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Abstract Objectives Biofilm production in extended spectrum β-lactamase (ESBL)-producing Enterobacteriaceae provides a favourable environment for the exchange of antibiotic-resistance genes and could facilitate widespread dissemination. We aimed to assess biofilm development in ESBL-producing E. coli and K. pneumoniae isolates and determine how development relates to microbiological characteristics and clinical outcomes. Methods 147 ESBL-producing E. coli and 82 K. pneumoniae were genetically characterized. Biofilm formation was measured at 1.5, 4, 6, and 24 h during culture in blood heart infusion using a microbead immobilization assay (BioFilm Ring test®). Results were given as biofilm formation index (BFI) with lower values indicating increased presence of biofilm (range = 0–21). Results In total, 57.1% of strains were strong producers of biofilm (BFI < 2), whereas 13.4% lacked biofilm production (BFI > 18). Standard biofilm production (BFI < 7) was common in E. coli isolates (61.9%). For E. coli , biofilm production was less frequently observed in ST131 clones (p = 0.03) but more frequently in strains harbouring toxin (p = 0.008) or adhesin (p = 0.008) virulence factor genes. Despite almost all K. pneumoniae having standard biofilm production (90.2%), there was a 2.4-times higher odds of observing biofilm in ST29/147/323 versus other ST-types (p = 0.13). Patients with standard biofilm producing isolates were not at increased risk of transfer to intensive-care (odds-ratio=2.80, 95%CI=0.59–13.21) or death within 12-months (odds-ratio=1.61, 95%CI=0.75–3.43). Conclusion In these ESBL-producing strains, biofilm production is linked to certain virulence factors in E. coli and is common in K. pneumoniae. Further exploration of whether biofilm production increases dissemination and risk of severe clinical outcomes is needed in larger collections of isolates. [ABSTRACT FROM AUTHOR]

The future of rural youth employment is a major issue in sub-Saharan Africa because structural transformation is slow. However, youth employment is rarely considered in terms of institutional economics or in a long-term perspective, both of which are essential for analysing structural change. Little research has focused on employment issues and how they are affected by the type of rural areas, and especially the farming potential. This paper seeks to identify the main institutional determinants of youth transitions. We examine the assets and opportunities, internal or external to the family, available to young people. Combining a theoretical framework based on institutional economics with a comparative and historical methodology can reveal structural change in rural areas. The research draws on original longitudinal data collected in four rural areas in Senegal and Zambia in order to analyse transitions to adulthood among three successive cohorts of youth. While the majority of youth employment policies focus on improving young people's skills, the long-term analysis reveals the institutional changes regarding access to land, capital and knowledge, as well as the redistribution of value and social protection. The analysis points to the importance of taking account of structural characteristics and related institutions when addressing the challenge of employment for future cohorts of youth. • The future of rural youth employment is a major issue in sub-Saharan Africa because the structural transformation in the region is slow. • Access to capital by youth is now more dependent on labour market institutions than on family especially in areas with low farming potential. • It is increasingly common for young men to gain access to land through informal rent agreements or purchase. • young women have access to land through their husband or women's collective organisations. • Rural youth employment challenge asks for structural policies while current public policies often boil down to facilitating the emergence of « agripreneurs » [ABSTRACT FROM AUTHOR]

Knowledge about lipid interventions in secondary prevention in HIV-infected individuals is limited; studies are sparse.Methods: A prospective observational multicenter study enrolled 282 patients on statin 1 month after first acute coronary syndrome (ACS) (95 HIV-infected individuals, 187 HIV-uninfected). Data on fasting lipids (total cholesterol [TC], low-density lipoprotein cholesterol, high-density lipoprotein cholesterol [HDL-C], non-HDL-C, triglycerides, TC/HDL-C ratio) were collected over 3 years. The evolution of lipid concentrations was analyzed using mixed-effects models. Achievement of National Cholesterol Education Program Adult Treatment Panel III lipid goals and prescribed statin intensity was assessed.Results: Mean age of patients was 49.0 years, and 94% were men. Baseline lipids were similar in the 2 groups. Six months after first ACS, less low-density lipoprotein cholesterol reduction was observed in the HIV-infected GROUP (adjusted mean change -10.13; 95% CI -20.63 to 0.37; P=.06) than in the HIV-uninfected group (Adjusted mean change -38.51; 95% CI -46.00 to -31.04; P<.0001) Similar trends were observed for TC and non-HDL-C. After ACS, initial statin prescription for HIV-infected compared with HIV-uninfected individuals was more frequently a moderate-intensity statin (66% vs 45%) and less frequently a high-intensity statin (15% vs 45%). Over 3 years of follow-up, the proportion of HIV-infected patients receiving high-intensity statin remained persistently lower than the proportion observed in the HIV-uninfected group.Conclusions: In this observational study, HIV-infected individuals after first ACS exhibited worse lipid profiles than controls particularly during the first 6 months while receiving less potent statins. Appropriate statin intensity should be prescribed in HIV-infected individuals with awareness of potential drug-drug interactions. [ABSTRACT FROM AUTHOR]

Nonalcoholic fatty liver disease (NAFLD) is a growing concern in the aging population with human immunodeficiency virus (HIV). Screening for NAFLD is recommended in patients with metabolic risk factors or unexplained transaminitis. This study aimed to prospectively assess the prevalence and associated factors of liver steatosis and advanced fibrosis (AF) in HIV-monoinfected patients at risk of NAFLD. We conducted a multicenter study in HIV-monoinfected patients, nonexcessive drinkers with metabolic syndrome, and/or persistently elevated liver enzymes, and/or clinical lipodystrophy. All participants had magnetic resonance imaging proton density fat fraction (MRI-PDFF), Fibroscan/controlled attenuation parameter (CAP), and cytokine and genetic analysis. From March 2014 to November 2015, we enrolled 442 participants and analyzed 402: male (85%); median age, 55 years (interquartile range [IQR], 50-61 years); body mass index, 27.0 kg/m2 (IQR, 23.6-28.7 kg/m2); metabolic syndrome (67%); and CD4 cell count, 630/mm3 (IQR, 510-832/mm3). Overall 257 of 402 (64%) had NAFLD (MRI-PDFF ≥5%). Among them, 11.3% had a liver stiffness ≥9.6 kPa, suggestive of AF. Multivariable analysis identified 7 factors of steatosis: high CD4-cell count (odds ratio [OR], 4.04; 95% confidence interval [CI], 1.92–8.51), high leptin level (OR, 2.12; 95% CI, 1.14–3.93), non-CC PNPLA3 s738409 genetic polymorphism (OR, 1.92; 95% CI, 1.11–3.33), low high-density lipoprotein (OR, 1.83; 95% CI, 1.03–3.27), high triglycerides (OR, 1.48; 95% CI, 1.18–1.84), elevated alanine transaminase (OR, 1.23; 95% CI, 1.16–1.31), and hyper ferritinemia (OR, 1.05; 95% CI, 1.03–1.07). Two factors were associated with AF: high body mass index (OR, 1.23 ; 95% CI, 1.07–1.42 ; P =.005, and elevated aspartate aminotransferase (OR, 1.03; 95% CI, 1.01–1.05; P =.001). Using MRI-PDFF as a reference, CAP (best cutoff, 280 dB/m) had good accuracy (area under the receiver operating characteristic curve = 0.86; 95% CI, 0.82–0.90) for the diagnosis of moderate to severe steatosis. In a large cohort of HIV-moninfected patients at risk of NAFLD, steatosis is present in two-thirds of cases, and around 10% have AF. The CAP technique is accurate for screening steatosis in this population. [Display omitted] [ABSTRACT FROM AUTHOR]

We introduce here the term “River Culture” to delineate an eco-social approach to mitigate the biological and cultural diversity crisis in riverscapes. It is based on the insight that current environmental change endangers both, biological and cultural diversities in rivers and their basins, and those activities to improve ecosystem functions, biodiversity and capacity of the biological species to evolve will have a similarly positive effect on human cultural diversity. “River Culture” has two dimensions, including (a) the influence of the biophysical setting of rivers (specifically, their pulsating flow regimes and their biological features) on the expression of elements of human culture in general and (b) the aspect of “learning from the river” for the development of technologies and management options that are targeted to maintain and improve ecosystem functions and diversity in a more sustainable way. The River Culture approach, as given in this concept and discussion paper, is preliminarily based on five tenets: (1) Reset values and priorities in riverscape management in favor of human wellbeing and a harmonious coexistence of man and riverscape; (2) Live in the rhythm of the waters, i.e. adapt management options in accordance with the hydrological dynamics rather than fighting against them; (3) Transform traditional use of rivers into modern cultural activities and management options; (4) ‘Ecosystem bionics’: by copying survival strategies of flood-pulse adapted organisms novel forms of human use can be developed; (5) Make the catchment (river basin) the geographical base unit for all kinds of political decisions in landscape management. [ABSTRACT FROM AUTHOR]

Summary Background Our objective was to assess therapeutic non-inferiority of dual treatment with lopinavir–ritonavir and lamivudine to triple treatment with lopinavir–ritonavir plus two nucleos(t)ides for maintenance of HIV-1 viral suppression. Methods In this randomised, open-label, non-inferiority trial, we recruited patients from 32 HIV units in hospitals in Spain and France. Eligible patients were HIV-infected adults (aged ≥18 years) with HIV-1 RNA of less than 50 copies per mL, for at least 6 months on triple treatment with lopinavir–ritonavir (twice daily) plus lamivudine or emtricitabine and a second nucleos(t)ide, with no resistance or virological failure to these drugs, and no positive hepatitis B serum surface antigen. Investigators at each centre randomly assigned patients (1:1; block size of four; stratified by time to suppression [<1 year or >1 year] and nadir CD4 cell count [<100 cells per μL or >100 cells per μL]; computer-generated random sequence) to continue triple treatment or switch to dual treatment (oral lopinavir 400 mg and oral ritonavir 100 mg twice daily plus oral lamivudine 300 mg once daily). The primary endpoint was response to treatment in the intention-to-treat population (all randomised patients) at 48 weeks. The non-inferiority margin was 12%. This study is registered with ClinicalTrials.gov , number NCT01471821 . Findings Between Oct 1, 2011, and April 1, 2013, we randomly assigned 250 participants to continue triple treatment (127 [51%] patients) or switch to dual treatment (123 [49%] patients). In the intention-to-treat population, 110 (86·6%) of 127 patients in the triple-treatment group responded to treatment versus 108 (87·8%) of 123 in the dual-treatment group (difference −1·2% [95% CI −9·6 to 7·3]; p=0·92), meeting the criteria for non-inferiority. Serious adverse events occurred in eight (7%) patients in the triple-treatment group and five (4%) in the dual-treatment group (p=0·515), and study drug discontinuations due to adverse events occurred in four (3%) in the triple-treatment group and one (1%) in the dual-treatment group (p=0·223). Interpretation Dual treatment with lopinavir–ritonavir plus lamivudine has non-inferior therapeutic efficacy and is similarly tolerated to triple treatment. Funding AbbVie and Red Temática Cooperativa de Investigación en Sida. [ABSTRACT FROM AUTHOR]

[Display omitted] • Spodoptera frugiperda genome contains two genes encoding prophenoloxidases. • Sf PPOs complement one another for hemolymph phenoloxidase activity. • Sf PPO1Δ,PPO2Δ double mutant presents morphological defects during pupal stage. • Sf PPO1Δ,PPO2Δ double mutant pupae are not able to develop into adults. Upon infection, the phenoloxidase system in arthropods is rapidly mobilized and constitutes a major defense system against invaders. The activation of the key enzymes prophenoloxidase (PPO) and their action in immunity through melanization and encapsulation of foreign bodies in hemolymph has been described in many insects. On the other hand, little is known about PPOs involvement in other essential functions related to insect development. In this paper, we investigated the function of the two PPOs of the crop pest, Spodoptera frugiperda (PPO1 and PPO2). We show that PPOs are mainly expressed in hemocytes with the PPO2 expressed at higher levels than the PPO1. In addition, these two genes are expressed in the same tissue and at the same stages of insect development. Through the generation of loss-of-function mutants by CRISPR/Cas9 method, we show that the presence of PPOs is essential for the normal development of the pupa and the survival of the insect. [ABSTRACT FROM AUTHOR]

Abstract: Lysozyme is an important component of the insect non-specific immune response against bacteria that is characterized by its ability to break down bacterial cell-walls. By searching an EST database from the fall armyworm, Spodoptera frugiperda (), we identified five sequences encoding proteins of the lysozyme family. The deduced protein sequences corresponded to three classical c-type lysozymes Sf-Lys1, Sf-Lys2 and Sf-Lys3, and two lysozyme-like proteins, Sf-LLP1 and Sf-LLP2. Sf-Lys1 was purified from the hemolymph of Escherichia coli-challenged S. frugiperda larvae. The mature protein had a molecular mass of 13.975Da with an isoelectric point of 8.77 and showed 98.3% and 96.7% identity with lysozymes from Spodoptera litura and Spodoptera exigua, respectively. As the other insect lysozymes, Sf-Lys1 was active against Gram positive bacteria such as Micrococcus luteus but also induced a slight permeabilization of the inner membrane of E. coli. Genes encoding these five Sf-Lys or Sf-LLPs were differentially up-regulated in three immune-competent tissues (hemocytes, fat body and gut) after challenges with non-pathogenic bacteria, E. coli and M. luteus, or entomopathogenic bacterium, Photorhabdus luminescens. Sf-Lys1 and Sf-Lys2 were mainly induced in fat body in the presence of E. coli or P. luminescens. Sf-Lys3, which had an acidic isoelectric point, was found to be the most up-regulated of all five Sf-Lys or Sf-LLPs in hemocytes and gut after challenge with P. luminescens. More molecular data are now available to investigate differences in physiological functions of these different members of the lysozyme superfamily. [Copyright &y& Elsevier]

Abstract: Ultraviolet A (UVA) radiation represents more than 90% of the UV spectrum reaching Earth''s surface. Exposure to UV light, especially the UVA part, induces the formation of photoexcited states of cellular photosensitizers with subsequent generation of reactive oxygen species (ROS) leading to damages to membrane lipids, proteins and nucleic acids. Although UVA, unlike UVC and UVB, is poorly absorbed by DNA, it inhibits cell cycle progression, especially during S-phase. In the present study, we examined the role of the DNA damage checkpoint response in UVA-induced inhibition of DNA replication. We provide evidence that UVA delays S-phase in a dose dependent manner and that UVA-irradiated S-phase cells accumulate in G2/M. We show that upon UVA irradiation ATM-, ATR- and p38-dependent signalling pathways are activated, and that Chk1 phosphorylation is ATR/Hus1 dependent while Chk2 phosphorylation is ATM dependent. To assess for a role of these pathways in UVA-induced inhibition of DNA replication, we investigated (i) cell cycle progression of BrdU labelled S-phase cells by flow cytometry and (ii) incorporation of [methyl-3H]thymidine, as a marker of DNA replication, in ATM, ATR and p38 proficient and deficient cells. We demonstrate that none of these pathways is required to delay DNA replication in response to UVA, thus ruling out a role of the canonical S-phase checkpoint response in this process. On the contrary, scavenging of UVA-induced reactive oxygen species (ROS) by the antioxidant N-acetyl-l-cystein or depletion of vitamins during UVA exposure significantly restores DNA synthesis. We propose that inhibition of DNA replication is due to impaired replication fork progression, rather as a consequence of UVA-induced oxidative damage to protein than to DNA. [Copyright &y& Elsevier]

Abstract: We report here the isolation in Spodoptera frugiperda (Lepidoptera) of an immune-related protein (hereafter named Spod-11-tox), characterized by imperfectly conserved tandem repeats of 11 cysteine-stabilized alpha beta motifs (CS-αβ), the structural scaffold characteristic of invertebrate defensins and scorpion toxins. Spod-11-tox orthologs were only found in Lepidopteran species, suggesting that this new protein family (named X-tox) is specific to this insect order. Moreover, phylogenetic analysis suggests that X-tox proteins represent a new class of proteins restricted to Lepidoptera and likely derived from Lepidopteran defensins. In S. frugiperda, analysis of gene expression revealed that spod-11-tox is rapidly induced by infection. However, and conversely to what is known for most insect antimicrobial peptides (AMP), spod-11-tox is mainly expressed in blood cells. Moreover, recombinant Spod-11-tox produced in the Sf9 cell line does not show any antimicrobial activity. Altogether, these results suggest that although X-tox proteins are derived from defensins, they may play a different and still unknown role in Lepidoptera immune response. [Copyright &y& Elsevier]

Aims: Natural history and prognosis of ACS in HIV-infected patients remain to be determined. The aim of our study was to compare coronary risk factors, angiographic features, acute results of PCI, in-hospital outcomes, and prespecified 1and 3-year prognosis of HIV-infected and HIV-uninfected patients with ACS. Methods and results: HIV-infected and HIV-uninfected patients with a first episode of ACS were matched for age (±5 years), sex, and type of ACS. The primary endpoint was the rate of major adverse cardiac and cerebral events (MACCE), comprising cardiac death, recurrent ACS, recurrent coronary revascularization, and stroke. Overall 103 HIV-infected and 195 HIV-uninfected patients were enrolled (mean age 49.0±9.4 years, 94% men). Coronary risk factors were well balanced, but HIV-infected patients more frequently used illicit drugs (23% vs 6%, P =0.001) and had higher triglyceride level (246±189 vs 170±139mg/dl, P =0.002) compared with HIV-uninfected patients. Angiographic features of CAD was similar (multivessel disease 41% vs 39%, p=0.96; ACC/AHA type culprit lesion≥B2, both 77%, P =0.83). At 1 year, the rate of occurrence of first MACCE did not differ between groups (hazard ratio [HR:] 1.4; 95% CI: 0.6 to 3.0). Recurrent ACS was more frequent in HIV-infected patients (HR: 4.6; 95% CI: 1.4 to 15.0) with no difference in the rate of clinical restenosis. Conclusion: These results suggest that acute management of ACS in HIV-infected patients can routinely be the same as that of HIV-uninfected patients but that specific secondary prevention measures are needed to alleviate this increased risk of recurrent ACS. The 3-year follow up will be obtained and analyzed before the end of 2010. [ABSTRACT FROM AUTHOR]

Objective: Human immunodeficiency virus (HIV)-infected patients receiving combined active antiretroviral therapy (cART) are at higher risk of cardiovascular disease, due in part to metabolic complications such as lipodystrophy syndrome, insulin resistance, and dyslipidemia. Whether lipodystrophy and cART impact on the vasculature is debated. We investigated the impact of lipodystrophy and protease inhibitors (PIs) on aortic stiffness. Methods: Aortic stiffness was evaluated using carotid-femoral pulse wave velocity (PWV) in consecutive HIV-infected patients without a history of cardiovascular disease referred to a cardiovascular clinic. Results: 175 patients were enrolled (mean age 48.2±8.7 years; 89% men). Eighty six per cent of patients were receiving cART. Dyslipidemia, tobacco, and hypertension were the most prevalent cardiovascular risk factors (39%, 38%, and 31%, respectively). Seventy-nine (45%) HIV-infected patients had lipodystrophy and 80 (46%) were on PIs. Aortic PWV was similar in patients with or without lipodystrophy (9.7±1.9 vs 9.8±2.5ms−1, respectively; P =0.81) and in patients on or not on PIs (9.8±2.6 vs 9.7±1.9ms−1; P =0.71). In univariate analysis, aortic PWV was associated with increasing age, waist/hip ratio, systolic and diastolic blood pressures, mean arterial and pulse pressures, but not with presence of lipodystrophy, PIs, or specific factors related to HIV infection. Linear regression analysis showed an association between aortic PWV and age (=0.49, P =0.001) and systolic arterial pressure (=0.21, P =0.006). Conclusions: Aortic stiffness is associated with traditional cardiovascular risk factors, particularly ageing and blood pressure. Hypertension is becoming an emerging complication in HIV-infected patients. [ABSTRACT FROM AUTHOR]

• Rapid evaluation of treatment efficacy is needed in case of empirical tuberculosis (TB) treatment. • Innate immune markers were measured in patients with TB who are HIV-negative and HIV-positive. • Interleukin-1 receptor antagonist and interferon-γ–induced protein-10 plasma levels decreased as soon as 7 days of TB treatment onset. • It is the first report of an early decrease of markers measured with standard enzyme-linked immunosorbent assay. • Interleukin-1 receptor antagonist-guided management deserves evaluation in empirically treated people with HIV. Monitoring tools that could provide quick predictions of tuberculosis (TB) treatment outcomes are urgently needed. Here, we assessed whether the evolution of selected biomarkers of innate immunity may help monitoring TB treatment response within 2 weeks of treatment initiation. ANRS12394-LILAC-TB was a proof-of-concept prospective study: adults with a rifampicin-susceptible TB who are HIV-negative and HIV-infected documented by a positive Xpert MTB/RIF test were enrolled in Cambodia and Côte d'Ivoire. Plasma concentrations of interleukin-1 receptor antagonist (IL-1Ra), interferon-γ–induced protein-10 and clusters of differentiation (CD) (scavenging CD163) were measured by commercial enzyme-linked immunosorbent assay kits. A Wilcoxon test for paired data was used for longitudinal comparisons. A total of 55 patients were enrolled (women: 31%, median age: 37 years; median CD4 count in the 10 of 13 participants with HIV: 53 cells/mm3). Overall, 83% were considered in TB treatment success. Compared with baseline, the IL-1Ra plasma levels significantly decreased as soon as week (W) 1, independent of HIV status (−71% in HIV-positive vs −33% in HIV-negative; P <0.001). The IP-10 plasma levels significantly decreased at W1 and W2 compared with baseline (P <0.0001); however, that decrease was less marked in participants with HIV. Our findings suggest that measuring IL-1Ra plasma levels with a standard enzyme-linked immunosorbent assay technique at baseline and then 1 week after TB treatment onset could help clinicians to quickly assess TB treatment response. [ABSTRACT FROM AUTHOR]

Abstract: The proteins of the X-tox family have imperfectly conserved tandem repeats of several defensin-like motifs known as cysteine-stabilized αβ (CS-αβ) motifs. These immune-related proteins are inducible and expressed principally in hemocytes, but they have lost the antimicrobial properties of the ancestral defensins from which they evolved. We compared x-tox gene structure and expression in three lepidopteran species (Spodoptera frugiperda, Helicoverpa armigera and Bombyx mori). Synteny and phylogenetic analyses showed that the x-tox exons encoding CS-αβ motifs were phylogenetically closely related to defensin genes mapping to chromosomal positions close to the x-tox genes. We were able to define two groups of paralogous x-tox exons (three in Noctuids) that each followed the expected species tree. These results suggest that the ancestor of the three species already possessed an x-tox gene with at least two proto-domains, and an additional duplication/fusion should have occurred in the ancestor of the two noctuid species. An expansion of the number of exons subsequently occurred in each lineage. Alternatively, the proto x-tox gene possessed more copy and each group of x-tox domains might undergo concerted evolution through gene conversion. Accelerated protein evolution was detected in x-tox domains when compared to related defensins, concomitantly to multiplication of exons and/or the possible activation of concerted evolution. The x-tox genes of the three species have similar structural organizations, with repeat motifs composed of CS-αβ-encoding exons flanked by introns in phase 1. Diverse mechanisms underlie this organization: (i) the acquisition of new repeat motifs, (ii) the duplication of preexisting repeat motifs and (iii) the duplication of modules. A comparison of gDNA and cDNA structures showed that alternative splicing results in the production of multiple X-tox protein isoforms from the x-tox genes. Differences in the number and sequence of CS-αβ motifs in these isoforms were found between species, but also between individuals of the same species. Thus, our analysis of the genetic organization and expression of x-tox genes in three lepidopteran species suggests a rapid evolution of the organization of these genes. [Copyright &y& Elsevier]

Abstract: Haemocytes are the main immunocompetent cells in insect cellular immune reactions. Here, we show that in Spodoptera littoralis, granular haemocytes are the primary phagocyte haemocytes, both in vivo and in vitro. The “trigger” and “zipper” modes of engulfment known in mammal macrophages are active, in vivo, in S. littoralis granular haemocytes, together with macropinocytosis. Lipopolysaccharide as well as lipoteichoic acid inhibit the binding of both Gram-positive (Corynebacterium xerosis) and Gram-negative (Escherichia coli) bacteria on granular haemocytes. In addition, different ligands can inhibit the binding of E. coli. Most of these inhibitors are known as ligands of scavenger receptors in mammal macrophages and we hypothesise that one of the receptors present on S. littoralis granular haemocytes could be a scavenger-like receptor. [Copyright &y& Elsevier]

The Achilles heel of anticancer treatments is intrinsic or acquired resistance. Among many targeted therapies, the DNA repair inhibitors show limited efficacy due to rapid emergence of resistance. We examined evolution of cancer cells and tumors treated with AsiDNA, a new DNA repair inhibitor targeting all DNA break repair pathways. Effects of AsiDNA or Olaparib were analyzed in various cell lines. Frequency of AsiDNA- and olaparib-resistant clones was measured after 2 weeks of continuous treatment in KBM7 haploid cells. Cell survivals were also measured after one to six cycles of 1-week treatment and 1-week recovery in MDA-MB-231 and NCI-H446. Transcriptomes of cell populations recovering from cyclic treatments or mock treatment were compared. MDA-MB-231 xenografted models were treated with three cycles of AsiDNA to monitor the effects of treatment on tumor growth and transcriptional modifications. No resistant clones were selected after AsiDNA treatment (frequency < 3x10−8) in treatment conditions that generate resistance to olaparib at a frequency of 7.2x10−7 resistant clones per treated cell. Cyclic treatments promote cumulative sensitivity characterized by a higher mortality of cells having undergone previous treatment cycles. This sensitization was stable, and transcriptome analysis revealed a major gene downregulation with a specific overrepresentation of genes coding for targets of DNA-PK. Such changes were also detected in tumor models which showed impaired growth after cycles of AsiDNA treatment. [ABSTRACT FROM AUTHOR]

The precore (pc) W28* mutation arises from immune-selective pressures during the hepatitis B “e” antigen (HBeAg)-positive phase of chronic hepatitis B virus (HBV) infection and has been linked to severe liver-related morbidity. Here, we examined the determinants of harboring this mutation and its rate of emergence in treated patients co-infected with human immunodeficiency virus (HIV) and HBV. In a three-year prospective cohort of 165 HIV-HBV co-infected patients, pcW28* mutation was determined via DNA-chip during yearly sampling. In a subgroup with liver biopsies, HBV covalently-closed circular (ccc)-DNA and total intrahepatic (IH)-DNA were quantified by real-time PCR. From respective inclusion to year-3 visits, median HBV-DNA levels decreased (5.88 log 10 IU/mL to <1.78 log 10 IU/mL, p < 0.001) and tenofovir-use increased (15.8%–71.4%, p < 0.001). At baseline, 47 of 162 (29.0%) patients had the pcW28* mutation and were more frequently HBeAg-negative (adjusted-OR = 4.37, 95%CI = 1.76–10.86) and had non-A HBV genotypes (adjusted-OR = 9.14, 95%CI = 4.05–20.66). No association with HIV-related factors was observed. In 114 patients without baseline mutation and available data, four developed incident pcW28* mutation by the end of follow-up (cumulative 3.5%, 95%CI = 1.3–9.1%). In the 32 patients with liver biopsies, 10 (31.3%) patients harboring the pcW28* mutation had significantly lower adjusted mean cccDNA (0.05 versus without = 0.43 copies/cell, p < 0.001) and total IH-DNA levels (2.31 versus without = 18.59 copies/cell, p = 0.006). In conclusion, the pcW28* mutation infrequently appeared in this co-infected study population with increased use of potent antivirals and suppressed levels of circulating virus. [ABSTRACT FROM AUTHOR]

Background & Aims In the presence of highly-potent antivirals, persistence of hepatitis B virus (HBV) is most well-characterized by covalently-closed circular DNA (cccDNA) and total intrahepatic DNA (IH-DNA). We sought to determine how antiviral therapy could affect their levels during human immunodeficiency virus (HIV)-HBV co-infection. Methods Sixty co-infected patients from a well-defined cohort with ⩾1 liver biopsy were studied. HBV cccDNA and total IH-DNA were extracted from biopsies and quantified by real-time PCR. Factors associated with intrahepatic viral load were determined using mixed-effect linear regression and half-life viral kinetics during reconstructed follow-up using non-linear exponential decay models. Results At biopsy, 35 (58.3%) patients were hepatitis B “e” antigen (HBeAg)-positive and 33 (55.0%) had detectable plasma HBV-DNA (median = 4.58 log 10 IU/ml, IQR = 2.95–7.43). Overall, median cccDNA was −0.95 log 10 copies/cell (IQR = −1.70, −0.17) and total IH-DNA was 0.27 log 10 copies/cell (IQR = −0.39, 2.00). In multivariable analysis, significantly lower levels of cccDNA and total IH-DNA were observed in patients with HBeAg-negative serology, nadir CD4 + cell counts >250/mm 3 , and longer cumulative TDF-duration, but not lamivudine- or adefovir-duration. In post-hoc analysis using reconstructed TDF-duration (median 29.6 months, IQR = 15.0–36.1, n = 31), average half-life of cccDNA was estimated at 9.2 months (HBeAg-positive = 8.6, HBeAg-negative = 26.2) and total IH DNA at 5.8 months (HBeAg-positive = 1.3, HBeAg-negative = 13.6). Intrahepatic viral loads remained detectable for all patients, even with prolonged TDF-exposure. Conclusions In co-infection, TDF-use is associated with lower levels of HBV replication intermediates and cccDNA. Slow decay of intrahepatic viral loads underscores that TDF is unable to completely block intracellular viral DNA synthesis, which possibly accounts for continuous replenishment of the cccDNA pool. Lay summary Chronic hepatitis B virus (HBV) is a persistent infection, while the only real way of knowing the extent of this persistence is through measuring levels of virus in the liver. In this study, we examine levels of HBV in the liver among patients with both HBV and human immunodeficiency virus, or HIV, infection. It would appear that the currently available medication, namely “tenofovir”, works well to decrease virus levels in the liver, but it remains at low levels despite long periods of treatment. [ABSTRACT FROM AUTHOR]

Background: Antiretroviral regimens containing tenofovir disoproxil fumarate have been associated with renal toxicity and reduced bone mineral density. Tenofovir alafenamide is a novel tenofovir prodrug that reduces tenofovir plasma concentrations by 90%, thereby decreasing off-target side-effects. We aimed to assess whether efficacy, safety, and tolerability were non-inferior in patients switched to a regimen containing tenofovir alafenamide versus in those remaining on one containing tenofovir disoproxil fumarate.Methods: In this randomised, actively controlled, multicentre, open-label, non-inferiority trial, we recruited HIV-1-infected adults from Gilead clinical studies at 168 sites in 19 countries. Patients were virologically suppressed (HIV-1 RNA <50 copies per mL) with an estimated glomerular filtration rate of 50 mL per min or greater, and were taking one of four tenofovir disoproxil fumarate-containing regimens for at least 96 weeks before enrolment. With use of a third-party computer-generated sequence, patients were randomly assigned (2:1) to receive a once-a-day single-tablet containing elvitegravir 150 mg, cobicistat 150 mg, emtricitabine 200 mg, and tenofovir alafenamide 10 mg (tenofovir alafenamide group) or to carry on taking one of four previous tenofovir disoproxil fumarate-containing regimens (tenofovir disoproxil fumarate group) for 96 weeks. Randomisation was stratified by previous treatment regimen in blocks of six. Patients and treating physicians were not masked to the assigned study regimen; outcome assessors were masked until database lock. The primary endpoint was the proportion of patients who received at least one dose of study drug who had undetectable viral load (HIV-1 RNA <50 copies per mL) at week 48. The non-inferiority margin was 12%. This study was registered with ClinicalTrials.gov, number NCT01815736.Findings: Between April 12, 2013 and April 3, 2014, we enrolled 1443 patients. 959 patients were randomly assigned to the tenofovir alafenamide group and 477 to the tenofovir disoproxil fumarate group. Viral suppression at week 48 was noted in 932 (97%) patients assigned to the tenofovir alafenamide group and in 444 (93%) assigned to the tenofovir disoproxil fumarate group (adjusted difference 4·1%, 95% CI 1·6-6·7), with virological failure noted in ten and six patients, respectively. The number of adverse events was similar between the two groups, but study drug-related adverse events were more common in the tenofovir alafenamide group (204 patients [21%] vs 76 [16%]). Hip and spine bone mineral density and glomerular filtration were each significantly improved in patients in the tenofovir alafenamide group compared with those in the tenofovir disoproxil fumarate group.Interpretation: Switching to a tenofovir alafenamide-containing regimen from one containing tenofovir disoproxil fumarate was non-inferior for maintenance of viral suppression and led to improved bone mineral density and renal function. Longer term follow-up is needed to better understand the clinical impact of these changes.Funding: Gilead Sciences. [ABSTRACT FROM AUTHOR]

Most research in sub-Saharan Africa establishes hepatitis B infection via one-time hepatitis B surface antigen (HBsAg) testing. Of 237 HIV-infected patients from two clinical trials testing HBsAg positive (MiniVidas®), 206 (86.9%) had validated serological tests using another assay (Architect). Discrepancies could be due to inactive infection, highlighting the importance of assessing hepatitis B virus infection phase. [ABSTRACT FROM AUTHOR]

Summary: Background: Coformulated elvitegravir, cobicistat, emtricitabine, and tenofovir disoproxil fumarate (tenofovir) might be a safe and efficacious switch option for virologically suppressed patients with HIV who have neuropsychiatric side-effects on a non-nucleoside reverse transcriptase inhibitor (NNRTI) or who are on a multitablet NNRTI-containing regimen and want a regimen simplification. We assessed the non-inferiority of such a switch compared with continuation of an NNRTI-containing regimen. Methods: STRATEGY-NNRTI is a 96 week, international, multicentre, randomised, open-label, phase 3b, non-inferiority trial enrolling adults (≥18 years) with HIV-1 and plasma HIV RNA viral load below 50 copies per mL for at least 6 months on an NNRTI plus emtricitabine and tenofovir regimen. With a computer-generated randomisation sequence, we randomly allocated participants (2:1; blocks of six, stratified by efavirenz use at screening) to switch to coformulated elvitegravir, cobicistat, emtricitabine, and tenofovir (switch group) or continue the NNRTI plus emtricitabine and tenofovir regimen (no-switch group). Key eligibility criteria included no history of virological failure and an estimated glomerular filtration rate of 70 mL per min or greater. The primary endpoint was the proportion of participants with plasma viral loads below 50 copies per mL at week 48 based on a snapshot algorithm with a non-inferiority margin of 12% (assessed by modified intention to treat). This trial is ongoing and is registered at ClinicalTrials.gov, number NCT01495702. Findings: Between Dec 29, 2011, and Dec 13, 2012, we randomly allocated 439 participants to treatment: 290 participants in the switch group and 143 participants in the no-switch group received treatment and were included in the modified intention-to-treat population. At week 48, 271 (93%) of 290 participants in the switch group and 126 (88%) of 143 participants in the no-switch group maintained plasma viral loads below 50 copies per mL (difference 5·3%, 95% CI −0·5 to 12·0; p=0·066). We detected no treatment-emergent resistance in either group. Safety events leading to discontinuation were uncommon in both groups: six (2%) of 291 participants in the switch group and one (1%) of 143 in the no-switch group. Interpretation: Coformulated elvitegravir, cobicistat, emtricitabine, and tenofovir seems to be efficacious and well tolerated in virologically suppressed adults with HIV and might be a suitable alternative for patients on an NNRTI with emtricitabine and tenofovir regimen considering a regimen modification or simplification. Funding: Gilead Sciences. [ABSTRACT FROM AUTHOR]

Abstract: Background: Among patients infected with hepatitis B virus (HBV), quantification of hepatitis B e antigen (HBeAg) is accruing substantial clinical relevance as a marker for HBeAg-loss during treatment. No direct comparison has been made between assays that quantify HBeAg. Objectives: To compare the performance of HBeAg quantification (qHBeAg) between Architect and Elecsys HBeAg assays among 183 patients with chronic HBV infection (94 treatment-naïve HBV-monoinfected and 89 antiretroviral-experienced HIV-HBV co-infected). Study design: qHBeAg was determined in Paul Erlich Institute Units (PEIU)/mL using previously designed protocols. Values were compared with correlation and linear regression. Bland–Altman analysis was used to compare mean differences between Elecsys and Architect assays and limits of agreement (LOA) (±2 standard deviations [SD]). Results: Between-assay correlation was significant overall (r =0.970), yet stronger for qHBeAg<1000 (n =131) versus >1000PEIU/mL (n =52) as determined by the Elecsys assay (r =0.969 vs. 0.880, respectively). On average, the Elecsys assay reported qHBeAg at 13.3PEIU/mL lower than the Architect assay (LOA: −415.9, 389.3), while LOA between assays were much wider at higher levels (<1000: −198.2, 147.9; ≥1000PEIU/mL: −688.4, 721.5). Further analysis indicated that did not change substantially with respect to HBV genotype, precore mutation, and CD4+ cell count, regardless of HBeAg-level. Nevertheless, seven of eight patients with highly divergent between-assay results had HBV-DNA>2000IU/mL. Conclusions: Elecsys and Architect assays report similar qHBeAg units with high correlation. Since qHBeAg was performed using an in-house approach, a commercially-available assay could reduce between-assay discrepancies, especially at higher HBeAg-levels. [Copyright &y& Elsevier]

Background & Aims: The systematic use of rapid tests performed at points-of-care may facilitate hepatitis B virus (HBV) screening and substantially increase HBV infection awareness. The aim of this study was to evaluate the effectiveness of such tests for HBsAg and anti-HBsAb detection among individuals visiting a variety of healthcare centers located in a low HBV-prevalent area. Methods: Three rapid tests for hepatitis B surface antigen (HBsAg) detection (VIKIA®, Determine™ and Quick Profile™) and one test for anti-hepatitis B surface antibody (anti-HBsAb) detection (Quick Profile™) were evaluated in comparison to ELISA serology. Sensitivity (Se), specificity (Sp), positive and negative predictive values (PPV and NPV, respectively) and area under the ROC curve were used to estimate test performance. Non-inferiority criteria of the joint Se, Sp were set at 0.80, 0.95. Results: Among the 3956 subjects screened, 85 (2.1%) were HBsAg-positive and 2225 (56.5%) had a protective anti-HBsAb titer. Test Se and Sp (lower bound of 97.5% CI) were as follows: 96.5% (89.0%), 99.9% (99.8%) for Vikia®; 93.6% (80.7%), 100.0% (99.8%) for Determine™; and 90.5% (80.8%), 99.7% (99.5%) for Quick Profile™; with all three tests achieving minimal non-inferiority criteria. False negatives were typically observed in inactive HBsAg carriers. The anti-HBsAb Quick Profile™ test had excellent specificity (97.8%) and PPV (97.8%) albeit low sensitivity (58.3%), thus failing to establish non-inferiority. Conclusions: All three HBsAg rapid tests could be considered ideal for HBV screening in low HBV-prevalent countries, given the ease of use, rapidity, and high classification probabilities. The anti-HBsAb Quick Profile™ could be considered reliable only for positive tests. [ABSTRACT FROM AUTHOR]

Abstract: Cycle inhibiting factors (Cif) constitute a broad family of cyclomodulins present in bacterial pathogens of invertebrates and mammals. Cif proteins are thought to be type III effectors capable of arresting the cell cycle at G2/M phase transition in human cell lines. We report here the first direct functional analysis of Cif Pl , from the entomopathogenic bacterium Photorhabdus luminescens, in its insect host. The cifPl gene was expressed in P. luminescens cultures in vitro. The resulting protein was released into the culture medium, unlike the well characterized type III effector LopT. During locust infection, cifPl was expressed in both the hemolymph and the hematopoietic organ, but was not essential for P. luminescens virulence. Cif Pl inhibited proliferation of the insect cell line Sf9, by blocking the cell cycle at the G2/M phase transition. It also triggered host cell death by apoptosis. The integrity of the Cif Pl catalytic triad is essential for the cell cycle arrest and pro-apoptotic activities of this protein. These results highlight, for the first time, the dual role of Cif in the control of host cell proliferation and apoptotic death in a non-mammalian cell line. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To evaluate the impact of highly active antiretroviral therapy (HAART) on survival in HIV infected patients with non-small-cell lung cancer (NSCLC). Patients and methods: All consecutive HIV infected patients with NSCLC diagnosed between 06/1996 and 03/2007 at two University hospitals in Paris (France) were prospectively followed until death. The association between survival and clinical and biological factors was analyzed by univariate and multivariate models. Survival analysis was performed by Kaplan–Meier estimates and the Cox proportional hazards regression model. Results: During the study period, NSCLC was diagnosed in 49 consecutive HIV infected patients (median age 46 years); 84% had advanced disease. Median survival was 8.1 months (range 5–10 months). In multivariate analysis, baseline parameters with significant positive impact on survival included performance status (PS) ≤1 (HR=0.2, 95%CI [0.09, 0.46], p =0.0001), stage I–II disease (HR=0.15, 95%CI [0.04, 0.53], p =0.003), and use of HAART (HR=0.4, 95%CI [0.2, 0.9], p =0.027). Conclusion: HAART is a good prognostic factor for survival in HIV infected patients with NSCLC. Stage of disease and PS are two other valid survival prognostic factors. [Copyright &y& Elsevier]

Background/Aims: The aim of this study was to compare the performance of 11 biochemical scores to estimate liver fibrosis in HIV/HBV co-infection. Methods: Performance was evaluated using the Receiver Operating Characteristics (ROC) curve method. The Kappa index was used to study overall agreement with liver biopsy results. Interpretative algorithms were established by optimizing sensitivity and specificity and the percentage of correctly classified patients. Results: One hundred and eight patients (F0–F1, n =47; F2, n =28; F3, n =17; F4, n =16) were considered for the evaluation of serum biomarker performance. The AUROCs of the Fibrotest®, Hepascore®, Fibrometer®, and Zeng’s scores ranged from 0.74 to 0.77 for significant fibrosis (⩾ F2), from 0.79 to 0.84 for advanced fibrosis (⩾ F3) and from 0.87 to 0.92 for cirrhosis (F4). Thresholds defined for each stage of fibrosis were close to those previously published for the Fibrotest® and Hepascore®. Strict concordance with biopsies correctly classified 50% of the patients. Conclusions: Fibrotest®, Fibrometer®, Hepascore®, and Zeng’s score were the most accurate non-invasive biochemical scores for liver fibrosis assessment in HIV/HBV co-infection. Global performance of biomarkers was not significantly improved by a decision tree combining the results of two biochemical scores. [Copyright &y& Elsevier]

Although cardiotoxic effects of highly active antiretroviral therapy (HAART) are a growing concern, there is a lack of prospective studies of subclinical involvement of the heart in human immunodeficiency virus (HIV)-infected patients. This study evaluated noninvasively cardiac morphologic characteristics and function in HIV-positive (HIV+) men receiving HAART for ≥2 years with no clinical evidence of cardiovascular disease. Echocardiography at rest, including tissue Doppler imaging and exercise testing, were performed in 30 HIV+ men (age 42.1 ± 4.7 years, duration of HIV infection 10.4 ± 4.7 years, duration of HAART 5.3 ± 2.1 years) and 26 age-matched healthy controls. At rest, HIV+ patients had similar left ventricular (LV) mass indexed to height2.7 (40.6 ± 9.5 vs 37.5 ± 9.3 g/m; p >0.05), but a higher prevalence of LV diastolic dysfunction (abnormal relaxation or pseudonormal filling pattern in 64% of patients vs 12% of controls; p <0.001). LV systolic function indexes were significantly lower (ejection fraction 60.4 ± 8.7% vs 66.9 ± 6.9%; p <0.01, and tissue Doppler imaging peak systolic velocity 11.4 ± 1.6 vs 13.5 ± 2.2 cm/s; p <0.001). Pulmonary artery pressure was higher in patients compared with controls (32.1 ± 5.4 vs 26.1 ± 6.5 mm Hg; p <0.001). Exercise testing showed decreased exercise tolerance in HIV+ patients, with no case of myocardial ischemia. In conclusion, subclinical cardiac abnormalities are frequently observed in HIV+ patients on HAART. The usefulness of systematic noninvasive screening in this population should be considered. GECEM study no. 30: National Agency for AIDS Research (ANRS). [Copyright &y& Elsevier]

Demonstrates that the efficiency of the DNA ligase IV·XRCC4 complex (LX) ligation of double-stranded ends is critically dependent upon the length of the DNA substrate, in contrast to ligation by T4 ligase. Overexpression and purification of LX and Ku70/80 complexes; Impact of Ku on LX ligation using different length substrates.