bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of E3 in pH Protection during Alphavirus Assembly and Exit
Authors: Uchime Onyinyechukwu , Fields Whitney , Kielian Margaret
Published: July 2013

Alphaviruses are small enveloped viruses whose surface is covered by spikes composed of trimers of E2/E1 glycoprotein heterodimers. During virus entry, the E2/E1 dimer dissociates within the acidic endosomal environment, freeing the E1 protein to mediate fusion of the viral and endosome membranes. E2 is synthesized as a precursor, p62, which is cleaved by furin in the late secretory pathway to produce mature E2 and a small peripheral glycoprotein, E3. The immature p62/E1 dimer is acid resistant, but since p62 is cleaved before exit from the acidic secretory pathway, low pH-dependent binding of E3 to the spike complex is believed to prevent premature fusion. Based on analysis of the structure of the Chikungunya virus E3/E2/E1 complex, we hypothesized that interactions of E3 residues Y47 and Y48 with E2 are important in this binding. We then directly tested the
in vivo
role of E3 in pH protection by alanine substitutions of E3 Y47 and Y48 (Y47/48A) in Semliki Forest virus. The mutant was nonviable and was blocked in E1 transport to the plasma membrane and virus production. Although the Y47/48A mutant initially formed the p62/E1 heterodimer, the dimer dissociated during transport through the secretory pathway. Neutralization of the pH in the secretory pathway successfully rescued dimer association, E1 transport, and infectious particle production. Further mutagenesis identified the critical contact as the cation-π interaction of E3 Y47 with E2. Thus, E3 mediates pH protection of E1 during virus biogenesis via interactions strongly dependent on Y47 at the E3-E2 interface.
Hydrophobic and Charged Residues in the Central Segment of the Measles Virus Hemagglutinin Stalk Mediate Transmission of the Fusion-Triggering Signal
Authors: Apte-Sengupta Swapna , Navaratnarajah Chanakha K. , Cattaneo Roberto
Published: July 2013

The pH-independent measles virus membrane fusion process begins when the attachment protein H binds to a receptor. Knowing that the central segment of the tetrameric H stalk transmits the signal to the fusion protein trimer, we investigated how. We document that exact conservation of most residues in the 92 through 99 segment is essential for function. In addition, hydrophobic and charged residues in the 104 through 125 segment, arranged with helical periodicity, are critical for F protein interactions and signal transmission.
Mode of Virus Rescue Determines the Acquisition of VHS Mutations in VP22-Negative Herpes Simplex Virus 1
Authors: Ebert Katja , Depledge Daniel P. , Breuer Judith , Harman Laura , Elliott Gillian
Published: July 2013

It has been proposed that herpes simplex virus 1 with VP22 deleted requires secondary mutation of VHS for viability. Here we show that a replication-competent Δ22 virus constructed by homologous recombination maintains a wild-type (Wt) VHS gene and has no other gross mutations. By contrast, Δ22 viruses recovered from a bacterial artificial chromosome contain multiple amino acid changes within a conserved region of VHS. Hence, the mode of virus rescue influences the acquisition of secondary mutations.
Functional Analysis of Claudin-6 and Claudin-9 as Entry Factors for Hepatitis C Virus Infection of Human Hepatocytes by Using Monoclonal Antibodies
Authors: Fofana Isabel , Zona Laetitia , Thumann Christine , Heydmann Laura , Durand Sarah C. , Lupberger Joachim , Blum Hubert E. , Pessaux Patrick , Gondeau Claire , Reynolds Gary M. , McKeating Jane A. , Grunert Fritz , Thompson John , Zeisel Mirjam B. , Baumert Thomas F.
Published: July 2013

The relevance of claudin-6 and claudin-9 in hepatitis C virus (HCV) entry remains elusive. We produced claudin-6- or claudin-9-specific monoclonal antibodies that inhibit HCV entry into nonhepatic cells expressing exogenous claudin-6 or claudin-9. These antibodies had no effect on HCV infection of hepatoma cells or primary hepatocytes. Thus, although claudin-6 and claudin-9 can serve as entry factors in cell lines, HCV infection into human hepatocytes is not dependent on claudin-6 and claudin-9.
The Recent Establishment of North American H10 Lineage Influenza Viruses in Australian Wild Waterfowl and the Evolution of Australian Avian Influenza Viruses
Authors: Vijaykrishna Dhanasekaran , Deng Yi-Mo , Su Yvonne C. F. , Fourment Mathieu , Iannello Pina , Arzey George G. , Hansbro Philip M. , Arzey K. Edla , Kirkland Peter D. , Warner Simone , O'Riley Kim , Barr Ian G. , Smith Gavin J. D. , Hurt Aeron C.
Published: July 2013

Influenza A H10N7 virus with a hemagglutinin gene of North American origin was detected in Australian chickens and poultry abattoir workers in New South Wales, Australia, in 2010 and in chickens in Queensland, Australia, on a mixed chicken and domestic duck farm in 2012. We investigated their genomic origins by sequencing full and partial genomes of H10 viruses isolated from wild aquatic birds and poultry in Australia and analyzed them with all available avian influenza virus sequences from Oceania and representative viruses from North America and Eurasia. Our analysis showed that the H10N7 viruses isolated from poultry were similar to those that have been circulating since 2009 in Australian aquatic birds and that their initial transmission into Australia occurred during 2007 and 2008. The H10 viruses that appear to have developed endemicity in Australian wild aquatic birds were derived from several viruses circulating in waterfowl along various flyways. Their hemagglutinin gene was derived from aquatic birds in the western states of the United States, whereas the neuraminidase was closely related to that from viruses previously detected in waterfowl in Japan. The remaining genes were derived from Eurasian avian influenza virus lineages. Our analysis of virological data spanning 40 years in Oceania indicates that the long-term evolutionary dynamics of avian influenza viruses in Australia may be determined by climatic changes. The introduction and long-term persistence of avian influenza virus lineages were observed during periods with increased rainfall, whereas bottlenecks and extinction were observed during phases of widespread decreases in rainfall. These results extend our understanding of factors affecting the dynamics of avian influenza and provide important considerations for surveillance and disease control strategies.
Magical NiTi expander
Authors: Katti Chandrika Girish , Katti Girish , Kallur Ravi , Ghali Srinivas Rao
Published: July 2013

A 24-year-old male patient was referred to our department for expansion of the constricted maxillary arch as a presurgical procedure for the correction of congenital facial disfigurement. On examination, the patient had a convex profile, increased interlabial gap, tongue thrust, limited mouth opening, posterior crossbite, asymmetric ‘V’-shaped maxillary arch with severe constriction, crowding of anterior teeth in the maxillary arch and a massive open bite. Radiographic investigations included orthopantomograph and occlusal radiographs. The patient photographs and models were analysed. On careful evaluation, the treatment for maxillary arch expansion was planned with a nickel titanium (NiTi) slow maxillary expander along with fixed mechanotherapy for alignment of teeth. An unexpectedly successful outcome was appreciated from the treatment. An emphasis should be laid on selecting and treating the case of constricted arches with a surgical or non-surgical approach, as expansion can be achieved orthodontically by using NiTi expanders.
Intravitreal ranibizumab in choroidal neovascularisation due to multifocal choroiditis and panuveitis syndrome
Authors: Dardabounis Doukas , Alvanos Efstratios , Gatzioufas Zisis , Panos Georgios D
Published: July 2013

We report the use of intravitreal ranibizumab as initial and only treatment in a case of peripapillary choroidal neovascularisation (CNV) in a patient with multifocal choroiditis and panuveitis (MCP) syndrome. A 54-year-old woman presented with sudden reduced vision in the right eye of 2 weeks duration. A full ophthalmological examination was performed including biomicroscopy, fluorescein angiography and optical coherence tomography. A peripapillary choroidal neovascular membrane with subretinal haemorrhage extending to the fovea was revealed due to MCP syndrome. Three intravitreal injections of ranibizumab were performed with monthly intervals. There was clearance of the subretinal haemorrhage and reduction of the leakage after the first injection. The visual acuity improved to 20/20 in 3 months and remained stable at 2-year follow-up without angiographical leakage. Intravitreal injections of ranibizumab could be tried in cases of CNV due to MCP syndrome with very good response.
The adult Tillaux fracture: one not to miss
Authors: Sharma Bishnulatpam , Reddy Ippala Sreenath , Meanock Colin
Published: July 2013

This case is a rare adult type Tillaux fracture. This fracture should be diagnosed in the emergency department. The fracture requires careful evaluation with awareness of associated injury. Standard radiological views (antero-posterior and lateral) of the ankle may not clearly show the fracture displacement hence, an oblique view is required. The fracture should be managed by the emergency physician, if the displacement is less than 2 mm, by immobilising with a non-weight bearing cast or ankle braces for 6 weeks. If the fracture fragment is displaced more than 2 mm, it should be treated by closed reduction failing which open reduction and internal fixation is the standard practice.
A core needle biopsy provides more malignancy-specific results than fine-needle aspiration biopsy in thyroid nodules suspicious for malignancy
Authors: Hakala T , Kholová I , Sand J , Saaristo R , Kellokumpu-Lehtinen P
Published: July 2013

Background and aims

The most commonly used diagnostic method for the preoperative diagnosis of thyroid nodules is ultrasound-guided fine-needle aspiration biopsy (FNA), which often yields non-diagnostic or non-definitive results and seldom produces definite malignant diagnoses. To improve upon the malignancy-specific sensitivity, we tested core needle biopsies (CNBs) of thyroid lesions taken from surgical specimens.

Material and methods

52 consecutive patients with malignant or malignant-suspicious thyroid nodules were referred to Tampere University Hospital between May 2010 and December 2011. Preoperative FNAs were categorised as follicular neoplasm (48%), suspicion for malignancy (46%) or malignancy (6%). Intraoperative FNA and CNB samples were acquired from surgical specimens removed during surgery. The results of the needle biopsies were compared with the final pathological diagnosis.

Results

CNBs had a high definitive sensitivity for malignancy (61%, CI 41% to 78%) whereas the definitive sensitivity for malignancy of FNAs was significantly lower (22%, CI 10% to 42%). CNB was not beneficial in the diagnosis of follicular thyroid lesions. When all suspected follicular tumours were excluded, the definitive sensitivity of CNB rose to 70% (CI 48% to 86%).

Conclusions

CNB may be beneficial for the diagnosis of papillary thyroid carcinoma and other non-follicular thyroid lesions. CNB may be considered as an additional diagnostic procedure in cases with FNA suspicious for malignancy.
PL02.1 The Dynamic Evolution of Sexual Behavior as It Impacts STD Spread
Authors: Aral S
Published: July 2013

The past three decades have been marked by significant progress in our understanding of the biological foundations of human behaviour, the evolutionary forces that drive human conduct and cognition and environmental factors that turn genes on and off. Concurrently, advances in communication technologies have drastically changed the way people meet and mate; demographic trends have modified patterns of supply and demand of potential mates while economic trends have affected men and women’s’ motivations for sexual partnering.

Consequent changes in sexual values, attitudes and norms; sexual behaviours including sexual practises; and concepts of marriage, commitment, love and sex have been remarkable. Major trends in sexual behavioural determinants of sexually transmitted infection spread include: exponential increases in the size of sexual core groups; increased acceptance of commercial and transactional sex; increases in male, female and underage sex tourism; and increased geographic mobility of sex workers. These trends result in greater connectedness among sexual core groups across national boundaries thereby limiting the effectiveness of national prevention programmes. Moreover, both increased sexual mixing between core groups and the general population and increased reporting about sexual core behaviours through social media facilitate behavioural contagion and enhance the impact of sexual core groups on the behaviours of the general population. It is surprising that STI are not spreading faster globally in light of these changes in their social and behavioural determinants.
PL03.1 STIs - 2013: Resurgence of Early Syphilis, Persistence of High STI Morbidity and Mortality, and Emergence of Antimicrobial Resistant STIs: Implications and Potential Responses
Authors: Holmes K
Published: July 2013

Recent directions in clinical-epidemiologic research on STIs include (1) population-level estimates of STI prevalence (e.g., WHO’s 2011 publication of global estimates), and formal population-level surveys of STI prevalence and risk determinants; (2) analysis of global mortality and disability attributable to major diseases, including STIs (the Global Burden of Disease Study, published in the December 2012 triple issue of The Lancet); (3) conceptualization and implications of the “Treatment and Care Cascade” and the “Prevention Cascade” currently focused on HIV infection; and (4) emerging interest in Program Science, linking programme implementers and scientists in needs assessments, conceptualization, design, advocacy for funding, implementation, evaluation, cost-effectiveness, and continuous strengthening of STI/HIV programmes. The global emergence and rapid spread of anti-microbial resistant pathogens, suggests the clinical mantra of “first do no harm” to the individual patient must be mirrored in a similar public health mantra - “first do no harm” to the population. This means selective use and more systematic evaluation of the impact of antimicrobial use on human and animal pathogens and microbiomes. The reemergence of syphilis and persistence of other STIs in vulnerable populations, and the limited implementation of cost-effective interventions for STI control reflects global neglect of STI programmes, and perhaps increasing failure to effectively integrate STI, HIV/AIDS, and reproductive health programmes globally (for example, the very limited integration of HIV PMTCT with elimination of congenital syphilis programmes). Nonetheless, progress in sexual health promotion and STI control can be made possible with cost-effective use by clinicians and public health leaders of effective, available tools, such as scale up of HPV and HBV vaccines, and linked delivery of other sociobehavioral and biomedical STI interventions in vulnerable populations. Rigorous evaluation of the impact of such programmes, with assessment of what is not working as well as what is working, is essential.
S01.1 An Update on Topical Microbicide Development
Authors: McGowan I
Published: July 2013

Microbicides are products that can be applied to vaginal or rectal mucosa with the intent of preventing, or at least significantly reducing, the transmission of sexually transmitted infections including HIV-1. Unfortunately, the last 5–10 years of microbicide research have generated a number of disappointments. Large scale phase 2B/3 studies or topical microbicides have failed to demonstrate product efficacy, have been stopped prematurely for futility, or in the worst case scenario have possibly demonstrated microbicide induced harm including increased risk of HIV acquisition. Although the CAPRISA 004 study of vaginal peri-coital tenofovir gel demonstrated a significant reduction in HIV acquisition (39%; p = 0.017), the most recently completed microbicide effectiveness study, MTN-003 (the VOICE study), did not demonstrate the effectiveness of daily tenofovir gel, largely due to product non-adherence by study participants. However, the ongoing FACTS-001 study has the potential to confirm the results of the CAPRISA 004 study and may lead to licensure of tenofovir gel. In addition, current microbicide research is increasingly focused on technology, such as intravaginal rings, that provide sustained release of antiretrovirals to the cervicovaginal mucosa and may minimise adherence problems associated with peri-coital or daily use of vaginal gels. Two Phase 3 studies of a dapivirine vaginal ring are currently ongoing. The development of combination products that might provide contraceptive and antiretroviral drug delivery is also gathering momentum. Rectal microbicide research has moved from Phase 1 to Phase development. The MTN-017 study, which will be conducted in the USA, Peru, Thailand, and South Africa, will evaluate the safety and acceptability of oral Truvada and rectal tenofovir gel in men who have sex with men and transgender women. This talk will provide a comprehensive update on the challenges and opportunities associated with microbicide development.
S05.2 Challenges and Opportunities: Potentials in Research on AMR Gonococci
Authors: Deal C
Published: July 2013

Although gonorrhoea has afflicted humans for centuries and the causative bacterium,Neisseria gonorrhoeae, was identified over a century ago, gonorrhoea remains a global public health problem. Stepwise acquisition of genetic mutations has conferred gradually increasing resistance to multiple antibiotics, thus limiting their usage for presumptive therapy. Until recently cephalosporins have remained the foundation of treatment, but growing evidence suggests that resistance is emerging. Even as researchers work to understanding the genetic mutations required for cephalosporin resistance, it is clear that new treatment options are needed. This presents a challenge as the number of new systemic antimicrobials evaluated and approved each year by regulatory agencies has steadily fallen over the past 30 years. Currently one new antimicrobial is undergoing clinical study as a potential treatment of gonorrhoea. An alternative strategy is to repurpose older antibiotics by studying the efficacy of dual therapy combinations of existing antimicrobials. Ultimately new antimicrobial development is needed now, since the development process can take more than a decade.

Detecting and responding to emergence of multidrug resistant gonorrhoea remains a challenge. Rapid detection of resistant infections is facilitated by local antimicrobial susceptibility testing, which requires live organisms isolated by culture. However, as the use of nucleic acid amplification tests (NAATs) have expanded, the number ofN. gonorrhoeaecultures performed by public health laboratories decreased rapidly and the capacity of laboratories to perform culture has declined. An alternative strategy is to develop molecular assays for detecting genetic mutations associated with resistance or susceptibility to specific antimicrobials to guide antibiotic selection by the clinician at the point of care. Even with this potential, molecular assays may not be able to supplant culture-based antimicrobial susceptibility testing for surveillance to detect novel resistance phenotypes and genotypes. Ultimately a gonococcal vaccine may be the most effective public health strategy.
S09.1 Molecular Detection of Antimicrobial Resistance in STI Pathogens
Authors: Whiley D , Chen M , Donovan B , Fairley C , Guy R , Kaldor J , Regan D , Trembizki E , Ward J , Lahra M
Published: July 2013

The emergence of antimicrobial resistance (AMR) among sexually transmitted infections (STI) is a cause for global concern, and is epitomised by the fact we are now running out of treatment options for gonorrhoea. The role of AMR surveillance is now more important than ever. Ideally, AMR surveillance should be fast, easy, inexpensive, accessible, reproducible across testing methods, and provide clinically meaningful information to inform treatment strategies. In reality this is not the case, with AMR surveillance activities for STIs typically weak or non-existent in many parts of the world. Molecular methods have the potential to enhance AMR surveillance, particularly for organisms that cannot easily or readily be characterised phenotypically; which is the case for most STIs. The challenges for molecular surveillance are however many and include factors such as; the mechanisms of resistance may be many or otherwise unknown, they may miss novel mutations, the technology can be expensive, they need specialised laboratories and trained staff, and that their specificity can be undermined where target sequences are shared across different species. Despite these challenges, such methods are being developed and are now finding their way into routine settings. Advances in molecular technology and expanding knowledge of resistance mechanisms continue to pave new directions in this important area.
S14.1 Testing For Syphilis in Pregnancy and Associated Adverse Outcomes in Mozambique
Authors: Mahomed M , Gimbel S , Hoek R , Rustagi A , Come C , Newman L , Faria F , Manuel J , Gloyd S , Broutet N
Published: July 2013

Background

Antenatal syphilis screening and treatment is an effective intervention to reduce perinatal, infant, and maternal morbidity and mortality. Prenatal syphilis screening increased in Manica and Sofala Provinces from < 5% in 1993 to over 90% since 2005. This study aimed to (1) estimate the prevalence of congenital syphilis in the cities of Beira and Chimoio, (2) determine differences in congenital syphilis among women treated early in pregnancy, late in pregnancy, and not treated, and (3) identify factors associated with syphilis screening among pregnant women.

Methods

Pregnant women presenting at one of six maternities in Beira and Chimoio cities, were recruited and screened with rapid syphilis tests (RST) and, if positive, further screened using Rapid Plasma Reagin (RPR). All live newborns whose mothers tested positive at any point during pregnancy or at birth were RPR screened to detect congenital syphilis. A newborn subsample also had blood and tissue samples taken for IgM and PCR testing to validate the RPR results.

Results

Between March 5, 2012 and March 15, 2013, 16,812 women were recruited out of the 19,821 births registered at the participating maternities. Overall, 611 women (3.6% total) tested positive for syphilis; 498 were syphilis positive by RST at the maternity. An additional 113 women had tested positive at ANC, but were RST negative at delivery. Of the total 405 women who had tested syphilis positive (RST or RPR) and been treated in ANC, their syphilis results at delivery were the following: 286 (71%) were RST positive, 192 (47%) were both RST and RPR positive, and 35 (9%) were RPR positive and RST negative. There was no significant difference in RST/RPR rates between women treated before or after 28 weeks gestation. Of the 16,322 women who were syphilis negative at ANC, 211 (1.26%) were RPR positive at birth (suggestive of recent infection); of these women, 152 (54%) were RPR positive. Estimates of the prevalence of and factors associated with congenital syphilis and association with gestational age of treatment are pending, awaiting results of confirmatory testing.

Conclusions

The congenital syphilis prevalence rate is substantially lower in this region than in other regions of Mozambique, likely related to years of antenatal syphilis screening. Lifetime positivity of RST was variable, as nearly 30% of women with positive RST tests in pregnancy were RST negative at birth. In addition, nearly half of women treated adequately remained RPR positive at delivery after treatment.
S15.1 Frequent Viral Introductions Sustain Local HIV Epidemics in Rural Africa
Authors: Grabowski M K , Lessler J , Redd A , Kagaayi J , Laeyendecker O , Nalugoda F , Serwadda D , Wawer M , Quinn T C , Gray R H
Published: July 2013

Background

It is often assumed that local sexual networks play a dominant role in HIV spread in sub-Saharan Africa. However, the extent to which continued HIV transmission in rural African communities, home to two-thirds of the continent’s population, is driven by intra-community sexual networks versus viral introductions from outside of communities remains unknown.

Methods

We analysed the spatial dynamics of HIV transmission in rural Rakai District, Uganda using population-based cohort data on 14594 individuals within 46 communities experiencing a generalised HIV epidemic (prevalence 12·%, incidence ∼2 per 100 person years). We applied spatial clustering statistics, phylogenetic analyses, and probabilistic transmission models to quantify the relative contributions of viral introductions versus local HIV transmission, including household transmission, to HIV incidence.

Results

Individuals in households with incident (n = 189) or prevalent (n = 1597) HIV-infected persons were 3·2 (95% CI: 2·7–3·7) times more likely to be HIV-infected, compared to the population in general, but spatial clustering outside of households was weak and confined to distances < 500m. There was high viral diversity: only 34% of infected persons phylogenetically clustered with at least one other participant. One-third of phylogenetic clusters were individuals sharing a household. Of the remaining two-thirds, 71% crossed community boundaries. We estimate that 39% (95% CI: 32–44%) of viral transmissions occurred within stable household partnerships and that 40% (95% CI: 34–42%) of transmissions were from named extra-household sexual contacts, of which 62% (95% CI: 55–70%) were non-stable partners from outside the community.

Interpretation: External HIV introductions into rural communities are common and account for a substantial proportion of new HIV infections in Rakai, Uganda. Our findings suggest that combination HIV prevention will most effectively control local HIV epidemics when implemented at broad spatial scales, and imply a need to identify key populations serving as sources of introduced infections into general populations.
S15.2 Social Networking and Diffusion of Risks and Interventions Among Youth
Authors: Young S D
Published: July 2013

Use of online social networking technologies has been growing rapidly, especially among groups at risk for sexually transmitted infections (STIs), such as men who have sex with men (MSM), homeless youth, and African American and Latino populations. International studies are beginning to suggest a number of important relationships between social networking technologies, at-risk populations, and risk for sexually transmitted infections: (1) At-risk populations are using social networking technologies to meet sexual partners, (2) Although at-risk populations who use social networking technologies are engaging in high rates of sexual intercourse, they are engaging in prevention behaviours that might mitigate their risk, and (3) Social networking technologies can be used as platforms for scaling and diffusing STI prevention and testing interventions. Online social networks are tools that can be used to rapidly spread information and social norms. These same technologies that can be used to potentially spread STI transmission must also be implemented as tools for preventing transmission. Recent research is discussed.
S15.3 Modification of Sexual Networks and Sexual Risk For Men Having Sex with Men (MSM) by the Internet
Authors: Marcus U , Schmidt A J
Published: July 2013

The Internet provides new opportunities for sexual minorities to communicate and interact. Access to internet may facilitate coming-out for young MSM, sexual identity formation, and community building. On the other side the internet partly substitutes social and sexual venues where MSM congregate. Population and individual level aspects of internet communication among MSM were analysed based on data collected in the European MSM Internet Survey (EMIS).

EMIS was a large collaborative project of public health, academia and community based organisations from 35 European countries. The survey was online from June through August 2010, advertised on a large range of MSM websites. The questionnaire anonymously collected data from MSM across Europe. Among others, detailed information on the last sexual intercourse with a non-steady partner was collected.

The analysis is based on 174,209 eligible respondents. Participation rates in 38 countries with more than 100 respondents varied considerably (0.3 – 6.8/10,000 inhabitants) and correlated with internet access. A comparison of self-reported new diagnoses of HIV and national surveillance data suggests different relative sizes of MSM populations in different countries. Among men reporting a non-steady sex partner in the last 12 months, 58% had met this partner on the internet. Compared with partners met in venues, serostatus communication with internet partners was more frequent, respondents more often already had sex with their internet partners before, and partners were more often presumed HIV seroconcordant. Contrastingly, refraining from anal sex was less often an option for partners met on the internet.

Improved access to internet may increase the relative size of MSM populations by involving a larger number of individuals into sexual networks. With broadening access to internet risk reducing aspects like increased communication before sexual encounters become more pronounced. Promoting protective and preventive behaviours can counter adverse effects of the internet on the HIV epidemic.
S16.3 Recurrent TV: The Potential of Molecular Techniques to Improve Clinical Practise
Authors: Hobbs M M
Published: July 2013

Infection with the widespread protozoan pathogenTrichomonas vaginalis(TV) does not result in lasting immunity, and recurrent infections are common. Whether due to unrecognised, inadequately treated or repeatedly acquired TV infections, recurrent vaginitis in women, nongonococcal urethritis and chronic prostatitis in men are well-recognised and challenging clinical conditions. Despite increased recognition in the STI research community of the adverse consequences of trichomoniasis in women and men and the potential for TV infection to increase transmission of HIV and other STIs,T. vaginalisinfection remains underappreciated by clinicians, public health professionals, policy makers and patients. Trichomoniasis is not a reportable STI in most countries, and TV infection is often asymptomatic, thus many infections are neither diagnosed nor treated. Symptomatic infection in women is the clinical presentation most likely to be recognised and treated, but treatment of male sexual partners of infected women is infrequent or inadequate, and testing and treatment of trichomoniasis in male patients is rare. Thus, reservoirs of infection persist.

Recent improvements in molecular diagnostics for detection of TV in women and men have the potential to improve clinical practise. Rapid antigen detection tests offer point of care testing and improved treatment options in settings where technically complex and costly nucleic acid amplification tests (NAATs) are not available, and NAATs offer highly sensitive and specific testing options for detection of TV in urogenital specimens commonly tested for other sexually transmitted pathogens includingN. gonorrhoeaeandC. trachomatis. With enhanced awareness, availability and application of these molecular tests, better detection and treatment of trichomoniasis in women and in their sexual partners can be achieved with eventual reduction of the adverse reproductive consequences associated withT. vaginalisinfection.
O01.5 Efficacy of RG1-VLP Vaccination Against Genital and Cutaneous Human Papillomavirusesin Vitroandin Vivo
Authors: Schellenbacher C , Kwak K , Fink D , Shafti-Keramat S , Huber B , Jindra C , Roden R , Kirnbauer R
Published: July 2013

Licensed human papillomavirus (HPV) vaccines, based on virus-like particles (VLP) self-assembled from major capsid protein L1, afford type-restricted protection against types 16/18/6/11 (or 16/18 for the bivalent vaccine), which cause 70% of cervical carcinomas (Cxca) and 90% of genital warts. However, they do not protect against less prevalent high-risk types causing 30% of CxCa, or cutaneous HPV. The minor capsid protein L2 confers low-level immunity to type-common epitopes.

Chimeric RG1-VLP presenting HPV16L2 amino acids 17–36 (RG1 epitope) within the DE-surface loop of HPV16L1 induce cross-neutralisationin vitro. We hypothesised, that RG1-VLP vaccination protects against a large spectrum of mucosal and cutaneous HPV infectionsin vivo.

L2-specific antibody and CTL responses in RG1-VLP vaccinated rabbits were determined by ELISA and ELISPOT assays. Cross-neutralisation was analysed using native or pseudovirion (PsV) assays. Vaccine efficacyin vivowas determined in a mouse genital challenge model.

Immunization with RG1-VLP adjuvanted with human-applicable alum-MPL induced robust L2 antibodies (ELISA titers 2,500–12,500), which cross-neutralised mucosal high-risk HPV26/33/35/39/68/59/68/73/69/53/34, low-risk HPV6/11/32/40/44/70, and cutaneous HPV2/27/3/76 (titers 25–1,000), and a vigorous CTL response.In vivo, mice were efficiently protected against experimental vaginal challenge with mucosal high-risk PsV types HPV16/18/45/31/33/52/58/35/39/51/59/68/56/73/26/53/66/34 and low-risk HPV6/43/44. Enduring protection was demonstrated 1 year after vaccination.

RG1-VLP is a promising next-generation vaccine with broad efficacy against all relevant mucosal and also cutaneous HPV types.
O02.3 Treatment Outcomes For Persistent Mycoplasma Genitalium-Associated NGU: Evidence of Moxifloxacin Treatment Failures
Authors: Manhart L E , Khosropour C M , Gillespie C W , Lowens M S , Golden M R , Totten P A
Published: July 2013

Background

Recent treatment trials have demonstrated low efficacy of doxycycline againstMycoplasma genitalium(MG) and increasing resistance to azithromycin. Treatment with azithromycin is recommended for persistent NGU if not used for the initial episode. We evaluated microbiologic cure rates for men with NGU and persistent detection of MG.

Methods

English-speaking men aged 16 years attending the STD clinic in Seattle, WA with NGU (urethral discharge or urethral symptoms plus ≥ 5 PMNs/HPF) were enrolled in a randomised trial of NGU therapy between January 2007 and July 2011. Urine was tested for MG by PCR. Men received 1g azithromycin plus placebo doxycycline or doxycycline (100mg bid x 7d) plus placebo azithromycin. Treatment failures after 3 weeks received ‘reverse therapy’ (active doxycycline if they first received active azithromycin and vice versa). Persistent failures after 6 weeks received moxifloxacin (400mg x 7d). After September 2010, microbiologic failures at 3 weeks received moxifloxacin.

Results

Of 606 enrolled men, 65 were positive for MG at enrollment and returned after 3 weeks. Microbiologic failure (positive MG test) occurred in 23/38 (60.5%) who received azithromycin and 19/27 (70.4%) who received doxycycline (p = 0.41). Of the 37 men with microbiologic treatment failure who received ‘reverse therapy’ and returned after 6 weeks, 19 (51.4%) had persistent detection of MG, including 14/20 (70.0%) retreated with doxycycline and 5/17 (29.4%) retreated with azithromycin (p = 0.02). All 19 men were prescribed moxifloxacin; 16 returned at 9 weeks and 2 (12.5%) had microbiologic failure, despite clinical cure. Four men received moxifloxacin after initial failure; 1 had microbiologic failure at 6 weeks, was retreated with moxifloxacin and microbiologically cured at 9 weeks.

Conclusion

One half of MG-positive men retreated with a second standard NGU treatment regimen experienced microbiologic treatment failure. Moxifloxacin treatment failure, while not common, did occur, suggesting antimicrobial susceptibility in MG merits careful monitoring.
O02.5 A Phase II, Dose Ranging Study to Evaluate the Efficacy and Safety of Single-Dose Oral Solithromycin (CEM-101) For Treatment of Patients with Uncomplicated Urogenital Gonorrhoea
Authors: Hook E W , Jamieson B D , Oldach D , Harbison H , Whittington A , Fernandes P
Published: July 2013

Objectives

Emerging resistance to available treatment creates an urgent need for new therapies for uncomplicated gonorrhoea. Solithromycin, a new 4th generation macrolide with 3 ribosomal targets, is highly active against most antibiotic-resistant strains ofNeisseria gonorrhoeae. A Phase II, dose ranging study to evaluate the efficacy and safety of single-dose oral solithromycin for uncomplicated urogenital gonorrhoea was conducted.

Methods

Consenting participants with suspectedNeisseria gonorrhoeaeinfection were cultured at the urethra/cervix, rectum, and pharynx at enrollment and Day 7.The primary outcome was bacterial eradication (conversion from positive baselineN. gonorrhoeaeurethral/cervical culture to negative) at Day 7. Secondary outcomes included eradication of rectal or pharyngeal gonorrhoea and the eradication of gonococcal and chlamydial nucleic acids. Initially, eligible patients received a single 1200 mg oral dose of solithromycin; following demonstration of bacteriologic efficacy, a second cohort was treated with a single 1000 mg dose.

Results

Of 41 (19 M, 22 F) participants enrolled, 28 were treated with a 1200 mg dose and, to date, 13 with 1000 mg. Gonococcal eradication rates in 22 evaluable 1200 mg patients were 100% (22/22) for urethral/cervical, pharyngeal (5/5), and rectal (2/2) infections. Of 9 evaluable 1000 mg patients enrolled to date, gonococcal eradication rates have been 100% (9/9) for urethral/cervical, pharyngeal (2/2), and rectal (1/1) infections. Susceptibility data from 25 isolates show the median MIC (range) for solithromycin was 0.06 µg/mL (0.015–0.125) and for azithromycin was 0.125 µg/mL (0.06–0.5).

Solithromycin was generally well-tolerated with mild dose-related gastrointestinal AEs (68%; 28/41). The most common AE was mild diarrhoea, occurring in 61% (17/28) of patients receiving the 1200 mg dose and 15% (2/13) of patients receiving the 1000 mg dose.

Conclusions

A single dose of 1200 or 1000 mg solithromycin appears to be well-tolerated and effective in eradicatingN. gonorrhoeae.
O02.6 Azithromycin Versus Doxycycline For the Treatment of Genital Chlamydia Infection - a Meta-Analysis of Randomised Controlled Trials
Authors: Hocking J , Kong F , Vodstrcil L , Guy R , Chen M , Fairley C , Bradshaw C , Tabrizi S
Published: July 2013

Introduction

There has been considerable debate questioning the efficacy of azithromycin for the treatment of genital chlamydia. We conducted a meta-analysis to compare the efficacy of 1 gramme azithromycin with 100mg doxycycline twice daily for seven days for the treatment of genital chlamydia infection.

Methods

Medline, PubMed, Embase and the Cochrane Controlled Trials Register were searched till end 2012. Inclusion criteria included (1) randomised controlled trial of azithromycin versus doxycycline for the treatment of urethral or cervical chlamydia, and; (2) evaluation of microbial cure within 3 months of treatment. Type of diagnostic test, duration of follow up, gender, patient status (all symptomatic versus both symptomatic/asymptomatic) and microbial cure were extracted. The primary outcome was difference in efficacy (doxycycline efficacy minus azithromycin efficacy) at final follow up. Meta-analysis calculated a pooled efficacy for each treatment and the difference in efficacy between treatments.

Results

Of 692 references identified, 23 trials met the inclusion criteria. 1065 individuals were treated with azithromycin and 850 with doxycycline; all studies reported efficacy within 6 weeks follow-up. Pooled cure rates were 96.2%(95% CI: 94.2%, 98.3%) for azithromycin and 98.1%(95% CI: 96.6%, 99.7%) for doxycycline. The pooled efficacy difference was 1.9%(95% CI: 0.4%, 3.4%) showing a small but significant difference in favour of doxycycline; there was negligible heterogeneity between studies (I2 = 1.9%, p = 0.44). There was no difference in efficacy in men (3.8%; 95% CI:-1.2%, 8.8%) or women (–0.9%; 95% CI: –5.3%, 3.6%). When stratified by type of test, efficacy was significantly higher for doxycycline in culture-based studies (1.8%; 95% CI: 0.4%, 3.3%), but not in NAAT-based studies (5.5%; 95% CI: –2.1%, 13.1%). Efficacy was higher for doxycycline in symptomatic men (6.3%; 95% CI: 3.0%, 12.3%), but not in symptomatic women (–4.5%; 95% CI: –14.9%, 5.9%).

Conclusion

These results suggest that doxycycline may be more effective than azithromycin for the treatment of urethral or cervical chlamydia infection, especially in symptomatic men.
O03.2 Antimicrobial Resistance of Neisseria Gonorrhoeae in the European Union: Response to the Threat of Multidrug Resistant Gonorrhoea
Authors: Spiteri G , Cole M , Unemo M , Hoffmann S , Ison C , Laar M van de
Published: July 2013

Background

Increasing reports of Neisseria gonorrhoeaein vitrodecreased susceptibility and resistance, and treatment failures with third generation cephalosporins are of major concern as these drugs are the last remaining options for effective antimicrobial therapy in many settings.

Methods

The European Gonococcal Antimicrobial Surveillance Programme (Euro-GASP), a sentinel surveillance programme funded by the European Centre for Disease Prevention and Control since 2009, monitors antimicrobial susceptibility patterns across the European Union (EU/EEA). Participating laboratories performed susceptibility testing by Etest or agar dilution breakpoint method, or sent isolates to reference laboratories in Denmark, Sweden or the United Kingdom for testing. Euro-GASP validated proficiency and result accuracy through an external quality assessment scheme.

Results

In 2011, 1902 isolates from 21 countries were collected and tested. The percentage of tested isolates with decreased susceptibility to cefixime (8.0%) remained stable compared to 2010 (8.7%), but was still significantly higher than in 2009 (5.1%, p < 0.01). Isolates with decreased susceptibility to cefixime were reported from 17 countries in 2011, the same as in 2010; however three countries reported such isolates for the first time in 2011. For the first time, ten isolates with decreased susceptibility to ceftriaxone were reported from two countries. Rates of ciprofloxacin (49%) and azithromycin (5.3%) resistance remained high. Minimum inhibitory concentration of gentamicin remained low (MIC50: 4 mg/L; MIC90: 8 mg/L).

Conclusions

Although the rapid increase and spread of decreased susceptibility to cefixime in 2010 has not continued, the detection of isolates with decreased susceptibility to ceftriaxone is concerning. ECDC has published a response plan which aims to strengthen surveillance of gonococcal antimicrobial susceptibility in the EU/EEA; ensure that capacity for culture and susceptibility testing is maintained; establish a system for collection and verification of data on clinical treatment failure; and to recommend public health actions at national and European level.
O05.1 High Grade Anal Intraepithelial Neoplasia: One Virus, One Lesion
Authors: Richel O , Quint K D , Lindenman J , Noesel C M Van , Koning M N C De , Munckhof H A M Van den , Vries H J C De , Prins J M , Quint W G V
Published: July 2013

Background

Prevention and treatment of anal intraepithelial neoplasia (AIN) in HIV+ men who have sex with men (MSM) is subject of discussion. Knowledge on causative HPV types is crucial in understanding AIN and in vaccination studies. However, data on AIN-specific HPV are limited and whole tissue sections (WTS) often show multiple HPV infections.

In this study, we analysed whether WTS and subsequent laser capture micro-dissection (LCM) with HPV PCR genotyping accurately detects type-specific HPV DNA in individual areas of high grade (HG)AIN.

Methods

31 WTS with HGAIN of 21 HIV+ MSM were analysed by the SPF10 PCR/LiPA25 (version 1) HPV genotyping system. In case of multiple HPV types, PCR was repeated in selected areas of AIN, isolated by LCM.

Results

WTS PCR showed a single HPV type in 17 (55%). In the remaining 14 WTS sections with multiple HPV types, PCR was repeated in LCM-isolated dysplastic areas (median: 4 per WTS). In 12 of 14 these samples, the number of HPV types could be reduced to single HPV types within discrete areas of a lesion, resulting in a total of 29 (17+12), in which (components of) HGAIN show a singe HPV type. HPV 16 was found in 14/29 (48%), HPV 18 in 3 and HPV 58 in 3. The remaining HPV types that could be linked to a lesional area were HPV 26, 31, 35, 39, 52, 53, 54, 59, 67, 68/73, 74, 91 and one indeterminate HPV type.

Conclusion

WTS PCR and subsequent LCM PCR is accurate in detecting lesion specific HPV types in AIN and it seems that 94% of the AIN-lesions (on macroscopic or microscopic level) are caused by a single HPV type. Apart from HPV 16, the predominant type, a wide range of other HPV types are responsible for HGAIN, which has consequences for vaccination development.
O05.3 More Than Meets the Eye: A Molecular Phylogenetic Analysis Reveals Importance of Novel Bacteria in Bacterial Vaginosis (BV)
Authors: Srinivasan S , Morgan M T , Liu C , Matsen F A , Hoffman N G , Fiedler T L , Agnew K J , Marrazzo J M , Fredricks D N
Published: July 2013

Background

BV is a highly prevalent dysbiotic condition associated with adverse reproductive and health outcomes in women. BV is marked by loss of certain lactobacilli and acquisition of complex communities of anaerobic bacteria. Gram stain is the gold standard for diagnosis, wherein the abundances of four bacterial morphotypes are assessed; Lactobacillus, Gardnerella, Bacteroides and Mobiluncus. Newly described uncultivated bacteria are highly specific for BV, but no studies have investigated the association between these bacteria and Gram stain morphotypes.

Methods

We examined the association of bacteria detected by broad-range 16S-rRNA gene PCR/pyrosequencing with bacterial morphotypes detected in Gram stains from 220 women with and without BV. We also used species-specific quantitative PCR and fluorescence in situ hybridization (FISH) methods to document concentrations of two bacteria with curved rod morphologies:Mobiluncusspecies and the uncultivated BV-associated bacterium-1 (BVAB1).

Results

We provide evidence that curved Gram-negative rods designated Mobiluncus morphotypes by Gram stain are more likely BVAB1. Rank abundance plots of vaginal bacteria in women with curved rods (Nugent 9–10) showed that BVAB1 was the dominant bacterium (26%), while relative abundance ofMobiluncuswas only 0.2%. BVAB1 sequence reads were also associated with Mobiluncus morphotypes (p = 7.4E-06). Among women with Nugent scores 9–10, the mean concentration of BVAB1 DNA was 2-log units greater thanMobiluncus(p < 0.001). FISH analyses also revealed that among women with Nugent scores of 10, the mean number of BVAB1 cells was greater thanMobiluncuscells (p < 0.001). In addition, we noted thatPrevotellaandPorphyromonasspp. are significantly associated with the Bacteroides morphotype, whereasBacteroidesspecies are rare.

Conclusions

Gram stain morphotype designations for BV need revision to account for novel vaginal bacteria. These findings have major implications for studies using Gram stains as a proxy to describe the vaginal microbiota.