group (100 mg/kg). 3. Blood samples were collected to determine the activity of AST (aspartate aminotransferase). At the end of the trial, broilers were killed and liver samples were collected to determine the activity of SOD (superoxide dismutase), GSH-Px (glutathione peroxidase) and MDA (malondialdehyde) content, while the right proximal tibiotarsi were dissected in longitudinal section for assessment of tibial dyschondroplasia (TD) incidence and TD score. 4. The results showed that thiram increased the incidence of TD and TD scores, increased serum AST activity and MDA content of liver, and decreased the activity of SOD and GSH-Px in the liver. 5. They suggest that thiram causes TD in broilers by reducing liver antioxidation capability and damaging liver function; this may be one of the mechanisms by which thiram causes TD in broilers.

Development of indirect enzyme-linked immunosorbent assay with nucleoprotein as antigen for detection and quantification of antibodies against avian influenza virus.

Avian influenza (AI) is a serious infectious disease caused by avian influenza virus (AIV) belonging to type A Orthomyxovirus. In the present study, we developed an indirect enzyme-linked immunosorbent assay (ELISA) employing E. coli-expressed full-length nucleoprotein (NP) of H9N2 avian influenza virus for the detection and quantification of antibodies against AIV nucleoprotein. The NP-ELISA was compared with the AI agar gel propagation (AGP) test, haemagglutination inhibition (HI) test, and IDEXX-FlockChek ELISA using 263 sera. The NP-ELISA was significantly more sensitive than the AGP and HI tests, and showed 96.2% agreement ratio with IDEXX-FlockChek ELISA. With results obtained using the NP-ELISA, an ELISA titre (ET) prediction equation, with which the ELISA titres of a flock or individual chickens can be determined, was derived from a positive/negative (P/N) ratio standard curve. The NP-ELISA enables an alternative rapid serological diagnosis and is suitable for influenza A antibody screening, especially in species that harbour several influenza subtypes.

Hypovirulence of Sclerotium rolfsii Caused by Associated RNA Mycovirus.

Mycoviruses associated with hypovirulence are potential biological control agents and could be useful to study the pathogenesis of fungal host pathogens. Sclerotium rolfsii, a pathogenic fungus, causes southern blight in a wide variety of crops. In this study, we isolated a series of dsRNAs from a debilitated S. rolfsii strain, BLH-1, which had pronounced phenotypic aberrations including reduced pathogenicity, mycelial growth and deficient sclerotia production. Virus-curing and horizontal transmission experiments that eliminated or transmitted, respectively, all dsRNA elements showed that the dsRNAs were involved in the hypovirulent traits of BLH-1. Ultrastructure examination also showed hyphae fracture and cytoplasm or organelle degeneration in BLH-1 hyphal cells compared to the virus-free strain. Three assembled cDNA contigs generated from the cDNA library cloned from the purified dsRNA indicated that strain BLH-1 was infected by at least three novel mycoviruses. One has similarity to the hypovirulence-associated Sclerotinia sclerotiorum hypovirus 2 (SsHV2) in the family Hypoviridae, and the other two are related to two different unclassified dsRNA mycovirus families. To our knowledge, this is the first report of S. rolfsii hypovirulence that was correlated with its associated dsRNA.

BRCA1 physically associates with p53 and stimulates its transcriptional activity.

Mutations of the BRCA1 tumor suppressor gene are the most commonly detected alterations in familial breast and ovarian cancer. Although BRCA1 is required for normal mouse development, the molecular basis for its tumor suppressive function remains poorly understood. We show here that BRCA1 increases p53-dependent transcription from the p21WAF1/CIP1 and bax promoters. We also show that BRCA1 and p53 proteins interact both in vitro and in vivo. The interacting regions map, in vitro, to aa 224-500 of BRCA1 and the C-terminal domain of p53. Tumor-derived transactivation-deficient BRCA1 mutants are defective in co-activation of p53-dependent transcription and a truncation mutant of BRCA1 that retains the p53-interacting region acts as a dominant inhibitor of p53-dependent transcription. BRCA1 and p53 cooperatively induce apoptosis of cancer cells. The results indicate that BRCA1 and p53 may coordinately regulate gene expression in their role as tumor suppressors.

Phase I and pharmacokinetic trial of weekly oral fluorouracil given with eniluracil and low-dose leucovorin to patients with solid tumors.

PURPOSE: Fluorouracil (5-FU) given as a weekly, high-dose 24-hour infusion is active and tolerable. We evaluated an oral regimen of eniluracil (which inactivates dihydropyrimidine dehydrogenase [DPD]), 5-FU, and leucovorin to simulate this schedule. PATIENTS AND METHODS: Patients received a single 24-hour infusion of 5-FU (2,300 mg/m(2) on day 2) with leucovorin (15 mg orally [PO] bid on days 1 through 3) to provide reference pharmacokinetic data. Two weeks later, patients began treatment with eniluracil (20 mg) and leucovorin (15 mg) (PO bid on days 1 through 3) and 5-FU (10 to 15 mg/m(2) PO bid on day 2). RESULTS: Dose-limiting toxicity (diarrhea, neutropenia, and fatigue) was seen with 5-FU 15 mg/m(2) PO bid on day 2 given weekly for either 6 of 8 weeks or 3 of 4 weeks, whereas five of seven patients tolerated 5-FU 10 mg/m(2) PO bid given weekly for 3 of 4 weeks. Eniluracil led to a 35-fold reduction in 5-FU clearance. Fluoro-beta-alanine, a 5-FU catabolite, was not detected in plasma during oral 5-FU-eniluracil therapy. DPD activity was markedly suppressed in all patients during eniluracil therapy; the inactivation persisted after the last eniluracil dose; percentages of baseline values were 1.8% on day 5, 4.5% on day 12, and 23.6% on day 19. CONCLUSION: The recommended oral dosage of 5-FU (10 mg/m(2) PO bid) given with eniluracil and leucovorin is approximately 115-fold lower than the reference dosage for 24-hour infusional 5-FU. This difference is greater than expected given the reduction in 5-FU clearance. DPD inactivation persisted for several weeks after completion of eniluracil therapy.