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1.  The Enhancement of Bone Regeneration by Gene Activated Matrix Encoding for Platelet Derived Growth Factor 
Biomaterials  2013;35(2):10.1016/j.biomaterials.2013.10.021.
Gene therapy using non-viral vectors that are safe and efficient in transfecting target cells is an effective approach to overcome the shortcomings of protein delivery of growth factors. The objective of this study was to develop and test a non-viral gene delivery system for bone regeneration utilizing a collagen scaffold to deliver polyethylenimine (PEI)-plasmid DNA (pDNA) [encoding platelet derived growth factor-B (PDGF-B)] complexes. The PEI-pPDGF-B complexes were fabricated at amine (N) to phosphate (P) ratio of 10 and characterized for size, surface charge, and in vitro cytotoxicity and transfection efficacy in human bone marrow stromal cells (BMSCs). The influence of the complex-loaded collagen scaffold on cellular attachment and recruitment was evaluated in vitro using microscopic techniques. The in vivo regenerative capacity of the gene delivery system was assessed in 5 mm diameter critical-sized calvarial defects in Fisher 344 rats. The complexes were ~100 nm in size with a positive surface charge. Complexes prepared at an N/P ratio of 10 displayed low cytotoxicity as assessed by a cell viability assay. Confocal microscopy revealed significant proliferation of BMSCs on complex-loaded collagen scaffolds compared to empty scaffolds. In vivo studies showed significantly higher new bone volume/total volume (BV/TV) % in calvarial defects treated with the complex-activated scaffolds following 4 weeks of implantation (14- and 44-fold higher) when compared to empty defects or empty scaffolds, respectively. Together, these findings suggest that non-viral PDGF-B gene-activated scaffolds are effective for bone regeneration and are an attractive gene delivery system with significant potential for clinical translation.
doi:10.1016/j.biomaterials.2013.10.021
PMCID: PMC3855224  PMID: 24161167
Plasmid DNA; Polyethylenimine; Gene Delivery; Scaffold; Platelet Derived Growth Factor; Collagen; Bone regeneration
3.  Crosswalk between DSM-IV Dependence and DSM-5 Substance Use Disorders for Opioids, Cannabis, Cocaine and Alcohol 
Drug and alcohol dependence  2013;132(0):387-390.
Background
Ascertaining agreement between DSM-IV and DSM-5 is important to determine the applicability of treatments for DSM-IV conditions to persons diagnosed according to the proposed DSM-5.
Methods
Data from a nationally representative sample of US adults were used to compare concordance of past-year DSM-IV Opioid, Cannabis, Cocaine and Alcohol Dependence with past-year DSM-5 disorders at thresholds of 3+, 4+ 5+ and 6+ positive DSM-5 criteria among past-year users of opioids (n=264), cannabis (n=1,622), cocaine (n=271) and alcohol (n=23,013). Substance-specific 2×2 tables yielded overall concordance (kappa), sensitivity, specificity, positive predictive values (PPV) and negative predictive values (NPV).
Results
For DSM-IV Alcohol, Cocaine and Opioid Dependence, optimal concordance occurred when 4+ DSM-5 criteria were endorsed, corresponding to the threshold for moderate DSM-5 Alcohol, Cocaine and Opioid Use Disorders. Maximal concordance of DSM-IV Cannabis Dependence and DSM-5 Cannabis Use Disorder occurred when 6+ criteria were endorsed, corresponding to the threshold for severe DSM-5 Cannabis Use Disorder. At these optimal thresholds, sensitivity, specificity, PPV and NPV generally exceeded 85% (>75% for cannabis).
Conclusions
Overall, excellent correspondence of DSM-IV Dependence with DSM-5 Substance Use Disorders was documented in this general population sample of alcohol, cannabis, cocaine and opioid users. Applicability of treatments tested for DSM-IV Dependence is supported by these results for those with a DSM-5 Alcohol, Cocaine or Opioid Use Disorder of at least moderate severity or Severe Cannabis Use Disorder. Further research is needed to provide evidence for applicability of treatments for persons with milder substance use disorders.
doi:10.1016/j.drugalcdep.2013.02.036
PMCID: PMC3748225  PMID: 23642316
DSM-IV; DSM-5; substance use disorder; concordance; kappa; diagnosis
4.  Phenotypic Diversity in Caucasian Adults with Moderate to Severe Class III Malocclusion 
INTRODUCTION
Class III malocclusion is characterized by a composite of dento-skeletal patterns that lead to the forward positioning of the mandibular teeth in relation to the maxillary teeth and a concave profile. Environmental and genetic factors are associated with this condition, which affects 1% of the US population and imposes significant esthetic and functional burdens on affected individuals. The purpose of this study was to capture the phenotypic variation present in a large sample of white adults with Class III malocclusion by using multivariate reduction methods.
METHODS
Sixty-three lateral cephalometric variables were measured from pre-treatment records of 292 Class II Caucasian adults (126 males, 166 females; ages 16-57 years). Principal component analysis and cluster analysis were used to capture the phenotypic variation and identify the most homogeneous groups of individuals to reduce genetic heterogeneity.
RESULTS
Principal component analysis resulted in 6 principal components that accounted for 81.2% of the variation. The first three components represented variations in mandibular horizontal and vertical position, maxillary horizontal position, and mandibular incisor angulation, respectively. The cluster model identified 5 distinct subphenotypes of Class III malocclusion.
CONCLUSIONS
A spectrum of phenotypic definitions was obtained replicating results of previous studies and supporting the validity of these phenotypic measures in future research of genetic and environmental etiology of Class III malocclusion.
doi:10.1016/j.ajodo.2013.02.019
PMCID: PMC3964133  PMID: 23810043
5.  High Risks of Losing Genetic Diversity in an Endemic Mauritian Gecko: Implications for Conservation 
PLoS ONE  2014;9(6):e93387.
Genetic structure can be a consequence of recent population fragmentation and isolation, or a remnant of historical localised adaptation. This poses a challenge for conservationists since misinterpreting patterns of genetic structure may lead to inappropriate management. Of 17 species of reptile originally found in Mauritius, only five survive on the main island. One of these, Phelsuma guimbeaui (lowland forest day gecko), is now restricted to 30 small isolated subpopulations following severe forest fragmentation and isolation due to human colonisation. We used 20 microsatellites in ten subpopulations and two mitochondrial DNA (mtDNA) markers in 13 subpopulations to: (i) assess genetic diversity, population structure and genetic differentiation of subpopulations; (ii) estimate effective population sizes and migration rates of subpopulations; and (iii) examine the phylogenetic relationships of haplotypes found in different subpopulations. Microsatellite data revealed significant population structure with high levels of genetic diversity and isolation by distance, substantial genetic differentiation and no migration between most subpopulations. MtDNA, however, showed no evidence of population structure, indicating that there was once a genetically panmictic population. Effective population sizes of ten subpopulations, based on microsatellite markers, were small, ranging from 44 to 167. Simulations suggested that the chance of survival and allelic diversity of some subpopulations will decrease dramatically over the next 50 years if no migration occurs. Our DNA-based evidence reveals an urgent need for a management plan for the conservation of P. guimbeaui. We identified 18 threatened and 12 viable subpopulations and discuss a range of management options that include translocation of threatened subpopulations to retain maximum allelic diversity, and habitat restoration and assisted migration to decrease genetic erosion and inbreeding for the viable subpopulations.
doi:10.1371/journal.pone.0093387
PMCID: PMC4070904  PMID: 24963708
6.  ORGANIZATION, BARRIER FUNCTION AND ANTIMICROBIAL LIPIDS OF THE ORAL MUCOSA 
Synopsis
As one moves from the skin across the vermilion region of the lip and into the oral cavity the oral mucosa is encountered. The oral mucosa consists of connective tissue known as the lamina propria covered by a stratified squamous epithelium. In the regions of the hard palate and gingiva the epithelium is keratinized like the epidermis. In the buccal region, the floor of the mouth and the underside of the tongue the epithelium is nonkeratinized. The epithelium on the dorsum of the tongue is a specialized epithelium but can be approximated as a mosaic of keratinized and nonkeratinized epithelia. The nonkeratinized epithelial regions do not produce a stratum corneum. Nuclei with intact DNA are retained in the superficial cells. In all regions the outer portions of the epithelium provides a protective permeability barrier, which varies regionally. Antimicrobial lipids at the surfaces of the oral mucosa are an integral part of innate immunity.
doi:10.1111/ics.12038
PMCID: PMC3640763  PMID: 23320785
mouth mucosa; keratinocytes; keratin; cell envelope; ceramide; permeability
7.  PROSPECTIVE CORRELATES OF DRINKING CESSATION: VARIATION ACROSS THE LIFE COURSE 
Addiction (Abingdon, England)  2013;108(4):712-722.
Aims
To assess age variation in correlates of drinking cessation.
Design
Prospective study of a U.S. general population sample.
Setting
Face-to-face household interviews.
Participants
Past-year ≥monthly drinkers interviewed at baseline and 3-year follow-up (n=14,885).
Measurements
Baseline values and selected changes over follow-up in alcohol consumption, alcohol use disorder (AUD), sociodemographic and health characteristics, other substance use and psychiatric comorbidity were used to predict drinking cessation in three age groups.
Findings
Correlates of drinking cessation varied over the life course, with pregnancy/presence of an infant, nicotine or drug use disorder, incident AUD, cluster A personality disorder, liver disease and incident retirement being important at younger ages and high-school graduation, family income ≥$70,000, volume of ethanol intake, Asian race/ethnicity, mood disorder and incident cardiovascular disease being significant at older ages. Age-invariant correlates included smoking cessation over follow-up, OR=2.82 (95% CI=1.62–4.92) to 3.45 (2.20–5.39); college education, OR=0.42 (0.27–0.65) to 0.54 (0.36–0.83); Black and Hispanic race/ethnicity, OR = 1.74 (1.18–2.29) to 1.88 (1.21–2.93) and 1.58 (1.11–1.25) to 1.73 (0.83–3.63), respectively, and months since last drink, OR=1.24 (1.13–1.36) to1.29 (1.19–1.39).
Conclusions
Factors associated with ceasing alcohol use in US adults appear to differ over the life course, reflecting age variation in both their prevalence and impact and supporting the importance of role transitions and health problems (the “sick quitter” effect). The most consistent correlates of drinking cessation included factors reflecting ability/inability to give up potentially addictive substances and factors associated with perceived acceptability of drinking and subgroup-specific drinking contexts that might facilitate/impede continued drinking.
doi:10.1111/add.12079
PMCID: PMC3602325  PMID: 23216848
drinking cessation; former drinkers; sick quitters
8.  Sphingoid bases are taken up by Escherichia coli and Staphylococcus aureus and induce ultrastructural damage 
Sphingoid bases found in the outer layers of the skin exhibit antimicrobial activity against Gram-positive and Gram-negative bacteria. We investigated the uptake of several sphingoid bases by Escherichia coli and Staphylococcus aureus, and assessed subsequent ultrastructural damage. E. coli and S. aureus were incubated with D-sphingosine, dihydrosphingosine, or phytosphingosine at ten times their MIC for 0.5 h and 4 h, respectively, to kill 50% of viable bacteria. Treated bacterial cells were immediately prepared for SEM, TEM, and analyzed for lipid content by QTLC. E. coli and S. aureus treated with sphingoid bases were distorted and their surfaces were concave and rugate. Significant differences were observed in the visual surface area relative to controls for both E. coli and S. aureus when treated with dihydrosphingosine and sphingosine (p<0.0001) but not phytosphingosine. While sphingoid base-treated S. aureus exhibited disruption and loss of cell wall and membrane, E. coli cytoplasmic membranes appeared intact and the outer envelope uncompromised. Both E. coli and S. aureus cells contained unique internal inclusion bodies, likely associated with cell death. QTLC demonstrated extensive uptake of sphingoid bases by the bacteria. Hence, sphingoid bases induce both extracellular and intracellular damage and cause intracellular inclusions that may reflect lipid uptake.
doi:10.1159/000343175
PMCID: PMC3634627  PMID: 23128426
Antimicrobial lipids; sphingoid bases; sphingolipids; Escherichia coli; Staphylococcus aureus; ultrastructure; electron microscopy
9.  DIFFERENCES IN THE PROFILES OF DSM-IV AND DSM-5 ALCOHOL USE DISORDERS: IMPLICATIONS FOR CLINICIANS 
Background
Existing information on consequences of the DSM-5 revision for diagnosis of alcohol use disorders (AUD) has gaps, including missing information critical to understanding implications of the revision for clinical practice.
Methods
Data from Wave 2 of the National Epidemiologic Survey on Alcohol and Related Conditions were used to compare AUD severity, alcohol consumption and treatment, sociodemographic and health characteristics and psychiatric comorbidity among individuals with DSM-IV abuse versus DSM-5 moderate AUD and DSM-IV dependence versus DSM-5 severe AUD. For each pair of disorders, we additionally compared three mutually exclusive groups: individuals positive solely for the DSM-IV disorder, those positive solely for the DSM-5 disorder and those positive for both.
Results
Whereas 80.5% of individuals positive for DSM-IV dependence were positive for DSM-5 severe AUD, only 58.0% of those positive for abuse were positive for moderate AUD. The profiles of individuals with DSM-IV dependence and DSM-5 severe AUD were almost identical. The only significant (p<.005) difference, more AUD criteria among the former, reflected the higher criterion threshold (≥4 vs. ≥3) for severe AUD relative to dependence. In contrast, the profiles of individuals with DSM-5 moderate AUD and DSM-IV abuse differed substantially. The former endorsed more AUD criteria, had higher rates of physiological dependence, were less likely to be White and male, had lower incomes, were less likely to have private and more likely to have public health insurance, and had higher levels of comorbid anxiety disorders than the latter.
Conclusions
Similarities between the profiles of DSM-IV and DSM-5 AUD far outweigh differences; however, clinicians may face some changes with respect to appropriate screening and referral for cases at the milder end of the AUD severity spectrum, and the mechanisms through which these will be reimbursed may shift slightly from the private to public sector.
doi:10.1111/j.1530-0277.2012.01930.x
PMCID: PMC3800556  PMID: 22974144
DSM-5; AUD; treatment; severity; clinical profile
10.  Comparative Performance of the AUDIT-C in Screening for DSM-IV and DSM-5 Alcohol Use Disorders* 
Drug and alcohol dependence  2012;126(3):384-388.
Objective
Under the proposed DSM-5 revision to the criteria for alcohol use disorder (AUD), a substantial proportion of DSM-IV AUD cases will be lost or shifted in terms of severity, with some new cases added. Accordingly, the performance of the AUDIT-C in screening for DSM-IV AUD cannot be assumed to extend to DSM-5 AUD. The objective of this paper is to compare the AUDIT-C in screening for DSM-IV and DSM-5 AUD.
Methods
Using a broad range of performance metrics, the AUDIT-C was tested and contrasted as a screener for DSM-IV AUD (any AUD, abuse and dependence) and DSM-5 AUD (any AUD, moderate AUD and severe AUD) in a representative sample of U.S. adults aged 21 and older and among past-year drinkers.
Results
Optimal AUDIT-C cutpoints were identical for DSM-IV and DSM-5 AUD: ≥4 for any AUD, ≥3 or ≥4 for abuse/moderate AUD and ≥4 or ≥5 for dependence/severe AUD. Screening performance was slightly better for DSM-5 severe AUD than DSM-IV dependence but did not differ for other diagnoses. At optimal screening cutpoints, positive predictive values were slightly higher for DSM-5 overall AUD and moderate AUD than for their DSM-IV counterparts. Sensitivities were slightly higher for DSM-5 severe AUD than DSM-IV dependence. Optimal screening cutpoints shifted upwards for past-year drinkers but continued to be identical for DSM-IV and DSM-5 disorders.
Conclusions
Clinicians should not face any major overhaul of their current screening procedures as a result of the DSM-5 revision and should benefit from fewer false positive screening results.
doi:10.1016/j.drugalcdep.2012.05.029
PMCID: PMC3469743  PMID: 22728044
AUDIT-C; screening; alcohol use disorder; DSM-IV; DSM-5
11.  Antisocial Behavioral Syndromes and Three-Year Quality of Life Outcomes in United States Adults 
Acta psychiatrica Scandinavica  2012;126(2):10.1111/j.1600-0447.2012.01848.x.
Objective
To examine 3-year quality-of-life (QOL) outcomes among United States adults with Diagnostic and Statistical Manual of Mental Disorders – Fourth Edition (DSM-IV) antisocial personality disorder (ASPD), syndromal adult antisocial behavior without conduct disorder (CD) before age 15 (AABS, not a DSM-IV diagnosis), or no antisocial behavioral syndrome at baseline.
Method
Face-to-face interviews (n= 34,653). Psychiatric disorders were assessed using the Alcohol Use Disorder and Associated Disabilities Interview Schedule – DSM-IV Version. Health-related QOL was assessed using the Short-Form 12-Item Health Survey, version 2 (SF-12v2). Other outcomes included past-year Perceived Stress Scale-4 (PSS-4) scores, employment, receipt of Supplemental Security Income (SSI), welfare, and food stamps, and participation in social relationships.
Results
ASPD and AABS predicted poorer employment, financial dependency, social relationship, and physical health outcomes. Relationships of antisociality to SSI and food stamp receipt and physical health scales were modified by baseline age. Both antisocial syndromes predicted higher PSS-4, AABS predicted lower SF-12v2 Vitality, and ASPD predicted lower SF-12v2 Social Functioning scores in women.
Conclusion
Similar prediction of QOL by ASPD and AABS suggests limited utility of requiring CD before age 15 to diagnose ASPD. Findings underscore the need to improve prevention and treatment of antisocial syndromes.
doi:10.1111/j.1600-0447.2012.01848.x
PMCID: PMC3837547  PMID: 22375904
Antisocial personality disorder; quality of life; epidemiology; longitudinal course
12.  Dental Caries in a Cohort of Very Young American Indian Children 
OBJECTIVES
This paper reports the prevalence and severity of caries in a group of 16-month-old American Indian children.
METHODS
The study is an ongoing longitudinal study of risk factors for caries in children from a Northern Plains Tribal community. Children were examined for caries and risk factor data collected at approximately 1, 4, 8, 12 and 16 months of age. Surface-specific caries data were collected and the presence of pre-cavitated “white spot” lesions was recorded at the subject level.
RESULTS
The mean age was 15.4 months for the sample of 232 children. Caries prevalence was 31.9%, while an additional 29.3% had white spot lesions only. Mean dmfs was 1.57, and ranged from 0 to 44 surfaces. Nearly 3% of all erupted tooth surfaces were affected and maxillary central incisors had the highest prevalence of caries (22%).
CONCLUSIONS
Among the very youngest children, dental caries prevalence was very high among these American Indian children.
doi:10.1111/j.1752-7325.2012.00372.x
PMCID: PMC3509261  PMID: 23017107
13.  Oral mucosal lipids are antibacterial against Porphyromonas gingivalis, induce ultrastructural damage, and alter bacterial lipid and protein compositions 
Oral mucosal and salivary lipids exhibit potent antimicrobial activity for a variety of Gram-positive and Gram-negative bacteria; however, little is known about their spectrum of antimicrobial activity or mechanisms of action against oral bacteria. In this study, we examine the activity of two fatty acids and three sphingoid bases against Porphyromonas gingivalis, an important colonizer of the oral cavity implicated in periodontitis. Minimal inhibitory concentrations, minimal bactericidal concentrations, and kill kinetics revealed variable, but potent, activity of oral mucosal and salivary lipids against P. gingivalis, indicating that lipid structure may be an important determinant in lipid mechanisms of activity against bacteria, although specific components of bacterial membranes are also likely important. Electron micrographs showed ultrastructural damage induced by sapienic acid and phytosphingosine and confirmed disruption of the bacterial plasma membrane. This information, coupled with the association of treatment lipids with P. gingivalis lipids revealed via thin layer chromatography, suggests that the plasma membrane is a likely target of lipid antibacterial activity. Utilizing a combination of two-dimensional in-gel electrophoresis and Western blot followed by mass spectroscopy and N-terminus degradation sequencing we also show that treatment with sapienic acid induces upregulation of a set of proteins comprising a unique P. gingivalis stress response, including proteins important in fatty acid biosynthesis, metabolism and energy production, protein processing, cell adhesion and virulence. Prophylactic or therapeutic lipid treatments may be beneficial for intervention of infection by supplementing the natural immune function of endogenous lipids on mucosal surfaces.
doi:10.1038/ijos.2013.28
PMCID: PMC3967327  PMID: 23867843
antimicrobial lipid; fatty acid; Porphyromonas gingivalis; sphingoid base; sphingolipid; ultrastructure
14.  CORRELATES OF RECOVERY FROM ALCOHOL DEPENDENCE: A PROSPECTIVE STUDY OVER A 3-YEAR FOLLOW-UP INTERVAL 
Background
Correlates of recovery from alcohol dependence have been identified through a variety of study designs characterized by different strengths and limitations. The goal of this study was to compare correlates of recovery based on a 3-year prospective design with those based on cross-sectional analyses of data from the same source.
Methods
Data from the 2001-2002 Wave 1 and 2004-2005 Wave 2 National Epidemiologic Survey on Alcohol and Related Conditions (NESARC) were used to examine baseline characteristics associated with Wave 2 recovery from alcohol dependence, among those who classified with past-year DSM-IV alcohol dependence at Wave 1 (n=1,172).
Results
Abstinent recovery (AR) was significantly associated with Black/Asian/Hispanic race/ethnicity, children <1 year of age in the household at baseline, attending religious services ≥weekly at follow-up, and having initiated help seeking that comprised/included 12-step participation within <3 years prior to baseline. Nonabstinent recovery (NR) was positively associated with being never married at baseline, having job problems or being unemployed in the year preceding baseline, attending religious services
Conclusions
Various aspects of study design must be considered when interpreting correlates of recovery. Cross-sectional analyses of lifetime correlates of recovery are highly subject to misinterpretation, but pseudo-prospective survival analyses with time-dependent covariates may yield results as valid as those from prospective studies.
doi:10.1111/j.1530-0277.2011.01729.x
PMCID: PMC3349820  PMID: 22309217
alcohol dependence; abstinent recovery; nonabstinent recovery; study design; prospective
BMC Genomics  2013;14:176.
Background
Microsatellites are widely used for many genetic studies. In contrast to single nucleotide polymorphism (SNP) and genotyping-by-sequencing methods, they are readily typed in samples of low DNA quality/concentration (e.g. museum/non-invasive samples), and enable the quick, cheap identification of species, hybrids, clones and ploidy. Microsatellites also have the highest cross-species utility of all types of markers used for genotyping, but, despite this, when isolated from a single species, only a relatively small proportion will be of utility. Marker development of any type requires skill and time. The availability of sufficient “off-the-shelf” markers that are suitable for genotyping a wide range of species would not only save resources but also uniquely enable new comparisons of diversity among taxa at the same set of loci. No other marker types are capable of enabling this. We therefore developed a set of avian microsatellite markers with enhanced cross-species utility.
Results
We selected highly-conserved sequences with a high number of repeat units in both of two genetically distant species. Twenty-four primer sets were designed from homologous sequences that possessed at least eight repeat units in both the zebra finch (Taeniopygia guttata) and chicken (Gallus gallus). Each primer sequence was a complete match to zebra finch and, after accounting for degenerate bases, at least 86% similar to chicken. We assessed primer-set utility by genotyping individuals belonging to eight passerine and four non-passerine species. The majority of the new Conserved Avian Microsatellite (CAM) markers amplified in all 12 species tested (on average, 94% in passerines and 95% in non-passerines). This new marker set is of especially high utility in passerines, with a mean 68% of loci polymorphic per species, compared with 42% in non-passerine species.
Conclusions
When combined with previously described conserved loci, this new set of conserved markers will not only reduce the necessity and expense of microsatellite isolation for a wide range of genetic studies, including avian parentage and population analyses, but will also now enable comparisons of genetic diversity among different species (and populations) at the same set of loci, with no or reduced bias. Finally, the approach used here can be applied to other taxa in which appropriate genome sequences are available.
doi:10.1186/1471-2164-14-176
PMCID: PMC3738869  PMID: 23497230
Drug and Alcohol Review  2011;31(2):141-150.
Introduction and Aims
This paper proposes an approach for evaluating the validity of alternative low-risk drinking guidelines.
Design and Methods
Twenty-seven alternative guidelines were evaluated in terms of their ability to predict 9 measures of concurrent and prospective alcohol-related harm, using longitudinal data from a nationally representative sample of U.S. adults (n=26,438 to 12,339 depending upon outcome). Parameters compared included sensitivity, specificity, adjusted odds ratios and measures of model fit.
Results
Performance varied by harm. The guidelines that best predicted concurrent alcohol-related harm comprised daily-only limits of 4/3 drinks for men/women, but gender-invariant limits of 4/4 drinks also performed well. Adding weekly limits did little to improve the prediction of concurrent harm. The guidelines that best predicted prospective harm comprised daily limits of 4/4 drinks combined with weekly limits of 14 drinks for men and 7 drinks for women, with weekly limits of 14/14 drinks running second. When concurrent and incident harms were aggregated, daily-only limits of 4/3 drinks performed nearly on a par with the combination of 14/14 drinks per week and 4/3 drinks per day.
Discussion
This paper supported gender-specific daily limits and suggested that optimal guidelines might take daily limits from analyses of concurrent harms and weekly limits from analyses of prospective harms.
Conclusions
This paper illustrates a mechanism for validating the ability of low-risk drinking guidelines to accurately predict a range of alcohol-related harms, whereby countries could use their own data on consumption and its association with harm to evaluate their low-risk drinking guidelines.
doi:10.1111/j.1465-3362.2011.00335.x
PMCID: PMC3252404  PMID: 21954858
drinking guidelines; alcohol-related harm; validity; prediction
PLoS ONE  2013;8(1):e51923.
Background
Alcohol has been linked to health disparities between races in the US; however, race-specific alcohol-attributable mortality has never been estimated. The objective of this article is to estimate premature mortality attributable to alcohol in the US in 2005, differentiated by race, age and sex for people 15 to 64 years of age.
Methods and Findings
Mortality attributable to alcohol was estimated based on alcohol-attributable fractions using indicators of exposure from the National Epidemiologic Survey on Alcohol and Related Conditions and risk relations from the Comparative Risk Assessment study. Consumption data were corrected for undercoverage (the observed underreporting of alcohol consumption when using survey as compared to sales data) using adult per capita consumption from WHO databases. Mortality data by cause of death were obtained from the US Department of Health and Human Services. For people 15 to 64 years of age in the US in 2005, alcohol was responsible for 55,974 deaths (46,461 for men; 9,513 for women) representing 9.0% of all deaths, and 1,288,700 PYLL (1,087,280 for men; 201,420 for women) representing 10.7% of all PYLL. Per 100,000 people, this represents 29 deaths (29 for White; 40 for Black; 82 for Native Americans; 6 for Asian/Pacific Islander) and 670 PYLL (673 for White; 808 for Black; 1,808 for Native American; 158 for Asian/Pacific Islander). Sensitivity analyses showed a lower but still substantial burden without adjusting for undercoverage.
Conclusions
The burden of mortality attributable to alcohol in the US is unequal among people of different races and between men and women. Racial differences in alcohol consumption and the resulting harms explain in part the observed disparities in the premature mortality burden between races, suggesting the need for interventions for specific subgroups of the population such as Native Americans.
doi:10.1371/journal.pone.0051923
PMCID: PMC3534703  PMID: 23300957
The Journal of clinical psychiatry  2011;72(11):1494-1502.
Background
Associations between overweight and obesity and medical conditions have been extensively studied, but little is known about their relationships to psychiatric disorders.
Objective
To present nationally representative findings on the prospective relationships between overweight and obesity and DSM-IV substance use, mood and anxiety disorders.
Design, Setting and Participants
Waves 1 and 2 of the National Epidemiologic Survey on Alcohol and Related Conditions, a nationally representative sample of 34,653 U.S. adults.
Main Outcome Measures
Incidence of DSM-IV substance use, mood and anxiety disorders and changes in BMI status during the 3-year follow-up.
Results
Regression analyses that controlled for a wide array of covariates showed that overweight and obese women were at increased risk for incident major depressive disorder (MDD) during the follow-up period. Overweight men and obese men were at decreased risk of incident drug abuse and alcohol dependence, respectively. Obese women had a decreased risk of incident alcohol abuse and drug dependence. Men with drug dependence and women with specific phobia had a decreased risk of becoming overweight or obese during the follow-up.
Limitations
The NESARC excluded adolescents and the homeless and weight was self-reported, but highly correlated with external validating data.
Conclusions
Increased risk of MDD among overweight and obese women could be attributed to stigma and greater body dissatisfaction among women in Western cultures. Overweight and obesity may serve as protective factors against developing incident substance use disorders possibly due to shared neural functions in the brain underlying addictions to numerous substances. Results are discussed in terms of their clinical implications including the need to update treatment guidelines for the management of overweight, obesity and MDD.
doi:10.4088/JCP.10m06077gry
PMCID: PMC3227748  PMID: 21457678
There is growing evidence that the role of lipids in innate immunity is more important than previously realized. How lipids interact with bacteria to achieve a level of protection, however, is still poorly understood. To begin to address the mechanisms of antibacterial activity, we determined MICs and minimum bactericidal concentrations (MBCs) of lipids common to the skin and oral cavity—the sphingoid bases d-sphingosine, phytosphingosine, and dihydrosphingosine and the fatty acids sapienic acid and lauric acid—against four Gram-negative bacteria and seven Gram-positive bacteria. Exact Kruskal-Wallis tests of these values showed differences among lipid treatments (P < 0.0001) for each bacterial species except Serratia marcescens and Pseudomonas aeruginosa. d-Sphingosine (MBC range, 0.3 to 19.6 μg/ml), dihydrosphingosine (MBC range, 0.6 to 39.1 μg/ml), and phytosphingosine (MBC range, 3.3 to 62.5 μg/ml) were active against all bacteria except S. marcescens and P. aeruginosa (MBC > 500 μg/ml). Sapienic acid (MBC range, 31.3 to 375.0 μg/ml) was active against Streptococcus sanguinis, Streptococcus mitis, and Fusobacterium nucleatum but not active against Escherichia coli, Staphylococcus aureus, S. marcescens, P. aeruginosa, Corynebacterium bovis, Corynebacterium striatum, and Corynebacterium jeikeium (MBC > 500 μg/ml). Lauric acid (MBC range, 6.8 to 375.0 μg/ml) was active against all bacteria except E. coli, S. marcescens, and P. aeruginosa (MBC > 500 μg/ml). Complete killing was achieved as early as 0.5 h for some lipids but took as long as 24 h for others. Hence, sphingoid bases and fatty acids have different antibacterial activities and may have potential for prophylactic or therapeutic intervention in infection.
doi:10.1128/AAC.05151-11
PMCID: PMC3294957  PMID: 22155833
PLoS ONE  2012;7(8):e43524.
The exceptional biodiversity of Reunion Island is threatened by anthropogenic landscape changes that took place during the 350 years of human colonization. During this period the human population size increased dramatically from 250 to 800,000. The arrival of humans together with the development of agriculture, invasive species such as rats and cats, and deforestation has lead to the extinction of more than half of the original vertebrate species of the island. For the remaining species, significant work is being carried out to identify threats and conservation status, but little genetic work has been carried on some of the most endangered species. In the last decade theoretical studies have shown the ability of neutral genetic markers to infer the demographic history of endangered species and identify and date past population size changes (expansions or bottlenecks). In this study we provide the first genetic data on the critically endangered species the Reunion cuckoo-shrike Coracina newtoni. The Reunion cuckoo-shrike is a rare endemic forest bird surviving in a restricted 12-km2 area of forested uplands and mountains. The total known population consists of less than one hundred individuals out of which 45 were genotyped using seventeen polymorphic microsatellite loci. We found a limited level of genetic variability and weak population structure, probably due to the limited geographic distribution. Using Bayesian methods, we identified a strong decline in population size during the Holocene, most likely caused by an ancient climatic or volcanic event around 5000 years ago. This result was surprising as it appeared in apparent contradiction with the accepted theory of recent population collapse due to deforestation and predator introduction. These results suggest that new methods allowing for more complex demographic models are necessary to reconstruct the demographic history of populations.
doi:10.1371/journal.pone.0043524
PMCID: PMC3423348  PMID: 22916272
Archives of oral biology  2011;56(6):588-591.
Objective
The purpose of this study was to determine the presence and relative composition of neutral lipids in human saliva.
Design
Whole unstimulated saliva was collected from 12 subjects ranging from 21 to 29 years old. Samples were lyophilized, and lipids were extracted using chloroform-methanol. Lipids were analyzed by thin-layer chromatography.
Results
Human saliva contains cholesterol, fatty acids, triglycerides, wax esters, cholesterol esters and squalene. The mean total neutral lipid content was 12.1 +/− 6.3 µg/ml.
Conclusions
This lipids in human saliva closely resemble the lipids found on the skin surface. These salivary lipids are most likely produced by the sebaceous follicles in the oral mucosa and sebaceous glands associated with major salivary glands.
doi:10.1016/j.archoralbio.2010.12.002
PMCID: PMC3095707  PMID: 21247555
The objective of this study was to assess the perceived oral health-related quality of life (OHQoL) of adolescents affected with one of the ectodermal dysplasias (EDs).
Data were collected from 2003 to 2007 in a cross-sectional study of a convenience sample of individuals affected by ED (n=35) using the Child Perceptions Questionnaire(CPQ11–14)for children and the Parent-Caregiver Perceptions Questionnaire (P-CPQ) for their caregivers. The main findings of this study were that individuals who were affected with ED in the older age group (15–19 years old) perceived more functional problems than younger individuals (11–14 years old) (p=0.04). Females with ED (n= 13) perceived more emotional problems than males (n=22; p=0.01). Although caregivers tended to report slightly higher OHQoL scores (indicating worse OHQoL), no significant differences were observed between children’s and parents’ total OHQoL and individual domains’ median scores (p>0.05). Thus, the perceptions of oral health and well-being may vary by age and gender for children who have ED. Caution is warranted concerning using parents as proxies for their children when assessing the child’s OHQoL.
doi:10.1111/j.1754-4505.2011.00189.x
PMCID: PMC3105356  PMID: 21592162
ectodermal dysplasias; oral health-related quality of life; children; teenagers; parents; perceptions
PLoS ONE  2012;7(2):e31667.
Hybridization is observed frequently in birds, but often it is not known whether the hybrids are fertile and if backcrossing occurs. The breeding ranges of the great reed warbler (Acrocephalus arundinaceus) and the clamorous reed warbler (A. stentoreus) overlap in southern Kazakhstan and a previous study has documented hybridization in a sympatric population. In the present study, we first present a large set of novel microsatellite loci isolated and characterised in great reed warblers. Secondly, we evaluate whether hybridization in the sympatric breeding population has been followed by backcrossing and introgression.
We isolated 181 unique microsatellite loci in great reed warblers. Of 41 loci evaluated, 40 amplified and 30 were polymorphic. Bayesian clustering analyses based on genotype data from 23 autosomal loci recognised two well-defined genetic clusters corresponding to the two species. Individuals clustered to a very high extent to either of these clusters (admixture proportions ≥0.984) with the exception of four previously suggested arundinaceus–stentoreus hybrid birds that showed mixed ancestry (admixture proportions 0.495–0.619). Analyses of simulated hybrids and backcrossed individuals showed that the sampled birds do not correspond to first–fourth-generation backcrosses, and that fifth or higher generation backcrosses to a high extent resemble ‘pure’ birds at this set of markers.
We conclude that these novel microsatellite loci provide a useful molecular resource for Acrocephalus warblers. The time to reach reproductive isolation is believed to be very long in birds, approximately 5 Myrs, and with an estimated divergence time of 2 Myrs between these warblers, some backcrossing and introgression could have been expected. However, there was no evidence for backcrossing and introgression suggesting that hybrids are either infertile or their progeny inviable. Very low levels of introgression cannot be excluded, which still may be an important factor as a source of new genetic variation.
doi:10.1371/journal.pone.0031667
PMCID: PMC3288047  PMID: 22384052
Drug and alcohol dependence  2010;112(1-2):9-17.
Objective
To determine whether internalizing and externalizing psychopathology were differentially associated with alcohol dependence in men and women.
Methods
Four categories of lifetime psychopathology were examined: neither internalizing nor externalizing (NINE), internalizing only (IO), externalizing only (EO) and both internalizing and externalizing (BIE). Multivariate models assessed gender differences in the adjusted associations of these categories with the odds of lifetime alcohol dependence in a representative sample of 43,093 U.S. adults 18 and older and with clinical course and expression in a subsample of 4,781 lifetime alcoholics.
Results
The excess odds of lifetime alcohol dependence associated with IO, EO and BIE were significantly greater for women than men, OR = 2.6, 8.8 and 10.7 versus 1.9, 4.0 and 6.5, respectively. Regardless of gender, the ORs were significantly higher for EO than IO and for BIE than EO. Gender differences in the expression and course of alcoholism were most pronounced for the categories of NINE and IO, with men having greater consumption, dependence severity and treatment but less familial alcoholism. Gender variation in the association of psychopathology with the expression and course of alcoholism most evident in the BIE category, where the associations were stronger for women. Lifetime externalizing psychopathology was associated with an increased likelihood of treatment utilization, especially among women.
Conclusions
Findings highlight the need to increase alcoholism screening, prevention and intervention among women with psychopathology, especially externalizing. The greater numbers of internalizing than externalizing alcoholics emphasizes the need to treat symptoms of depression and anxiety in alcohol treatment settings.
doi:10.1016/j.drugalcdep.2010.04.019
PMCID: PMC2953598  PMID: 20558014
alcohol dependence; psychopathology; gender; externalizing; internalizing

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