Junjia Zhu

Efficient replication of Epstein–Barr virus in stratified epithelium in vitro

Significance Epstein–Barr virus establishes a life-long latent infection (i.e., no virus is produced) in B cells in most people worldwide. A specific group of latency-associated proteins is expressed in B-cell and epithelial malignancies and likely contributes to tumorigenesis, but the role of epithelial cells in the virus life cycle is not well understood. We grew epithelial cells in organotypic cultures, allowing the cells to differentiate and stratify as they do in vivo. Unlike B-cell infection, EBV infection of epithelial cultures resulted in spontaneous production of high levels of virus, likely expanding the virus pool and increasing efficiency of transmission. This model of EBV-infected epithelium will enhance our understanding of the role of epithelial cells in the EBV life cycle. Epstein–Barr virus is a ubiquitous human herpesvirus associated with epithelial and lymphoid tumors. EBV is transmitted between human hosts in saliva and must cross the oral mucosal epithelium before infecting B lymphocytes, where it establishes a life-long infection. The latter process is well understood because it can be studied in vitro, but our knowledge of infection of epithelial cells has been limited by the inability to infect epithelial cells readily in vitro or to generate cell lines from EBV-infected epithelial tumors. Because epithelium exists as a stratified tissue in vivo, organotypic cultures may serve as a better model of EBV in epithelium than monolayer cultures. Here, we demonstrate that EBV is able to infect organotypic cultures of epithelial cells to establish a predominantly productive infection in the suprabasal layers of stratified epithelium, similar to that seen with Kaposi’s-associated herpesvirus. These cells did express latency-associated proteins in addition to productive-cycle proteins, but a population of cells that exclusively expressed latency-associated viral proteins could not be detected; however, an inability to infect the basal layer would be unlike other herpesviruses examined in organotypic cultures. Furthermore, infection did not induce cellular proliferation, as it does in B cells, but instead resulted in cytopathic effects more commonly associated with productive viral replication. These data suggest that infection of epithelial cells is an integral part of viral spread, which typically does not result in the immortalization or enhanced growth of infected epithelial cells but rather in efficient production of virus.

Mode of First Delivery and Women's Intentions for Subsequent Childbearing: Findings from the First Baby Study

BACKGROUND More than a dozen studies have reported a reduced rate of childbearing after caesarean delivery (CD). It has been hypothesised that this is because women who deliver by CD are less likely to intend to have subsequent children than women who deliver vaginally - either before childbirth or as a consequence of CD. Little research has addressed either of these hypotheses. METHODS As part of an ongoing prospective study, we interviewed 3006 women in their third trimester and 1 month after first childbirth to assess subsequent childbearing intentions. RESULTS Women who delivered by CD were similar to those who delivered vaginally in intent to have at least one additional child, both before childbirth (90.1% vaginal, 89.9% CD; P = 0.97) and after (87.8% vaginal, 87.1% CD; P = 0.87); however, women who had CD were less likely to intend two or more additional children, both before childbirth (34.7% vaginal, 29.2% CD; P = 0.03) and after (32.2% vaginal, 26.1% CD; P = 0.01). Among women who intended to have at least one additional child before childbirth, 5.0% reported intending to have no additional children 1 month after delivery (5.1% vaginal, 4.6% CD; P = 0.52). CONCLUSIONS Women whose first delivery is by CD are less likely to intend a relatively large family of three or more children than those who deliver vaginally, but delivery by CD does not decrease womens intentions to have at least one more child any more than does vaginal delivery, at least in the short term.

Association of prenatal physical activity and gestational weight gain: results from the first baby study.

BACKGROUND In response to increasing rates of excessive gestational weight gain (GWG) and evidence of postpartum weight retention and long-term overweight and obesity, the Institute of Medicine (IOM) revised their guidelines for GWG in 2009. Prenatal physical activity is recommended, although its role in preventing excessive GWG is unclear. We sought to understand the association between prenatal physical activity and GWG in a longitudinal cohort. METHODS During a baseline survey at 34 weeks, women (n = 3,006) reported their height, prepregnancy weight, and physical activity during pregnancy. GWG was self-reported at 1-month postpartum. Multivariable logistic regression adjusting for age, race/ethnicity, education, poverty status, marital status, gestational age at the time of delivery, and smoking was used to model the association between adequate physical activity during pregnancy and exceeding the IOM recommendations for GWG. FINDINGS Overweight women were most likely to exceed the IOM recommendations for GWG (78.7%), followed by obese women and normal weight women (65.0% and 42.4%, respectively). The majority of women participated in some physical activity during pregnancy, with 41.2% engaging in 60 to 149 minutes and 32.1% engaging in at least 150 minutes of physical activity per week. In adjusted analysis, meeting the physical activity guidelines was associated with a 29% (confidence interval, 0.57-0.88) lower odds of exceeding the IOM recommendations for GWG compared with inactive women. CONCLUSIONS Findings of high rates of excessive GWG, especially among women with overweight and obesity, are concerning given the associated health burdens. The association of guideline-concordant physical activity with appropriate GWG suggests this is an important target for future interventions.

The effect of copy number variation in the phase II detoxification genes UGT2B17 and UGT2B28 on colorectal cancer risk

Genetic polymorphisms in combination with the Western‐style diet, physical inactivity, smoking, excessive alcohol consumption, and obesity have been hypothesized to affect colorectal cancer (CRC) risk. Metabolizers of environmental carcinogenic and endogenous compounds affecting CRC risk include the phase II detoxification UDP‐glucuronosyltransferase (UGT) enzymes UGT2B17 and UGT2B28, which are 2 of the most commonly deleted genes in the genome.

Olanzapine metabolism and the significance of UGT1A448V and UGT2B1067Y variants.

Objectives Olanzapine is an antipsychotic used in the treatment of schizophrenia, bipolar disorder, and treatment-resistant depression. Glucuronidation by the UDP-glucuronosyltransferase (UGT) family of enzymes is the major mode of olanzapine metabolism, and polymorphisms in these enzymes could contribute to interindividual variability in olanzapine metabolism and therapeutic response. Methods Cell lines overexpressing individual UGT enzymes were used to determine which UGTs have enzymatic activity against olanzapine, characterize the kinetics of this reaction, and examine the effects of UGT variants on olanzapine metabolism. A bank of 105 human liver microsomes (HLM) were used to perform a phenotype–genotype study comparing glucuronidation activity against UGT genotype. Results Cell lines overexpressing the individual UGTs 1A4 and 2B10 exhibited glucuronidation activity against olanzapine. The UGT1A424Pro/48Val variant exhibited a 3.7-fold (P<0.0001) higher Vmax/KM for the formation of the olanzapine-10-N-glucuronide isomer 1, and a 4.3-fold (P<0.0001) higher Vmax/KM for the formation of the olanzapine-10-N-glucuronide isomer 2 than wild-type UGT1A424Pro/48Leu. The UGT2B1067Y variant exhibited no glucuronidation activity against olanzapine. In a screening of 105 HLM specimens, there was a 2.1-fold (P=0.04) and 1.6-fold (P=0.0017) increase in the rate of olanzapine-10-N-glucuronide isomer 1 and olanzapine-4′-N-glucuronide formation, and a 2-fold (P=0.02) increase in the overall olanzapine glucuronidation formation, in HLM with the UGT1A4 (*3/*3)/UGT2B10 (*1/*1) genotype compared with HLM with the UGT1A4 (*1/*1)/UGT2B10 (*1/*1) genotype. There was a 1.9-fold (P<0.003) decrease in the formation of both isomers of the olanzapine-10-N-glucuronide, a 2.7-fold (P<0.0001) decrease in olanzapine-4′-N-glucuronide formation, and a 2.1-fold (P=0.0002) decrease in the overall olanzapine glucuronide formation in HLM with at least one UGT2B10*2 allele. In regression analysis, the UGT1A4*3 (P<0.02) and UGT2B10*2 (P<0.002) alleles were significant predictors of the formation of all olanzapine glucuronide isomers. Conclusion The UGTs 1A4 and 2B10 glucuronidate olanzapine and functional variants of these UGTs significantly alter olanzapine glucuronidation in vitro. These data suggest that the UGT1A4*3 and UGT2B10*2 alleles contribute significantly to interindividual variability in olanzapine metabolism.

Cost-effectiveness of the National Surgical Quality Improvement Program.

Objective:The purpose of this study was to compare the cost-effectiveness of the National Surgical Quality Improvement Program (NSQIP) at an academic medical center between the first 6 months and through the first and second years of implementation. Background:The NSQIP has been extended to private-sector hospitals since 1999, but little is known about its cost-effectiveness. Methods:Data included 2229 general or vascular surgeries, 699 of which were conducted after NSQIP was in place for 6 months. We estimated an incremental cost-effectiveness ratio (ICER) comparing costs and benefits before and after the adoption of NSQIP. Costs were estimated from the perspective of the hospital and included hospital costs for each admission plus the total annual cost of program adoption and maintenance, including administrator salary, training, and information technology costs. Effectiveness was defined as events avoided. Confidence intervals and a cost-effectiveness acceptability curve were computed by using a set of 10,000 bootstrap replicates. The time periods we compared were (1) July 2007 to December 2007 to July 2008 to December 2008 and (2) July 2007 to June 2008 to July 2008 to June 2009. Results:The incremental costs of the NSQIP program were

Unintended pregnancy and postpartum depression among first-time mothers.

832 and

Ten-year weight gain in smokers who quit, smokers who continued smoking and never smokers in the United States, NHANES 2003-2012.

266 for time periods 1 and 2, respectively, yielding ICERs of

How to cluster gene expression dynamics in response to environmental signals

25,471 and

Nocturnal phenotypical features of obstructive sleep apnea (OSA) in asthmatic children

7319 per event avoided. The cost-effectiveness acceptability curves suggested a high probability that NSQIP was cost-effective at reasonable levels of willingness to pay. Conclusions:In these data, not only did NSQIP appear cost-effective, but also its cost-effectiveness improved with greater duration of participation in the program, resulting in a decline to 28.7% of the initial cost.