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CD8(+) T cells are known to be important in clearing herpes simplex virus (HSV) infections. However, investigating the specific antiviral mechanisms employed by HSV-2-specific T cell populations is limited by a lack of reagents such as CD8(+) T cell epitopes and specific tetramers. Using a combination of intracellular cytokine staining flow cytometry and ELISpot methods, we functionally characterized peripheral HSV-2-specific CD8(+) T cells from peripheral blood mononuclear cell (PBMC) that recognize 14 selected HSV-2 open-reading frames (ORFs) from 55 HSV-2 seropositive persons; within these ORFs, we subsequently identified more than 20 unique CD8(+) T cell epitopes. CD8(+) T cells to HSV-2 exhibited significant heterogeneity in their functional characteristics, proliferation, production of inflammatory cytokines, and potential to degranulate ex vivo. The diversity in T cell response in these ex vivo assessments offers the potential of defining immune correlates of HSV-2 reactivation in humans.

Address: 1Department of Epidemiology, University of Washington; Seattle, Washington, USA, 2Department of Laboratory Medicine, University of Washington; Seattle, Washington, USA, 3Deparment of Medicine, University of Washington; Seattle, Washington, USA, 4Program in Biostatistics, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA, 5Program in Vaccine and Infectious Disease Institute, Fred Hutchinson Cancer Research Center, Seattle, Washington; USA, 6Uganda Cancer Institute, Kampala, Uganda and 7Infectious Disease Institute, Kampala, Uganda * Corresponding author

A 55-yr-old woman with history of Marfan syndrome presented with an enlarging aortic arch aneurysm. Fifteen years ago, she had undergone repair of an aortic root aneurysm. A preoperative transthoracic echocardiogram (TTE) revealed normal biventricular function, 3+ tricuspid regurgitation, a small nonobstructive mechanical aortic prosthesis, and an aortic root graft. The distal ascending aorta and aortic arch were dilated, measuring 5 cm in diameter. Left heart catheterization showed normal coronary arteries originating from the aortic root graft, with the left adjacent to the right. The patient was scheduled for aortic arch reconstruction and tricuspid valve repair.

After a major serials cancellation project in 1986 forced the Chemistry Library at The University of Texas to cancel 10% of its serial titles, a project was planned to assess its remaining serial titles. The project used the computer based Chemical Ahstracts Service Source Index as its basis. The chemistry serials collections of The University of Texas and University of California at Berkeley and Massachusetts Institute of Technology were compared. Preliminary collection overlap comparisons are made of the three collections.

Introduction Kaposi sarcoma (KS) is the leading cause of cancer in Uganda and occurs in people with and without HIV. Human herpesvirus-8 (HHV-8) replication is important both in transmission of HHV-8 and progression to KS. We characterized the sites and frequency of HHV-8 detection in Ugandans with and without HIV and KS. Methods Participants were enrolled into one of four groups on the basis of HIV and KS status (HIV negative/KS negative, HIV positive/KS negative, HIV negative/KS positive, and HIV positive/KS positive). Participants collected oral swabs daily and clinicians collected oral swabs, anogenital swabs, and plasma samples weekly over 4 weeks. HHV-8 DNA at each site was quantified by polymerase chain reaction (PCR). Results 78 participants collected a total of 2063 orals swabs and 358 plasma samples. Of these, 428 (21%) oral swabs and 96 (27%) plasma samples had detectable HHV-8 DNA. HHV-8 was detected more frequently in both the oropharynx of persons with KS (24 (57%) of 42 persons with KS vs. 8 (22%) of 36 persons without, p = 0.002) and the peripheral blood (30 (71%) of 42 persons with KS vs. 8 (22%) of 36 persons without, p

Primary genital herpes simplex virus (HSV) infection is commonly associated with systemic symptoms. A systematic evaluation of HSV viremia during primary genital infection has not been performed previously. Plasma samples from adults with a first clinical episode of genital HSV infection were assayed for HSV DNA by polymerase chain reaction. One hundred sixty-four adults had confirmed primary genital HSV infection. Of these, 40 (24%) had HSV DNA detected in plasma. Thirty-seven (93%) of 40 were infected with HSV-2. Viremic participants were more likely to be women, compared with aviremic participants (83% vs. 61%; P = .01). Viremia was detected more frequently and at a higher number of copies per milliliter early in infection. We conclude that HSV viremia during primary genital HSV infection is common, especially among women.

Introduction Quantitative estimation of the protective effect of HSV-2 infection against reinfection with other HSV-2 strains is an important parameter for HSV-2 vaccine development. We determined the prevalence of and risk factors for HSV-2 superinfection using a novel genotyping tool. Methods We first identified 96 high quality HSV SNPs that could determine whether HSV-2 strains were matched with >90% probability via next generation sequencing of 39 genital HSV-2 lesion swabs. These SNPs were then used to create a customised high throughput genotyping assay (GoldenGate, Illumina ® ). Two genital specimens collected from the same participant, each containing ≥5 log 10 copies HSV DNA/ml, were genotyped. HIV-infected and HIV-uninfected persons participating in studies in the USA, Africa, and Peru were included. Sample pairs were excluded if Results Paired genital swab specimens from 123 persons were analysed; 113 (92%) had the same strain detected at the two time points; 93 (76%) had identical SNP patterns, 18 (15%) had disagreements at one SNP, and 2 (2%) had disagreements at 2 SNPs. Ten persons (8%) were infected with more than one strain, with paired samples disagreeing at a median of 23 SNPs (range 5–33), for a minimum estimated superinfection prevalence of 8%. Of the 10 persons with HSV-2 superinfection, 7 (70%) were women and 7 (70%) were HIV infected; 6 were from Africa, one was from the USA, and 3 were from Peru. Conclusion We developed a custom genotyping assay that provides a high throughput method for genotyping HSV-2. HSV-2 superinfection was detected in 8% of paired samples, suggesting that naturally occurring immunity to HSV-2 may not be highly efficient to protect against reinfection, especially among HIV-infected persons. Disclosure of interest statement This study was funded by the US National Institutes of Health. No pharmaceutical grants were received for the conduct of this study.

Genital herpes simplex virus (HSV) infection causes recurrent genital ulcers, neonatal herpes, and increases the risk of HIV acquisition. HSV-2, the most common cause of genital herpes, is highly prevalent worldwide, with an estimated 417 million people infected between the ages of 15–49. The urgent need for a prophylactic vaccine against genital HSV has been long recognized. Multiple glycoprotein subunit vaccines candidates have been tested but none have successfully prevented HSV-2 genital ulcer disease in phase III trials. Despite these findings, there is strong interest in pursuing novel vaccine platforms to induce immune responses to protect against HSV acquisition. Therapeutic vaccines to reduce genital symptoms and viral shedding in persons already infected with HSV-2 are also being tested in early phase clinical trials. The global STI vaccine roadmap provided a framework for research priorities to move the HSV vaccine field forward. This presentation will review 1) lessons from prior clinical trials of HSV vaccines, 2) new insights into immunology of HSV infection, 3) current status of HSV vaccine pipeline, with an emphasis on candidates currently in clinical trials and 4) discussion of clinical trial design issues unique to HSV.

The innate immune response, and in particular the alpha/beta interferon (IFN-α/β) system, plays a critical role in the control of viral infections. Interferons α and β exert their antiviral effects through the induction of hundreds of interferon-induced (or -stimulated) genes (ISGs). While several of these ISGs have characterized antiviral functions, their actions alone do not explain all of the effects mediated by IFN-α/β. To identify additional IFN-induced antiviral molecules, we utilized a recombinant chimeric Sindbis virus to express selected ISGs in IFN-α/β receptor (IFN-α/βR)−/−mice and looked for attenuation of Sindbis virus infection. Using this approach, we identified a ubiquitin homolog, interferon-stimulated gene 15 (ISG15), as having antiviral activity. ISG15 expression protected against Sindbis virus-induced lethality and decreased Sindbis virus replication in multiple organs without inhibiting the spread of virus throughout the host. We establish that, much like ubiquitin, ISG15 requires its C-terminal LRLRGG motif to form intracellular conjugates. Finally, we demonstrate that ISG15's LRLRGG motif is also required for its antiviral activity. We conclude that ISG15 can be directly antiviral.

Several different mammalian neurotropic viruses produce an age-dependent encephalitis characterized by more severe disease in younger hosts. To elucidate potential factors that contribute to age-dependent resistance to lethal viral encephalitis, we compared central nervous system (CNS) gene expression in neonatal and weanling mice that were either mock infected or infected intracerebrally with a recombinant strain, dsTE12Q, of the prototype alphavirus Sindbis virus. In 1-day-old mice, infection with dsTE12Q resulted in rapidly fatal disease associated with high CNS viral titers and extensive CNS apoptosis, whereas in 4-week-old mice, dsTE12Q infection resulted in asymptomatic infection with lower CNS virus titers and undetectable CNS apoptosis. GeneChip expression comparisons of mock-infected neonatal and weanling mouse brains revealed developmental regulation of the mRNA expression of numerous genes, including some apoptosis regulatory genes, such as the proapoptotic molecules caspase-3 and TRAF4, which are downregulated during development, and the neuroprotective chemokine, fractalkine, which is upregulated during postnatal development. In parallel with increased neurovirulence and increased viral replication, Sindbis virus infection in 1-day-old mice resulted in both a greater number of host inflammatory genes with altered expression and greater changes in levels of host inflammatory gene expression than infection in 4-week-old mice. Only one inflammatory response gene, an expressed sequence tag similar to humanISG12, increased by a greater magnitude in infected 4-week-old mouse brains than in infected 1-day-old mouse brains. Furthermore, we found that enforced neuronal ISG12 expression results in a significant delay in Sindbis virus-induced death in neonatal mice. Together, our data identify genes that are developmentally regulated in the CNS and genes that are differentially regulated in the brains of different aged mice in response to Sindbis virus infection.

Background Genital herpes simplex virus-2 (HSV-2) infection consists of shedding episodes that correlate with transmission and genital lesion formation. Episodes are heterogeneous in terms of viral production and duration. Methods Using quantitative PCR data from 14 685 daily genital swabs performed daily in 531 persons, we summarised 1020 episodes according to their frequency, duration, peak HSV DNA copy number, first and last swab copy number, expansion and decay slopes, and morphology. We designed competing dynamical stochastic mathematical models and attempted to fully reproduce result ranges for each of these episode features. The models assumed the possibility of concurrent, spatially distinct plaques of viral infection and immunologic response. Results Our spatial model reproduced all kinetic features of the empirical shedding data. The model and parameter set that achieved best fit highlights three distinct mechanisms of viral spread. First, HSV-2 is constantly dispersed from neurons to epidermal cells throughout the genital tract at a rate of ∼100 HSV DNA copies/day. Approximately 75 times each year, a neuron-derived viral particle infects an epidermal cell, initiating an infectious plaque of epidermal cells and a detectable shedding episode. Second, within each plaque, cell-to-cell spread of HSV-2 particles occurs extremely rapidly between epidermal cells. One cell can produce 105 cell-associated HSV genomic copies/day, with 3% of viral particles infecting a neighbouring cell each day. In large plaques, >105 cells may be infected within ∼12 h producing up to 109 HSV DNA copies/ml. Third, cell-free viruses, despite being 3000-times less infectious than cell-associated viruses, decay more slowly and initiate spatially distinct secondary plaques. We use video simulations to display that episodes longer than 3 days, consist of dozens of concurrent viral plaques. Despite rapid cell-to-cell spread of HSV-2, infected cells are eliminated by localised CD8+ T-cells within 24 h of plaque initiation. Moreover, the extent of secondary plaque formation prior to episode termination is determined by spatial CD8+ T-cell density surrounding the site of infection. Conclusions Genital HSV-2 utilises three kinetically distinct methods of spread to initiate and sustain prolonged shedding episodes. The extent and severity of secondary plaque formation is determined by spatial immune cell density.

Background Serologic testing for herpes simplex virus type-2 (HSV-2) in persons without a history of genital herpes is currently not recommended partly due to the concern that HSV-2 diagnosis would lead to negative psychosocial sequelae, such as anxiety and depression. We conducted a systematic review to assess the evidence as to whether HSV-2 serologic testing among asymptomatic persons results in persistent negative psychosocial consequences. Methods Eight electronic databases and unpublished data sources were searched to identify studies measuring the psychosocial impact of HSV-2 serologic testing in persons without a history of genital herpes. To be included, studies had to test for HSV-2 using an HSV type-specific serologic test and to perform at least one psychosocial assessment of participants after they received HSV serologic results. We compared psychosocial responses in HSV-2 positive persons over time and vsHSV-2 negative persons (when available). Results Nine studies satisfied the inclusion criteria. Studies were published from the years 2000–2008 and were conducted in the USA (N=6), Australia (N=2), and the UK (N=1). In total, 1355 participants were included; 596 (44%) participants were HSV-2 positive, and of these 341 (57%) lacked prior history of genital herpes. Participants were recruited from a variety of settings (ie, STD clinics, HMO enrollees, college campuses). Follow-up ranged from immediately after diagnosis to 1 year afterwards. Seven studies reported that HSV-2 diagnosis by serologic test did not have a persistent negative impact on participants9 mental health (anxiety, depression, self-esteem) or sexual attitude and satisfaction. Two studies reported a negative impact of testing; one found that 5 HSV-2 seropositive college students had increased distress 3 months post-testing as compared to HSV-2 negatives, and the other found self-reports of sexual undesirability up to one year after diagnosis. A genital herpes diagnosis was perceived as moderately severe for participants prior to testing; however, after HSV-2 testing, the perceived severity of a herpes diagnosis was lower among those testing HSV-2 positive. Conclusions Diagnosis of HSV-2 by type-specific serologic testing did not result in long-term psychosocial harm in most asymptomatic persons. Concerns about sustained emotional impact should not deter clinicians from testing individuals without a history of genital herpes for HSV-2.

Background Current treatments for HSV infection are imperfect, do not completely abrogate viral shedding or transmission, and do not interrupt HSV-2—HIV interactions. AIC316 is a helicase-primase inhibitor, a new class of anti-HSV compounds that has a distinct mode of action from currently available nucleoside analogues. Methods We investigated the safety and efficacy of AIC316 in patients with genital HSV-2 infection in a Phase 2, randomised, multicenter, parallel, double-blind, placebo-controlled trial of 4 different doses of AIC316 administered orally for 4 weeks. Participants were randomised 1:1:1:1:1 to one of the following dose groups: 5 mg given once daily (qd), 25 mg qd, 75 mg qd, 400 mg given once-a-week, or matching placebo. Participants in the once daily dose groups received a loading dose. During the trial period participants obtained a swab of genital secretions daily and maintained a diary of genital lesions. The primary endpoint was the rate of genital HSV shedding measured by HSV DNA PCR in the AIC316 groups vs placebo. Results 156 (105 women and 51 men) healthy, immunocompetent, HSV-2 positive participants with a history of 1–9 recurrences per year prior to trial entry, or previous suppressive therapy, were randomised by seven US sites between May 2010 and October 2010. 147 completed the trial. Overall, about 9000 swabs for HSV PCR were collected and assayed for HSV DNA by a sensitive and accurate assay that can detect >150 copies/ml. The first results of these assessments will be presented. Conclusion The trial will provide insight into the antiviral activity of the novel agent AIC316 for genital HSV infections. This trial design presents a robust and efficient method for evaluating antiviral activity of candidate agents for mucocutaneous HSV infections. These initial efficacy and safety results will lead to selecting the dose for further trials with AIC316.

Life-long latent neuronal infection with varicella zoster virus (VZV) is a legacy of childhood chickenpox. VZV reactivation occurs in about 1/3 of persons to cause shingles. Live, attenuated VZV (vOka)—the only licensed therapeutic vaccine—moderately reduces both the incidence and the severity of shingles. The partial level of protection is poorly understood but may plausibly derive from variation in T cell boosting. Twelve subjects (>50 years old) were vaccinated with vOka and blood obtained on days 0, 14, and 28. Ex vivo ICC analyses confirmed boosting of vOka-reactive T cells: the mean increase in frequency of IFNγ+IL2+ CD4+ T cells was 74% by day 14 (p=0.02). We enriched and expanded VZV-specific CD4+ T cells in an unbiased manner and defined the specificity and breadth of the resulting polyclonal CD4+ T cell lines using each of the 70 known VZV proteins. Prior to boosting, the median number of detectable responses was 9 ORFs (range 3-10). Vaccination increased the median breadth to 17 ORFs (range 9-20) at day 28. Prevalent targets before and after boosting were glycoproteins gE, gI, and gH, and the tegument proteins encoded by ORF12 and ORF44. While new responses were noted post-vaccination, baseline responses rarely disappeared. T cell breadth was well-maintained during VZV latency and further increased by therapeutic vaccination. In addition to CD4+ T cell magnitude, specificity and breadth are now accessible as candidate immune correlates of vaccine efficacy.

Peptide-HLA diversity exceeds the TCR repertoire. The resultant T-cell cross-reactivity implies that prior antigen exposure could modulate the outcome of heterologous infections. The alphaherpesviruses HSV-1, HSV-2, and VZV have unexplained ranges of infection severity, and the reasons for partial efficacy of therapeutic live VZV vaccination are unknown. The viruses share 60 homologous ORFs. Using whole viruses, protein sets covering the viral ORFeomes, and peptides, we studied the cross-reactivity of polyclonal and mono-specific CD4 and CD8 T-cells. Effectors were enriched from PBMC using HSV or VZV by direct or cross-presentation (J. Clin. Invest. 122:654) or with tetramers. Cross-reactivity between HSV and VZV was readily detectable. For example, VZV-driven polyclonal CD4 T-cells lines showed a mean of 31% IFN-gamma/IL-2 reactivity to whole VZV and a mean of 11% reactivity to HSV-1, implying that 35% of T-cells were cross-reactive. Cross-reactivity was confirmed at the protein and peptide levels for both CD4 and CD8 T-cells. Cross-reactive CD8 T-cells directly recognized infected target cells, including non-IFN-treated VZV-infected dermal fibroblasts, despite VZV encoding a TAP-blocking protein. We hypothesize that cross-reactive alphaherpesvirus ORFs are rational subunit vaccine candidates to cover HSV and VZV, and that heterologous infections and resulting T-cell cross-reactivity may contribute to variability in infection severity and vaccine responses.

Single-amino-acid mutations in Sindbis virus proteins can convert clinically silent encephalitis into uniformly lethal disease. However, little is known about the host gene response during avirulent and virulent central nervous system (CNS) infections. To identify candidate host genes that modulate alphavirus neurovirulence, we utilized GeneChip Expression analysis to compare CNS gene expression in mice infected with two strains of Sindbis virus that differ by one amino acid in the E2 envelope glycoprotein. Infection with Sindbis virus, dsTE12H (E2-55 HIS), resulted in 100% mortality in 10-day-old mice, whereas no disease was observed in mice infected with dsTE12Q (E2-55 GLN). dsTE12H, compared with dsTE12Q, replicated to higher titers in mouse brain and induced more CNS apoptosis. Infection with the neurovirulent dsTE12H strain was associated with both a greater number of host genes with increased expression and greater changes in levels of host gene expression than was infection with the nonvirulent dsTE12Q strain. In particular, dsTE12H infection resulted in greater increases in the levels of mRNAs encoding chemokines, proteins involved in antigen presentation and protein degradation, complement proteins, interferon-regulated proteins, and mitochondrial proteins. At least some of these increases may be beneficial for the host, as evidenced by the demonstration that enforced expression of the antiapoptotic mitochondrial protein peripheral benzodiazepine receptor (PBR) protects neonatal mice against lethal Sindbis virus infection. Thus, our findings identify specific host genes that may play a role in the host protective or pathologic response to neurovirulent Sindbis virus infection.

Corrigendum: Immune surveillance by CD8αα + skin-resident T cells in human herpes virus infection

Background Short, rapidly cleared, subclinical shedding episodes are the predominant form of HSV-2 reactivation in the genital tract. Valacyclovir 500 mg once daily (SD-VAL) reduces the risk of sexual transmission of herpes simplex virus type 2 (HSV-2) by only 48%. We hypothesised that short HSV-2 shedding episodes occur frequently on SD-VAL and that high dose (HD)-VAL could suppress such episodes of genital HSV-2 shedding. Methods A randomised open-label crossover study using valacyclovir 500 mg daily (SD-VAL) vs valacyclovir 1 gm three times daily (HD-VAL) was conducted in HSV-2 seropositive, HIV seronegative persons with four or more genital herpes recurrences per year or laboratory confirmed primary genital HSV-2 infection in the previous 6 months. Each study arm lasted for 5 weeks, separated by 1 week wash out. Participants obtained genital swabs four times daily, which were assayed for HSV by quantitative PCR. The primary outcome was frequency of genital HSV shedding on each study arm; secondary outcomes included number and duration of HSV-2 shedding episodes and quantity of virus detected. Results Forty-three participants collected 9981 genital swabs during the study period. 292 (5.8%) of 5008 swabs had HSV detected during SD-VAL, compared to 164 (3.3%) of 4973 on HD-VAL (IRR=0.52, 95% CI=0.43% to 0.63%, p Conclusion Short bursts of genital HSV-2 reactivation persist during SD-VAL and HD-VAL. Compared to SD-VAL, HD-VAL decreased shedding frequency and episode duration but did not alter episode rate. These data may explain why risk of HSV transmission and HSV-specific genital inflammation persist even in the presence of antiviral therapy. More potent therapies are needed to completely suppress HSV-2 reactivation ClinicalTrials.gov number NCT00362297.

Background Standard doses of HSV suppressive therapy reduce plasma HIV-1 levels (0.25-0.5 log10 copies/mL) among HIV-1/HSV-2 co-infected persons, and modestly slow disease progression. Putative mechanisms for this effect include direct inhibition of HIV-1 by acyclovir or indirect reduction by decreasing HSV-associated inflammation. We hypothesised that higher-dose anti-HSV therapy would result in greater reduction in plasma HIV-1 RNA, and that the effect would be mediated by greater suppression of HSV shedding. Methods 34 participants with HIV-1 and HSV-2 who were not on antiretroviral therapy were enrolled into a randomised, open-label cross-over trial with valacyclovir 1000 mg twice daily or acyclovir 400 mg twice daily for 12 weeks. After a 2 week wash-out, they were crossed over to the alternate treatment arm for 12 weeks. HSV PCR was performed on self-collected genital swabs obtained daily during the first 4 weeks of each treatment period. Plasma HIV-1 RNA was measured weekly throughout the study. Results Among the 26 participants who completed both arms of the study, the mean age was 44; 21 were men. At entry, mean CD4 count was 525 cells/mm3 (range, 242–1055) and mean plasma HIV-1 RNA 3.9 log10 copies/ml (range 1.2–5.5). The mean plasma HIV RNA was 3.86 log10 copies/ml during acyclovir administration compared with 3.57 log10 copies/ml on valacyclovir; a 0.29 log10 copies/ml reduction (p=0.002). One week after initiation of valacyclovir, plasma HIV RNA decreased by a mean of 0.40 log10 copies/ml. Valacyclovir reduced HIV-1 RNA by ≥0.25 log10 copies/ml in 14 (54%) participants, compared with 3 (12%) on acyclovir. Neither the HSV shedding rate (8.92% vs 8.98% of days, p=0.94), nor the genital lesion rate (4.3% vs 1.1%; p=0.18) differed on acyclovir vs valacyclovir. Conclusions High-dose valacyclovir reduces plasma HIV-1 RNA levels more effectively than standard dose acyclovir in HIV-1, HSV-2 seropositive persons not receiving antiretroviral therapy. High dose valacyclovir does not provide more potent suppression of HSV reactivation in HIV-1 infected persons than acyclovir, suggesting that the effect of valacyclovir on HIV-1 RNA may not be mediated via HSV suppression.

In a sand-dune system at Tentsmuir Point, Fifeshire, Scotland, Glomus fasciculatus was the only endophyte present on roots of pioneer colonizing grasses. This fungus formed sporocarps and spore aggregates in the soil and in association with organic fragments, particularly root tissues. Amounts of infection and external mycelium varied with vegetation zone and season, but both became greater with dune stabilization and infection increased during summer and autumn. Growth of Ammophila arenaria and Agropyron junceiforme in unsterile foreshore sand was improved when mycorrhizal with G. fasciculatus and G. macrocarpus var. geosporus . Experiments in dune sands, with maize as a test plant, gave better growth with both these endophytes than with either alone, but indicated the complexity of interaction between host, endophyte and edaphic factors. The possible significance of mycorrhizas in nutrientpoor and unstable habitats is discussed.

Summary Background Skin and mucosal herpes simplex virus type 2 (HSV-2) shedding predominantly occurs in short subclinical episodes. We assessed whether standard-dose or high-dose antiviral therapy reduces the frequency of such shedding. Methods HSV-2-seropositive, HIV-seronegative people were enrolled at the University of Washington Virology Research Clinic (WA, USA). We did three separate but complementary open-label cross-over studies comparing no medication with aciclovir 400 mg twice daily (standard-dose aciclovir), valaciclovir 500 mg daily (standard-dose valaciclovir) with aciclovir 800 mg three times daily (high-dose aciclovir), and standard-dose valaciclovir with valaciclovir 1 g three times daily (high-dose valaciclovir). The allocation sequence was generated by a random number generator. Study drugs were supplied in identical, numbered, sealed boxes. Study periods lasted 4–7 weeks, separated by 1 week wash-out. Participants collected genital swabs four times daily for quantitative HSV DNA PCR. Clinical data were masked from laboratory personnel. The primary endpoint was within-person comparison of shedding rate in each study group. Analysis was per protocol. The trials are registered at ClinicalTrials.gov (NCT00362297, NCT00723229, NCT01346475). Results Of 113 participants randomised, 90 were eligible for analysis of the primary endpoint. Participants collected 23 605 swabs; 1272 (5·4%) were HSV-positive. The frequency of HSV shedding was significantly higher in the no medication group (n=384, 18·1% of swabs) than in the standard-dose aciclovir group (25, 1·2%; incidence rate ratio [IRR] 0·05, 95% CI 0·03–0·08). High-dose aciclovir was associated with less shedding than standard-dose valaciclovir (198 [4·2%] vs 209 [4·5%]; IRR 0·79, 95% CI 0·63–1·00). Shedding was less frequent in the high-dose valaciclovir group than in the standard-dose valaciclovir group (164 [3·3%] vs 292 [5·8%]; 0·54, 0·44–0·66). The number of episodes per person-year did not differ significantly for standard-dose valaciclovir (22·6) versus high-dose aciclovir (20·2; p=0·54), and standard-dose valaciclovir (14·9) versus high-dose valaciclovir (16·5; p=0·34), but did for no medication (28·7) and standard-dose aciclovir (10·0; p=0·001). Median episode duration was longer for no medication than for standard-dose aciclovir (13 h vs 7 h; p=0·01) and for standard-dose valaciclovir than for high-dose valaciclovir (10 h vs 7 h; p=0·03), but did not differ significantly between standard-dose valaciclovir and high-dose aciclovir (8 h vs 8 h; p=0·23). Likewise, maximum log 10 copies of HSV detected per mL was higher for no medication than for standard dose aciclovir (3·3 vs 2·9; p=0·02), and for standard-dose valaciclovir than for high-dose valaciclovir (2·5 vs 3·0; p=0·001), but no significant difference was recorded for standard-dose valaciclovir versus high-dose aciclovir (2·7 vs 2·8; p=0·66). 80% of episodes were subclinical in all study groups. Except for a higher frequency of headaches with high-dose valaciclovir (n=13, 30%) than with other regimens, all regimens were well tolerated. Interpretation Short bursts of subclinical genital HSV reactivation are frequent, even during high-dose antiherpes therapy, and probably account for continued transmission of HSV during suppressive antiviral therapy. More potent antiviral therapy is needed to eliminate HSV transmission. Funding NIH. Valaciclovir was provided for trial 3 for free by GlaxoSmithKline.

The ethics of studying or collecting unprovenanced material is intrinsically tied to the issue of looting. In recent years looting has become increasingly facilitated by the current political instability in various parts of the world, with looted objects often serving as a highly lucrative source of income for insurgent or even terrorist groups. As international statutes and industry protocols aim to alleviate further threats to cultural heritage, consideration is also due to the future of the vast collections of unprovenanced and subsidiary material acquired through decades of cultural tourism and incipient archaeological exploration. A survey of Canadian and American academic institutions was conducted to determine the extent of department-held material collections, as well as the current pedagogical and research purposes to which they are directed.

40,000 spectra from the EPA / NIH database and over 70,000 spectra from the Wiley collection.

Contains chapters and sections on all the main types of chemical literature, including paper and computerized information.