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Dansk Selskab for Infektionsmedicin
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1 Case conferences for infective endocarditis: A quality improvement initiativeCharlie Tan, Mark S. Hansen, Gideon Cohen, Karl Boyle, Alvin Yang, Asgar Rishu, Ruxandra Pinto, Neill K. J. Adhikari, Nick Daneman by Charlie Tan, Mark S. Hansen, Gideon Cohen, Karl Boyle, Alvin Yang, Asgar Rishu, Ruxandra Pinto, Neill K. J. Adhikari, Nick Daneman Background A multidisciplinary approach has been recommended for the management of patients with infective endocarditis. We evaluated the impact of multidisciplinary case conferences on morbidity, mortality, and quality of care for these patients. Methods We conducted a quasi-experimental study of consecutive patients admitted for infective endocarditis before (2013/10/1–2015/10/12, n = 97) and after (2015/10/13–2017/11/30, n = 80) implementation of case conferences to discuss medical and surgical management. These occurred as face-to-face discussions or electronically (for non-complex patients), and included physicians from cardiac surgery, cardiology, critical care, infectious diseases and neurology. We assessed process-of-care and clinical outcomes, with the primary outcome being complications up to 90 days after hospital discharge. Results A case conference was held for 80/80 (100%) of patients in the post-intervention group. After the intervention, more patients received inpatient cardiology assessment (81.3% [post-intervention] vs. 63.9% [pre-intervention], p = 0.01), and more patients with definite infective endocarditis underwent cardiac surgery treatment (44.6% vs. 21.7%, p = 0.007). All pre-intervention and post-intervention patients received guideline-concordant antimicrobial therapy. There was no difference in rates of complications (40.0% vs. 51.5%, p = 0.13) or mortality up to 90 days after hospital discharge (26.3% vs. 17.5%, p = 0.20). In multivariable analyses, the intervention was not associated with differences in mortality (odds ratio 1.87, 95% confidence interval 0.88–3.99) or a composite measure of complications and mortality (odds ratio 0.86, 95% confidence interval 0.46–1.58). Conclusion We successfully implemented a standardized multidisciplinary case conference protocol for patients with infective endocarditis. This intervention had no detectable effect on complications or mortality. 2 Comparative analysis of different preservation techniques for the storage of Staphylococcus phages aimed for the industrial development of phage-based antimicrobial productsEva González-Menéndez, Lucía Fernández, Diana Gutiérrez, Ana Rodríguez, Beatriz Martínez, Pilar García by Eva González-Menéndez, Lucía Fernández, Diana Gutiérrez, Ana Rodríguez, Beatriz Martínez, Pilar García Bacteriophages have been proven as effective antimicrobial agents in the treatment of infectious diseases and in other biocontrol applications including food preservation and disinfection. The extensive use of bacteriophages requires improved methodologies for medium- and long-term storage as well as for easy shipping. To this aim, we have determined the stability of four Staphylococcus phages (phiIPLA88, phiIPLA35, phiIPLA-RODI and phiIPLA-C1C) with antimicrobial potential at different temperatures (20°C/25°C, 4°C, -20°C, -80°C, -196°C) and during lyophilization (freeze drying) using several stabilizing additives (disaccharides, glycerol, sorbitol and skim milk). Differences between phages were observed at different temperatures (20°C/25°C, 4°C and -20°C), where phages were less stable. At lower temperatures (-80°C and -196°C), all phages showed good viability after 24 months regardless of the stabilizer. Differences between phages were also observed after lyophilization although the addition of skim milk yielded a dry powder with a stable titer after 24 months. As an alternative to facilitate storage and transportation, phage encapsulation has been also explored. Phage phiIPLA-RODI encapsulated in alginate capsules retained high viability when stored at 4°C for 6 months and at 20°C for 1 month. Moreover, the spray-dryer technique allowed obtaining dry powders containing viable encapsulated phages (phiIPLA-RODI and phiIPLA88) in both skim milk and trehalose for 12 months at 4°C. Storage of phages at 20°C was less effective; in fact, phiIPLA88 was stable for at least 12 months in trehalose but not in skim milk, while phiIPLA-RODI was stable only for 6 months in either stabilizer. These results suggest that encapsulated phages might be a suitable way for shipping phages. Staphylococcus phages aimed for the industrial development of phage-based antimicrobial products" som kan hentes fra Dansk Selskab for Infektionsmedicin's hjemmeside via linket vist nedenfor:%0D%0A%0D%0Ahttp%3A%2F%2Finfmed.dk%2Fnyheder-udefra%3Fsetpoint%3D99138%2399145"> 3 Controlled crack propagation for atomic precision handling of wafer-scale two-dimensional materialsShim, J., Bae, S.-H., Kong, W., Lee, D., Qiao, K., Nezich, D., Park, Y. J., Zhao, R., Sundaram, S., Li, X., Yeon, H., Choi, C., Kum, H., Yue, R., Zhou, G., Ou, Y., Lee, K., Moodera, J., Zhao, X., Ahn, J.-H., Hinkle, C., Ougazzaden, A., Kim, J. Although flakes of two-dimensional (2D) heterostructures at micrometer scale can be formed with adhesive-tape methods, isolation of 2D flakes into monolayers is extremely time-consuming as it is a trial-and-error process. Controlling the number of 2D layers through direct growth also presents difficulty because of the high nucleation barrier on 2D materials. We demonstrate a layer-resolved 2D material splitting technique that permits the high-throughput production of multiple monolayers of wafer-scale (5 centimeter diameter) 2D materials by splitting single stacks of thick 2D materials grown on a single wafer. Wafer-scale uniformity of h-BN, WS2, WSe2, MoS2, and MoSe2 monolayers was verified by photoluminescence (PL) response and by substantial retention of electronic conductivity. We fabricated wafer-scale van der Waals heterostructures, including field-effect transistors, with single-atom thickness resolution. 4 Costs and cost-effectiveness of LEEP versus cryotherapy for treating cervical dysplasia among HIV-positive women in Johannesburg, South AfricaNaomi Lince-Deroche, Craig van Rensburg, Jaqueline Roseleur, Busola Sanusi, Jane Phiri, Pam Michelow, Jennifer S. Smith, Cindy Firnhaber by Naomi Lince-Deroche, Craig van Rensburg, Jaqueline Roseleur, Busola Sanusi, Jane Phiri, Pam Michelow, Jennifer S. Smith, Cindy Firnhaber Background Cervical cancer incidence is significant in countries, such as South Africa, with high burdens of both HIV and human papillomavirus (HPV). Cervical cancer is largely preventable if dysplasia is diagnosed and treated early, but there is debate regarding the best approaches for screening and treatment, especially for low-resource settings. Currently South Africa provides Pap smears followed by colposcopic biopsy and LEEP if needed in its public health facilities. We estimated the costs and cost-effectiveness of two approaches for treating cervical intraepithelial neoplasia grade 2 or higher (CIN2+) among HIV-infected women, most of whom were taking antiretroviral treatment, at a public HIV treatment facility in Johannesburg, South Africa. Methods Method effectiveness was derived from an intention-to-treat analysis of data gathered in a clinical trial completed previously at the study facility. In the trial, women who were diagnosed with CIN2+ and eligible for cryotherapy were randomized to cryotherapy or LEEP. If women were CIN2+ at six months as determined via Pap smear and colposcopic biopsy, all women—regardless of their original treatment assignment—received LEEP. “Cure” was then defined as the absence of disease at 12 months based on Pap smear and colposcopic biopsy. Health service costs were estimated using micro-costing between June 2013 and April 2014. Capital costs were annualized using a discount rate of 3%. Two different service volume scenarios were considered, and results from an as-treated analysis were considered in sensitivity analysis. Results In total, 166 women with CIN2+ were enrolled (86 had LEEP; 80 had cryotherapy). At 12 months, cumulative loss to follow-up was 12.8% (11/86) for the LEEP group and 13.8% (11/80) for cryotherapy. Based on the unadjusted intention-to-treat analysis conducted for this economic evaluation, there was no significant difference in efficacy. At 12 months, 83.8% (95% CI 73.8–91.1) of women with CIN2+ at baseline and randomized to cryotherapy were free of CIN2+ disease. In contrast, 76.7% (95% CI 66.4–85.2) of women assigned to LEEP were free from disease. On average, women initially treated with cryotherapy were less costly per patient randomized at US$ 118.00 (113.91–122.10), and per case “cured” at US$ 140.90 (136.01–145.79). Women in the LEEP group cost US$ 162.56 (157.90–167.22) per patient randomized and US$ 205.59 (199.70–211.49) per case cured. In the as-treated analysis, which was based on trial data, LEEP was more efficacious than cryotherapy; however, the difference was not significant. Cryotherapy remained more cost-effective than LEEP in all sensitivity and scenario analyses. Conclusions For this cost-effectiveness analysis, using an intention-to-treat approach and taking into consideration uncertainty in the clinical and cost outcomes, a strategy involving cryotherapy plus LEEP if needed at six months was dominant to LEEP plus LEEP again at six months if needed for retreatment. However, compared to other studies comparing LEEP and cryotherapy, the efficacy results were low in both treatment groups–possibly due to the HIV-positivity of the participants. Further research is needed, but at present choosing the “right” treatment option may be less important than ensuring access to treatment and providing careful monitoring of treatment outcomes. 5 Epstein‐Barr virus, but not human cytomegalovirus, is associated with a high grade human papillomavirus‐associated cervical lesions among women in North CarolinaTamina Kienka, Matthew G. Varga, Josie Caves, Jennifer S. Smith, Vijay Sivaraman Abstract Statement of the Problem Human papillomavirus (HPV) infection is known to contribute to cervical carcinogenesis, yet other co‐factors that may contribute to oncogenesis are poorly understood. Herein, we examine whether Epstein‐Barr virus (EBV) and human cytomegalovirus (CMV), two onco‐modulatory viruses, are associated with HPV‐mediated cervical neoplastic progression. Methods of Study Sixty patient cervical brush samples from a study of North Carolina women were obtained. HPV RNA positivity was determined by Aptima testing (Hologic Corporation). The level of viral transcripts for EBV and CMV were quantified (RT‐PCR analysis), and co‐infection status with HPV was then compared to the patient’s cervical cytology grade. Results Over one third (38.3%) of the study population was CMV‐positive, while 43.3% was EBV‐positive. When sample data were stratified by cytology grade, 36.5% (19/52) of normal patients, 75% (3/4) of low‐grade squamous intraepithelial lesions (LSIL) and 100% (4/4) of patients with high‐grade SIL (HSIL) were EBV positive. Conversely, 35.2% (18/52) of normal patients, 25% (1/4) of patients with LSIL and 50% (2/4) of patients with HSIL were CMV positive. When examining only HPV‐positive associated HSIL,100% (4/4) were positive for both HPV and EBV detection. This suggests co‐viral detection with HPV and EBV is associated with more advanced HSIL cervical lesions, while CMV displayed no clear association with higher grade of cervical cytology. Conclusions Co‐viral detection with EBV may increase the oncogenicity and/or serve as a viral marker of progression to HPV‐associated high‐grade cervical dysplasia. This article is protected by copyright. All rights reserved. 6 High-resolution plasma metabolomics analysis to detect Mycobacterium tuberculosis-associated metabolites that distinguish active pulmonary tuberculosis in humansJeffrey M. Collins, Douglas I. Walker, Dean P. Jones, Nestani Tukvadze, Ken H. Liu, ViLinh T. Tran, Karan Uppal, Jennifer K. Frediani, Kirk A. Easley, Neeta Shenvi, Manoj Khadka, Eric A. Ortlund, Russell R. Kempker, Henry M. Blumberg, Thomas R. Ziegler by Jeffrey M. Collins, Douglas I. Walker, Dean P. Jones, Nestani Tukvadze, Ken H. Liu, ViLinh T. Tran, Karan Uppal, Jennifer K. Frediani, Kirk A. Easley, Neeta Shenvi, Manoj Khadka, Eric A. Ortlund, Russell R. Kempker, Henry M. Blumberg, Thomas R. Ziegler Introduction Pulmonary tuberculosis (TB) is a major worldwide health problem that lacks robust blood-based biomarkers for detection of active disease. High-resolution metabolomics (HRM) is an innovative method to discover low-abundance metabolites as putative blood biomarkers to detect TB disease, including those known to be produced by the causative organism, Mycobacterium tuberculosis (Mtb). Methods We used HRM profiling to measure the plasma metabolome for 17 adults with active pulmonary TB disease and 16 of their household contacts without active TB. We used a suspect screening approach to identify metabolites previously described in cell culture studies of Mtb based on retention time and accurate mass matches. Results The association of relative metabolite abundance in active TB disease subjects compared to their household contacts predicted three Mtb-associated metabolites that were significantly increased in the active TB patients based on accurate mass matches: phosphatidylglycerol (PG) (16:0_18:1), lysophosphatidylinositol (Lyso-PI) (18:0) and acylphosphatidylinositol mannoside (Ac1PIM1) (56:1) (p Mycobacterium tuberculosis-associated metabolites that distinguish active pulmonary tuberculosis in humans" som kan hentes fra Dansk Selskab for Infektionsmedicin's hjemmeside via linket vist nedenfor:%0D%0A%0D%0Ahttp%3A%2F%2Finfmed.dk%2Fnyheder-udefra%3Fsetpoint%3D99138%2399147"> 7 Human and entomologic investigations of chikungunya outbreak in Mandera, Northeastern Kenya, 2016Samson Limbaso Konongoi, Albert Nyunja, Victor Ofula, Samuel Owaka, Hellen Koka, Edith Koskei, Fredrick Eyase, Daniel Langat, James Mancuso, Joel Lutomiah, Rosemary Sang by Samson Limbaso Konongoi, Albert Nyunja, Victor Ofula, Samuel Owaka, Hellen Koka, Edith Koskei, Fredrick Eyase, Daniel Langat, James Mancuso, Joel Lutomiah, Rosemary Sang Chikungunya is a reemerging vector borne pathogen associated with severe morbidity in affected populations. Lamu, along the Kenyan coast was affected by a major chikungunya outbreak in 2004. Twelve years later, we report on entomologic investigations and laboratory confirmed chikungunya cases in northeastern Kenya. Patient blood samples were received at the Kenya Medical Research Institute (KEMRI) viral hemorrhagic fever laboratory and the immunoglobulin M enzyme linked immunosorbent assay (IgM ELISA) was used to test for the presence of IgM antibodies against chikungunya and dengue. Reverse transcription polymerase chain reaction (RT-PCR) utilizing flavivirus, alphavirus and chikungunya specific primers were used to detect acute infections and representative PCR positive samples sequenced to confirm the circulating strain. Immature mosquitoes were collected from water-holding containers indoors and outdoors in the affected areas in northeastern Kenya. A total of 189 human samples were tested; 126 from Kenya and 63 from Somalia. 52.9% (100/189) tested positive for Chikungunya virus (CHIKV) by either IgM ELISA or RT-PCR. Sequence analysis of selected samples revealed that the virus was closely related to that from China (2010). 29% (55/189) of the samples, almost all from northeastern Kenya or with a history of travel to northern Kenya, tested positive for dengue IgM antibodies. Entomologic risk assessment revealed high house, container and Breteau indices of, 14.5, 41.9 and 17.1% respectively. Underground water storage tanks were the most abundant, 30.1%, of which 77.4% were infested with Aedes aegypti mosquitoes. These findings confirm the presence of active chikungunya infections in the northeastern parts of Kenya. The detection of dengue IgM antibodies concurrently with chikungunya virus circulation emphasizes on the need for improved surveillance systems and diagnostic algorithms with the capacity to capture multiple causes of arbovirus infections as these two viruses share common vectors and eco-systems. In addition sustained entomological surveillance and vector control programs targeting most productive containers are needed to monitor changes in vector densities, for early detection of the viruses and initiate vector control efforts to prevent possible outbreaks. 8 Identity inference of genomic data using long-range familial searchesErlich, Y., Shor, T., Peer, I., Carmi, S. Consumer genomics databases have reached the scale of millions of individuals. Recently, law enforcement authorities have exploited some of these databases to identify suspects via distant familial relatives. Using genomic data of 1.28 million individuals tested with consumer genomics, we investigated the power of this technique. We project that about 60% of the searches for individuals of European-descent will result in a third cousin or closer match, which can allow their identification using demographic identifiers. Moreover, the technique could implicate nearly any US-individual of European-descent in the near future. We demonstrate that the technique can also identify research participants of a public sequencing project. Based on these results, we propose a potential mitigation strategy and policy implications to human subject research. 9 In vivo modeling of human neuron dynamics and Down syndromeReal, R., Peter, M., Trabalza, A., Khan, S., Smith, M. A., Dopp, J., Barnes, S. J., Momoh, A., Strano, A., Volpi, E., Knott, G., Livesey, F. J., De Paola, V. Harnessing the potential of human stem cells for modelling the physiology and diseases of cortical circuitry requires monitoring cellular dynamics in vivo. Here, we show that human iPSC–derived cortical neurons transplanted in the adult mouse cortex consistently organized in large (up to ~100 mm3) vascularized neuron-glia territories with complex cytoarchitecture. Longitudinal imaging of >4000 grafted developing human neurons revealed that neuronal arbors refined via branch-specific retraction; human synaptic networks substantially restructured over 4 months, with balanced rates of synapse formation and elimination; and oscillatory population activity mirrored the patterns of fetal neural networks. Finally, we found increased synaptic stability and reduced oscillations in transplants from two individuals with Down syndrome, demonstrating the potential of in vivo imaging in human tissue grafts for patient-specific modelling of cortical development, physiology, and pathogenesis. 10 Mathematical modelling of vancomycin-resistant enterococci transmission during passive surveillance and active surveillance with contact isolation highlights the need to identify and address the source of acquisitionAbstract Background Clinical studies and mathematical simulation suggest that active surveillance with contact isolation is associated with reduced vancomycin-resistant enterococci (VRE) prevalence compared to passive surveillance. Models using pre- and post-intervention data that account for the imperfect observation and serial dependence of VRE transmission events can better estimate the effectiveness of active surveillance and subsequent contact isolation; however, such analyses have not been performed. Methods A mathematical model was fitted to surveillance data collected pre- and post-implementation of active surveillance with contact isolation in the haematology-oncology ward. We developed a Hidden Markov Model to describe undetected and observed VRE colonisation/infection status based on the detection activities in the ward. Bayesian inference was used to estimate transmission rates. The effectiveness of active surveillance was assumed to be via increased detection and subsequent contact isolation of VRE positive patients. Results We estimated that 31% (95% credible interval: 0.33–85%) of the VRE transmissions were due to cross-transmission between patients. The ratio of transmission rates from patients with contact isolation versus those without contact isolation was 0.33 (95% credible interval: 0.050–1.22). Conclusions The majority of the VRE acquisitions in the haematology-oncology ward was estimated to be due to background rates of VRE, rather than within ward patient to patient acquisition. The credible interval for cross-transmission was wide which results in a large degree of uncertainty in the estimates. Factors that could account for background VRE acquisition include endogenous acquisition from antibiotic selection pressure and VRE in the environment. Contact isolation was not significantly associated with reduced VRE transmission in settings where the majority of VRE acquisition was due to background acquisition, emphasising the need to identify and address the source of acquisition. As the credible interval for the ratio of VRE transmission in contact isolated versus non-contact isolated patients crossed 1, there is a probability that the transmission rate in contact isolation was not lower. Our finding highlights the need to optimise infection control measures other than active surveillance for VRE and subsequent contact isolation to reduce VRE transmission. Such measures could include antimicrobial stewardship, environmental cleaning, and hand hygiene. 11 Methylammonium-free, high-performance and stable perovskite solar cells on a planar architectureTurren-Cruz, S.-H., Hagfeldt, A., Saliba, M. Currently, perovskite solar cells (PSCs) with high performances >20% contain Br, causing a suboptimal bandgap, and the thermally unstable methylammonium (MA) molecule. Avoiding Br and especially MA can, therefore, result in more optimal bandgaps and stable perovskites. We show that inorganic cation tuning, using Rb and Cs, enables highly crystalline formamidinium-based perovskites without Br or MA. On a conventional, planar device architecture, using polymeric interlayers at the electron and hole transporting interface, we demonstrate an efficiency of 20.35% (stabilized), one of the highest for MA-free perovskites, with a drastically improved stability reached without the stabilizing influence of mesoporous interlayers. The perovskite is not heated beyond 100°C. Going MA-free is a new direction for perovskites that are inherently stable and compatible with tandems or flexible substrates which are the main routes commercializing PSCs. 12 Multidrug resistant Pseudomonas aeruginosa in Estonian hospitalsAbstract Background We aimed to identify the main spreading clones, describe the resistance mechanisms associated with carbapenem- and/or multidrug-resistant P. aeruginosa and characterize patients at risk of acquiring these strains in Estonian hospitals. Methods Ninety-two non-duplicated carbapenem- and/or multidrug-resistant P. aeruginosa strains were collected between 27th March 2012 and 30th April 2013. Clinical data of the patients was obtained retrospectively from the medical charts. Clonal relationships of the strains were determined by whole genome sequencing and analyzed by multi-locus sequence typing. The presence of resistance genes and beta-lactamases and their origin was determined. Combined-disk method and PCR was used to evaluate carbapenemase and metallo-beta-lactamase production. Results Forty-three strains were carbapenem-resistant, 11 were multidrug-resistant and 38 were both carbapenem- and multidrug-resistant. Most strains (54%) were isolated from respiratory secretions and caused an infection (74%). Over half of the patients (57%) were ≥ 65 years old and 85% had ≥1 co-morbidity; 96% had contacts with healthcare and/or had received antimicrobial treatment in the previous 90 days. Clinically relevant beta-lactamases (OXA-101, OXA-2 and GES-5) were found in 12% of strains, 27% of which were located in plasmids. No Ambler class B beta-lactamases were detected. Aminoglycoside modifying enzymes were found in 15% of the strains. OprD was defective in 13% of the strains (all with CR phenotype); carbapenem resistance triggering mutations (F170 L, W277X, S403P) were present in 29% of the strains. Ciprofloxacin resistance correlated well with mutations in topoisomerase genes gyrA (T83I, D87N) and parC (S87 L). Almost all strains (97%) with these mutations showed ciprofloxacin-resistant phenotype. Multi-locus sequence type analysis indicated high diversity at the strain level – 36 different sequence types being detected. Two sequence types (ST108 (n = 23) and ST260 (n = 18)) predominated. Whereas ST108 was associated with localized spread in one hospital and mostly carbapenem-resistant phenotype, ST260 strains occurred in all hospitals, mostly with multi-resistant phenotype and carried different resistance genotype/machinery. Conclusions Diverse spread of local rather than international P. aeruginosa strains harboring multiple chromosomal mutations, but not plasmid-mediated Ambler class B beta-lactamases, were found in Estonian hospitals. Trial registration This trial was registered retrospectively in ClinicalTrials.gov (NCT03343119). 13 Observation of the topological Anderson insulator in disordered atomic wiresMeier, E. J., An, F. A., Dauphin, A., Maffei, M., Massignan, P., Hughes, T. L., Gadway, B. Topology and disorder have a rich combined influence on quantum transport. In order to probe their interplay, we synthesized one-dimensional chiral symmetric wires with controllable disorder via spectroscopic Hamiltonian engineering, based on the laser-driven coupling of discrete momentum states of ultracold atoms. Measuring the bulk evolution of a topological indicator following a sudden quench, we observed the topological Anderson insulator phase, in which added disorder drives the band structure of a wire from topologically trivial to non-trivial. In addition, we observed the robustness of topologically non-trivial wires to weak disorder and measured the transition to a trivial phase in the presence of strong disorder. Atomic interactions in this quantum simulation platform may enable realizations of strongly interacting topological fluids. 14 Pancoast’s syndrome: an unusual presentation of invasive pneumococcal disease15 Pre- and post-natal macronutrient supplementation for HIV–positive women in Tanzania: Effects on infant birth weight and HIV transmissionAlbert Magohe, Todd Mackenzie, Josephine Kimario, Zohra Lukmanji, Kristy Hendricks, John Koethe, Nyasule Majura Neke, Susan Tvaroha, Ruth Connor, Richard Waddell, Isaac Maro, Mecky Matee, Kisali Pallangyo, Muhammad Bakari, C. Fordham von Reyn, DarDar-2 Study Team by Albert Magohe, Todd Mackenzie, Josephine Kimario, Zohra Lukmanji, Kristy Hendricks, John Koethe, Nyasule Majura Neke, Susan Tvaroha, Ruth Connor, Richard Waddell, Isaac Maro, Mecky Matee, Kisali Pallangyo, Muhammad Bakari, C. Fordham von Reyn, DarDar-2 Study Team Objective To determine if a protein-calorie supplement (PCS) plus a micronutrient supplement (MNS) improves outcomes for HIV-infected lactating women and their infants. Design Randomized, controlled trial. Setting Dar es Salaam, Tanzania Subjects, participants Pregnant HIV-infected women enrolled in PMTCT programs who intended to breastfeed for 6 months. Intervention Randomization 1:1 to administration of a PCS plus MNS versus MNS alone among 96 eligible women beginning in the third trimester and continuing for 6 months of breast-feeding. Main outcome measure(s) Primary: infant weight at 3 months. Secondary: maternal BMI at 6 months. Results PCS resulted in significant increases in daily energy intake compared to MNS at all time points (range of differences: +388–719 Kcal); and increases in daily protein intake (range of differences: +22–33 gm). Infant birth weight (excluding twins) was higher in the PCS than MNS groups: 3.30 kg vs 3.04 kg (p = 0.04). Infant weight at 3 months did not differ between PCS and MNS groups: 5.63 kg vs 5.99 kg (p = 0.07). Maternal BMI at 6 months did not differ between PCS and MNS groups: 24.3 vs 23.8 kg/m2 (p = 0.68). HIV transmission occurred in 0 infants in the PCS group vs 4 in the MNS group (p = 0.03). Conclusions In comparison to MNS the PCS + MNS intervention was well tolerated, increased maternal energy and protein intake, and increased infant birth weight, but not weight at 3 months or maternal BMI at 6 months. Reduced infant HIV transmission in the PCS + MNS group was observed. Trial registration Clinical Trials.Gov NCT01461863. 16 Quantifying user preferences for sanitation construction and use: Application of discrete choice experiments in Amhara, EthiopiaFrederick G B Goddard, Maryann G Delea, Gloria D Sclar, Mulat Woreta, Kassahun Zewudie, Matthew C Freeman Abstract Objective Discrete choice experiments (DCEs) are stated preference methods that allow for the quantification of preferences by presenting respondents with hypothetical choices. We conducted image‐based DCEs to assess preferences for latrine use (stratified by gender) and construction (among men only) in Amhara, Ethiopia. Methods Preference was quantified using a conditional logistic model to estimate utilities and corresponding odds ratios associated with a set of latrine attributes. Results For latrine use, tin roofing, handwashing stations, and clean latrines had the highest relative utility coefficients. Tin roofing was preferred to no roof for use (Women: OR 3.68, 95% CI 3.18‐4.25; Men: OR 3.75, 95% CI 3.21‐4.39) and new latrine construction (5.92, 5.04‐6.95). Concrete slabs, a critical aspect of improved sanitation, was not preferred to dirt floors for use (Women: 0.87, 0.75‐1.00; Men: 1.03, 0.88‐1.20), but was preferred for new construction (1.52, 1.30‐1.78). We did not observe any trends in preference for direct (monetary) or indirect cost (labour days), so we were not able to elicit trade‐offs between latrine attributes and these costs for the construction of new latrines. Conclusion Our findings suggest similar latrine use preferences between men and women. We found that tin roofs are the most strongly preferred latrine characteristic, but concrete slabs, commonly promoted in sanitation programmes, were not preferred for use. We demonstrate the utility of DCEs to elicit stated preferences for latrine use and construction among community members who have myriad motivations for using and making improvements to their sanitation facilities, including the ease of cleaning and hygiene, durability, or privacy and comfort. This article is protected by copyright. All rights reserved. 17 Spatio‐temporal distribution of vertically transmitted dengue viruses by Aedes aegypti (Diptera: Culicidae) from Arroyo Naranjo, Havana, CubaGladys Gutiérrez‐Bugallo, Rosmari Rodríguez‐Roche, Gisell Díaz, Magaly Pérez, María Elena Mendizábal, Iris Peraza, Antonio A Vázquez, Mayling Alvarez, Magdalena Rodríguez, Juan A. Bisset, María G. Guzmán Abstract Objective To study the distribution of vertical transmission of dengue viruses in field‐collected Aedes aegypti larvae in the municipality of Arroyo Naranjo in Havana, Cuba. Methods Ae. aegypti larvae and pupae were collected monthly between September 2013 and July 2014 in the seven Municipal Health Areas of Arroyo Naranjo. Pools formed of 30‐55 larvae were examined through PCR and sequencing to detect the presence of each serotype. Results We analyzed 111 pools of larvae and pupae (4102 individuals) of which 37 tested positive for at least one DENV. More than one DENV type was observed in 10 of the 37 positive pools. Infected pools were detected every month, except in January, suggesting a sustained circulation of DENV in the vector populations. DENV‐1 and DENV‐3 were the most frequent and dispersed, though all four DENV types were detected. Nucleotide sequencing from positive pools confirmed RT‐PCR results for DENV‐1 (genotype V), DENV‐3 (genotype III) and DENV‐4 (genotype II). DENV‐2 was detected by RT‐PCR but could not be confirmed by nucleotide sequencing. Conclusion Our study of the distribution of natural vertical transmission of dengue virus types highlights extrinsic virus activity patterns in the area and could be used as a new surveillance tool. This article is protected by copyright. All rights reserved. 18 Substrate-engaged 26S proteasome structures reveal mechanisms for ATP-hydrolysis-driven translocationde la Pena, A. H., Goodall, E. A., Gates, S. N., Lander, G. C., Martin, A. The 26S proteasome is the primary eukaryotic degradation machine and thus critically involved in numerous cellular processes. The hetero-hexameric ATPase motor of the proteasome unfolds and translocates targeted protein substrates into the open gate of a proteolytic core, while a proteasomal deubiquitinase concomitantly removes substrate-attached ubiquitin chains. However, the mechanisms by which ATP hydrolysis drives the conformational changes responsible for these processes have remained elusive. Here we present the cryo-EM structures of four distinct conformational states of the actively ATP-hydrolyzing, substrate-engaged 26S proteasome. These structures reveal how mechanical substrate translocation accelerates deubiquitination, and how ATP-binding, hydrolysis, and phosphate-release events are coordinated within the AAA+ motor to induce conformational changes and propel the substrate through the central pore. 19 Using simultaneous amplification and testing method for evaluating the treatment outcome of pulmonary tuberculosisAbstract Background To evaluate the utility of Simultaneous Amplification and Testing (SAT-TB) Method for monitoring anti-TB treatment response. Methods Serial morning sputum specimens were obtained from 377 active pulmonary tuberculosis (PTB) cases at baseline, weeks 2, months 2, 5 and 6 (newly diagnosed patients) or 8 (previously treated patients) for AmpSure assay, smear fluorescence microscopy (FM) and BACTEC MGIT 960 culture assay. Results After treatment of 2 weeks, sputum culture was positive in 280 patients (74.27%). Among whom, 219 patients tested positive for SAT-TB assay and 143 patients smear FM positive. The detection rate of SAT-TB (78.21%) was significantly higher than sputum FM (51.07%, χ2 = 45.128, P
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