Biomedical subjects

C P Chou

Publications and source records attributed to C P Chou.

At least 19 recordsLinked to original sources

PubMed2001

DegP-coexpression minimizes inclusion-body formation upon overproduction of recombinant penicillin acylase in Escherichia coli.

We demonstrated the enhancement of recombinant penicillin acylase (PAC) production in Escherichia coli by increasing the intracellular concentration of the periplasmic protease DegP. Using appropriate host/vector systems (e.g., HB101 harboring pTrcKnPAC2902 or MDDeltaP7 harboring pTrcKnPAC2902) in which the expression of the pac gene was regulated by the strong trc promoter, the overproduction of PAC was often limited by periplasmic processing and inclusion bodies composed of protein aggregates of PAC precursors were formed in the periplasm. The amount of these periplasmic inclusion bodies was significantly reduced and PAC activity was significantly increased upon coexpression of DegP. The specific PAC activity reached an extremely high level of 674 U/L/OD(600) for MDDeltaP7 harboring pTrcKnPAC2902 and pKS12 under optimum culture conditions. However, such improvement in the production of PAC was not observed for the expression systems (e.g., MDDeltaP7 harboring pCLL2902) in which the periplasmic processing was not the step limiting the production of PAC. The results suggest that DegP could in vivo assist the periplasmic processing though the enzyme is shown to be not absolutely required for the formation of active PAC in E. coli. In addition, the steps limiting the production of PAC are identified and the reasons for the formation of PAC inclusion bodies are discussed here.

Bacterial Proteins↗

PubMed2001

Adolescent smoking in Wuhan, China: baseline data from the Wuhan Smoking Prevention Trial.

BACKGROUND: This study reports the prevalence of adolescent smoking in the urban and rural areas of Wuhan, China, the capital of Hubei Province, on the Yangtze River in central China. METHODS: Smoking behavior was examined by age, gender, and urbanicity as part of the Wuhan Smoking Prevention Trial. Subjects included 6994 seventh- to ninth-grade students attending 22 randomly selected schools in urban and rural districts. Outcome measures included lifetime smoking, past-30-day smoking, established smoking (>100 cigarettes in lifetime), and susceptibility to smoking (absence of a firm commitment not to smoke). RESULTS: Lifetime smoking prevalence was 47% among boys and 18% among girls. Past-30-day smoking prevalence was 16% among boys and 4% among girls. Established smoking prevalence was 2% among boys and 0% among girls. The prevalence of susceptibility to smoking was 31% among boys and 10% among girls. Smoking increased significantly with age (p<.0005). Susceptibility was more prevalent in rural areas than in urban areas (p<.05), but there were no urban-rural differences in lifetime, past 30-day smoking, or established smoking. Trend analyses revealed that smoking increased with age more rapidly among boys than among girls (p<.05). Smoking was more prevalent among rural boys than among urban boys, but it was more prevalent among urban girls than among rural girls (p<.05). CONCLUSIONS: Adolescent smoking is a significant public health problem in China. Boys are at particularly high risk, as are girls living in urban areas. Effective smoking prevention programs for adolescents, as well as restrictions on tobacco industry marketing and youth access to tobacco, are needed to prevent tobacco-related morbidity and mortality in China.

Adolescent↗

PubMed2001

Analytic approaches for assessing long-term treatment effects. Examples of empirical applications and findings.

Analytic approaches, including the structural equation model (autoregressive panel model), hierarchical linear model, latent growth curve model, survival/event history analysis, latent transition model, and time-series analysis (interrupted time series, multivariate time-series analysis) are discussed for their applicability to data of different structures and their utility in evaluating temporal effects of treatment. Methods are illustrated by presenting applications of the various approaches in previous studies examining temporal patterns of treatment effects. Recent advancements in these longitudinal modeling approaches and the accompanying computer software development offer tremendous flexibility in examining long-term treatment effects through longitudinal data with varying numbers and intervals of assessment and types of measures. A multimethod assessment will contribute to a more complete understanding of the complex phenomena of the long-term courses of substance use and its treatment.

Alcoholism↗

PubMed1999

Novel strategy for efficient screening and construction of host/vector systems to overproduce penicillin acylase in Escherichia coli.

A novel and simple method of using penicillin for screening of mutant strains with a high penicillin acylase (PAC) activity was developed. Random mutagenesis was conducted using a PAC-producing strain resistant to 6-aminopenicillanic acid (6-APA) as the parent strain and mutants were screened with penicillin at a high concentration. Results suggest that mutants with a high minimum inhibitory concentration for penicillin (MIC(penG)) usually overproduce PAC. Both volumetric and specific PAC activities of a mutant, MD7, were significantly higher than those of the parent strain, HBPAC101 harboring pCLL2902. The mutation(s) resulting in the enhanced expression was mapped on the host chromosome rather than the plasmid. In addition, the mutant strain of MDDeltaP7, derived by elimination of the harbored plasmid in MD7, was demonstrated to be efficient in production of PAC by using the expression plasmids for which expression of the pac gene is limited by translation. An extremely high specific PAC activity of more than 350 U/L/OD(600) was reached upon cultivation of MDDeltaP7 harboring pTrcKnPAC2902 in a bioreactor. As such, the strategy is effective in terms of constructing PAC overproducers and improving the process yield for production of PAC.

Bioreactors↗

PubMed1999

Genetic manipulation to identify limiting steps and develop strategies for high-level expression of penicillin acylase in Escherichia coli.

We have identified the bottleneck steps limiting expression of penicillin acylase (PAC) through comparison of the expression performance for various PAC-expression vectors constructed by genetically modulating the efficiencies of transcription and/or translation of the pac gene. To our knowledge, this is the first report demonstrating that expression of PAC could be limited by various steps, such as transcription, translation, and post-translational steps (i.e. translocation and periplasmic processing), depending on the host/vector systems. Results also indicate that the structure of the wild-type pac gene might not be optimal for direct use in production of PAC using recombinant DNA technology. To improve the gene expression, transcription was enhanced by manipulating certain DNA bases in the pac regulatory region, whereas translation was enhanced by enlarging the spacing between the ribosome binding site and the ATG initiation codon to increase the initiation efficiency. The information is useful in terms of developing genetic strategies for overproduction of recombinant PAC in Escherichia coli.

Cloning, Molecular↗

PubMed1999

Manipulation of carbon assimilation with respect to expression of the pac gene for improving production of penicillin acylase in Escherichia coli.

A strategy of genetically manipulating carbon assimilation with respect to expression of the pac gene was employed for overproduction of recombinant penicillin acylase (PAC). Two expression plasmids of pCLL2902 and pCLL3201, which contain the pac coding region but differ in the pac regulatory region, were constructed for the production experiments. Expression of the pac gene was subjected to phenyl acetic acid (PAA-) induction and glucose catabolite repression for pCLL3201, whereas it was subjected to neither of the two transcriptional regulations for pCLL2902. The specific PAC activity for strains harboring pCLL2902 was significantly higher than that for strains harboring pCLL3201 due to an improved transcription efficiency. In addition, no inclusion bodies were observed upon production of PAC using the current expression systems. The results suggest that using the native pac promoter instead of a strong promoter such as tac for regulation is a feasible approach for production of PAC. The impact of the current expression systems is also significant from a process viewpoint since, using strains harboring pCLL2902, not only could glucose replace PAA as a carbon source of Escherichia coli cultures for production of PAC but also the volumetric PAC activity was highly improved.

Cloning, Molecular↗

PubMed1999

Optimization of the host/vector system and culture conditions for production of penicillin acylase in Escherichia coli.

Culture performance for the production of penicillin acylase (PAC) in a bioreactor was investigated using HB101 or ATCC11105 as the host and pCLL2902, pCLL3201 or pTrcKnPAC2902 as the expression plasmid. We observed that the production of PAC by HB101 harboring pCLL3201 was, similar to ATCC11105, induced by phenyl acetic acid (PAA) and catabolicaily repressed by glucose, whereas the production of PAC by HB101 harboring pCLL2902 did not require PAA for induction and was not repressed by glucose. PAC activity of HB101 harboring pCLL2902 was significantly higher than that of HB101 harboring pCLL3201. There was no significant effect of host or carbon source on the production of PAC using pCLL2902. The production of PAC by HB101 harboring pTrcKnPAC2902, in which the pac gene expression was controlled by the trc promoter system, was about the same as that by HB101 harboring pCLL2902, when the culture was appropriately induced with isopropyl beta-d-thiogalactopyranoside (IPTG). Therefore, the use of both pCLL2902 and pTrcKnPAC2902 could be expected to be feasible for industrial applications. However, optimization of IPTG induction for HB101 harboring pTrcKnPAC2902 might be required, since formation of inclusion bodies tends to limit the production of PAC in some cases.

Journal Article↗

PubMed1999

Cocaine use and high-risk sexual behavior among STD clinic patients.

BACKGROUND AND OBJECTIVES: Crack-smoking sexually transmitted disease (STD) patients are at high-risk for contracting HIV. GOAL OF THE STUDY: To examine the effects of cocaine use and other correlates on high-risk sexual behavior among STD clinic patients. STUDY DESIGN: This was a cross-sectional study of 1,490 consecutive patients attending three Los Angeles County STD clinics between 1992 and 1994. RESULTS: Logistic regression analysis found high-risk sexual activity was associated with being a male and being of younger age. Among women, high-risk sexual behavior was associated with crack cocaine use and a perceived need for help. Among the men in the study, ethnicity (being black) and having an arrest history were associated with high-risk behavior. CONCLUSIONS: Effective intervention strategies should address cocaine use among STD patients and provide them with referrals to drug treatment.

Adolescent↗

PubMed1998

Effects of a community-based prevention program on decreasing drug use in high-risk adolescents.

OBJECTIVES: This study investigated the secondary prevention effects of a substance abuse primary prevention program. METHODS: Logistic regression analyses were conducted on 4 waves of follow-up data from sixth- and seventh-grade baseline users of cigarettes, alcohol, and marijuana taking part in a school-based program in Indianapolis. RESULTS: The program demonstrated significant reductions in cigarette use at the initial follow-up (6 months) and alcohol use at the first 2 follow-ups (up to 1.5 years). Models considering repeated measures also showed effects on all 3 substances. CONCLUSIONS: Primary prevention programs are able to reach and influence high-risk adolescents in a nonstigmatizing manner.

Adolescent↗

PubMed1998

Prospective psychosocial, interpersonal, and behavioral predictors of handgun carrying among adolescents.

OBJECTIVES: This study identified behavioral and psychosocial/interpersonal factors in young adolescence that are associated with handgun carrying in later adolescence. METHODS: A sample of 2200 high school students was surveyed at 9th grade and again at 12th grade. RESULTS: Multivariate logistic regression analyses indicated that measures of risk-taking preference, depression, stress, temper, and drug use assessed while the students were in 9th grade were predictive of handgun carrying in 12th grade for both male and female students. CONCLUSIONS: These findings suggest the need for a comprehensive approach to prevention that focuses on both individual and interpersonal factors associated with adolescents' decision to carry a handgun.

Adolescent↗

PubMed1998

Interaction effects of client and treatment program characteristics on retention: an exploratory analysis using hierarchical linear models.

This paper applied a hierarchical linear modeling approach to explore the interaction effects of treatment program and client characteristics on client retention in treatment for drug users. Program characteristics included services provision, funding sources, and staff-client gender congruence, and client characteristics included gender, age at admission, and drug use level prior to admission. The same model was applied separately to three modalities: residential, methadone maintenance, and outpatient drug-free programs. Data were obtained from 59 treatment programs and 3,764 of their clients who had discharge records. The most noteworthy significant interaction effect detected was program's funding source and client's gender on treatment retention in the outpatient drug-free modality. For example, female clients remained less time in the programs that accepted only public funding than in the programs that accepted both public and private funding. Male clients remained in the treatment an average of 25.3 fewer days than female clients in drug-free programs that only accepted public fund, but stayed about the same time as females if the programs received mixed funding.

Adult↗

PubMed1996

Pattern reliability of narcotics addicts' self-reported data: a confirmatory assessment of construct validity and consistency.

Pattern reliability, or the invariance of relationships among variables, was investigated in this study. The consistency of theoretical constructs reflected by measures taken at two separate occasions can be tested using confirmatory factor analysis. Self-report data were obtained from 323 narcotics addicts in two face-to-face interviews conducted in 1974/75 and 1985/86. The two interviews overlapped approximately 4 years between 1970 and 1974/75. Through the testing of the invariance of measurement and structural models, pattern reliability was confirmed in one of the models developed. Explication of pattern reliability offers an alternative means of assessing validity of self-report data.

Adult↗

PubMed1994

Measurement invariance in longitudinal clinical research assuming change from development and intervention.

Valid and reliable measures of psychological and behavioral constructs are critical to clinical research, particularly longitudinal treatment research, in which multiple groups are compared over time for possible changes within and between constructs as a result of intervention or development. Structural equation modeling (SEM) analysis is a class of statistical procedures that can be used to test multiple hypotheses about these relationships simultaneously while controlling for measurement error. The procedures have been applied primarily to testing between-construct relationships in nonexperimental studies, with relatively little emphasis on establishing whether measures are sufficiently invariant across groups and across time to permit these tests. This article uses an empirical example of a longitudinal experimental prevention study with two groups to illustrate the use of SEM, first, to systematically test measurement invariance across groups at each wave of measurement, and second, after establishing measurement invariance, to test structural invariance longitudinally.

Adolescent↗

PubMed1992

Dual stable point model of muscle activation and deactivation.

Two dynamic models of muscle activation and deactivation based on the concepts of ion transport, reaction rates, and muscle mechanics are proposed. Storage release and uptake of calcium by the sarcoplasmic reticulum, and a two-step chemical reaction of calcium and troponin are included in the first model. This is a concise version of the complex chemical reactions of muscle activation and deactivation in sarcoplasm. The second model is similar to the first, but calcium-troponin reactions are simplified into two nonlinear rates functions. Due to these nonlinear dynamics, the second model can explain the catch-like enhancement of isometric force response. Simulation results which match experimental data are shown. Also, two new phenomena which need further experiment to verify are predicted by the second model.

Animals↗

PubMed1991

Scaled test statistics and robust standard errors for non-normal data in covariance structure analysis: a Monte Carlo study.

Research studying robustness of maximum likelihood (ML) statistics in covariance structure analysis has concluded that test statistics and standard errors are biased under severe non-normality. An estimation procedure known as asymptotic distribution free (ADF), making no distributional assumption, has been suggested to avoid these biases. Corrections to the normal theory statistics to yield more adequate performance have also been proposed. This study compares the performance of a scaled test statistic and robust standard errors for two models under several non-normal conditions and also compares these with the results from ML and ADF methods. Both ML and ADF test statistics performed rather well in one model and considerably worse in the other. In general, the scaled test statistic seemed to behave better than the ML test statistic and the ADF statistic performed the worst. The robust and ADF standard errors yielded more appropriate estimates of sampling variability than the ML standard errors, which were usually downward biased, in both models under most of the non-normal conditions. ML test statistics and standard errors were found to be quite robust to the violation of the normality assumption when data had either symmetric and platykurtic distributions, or non-symmetric and zero kurtotic distributions.

Analysis of Variance↗

PubMedDate not supplied

The anthroform biorobotic arm: a system for the study of spinal circuits.

This paper reports the design, construction, and testing of a replica of the human arm, which aims to be dynamically as well as kinematically accurate. The arm model is actuated with McKibben pneumatic artificial muscles, and controlled by a special purpose digital signal processing system designed to simulate spinal neural networks in real time. An artificial muscle spindle has also been designed and tested. Design and test data are reviewed, and the paper describes how we hope to use the system to improve our understanding of the reflexive control of human movement and posture.

Arm↗

PubMedDate not supplied

Identification of adolescents at risk for smoking initiation: validation of a measure of susceptibility.

Primary prevention of smoking in adolescents requires effective screening instruments for identifying those adolescents who are most likely to experiment with cigarettes. This study investigated the predictive value of a measure of susceptibility to smoking (the lack of a firm commitment not to smoke) for predicting smoking initiation 1 and 2 years later among 687 seventh-grade nonsmokers. Results showed that susceptible adolescents were approximately two to three times more likely to experiment with cigarettes during the ensuing 2 years than were nonsusceptible adolescents. At the lower levels of smoking, these relationships persisted even after controlling for psychosocial variables. Measures of susceptibility to smoking could be an effective tool for identifying adolescents at increased risk of experimenting with cigarettes or assessing their readiness for smoking-prevention programs.

Adolescent↗

PubMedDate not supplied

An approach for enhancing heterologous production of Providencia rettgeri penicillin acylase in Escherichia coli.

Heterologous production of Providencia rettgeri penicillin acylase (PAC) was optimized in Escherichia coli. Several factors, including carbon, temperature, and host effects, were identified to be critical for the enzyme overproduction. The optimum culture conditions for the enzyme production vary for different host/vector systems. With the optimization, both volumetric and specific PAC activities could be significantly improved by more than 50-fold compared to the native expression in P. rettgeri. The heterologous production could be possibly limited by translation or posttranslational steps, depending on the culture temperature and host/vector system. To our knowledge, this is the first evidence demonstrating the limiting step for the production of P. rettgeri PAC and the existence of the P. rettgeri PAC precursor.

Culture Media↗