Jessica Mao

Total Synthesis of a Functional Designer Eukaryotic Chromosome

Designer Chromosome One of the ultimate aims of synthetic biology is to build designer organisms from the ground up. Rapid advances in DNA synthesis has allowed the assembly of complete bacterial genomes. Eukaryotic organisms, with their generally much larger and more complex genomes, present an additional challenge to synthetic biologists. Annaluru et al. (p. 55, published online 27 March) designed a synthetic eukaryotic chromosome based on yeast chromosome III. The designer chromosome, shorn of destabilizing transfer RNA genes and transposons, is ∼14% smaller than its wild-type template and is fully functional with every gene tagged for easy removal. A synthetic version of yeast chromosome III with every gene tagged can substitute for the original. Rapid advances in DNA synthesis techniques have made it possible to engineer viruses, biochemical pathways and assemble bacterial genomes. Here, we report the synthesis of a functional 272,871–base pair designer eukaryotic chromosome, synIII, which is based on the 316,617–base pair native Saccharomyces cerevisiae chromosome III. Changes to synIII include TAG/TAA stop-codon replacements, deletion of subtelomeric regions, introns, transfer RNAs, transposons, and silent mating loci as well as insertion of loxPsym sites to enable genome scrambling. SynIII is functional in S. cerevisiae. Scrambling of the chromosome in a heterozygous diploid reveals a large increase in a-mater derivatives resulting from loss of the MATα allele on synIII. The complete design and synthesis of synIII establishes S. cerevisiae as the basis for designer eukaryotic genome biology.

Catheter ablation of accessory pathways near the coronary sinus: Value of defining coronary arterial anatomy

BACKGROUND Accessory pathways can lie near or within the coronary sinus (CS). Radiofrequency catheter ablation of accessory pathways is a well-established treatment option, but this procedure can cause damage to adjacent coronary arteries. OBJECTIVE The purpose of this study was to evaluate the anatomic relationship between the coronary arteries and the CS. METHODS Retrospective data of patients who underwent catheter ablation of supraventricular tachycardia between June 2011 and August 2013 was reviewed. In addition, detailed analysis of coronary computed tomographic angiography (CTA) data from 50 patients was performed. RESULTS Between June 2011 and August 2013, 427 patients underwent catheter ablation of supraventricular tachycardia, of whom 105 (age 28 ± 17 years, 60% male) had accessory pathway-mediated tachycardia. Of these, 23 patients had accessory pathways near the CS, and 60% (N = 14) underwent concurrent coronary angiography. In 4 patients, the posterolateral (inferolateral) branch (PLA) of the right coronary artery was in close proximity to the CS, and 2 patients (18%) had stenosis of the PLA at the site of ablation. On CTA at their closest proximity, the PLA was 1.9 ± 1.3 mm and the left circumflex artery (LCx) was 2.0 ± 0.8 mm from the body of the CS, in right and left coronary artery-dominant patients, respectively. CS ostium and PLA were 3.6 ± 1.9 mm apart. In left-dominant patients, LCx and CS ostium were 3.8 ± 1.2 mm apart. CONCLUSION The PLA and LCx are in close proximity to the anteroinferior aspect of the CS ostium and proximal CS. The relationship of the CS and coronary arteries should be evaluated before ablation at these sites.

Comparing the effectiveness of a crowdsourced video and a social marketing video in promoting condom use among Chinese men who have sex with men: a study protocol.

Introduction Crowdsourcing has been used to spur innovation and increase community engagement in public health programmes. Crowdsourcing is the process of giving individual tasks to a large group, often involving open contests and enabled through multisectoral partnerships. Here we describe one crowdsourced video intervention in which a video promoting condom use is produced through an open contest. The aim of this study is to determine whether a crowdsourced intervention is as effective as a social marketing intervention in promoting condom use among high-risk men who have sex with men (MSM) and transgender male-to-female (TG) in China. Method We evaluate videos developed by crowdsourcing and social marketing. The crowdsourcing contest involved an open call for videos. Entries were judged on capacity to promote condom use, to be shareable or ‘go viral’ and to give value to the individual. 1170 participants will be recruited for the randomised controlled trial. Participants need to be MSM age 16 and over who have had condomless anal sex in the last 3 months. Recruitment will be through an online banner ad on a popular MSM web page and other social media platforms. After completing an initial survey, participants will be randomly assigned to view either the social marketing video or the crowdsourcing video. Follow-up surveys will be completed at 3 weeks and 3 months after initial intervention to evaluate condomless sex and related secondary outcomes. Secondary outcomes include condom social norms, condom negotiation, condom self-efficacy, HIV/syphilis testing, frequency of sex acts and incremental cost. Ethics and dissemination Approval was obtained from the ethical review boards of the Guangdong Provincial Center for Skin Diseases and STI Control, UNC and UCSF. The results of this trial will be made available through publication in peer-reviewed journals. Trial registration number NCT02516930.

Facilitators of HCV treatment adherence among people who inject drugs: a systematic qualitative review and implications for scale up of direct acting antivirals

BackgroundWhile the public health benefits of new HCV treatments depend on treatment adherence, particularly among people who inject drugs (PWID), several social and medical factors can jeopardize treatment adherence. The aim of this study is to examine the qualitative literature on facilitators to HCV treatment adherence among PWID.MethodsWe searched six databases to identify qualitative research studies on HCV treatment adherence facilitators among PWID. Two reviewers independently extracted and analyzed data using PRISMA guidelines and the CASP tool to evaluate study quality.ResultsFrom ten studies representing data from 525 participants, three major themes emerged across studies: logistical facilitators within health systems enhanced HCV treatment adherence, positive social interactions between PWID and staff provided positive feedback during treatment, and HCV treatment may complicate the addiction recovery process.ConclusionsAlthough PWID face several barriers to adherence, we identified treatment adherence facilitators that could be incorporated into clinical practice.

Disclosure of sexual orientation to health professionals in China: results from an online cross-sectional study

Background: Many men who have sex with men (MSM) in China are “in the closet.” The low rate of disclosure may impact sexual behaviours, testing for HIV and other sexually transmitted infections (STIs), and diseases transmission. This study examines factors associated with overall sexual orientation disclosure and disclosure to healthcare professionals.

HIV self-testing among high-risk men who have sex with men in China: a cross-sectional study

BACKGROUND HIV self-testing holds great promise for reaching high-risk key populations who do not access facility-based services, but it has rarely been evaluated outside of structured research settings. HIV self-testing is a process by which a person takes and interprets a test. We aimed to characterise implementation of unsupervised HIV self-testing among men who have sex with men (MSM) in China. METHODS We undertook a nationwide online survey of MSM in November, 2015, in China. Men were invited to participate by clicking through banner adverts on social network sites. Eligible men were at least 16 years of age, had ever had anal sex with a man, and had had sex without use of a condom in the past 3 months. Among MSM who ever self-tested for HIV, we assessed benefits (eg, first-time HIV test, increased testing frequency, confirmatory testing) and adverse outcomes, (eg, coercion, violence, suicidality). First-time HIV test was defined by what an individual reported was their first ever test for HIV (a self-test or a facility test). Among MSM who reported ever testing for HIV, we identified correlates of HIV self-testing as first-time HIV test with use of multivariable logistic regression. The institutional review boards of the Guangdong Provincial Center for Skin Diseases and STI Control, the University of North Carolina at Chapel Hill, and the University of California San Francisco approved this study. FINDINGS Of 1610 eligible men, 1189 (73·9%) completed the survey. 28·7% (341/1189) reported ever self-testing for HIV. The HIV prevalence among self-testers was 7·0% (24/341) and among non-self-testers was 4·9% (15/306). Among self-testers, 58·7% (200/341) reported HIV self-testing as a first-time HIV test and 22·6% (77/341) had increased testing frequency. 77·5% (31/40) of those with a positive HIV self-test received confirmatory facility-based testing. Minimal adverse outcomes were noted. Multivariable analysis showed that self-testing as first-time HIV test was associated with younger age (odds ratio 0·95, 95% CI 0·91-0·99), not telling health-care providers about having sex with men (2·22, 1·56-3·17), not using the internet to meet sex partners (2·53, 1·45-4·43), and having group sex (1·76, 1·03-2·98). INTERPRETATION HIV self-testing increases first-time HIV testing and testing frequency among those with previous testing. This tool could be useful to scale up HIV testing among high-risk men in China and other settings. FUNDING National Institutes of Health (NIAID 1R01AI114310-01), UNC-South China STD Research Training Center (FIC 1D43TW009532-01), UNC Center for AIDS Research (NIAID 5P30AI050410-13), UCSF Center for AIDS Research (NIAID P30 AI027763), NIMH (R00MH093201), Fulbright-Fogarty Fellowship (FIC R25TW0093), and SESH Global.

Crowdsourcing health communication about condom use in men who have sex with men in China: a randomised controlled trial.

BACKGROUND The Chinese government now encourages innovation, and more specifically, crowdsourcing. Crowdsourcing, the process of shifting individual tasks to a large group, might reimagine health communication, making it more people-centred. We aimed to compare the effectiveness of a crowdsourced versus social marketing condom promotion video to promote condom use. METHODS A non-inferiority randomised controlled trial was conducted in November 2015 in China. Men who have sex with men (MSM) aged 16 years or older who reported having had sexual contact without use of a condom in the preceding 3 months were recruited through a nationwide MSM website. Men were randomly allocated to one of the two arms in a 1:1 ratio using a computer algorithm and then watched one of two videos. The crowdsourced video was developed through an open contest and the social marketing video was designed by a company. Participants completed a baseline survey and follow-up surveys at 3 weeks and 3 months post-intervention. The primary outcome was incidence of sexual contact without a condom. Intention-to-treat analysis was used, with a non-inferiority margin of 10%. The trial is registered with ClinicalTrials.gov, number NCT02516930. All participants gave informed consent and the protocol was approved by Chinese (Guangdong Provincial Centre for Skin Diseases and STI Control) and American (University of North Carolina at Chapel Hill and the University of California San Francisco) institutional review boards. FINDINGS 1173 participants were recruited to the study; 907 (77%) completed the 3-week follow up and 791 (67%) the three-month follow up. At three weeks, 146/434 (33·6%) participants in the crowdsourced group and 153/473 (32·3%) in the social marketing group reported having sexual contact without a condom. The crowdsourced intervention achieved our pre-specified non-inferiority criterion (estimated difference: +1·3% [95%CI: 4·8-7·4]). At 3 months, 196/376 (52·1%) men in the crowdsourced group and 206/415 (49·6%) in the social-marketing group reported having sexual contact without a condom (estimated difference: +2·5% [95%CI -4·5-9·5%]). The two groups reported similar HIV testing rates and other condom-related secondary outcomes. The cost of the crowdsourcing intervention was substantially lower than the social marketing intervention (16 686 vs 26 845 USD). No adverse outcomes were reported. INTERPRETATION Crowdsourcing could be a useful tool in China, which has a large population and government support. Crowdsourcing provides a structured system for multi-sectoral input into health policy in China. FUNDING US National Institute of Allergy and Infectious Diseases (NIAID; 1R01AI114310); University of North Carolina (UNC)-South China Sexually Transmitted Disease Research Training Centre (Fogarty International Centre; 1D43TW009532); UNC Center for AIDS Research (NIAID; 5P30AI050410); University of California San Francisco Centre for AIDS Research (NIAID; P30 AI027763); US National Institute of Mental Health (R00MH093201 to CW); UNC Chapel Hill, Johns Hopkins University, Morehead School of Medicine and Tulane University (UJMT) Fogarty Fellowship (FIC R25TW0093); Doris Duke International Clinical Research Fellowship; US National Center for Advancing Translational Sciences (UL1TR001111).

Disclosure of sexual orientation in men who have sex with men in China: a cross-sectional survey

BACKGROUND Most men who have sex with men (MSM) in China do not publicly disclose their sexual orientation. This low rate of disclosure may affect sexual behaviours, testing for HIV or other sexually transmitted infections (STIs), and disease transmission. This study examined factors associated with overall sexual orientation disclosure and disclosure to a healthcare professional. METHODS We did a cross-sectional online survey of MSM in China from September 23, 2014, to October 8, 2014, through three national websites. Participants completed questions covering sociodemographic information, sexual behaviors, HIV and syphilis test history, and self-reported HIV status. We defined overall disclosure as having ever disclosed sexual orientation to anyone other than a sexual partner, and healthcare professional disclosure as disclosing to a doctor or other medical provider. Eligible participants gave informed consent electronically before beginning the survey. FINDINGS A total of 1819 men started the survey and 1424 (78%) completed it. Of 1424 participants, 62% (886/1424) reported overall disclosure, and 16% (232/1424) had disclosed to healthcare providers. In multivariate analyses, the odds of overall sexual orientation disclosure were 56% higher among MSM who used smartphone-based sex-seeking applications (adjusted OR 1·56 [95% CI 1·25-2·95]), but were lower among MSM reporting high-risk sexual risk behaviors such as sex while drunk (0·55 [0·37-0·81]) and sex while using drugs (0·66 [0·50-0·87]). The odds of disclosure to a healthcare professional were greater among MSM who had ever been tested for HIV (3·36 [2·50-4·51]) or STIs 4·92 [3·47-6·96]) or self-reported as HIV positive (1·59 [0·93-2·72]). INTERPRETATION More than 80% of MSM surveyed had not disclosed their sexual orientation to health professionals. This low level of disclosure probably represents a major obstacle to serving the unique needs of MSM in clinical settings. Further research and action to facilitate disclosure of sexual orientation by MSM, especially to healthcare professionals, is urgently needed. FUNDING US National Institute of Allergy and Infectious Diseases (NIAID; 1R01AI114310); University of North Carolina (UNC)-South China STD Research Training Centre (Fogarty International Centre 1D43TW009532); UNC Center for AIDS Research (NIAID; 5P30AI050410); University of California San Francisco Centre for AIDS Research (NIAID; P30 AI027763]; US National Institute of Mental Health (R00MH093201); UNC Chapel Hill, Johns Hopkins University, Morehead School of Medicine and Tulane University (UJMT) Fogarty Fellowship (FIC R25TW0093); Doris Duke International Clinical Research Fellowship; US National Center for Advancing Translational Sciences (UL1TR001111).

Sex tourism among Chinese men who have sex with men: a cross-sectional observational study

BACKGROUND Sex tourism among men who have sex with men (MSM) provides a risk for spread of HIV and other sexually transmitted infections. Sex tourism is defined as travelling outside of ones home town and purchasing sex with gifts or money. Our objective was to characterise the frequency, sociodemographic characteristics, and sexual risk behaviours among Chinese MSM sex tourists. METHODS We undertook an online, cross-sectional survey of high-risk MSM throughout China in November, 2015. Men were invited to participate by clicking through banner adverts on social network sites. Eligible men were at least 16 years of age, had ever had anal sex with a man, and had had sex without use of a condom in the past 3 months. Questions covered sociodemographic characteristics and sexual risk behaviours. Univariate and multivariable logistic regressions were done to identify correlates of sex tourism. We also compared the mean MSM HIV prevalence in each sex tourists origin and destination locations. Ethics approval was obtained from the ethics review boards of the Guangdong Provincial Center for Skin Diseases and STI control, the University of North Carolina at Chapel Hill, and the University of California at San Francisco. FINDINGS Among 1610 eligible men, 1189 (74%) completed the survey and 62 (5%) were identified as sex tourists. 20 (32%) of these men travelled primarily to purchase sex and the remainder purchased sex while travelling for another purpose. We noted little difference in sociodemographics and sexual risk behaviours between the two groups. 45 (73%) of sex tourists used a mobile app or website to find sex partners. In multivariable analyses, adjusting for age and income, sex tourism was correlated with high-risk sexual behaviours (adjusted odds ratio 1·83, 95% CI 1·06-3·17), higher income (4·44, 1·77-11·18), and living with HIV (2·79, 1·03-7·55). Sex tourism was more often from a location with a lower MSM HIV prevalence (mean prevalence 4·47, SD 2·01) to a location with a higher MSM HIV prevalence (6·86, 5·24; p<0·0001). INTERPRETATION MSM sex tourists were more likely to have risky sexual behaviours and travel from locations with low to high HIV prevalence rather than high to low HIV prevalence. MSM sex tourists might be part of core groups that disproportionately contribute to MSM HIV transmission. Enhanced surveillance and interventions tailored to MSM sex tourists should be considered. FUNDING National Institutes of Health, Doris Duke Charitable Foundation.

Reply to the Editor—More Awarenessless Risk—Interpretation of Ablation Risk Caused by Coronary Arterial Anatomy

We are grateful to Dr. Wu and colleagues for their interest in our article and the fact that they found it a valuable study. We agree with them that the heart is in constant motion, and therefore, measuring distances from still images has limitations. In patients who underwent coronary angiography during the electrophysiological study in this series, it was the closest distance on two views (left and right anterior oblique) that was used for the measurement of the proximity of the vessels to the coronary sinus (CS) and the decision to ablate at a specific site within the CS. Furthermore, although in diastole the coronary vessels may be pulled further away from the CS, they also dilate, bringing them closer to the CS. The reverse would be true in systole when venous filling of the CS occurs, moving the CS closer to the arteries. Given the location of these vessels at the fibrous trigone of the heart, their motion may indeed be less than other cardiac structures. We completely agree that the proximity of the vessels to the CS warrants coronary angiography before ablation within the proximal CS. Cooling within the coronary arteries can theoretically decrease the risk of complications, and perfusion of cold saline through coronary arteries is an option. Furthermore, the option of cryoablation is an important one. But we presume that the safety concerns about coronary proximity would still be relevant. Although not quantified in this study, mechanical perturbation of coronary arteries causing spasm is also a concern (we have noted coronary artery spasm with percutaneous epicardial catheter manipulation during mapping and ablation). With regard to the valuable point made regarding the anatomy of inferoparaseptal pathways, we concur that the endocardial aspect of the left atrium should be mapped via transseptal or retrograde approaches before mapping and ablation in the CS. In fact, the patients reported in this series had mapping performed in regions corresponding to the CS in the left atrium via a transseptal approach, and the endocardial sites were either late during activation mapping or unsuccessful in terminating arrhythmia.