Abstract Background The burden of malaria and anaemia in pregnancy remains high despite the availability of proven efficacious antenatal care interventions. Sub-optimal uptake of the interventions may be due to inadequate active participation of pregnant women in their antenatal care. It was hypothesized that providing opportunities for pregnant women to improve upon active participation in their antenatal care through malaria and anaemia point-of-care testing would improve adherence to ANC recommendations and interventions and lead to better pregnancy outcomes. Methods Fourteen antenatal clinics in the Ashanti region of Ghana were randomized into intervention (pregnant women participating in their care plus current routine care) and control (current routine care) arms. Pregnant women attending the clinics for the first time were recruited and followed up until delivery. Haemoglobin levels and malaria parasitaemia were measured at baseline, 4–8 weeks after recruitment and at 36–40 weeks gestation. Birth weight and pregnancy outcomes were also recorded. Results The overall mean age, gestational age and haemoglobin at baseline were 26.4 years, 17.3 weeks and 110 g/l, respectively, with no significant differences between groups; 10.7% had asymptomatic parasitaemia; 74.6% owned an ITN but only 48.8% slept under it the night before enrolment. The adjusted risk ratio by 8 weeks follow up and at 36–40 weeks gestation in the intervention versus the control was 0.97 (95% CI 0.78–1.22) and 0.92 (95% CI 0.63–1.34) for anaemia and 1.17 (95% CI 0.68–2.04) and 0.83 (95% CI 0.27–2.57) for parasitaemia. The adjusted risk ratio for low birth weight was 0.93 (95% CI 0.44–1.97) and for pregnancy complications (abortions, intrauterine fetal deaths and still births) was 0.77 (95% CI 0.17–3.52) in the intervention group versus controls. Conclusion Although its potential was evident, this study found no significant beneficial effect of women participating in their malaria and haemoglobin tests on pregnancy outcomes. Exploring factors influencing health worker compliance to health intervention implementation and patient adherence to health interventions within this context will contribute in future to improving intervention effectiveness. Trial registration ISRTCTN88917252…

Bernigaud, C., Fang, F., Fischer, K., Lespine, A., Aho, L. S., Mullins, A. J., Tecle, B., Kelly, A., Sutra, J.-F., Moreau, F., Lilin, T., Beugnet, F., Botterel, F., Chosidow, O., Guillot, J.

Scabies is a major and potentially growing public health problem worldwide with an unmet need for acaricidal agents with greater efficacy and improved pharmacological properties for its treatment. The objective of the present study was to assess the efficacy and describe the pharmacokinetics profile of a novel acaricide, afoxolaner (AFX), in a relevant experimental porcine model. Twelve pigs were experimentally infested and treated either with 2.5 mg/kg single dose oral AFX (n = 4), 0.2 mg/kg two-doses 8 days apart oral ivermectin (IVM, n = 4), or no treatment against scabies (n = 4). Response to treatment was assessed by reduction of mite counts in skin scrapings as well as clinical and pruritus scores over time. Plasma and skin pharmacokinetic profiles for both AFX and IVM were evaluated. AFX efficacy was 100% at days 8 and 14 post-treatment and remained unchanged until the study-end (day 45). IVM efficacy was 86% and 97% on days 8 and 14, respectively, with a few mites recovered at study-end. Clinical and pruritus scores decreased in both treated groups and remained constant in the control group. Plasma mean residence times (MRT) were 7.1±2.4 and 1.1±0.2 days for AFX and IVM, respectively. Skin MRT values were 16.2±16.9 and 2.7±0.5 days for AFX and IVM, respectively. Overall, a single oral dose of AFX was efficacious for the treatment of scabies in experimentally infested pigs and showed a remarkably long MRT in plasma and notably in the skin.

Mathieu Nacher, Antoine Adenis, Florence Huber, Edouard Hallet, Philippe Abboud, Emilie Mosnier, Bastien Bideau, Christian Marty, Aude Lucarelli, Vanessa Morel, François Lacapère, Loïc Epelboin, Pierre Couppié

by Mathieu Nacher, Antoine Adenis, Florence Huber, Edouard Hallet, Philippe Abboud, Emilie Mosnier, Bastien Bideau, Christian Marty, Aude Lucarelli, Vanessa Morel, François Lacapère, Loïc Epelboin, Pierre Couppié Background Given the great efforts put into the strategic objective of reducing the proportion of HIV-infected patients that are undiagnosed, the aim of the present study was to review the temporal trends between 1997 and 2016 for median estimates of infection duration and median CD4 count at diagnosis for the main patient origins in French Guiana. Methods CD4 cell count at HIV sero-conversion and square root of CD4 cell decline were obtained using the CD4 decline in a cohort of HIV-infected persons in the UK, fitting random effect (slope and intercept) multilevel linear regression models. Multivariate analysis used robust regression for modeling the delay between estimated HIV seroconversion and diagnosis and quantile regression for CD4 at HIV diagnosis. Results The median interval between the estimated HIV seroconversion and HIV diagnosis was 8 years for patients fromBrazil, 4.5 years for those from Haiti, 6.6 years for those from Suriname, 3.3 years for patients from Guyana, and 3.1 years for French patients. A simple robust regression model with French patients as reference group adjusting for sex and age at the time of diagnosis showed that the interval was significantly longer for Brazilian (β = +3.7 years, P = 0.001), Surinamese (β = +4.2 years, p

Katz, Jeffrey; Greenberg, Steven

No abstract available…

Lynde, Grant C.; Jabaley, Craig S.

No abstract available…

Horita, Y., Alsultan, A., Kwara, A., Antwi, S., Enimil, A., Orstin, A., Dompreh, A., Yang, H., Wiesner, L., Peloquin, C. A.

Optimal doses for anti-tuberculosis (TB) drugs in children have yet to be established. In 2010, the World Health Organization (WHO) recommended revised dosages of the first-line anti-TB drugs for children. Pharmacokinetic (PK) studies that investigated the adequacy of the WHO revised dosages to date have yielded conflicting results. We performed population PK modeling using data from one of these studies to identify optimal dosages ranges. Ghanaian children with tuberculosis on recommended therapy with rifampin (RIF), isoniazid (INH), pyrazinamide (PZA) and ethambutol (EMB) for at least 4 weeks had blood samples collected at pre-dose, 1, 2, 4, and 8-hours post-dose. Drug concentrations were determined by validated liquid chromatography mass spectrometry methods. Nonlinear mixed-effects models were applied to describe the population PK of those drugs using MonolixSuite2016R1 (Lixoft, France). Bayesian estimation was performed, and the correlation coefficient, bias, and precision between the observed and predicted areas under the concentration-time curve (AUCs) were calculated, and Bland-Altman plots were analyzed. The population PK of RIF and PZA was described by one-compartment model and that for INH and EMB by two-compartment model. Cmax and AUC targets were based on published results in children from India. The lowest target values for pediatric TB patients were attainable at the WHO-recommended dosage schedule for RIF and INH, except for N-acetyl transferase 2 non-slow (rapid and intermediate) acetylators in the lower weight bands. However, higher published adult targets were not attainable for RIF and INH. The targets were not achieved for PZA and EMB.

Tatiana Metcalf, Jaime Soria, Silvia M. Montano, Eduardo Ticona, Carlton A. Evans, Luz Huaroto, Matthew Kasper, Eric S. Ramos, Nicanor Mori, Podjanee Jittamala, Kesinee Chotivanich, Irwin F. Chavez, Pratap Singhasivanon, Sasithon Pukrittayakamee, Joseph R. Zunt

by Tatiana Metcalf, Jaime Soria, Silvia M. Montano, Eduardo Ticona, Carlton A. Evans, Luz Huaroto, Matthew Kasper, Eric S. Ramos, Nicanor Mori, Podjanee Jittamala, Kesinee Chotivanich, Irwin F. Chavez, Pratap Singhasivanon, Sasithon Pukrittayakamee, Joseph R. Zunt Background Meningitis caused by Mycobacterium tuberculosis is a major cause of morbidity and mortality worldwide. We evaluated the performance of cerebrospinal fluid (CSF) testing with the GeneXpert MTB/RIF assay versus traditional approaches for diagnosing tuberculosis meningitis (TBM). Methods Patients were adults (n = 37) presenting with suspected TBM to the Hospital Nacional Dos de Mayo, Lima, Peru, during 12 months until 1st January 2015. Each participant had a single CSF specimen that was divided into aliquots that were concurrently tested for M. tuberculosis using GeneXpert, Ziehl-Neelsen smear and culture on solid and liquid media. Drug susceptibility testing used Mycobacteria Growth Indicator Tube (MGIT 960) and the proportions method. Results 81% (30/37) of patients received a final clinical diagnosis of TBM, of whom 63% (19/30, 95% confidence intervals, CI: 44–80%) were HIV-positive. 22% (8/37, 95%CI: 9.8–38%), of patients had definite TBM. Because definite TBM was defined by positivity in any laboratory test, all laboratory tests had 100% specificity. Considering the 30 patients who had a clinical diagnosis of TBM: diagnostic sensitivity was 23% (7/30, 95%CI: 9.9–42%) for GeneXpert and was the same for all culture results combined; considerably greater than 7% (2/30, 95%CI: 0.82–22%) for microscopy; whereas all laboratory tests had poor negative predictive values (20–23%). Considering only the 8 patients with definite TBM: diagnostic sensitivity was 88% (7/8, 95%CI: 47–100%) for GeneXpert; 75% (6/8, 95%CI: 35–97%) for MGIT culture or LJ culture; 50% (4/8, 95%CI 16–84) for Ogawa culture and 25% (2/8, 95%CI: 3.2–65%) for microscopy. GeneXpert and microscopy provided same-day results, whereas culture took 20–56 days. GeneXpert provided same-day rifampicin-susceptibility results, whereas culture-based testing took 32–71 days. 38% (3/8, 95%CI: 8.5–76%) of patients with definite TBM with data had evidence of drug-resistant TB, but 73% (22/30) of all clinically diagnosed TBM (definite, probable, and possible TBM) had no drug-susceptibility results available. Conclusions Compared with traditional culture-based methods of CSF testing, GeneXpert had similar yield and faster results for both the detection of M. tuberculosis and drug-susceptibility testing. Including use of the GeneXpert has the capacity to improve the diagnosis of TBM cases.

Ana Rivero, Sylvain Gandon

Avian malaria is the oldest experimental system for investigating the biology and transmission of Plasmodium parasites. Recent molecular protocols for detecting and characterizing avian malaria lineages in the field are providing an ever-growing picture of the prevalence, distribution, host range, and diversity hotspots of avian malaria across the world. The unparalleled genetic diversity uncovered rivals anything that has been found in other vertebrate malarias and seems to be matched by an equally rich phenotypic diversity, providing endless opportunities for exploring the selective pressures under which hosts and parasites evolve.

Ge, S., Beechinor, R. J., Hornik, C. P., Standing, J. F., Zimmerman, K., Cohen-Wolkowiez, M., Laughon, M. M., Clark, R., Gonzalez, D.

Gentamicin is a common antibiotic used in neonates and infants. A recently published population pharmacokinetic (PK) model was developed using data from multiple studies, and the objective of our analyses is to evaluate the feasibility of using a national electronic health record (EHR) database to further externally evaluate this model. Our results suggest that with proper data capture procedures, EHR data can serve as a potential data source for external evaluation of PK models.

Abstract Background Accurate early diagnosis and prompt treatment are one of the key strategies to control and prevent malaria disease. External quality assessment is the most effective method for evaluation of the quality of malaria microscopy diagnosis. The aim of this study was to assess the quality of malaria microscopy diagnosis and its associated factors in selected public health facility laboratories in East Wollega Zone, Western Ethiopia. Methods Facility-based cross-sectional study design was conducted in 30 randomly selected public health facility laboratories from November 2014 to January 2015 in East Wollega Zone, Western Ethiopia. Ten validated stained malaria panel slides with known Plasmodium species, developmental stage and parasite density were distributed. Data were captured; cleaned and analyzed using SPSS version 20 statistical software-multivariate logistic regressions and the agreement in reading between the peripheral diagnostic centers and the reference laboratory were done using kappa statistics. Results A total of 30 health facility laboratories were involved in the study and the overall quality of malaria microscopy diagnosis was poor (62.3%). The associated predictors of quality in this diagnosis were in-service training [(AOR = 16, 95% CI (1.3, 1.96)], smearing quality [(AOR = 24, 95% CI (1.8, 3.13)], staining quality [(AOR = 15, 95% CI (2.35, 8.61), parasite detection [(AOR = 9, 95% CI (1.1, 8.52)] and identification skills [(AOR = 8.6, 95% CI (1.21, 1.63)]. Eighteen (60%) of health facility laboratories had in-service trained laboratory professionals on malaria microscopy diagnosis. Conclusion Overall quality of malaria microscopy diagnosis was poor and a significant gap in this service was observed that could impact on its diagnostic services.

Bassi, M. R., Sempere, R. N., Meyn, P., Polacek, C., Arias, A.

Flaviviruses constitute an increasing source of public health concern with growing numbers of pathogens causing disease, and a geographic spread to temperate climates. Despite a large body of evidence supporting mutagenesis as a conceivable antiviral strategy, there is currently no data on the sensitivity to increased mutagenesis for Zika virus (ZIKV) and Usutu virus (USUV), two emerging flaviviral threats. In this study, we demonstrate that both viruses are sensitive to three ribonucleosides that have shown mutagenic activity against other RNA viruses – favipiravir, ribavirin and 5-fluorouracil – while they remain unaffected by a mutagenic deoxyribonucleoside. Serial cell culture passages of ZIKV in the presence of these compounds resulted in the rapid extinction of infectivity, suggesting elevated sensitivity to mutagenesis. USUV extinction was achieved when a 10-fold dilution was applied between every passage, but not in experiments involving undiluted virus, indicating an overall lower susceptibility than ZIKV. Although both viruses are inhibited by the same three drugs, ZIKV is relatively more susceptive to serial passage in the presence of purine analogues (favipiravir and ribavirin) while USUV replication is suppressed more efficiently by 5-fluorouracil. These differences in sensitivity typically correlate with the increases in the mutation frequencies observed in each nucleoside treatment. These results are relevant to the development of efficient therapies based on lethal mutagenesis, and support the rational selection of different mutagenic nucleosides for each pathogen. We will discuss the implications of these results to the fidelity of flavivirus replication, and the design of antiviral therapies based on lethal mutagenesis.

Ayehu Mekonen, Yeshi Ayele, Yifru Berhan, Desalegn Woldeyohannes, Woldaregay Erku, Solomon Sisay

by Ayehu Mekonen, Yeshi Ayele, Yifru Berhan, Desalegn Woldeyohannes, Woldaregay Erku, Solomon Sisay Objectives Quality of tuberculosis (TB) microscopy diagnosis is not a guarantee despite implementation of External Quality Assurance (EQA) service in all laboratories of health facilities. Hence, we aimed at evaluating the technical quality and the findings of sputum smear microscopy for acid fast bacilli (AFB) at health centers in Hararge Zone, Oromia Region, Ethiopia. Methods A cross-sectional study was carried out between July 8, 2014 and July 7, 2015.A pre-tested structured questionnaire was used to collect data. Lot Quality Assurance Sampling (LQAS) method was put into practice for collecting all necessary sample slides. Data were analyzed by using SPSS (Statistical Package for Social Sciences) version 20 software. P-value < 0.05 was considered as statistically significant. Results Of the total55 health center laboratories which had been assessed during the study period, 20 (36.4%) had major technical errors; 13 (23.6%) had 15 false negative results and 17 (30.9%) had 22 false positive results. Moreover, poor specimen quality, smear size, smear thickness, staining and evenness were indicated in 40 (72.7%), 39 (70.9%), 37 (67.3%), 27(49.1%) and 37 (67.3%) of the collected samples, respectively. False negative AFB findings were significantly associated with lack of Internal Quality Control (IQC) measures (AOR (Adjusted Odds Ratio): 2.90 (95% CI (Confidence Interval): 1.25,6.75) and poor staining procedures (AOR: 2.16(95% CI: 1.01, 5.11). Conclusions The qualities of AFB smear microscopy reading and smearing were low in most of the laboratories of the health centers. Therefore, it is essential to strength EQA program through building the capacity of laboratory professionals.

E. Bottieau, C.P. Yansouni

Frédéric Debellut, Nathaniel Hendrix, Justin R. Ortiz, Philipp Lambach, Kathleen M. Neuzil, Niranjan Bhat, Clint Pecenka

by Frédéric Debellut, Nathaniel Hendrix, Justin R. Ortiz, Philipp Lambach, Kathleen M. Neuzil, Niranjan Bhat, Clint Pecenka Immunization of pregnant women against seasonal influenza remains limited in low- and lower-middle-income countries despite being recommended by the World Health Organization (WHO). The WHO/PATH Maternal Influenza Immunization Project was created to identify and address obstacles to delivering influenza vaccines to pregnant women in low resource setting. To gain a better understanding of potential demand from this target group, we developed a model simulating pregnant women populations eligible for vaccination during antenatal care (ANC) services in all low- and lower-middle-income countries. We assessed potential vaccine demand in the context of both seasonal and year-round vaccination strategies and identified the ways that immunization programs may be affected by availability gaps in supply linked to current vaccine production cycles and shelf life duration. Results of our analysis, which includes 54 eligible countries in 2015 for New Vaccine Support from Gavi, the Vaccine Alliance, suggest the demand for influenza vaccines could be 7.7 to 16.0 million doses in 2020, and 27.0 to 61.7 million doses by 2029. If current trends in production capacity and actual production of seasonal influenza vaccines were to continue, global vaccine supply would be sufficient to meet this additional demand—although a majority of countries would face implementation issues linked to timing of supply.

Alejandro Ortega-Beltran, Juan Moral, Ryan D. Puckett, David P. Morgan, Peter J. Cotty, Themis J. Michailides

by Alejandro Ortega-Beltran, Juan Moral, Ryan D. Puckett, David P. Morgan, Peter J. Cotty, Themis J. Michailides Interactions between pathogenic and nonpathogenic fungal species in the tree canopy are complex and can determine if disease will manifest in the plant and in other organisms such as honey bees. Seasonal dynamics of fungi were studied in an almond orchard in California where experimental release of the atoxigenic biopesticide Aspergillus flavus AF36 to displace toxigenic Aspergillus strains has been conducted for five years. The presence of the vegetative compatibility group (VCG) YV36, to which AF36 belongs, in the blossoms, and the honey bees that attend these blossoms, was assessed. In blossoms, A. flavus frequencies ranged from 0 to 4.5%, depending on the year of study. Frequencies of honey bees carrying A. flavus ranged from 6.5 to 10%. Only one A. flavus isolate recovered from a blossom in 2016 belonged to YV36, while members of the VCG were not detected contaminating honey bees. Exposure of pollinator honey bees to AF36 was detected to be very low. The density of several Aspergillus species was found to increase during almond hull split and throughout the final stages of maturation; this also occurred in pistachio orchards during the maturation period. Additionally, we found that AF36 effectively limited almond aflatoxin contamination in laboratory assays. This study provides knowledge and understanding of the seasonal dynamics of Aspergillus fungi and will help design aflatoxin management strategies for almond. The evidence of the low levels of VCG YV36 encountered on almond blossoms and bees during pollination and AF36’s effectiveness in limiting aflatoxin contamination in almond provided additional support for the registration of AF36 with USEPA to use in almond in California.

Monarrez, R., Wang, Y., Fu, Y., Liao, C.-H., Okumura, R., Braun, M. R., Jacoby, G. A., Hooper, D. C.

Expression of the quinolone resistance gene qnrS1 is increased by quinolones, but, unlike induction of some other qnr genes, the bacterial SOS system is not involved and no lexA box is found upstream. Nonetheless, at least 205-bp of upstream sequence is required for induction to take place. An upstream sequence bound to beads trapped potential binding proteins from cell extracts that were identified by mass spectrometry as Dps, Fis, Ihf, Lrp, CysB, and YjhU. To further elucidate their role a reporter plasmid linking the qnrS1 upstream sequence to lacZ was introduced into cells of the Keio collection with single gene deletions and screened for lacZ expression. Mutants in ihfA and ihfB had decreased lacZ induction while induction in a cysB mutant was increased and dps, fis, lrp, yjhU and other mutants showed no change. The essential upstream sequence contains potential binding sites for Ihf and DnaA. A dnaA deletion could not be tested because it provides essential functions in cell replication, but increased dnaA expression decreased qnrS1 induction while decreased dnaA expression enhanced it implying a role for DnaA as a repressor. In a mobility shift assay purified IhfA, IhfB, and DnaA proteins (but not CysB) were shown to bind to the upstream segment. Induction decreased in a gyrA quinolone-resistant mutant indicating that GyrA also has a role. Thus, quinolones acting through proteins DnaA, GyrA, IhfA, and IhfB regulates expression of qnrS1.

Shipley M, Renner D, Ott M, et al.

AbstractHere we present genomic and in vitro analyses of temporally separated episodes of herpes simplex virus type 1 (HSV-1) shedding by an HSV-1–seropositive and human immunodeficiency virus (HIV)/HSV-2–seronegative individual who has frequent recurrences of genital HSV-1. Using oligonucleotide enrichment, we compared viral genomes from uncultured swab specimens collected on different days and from distinct genital sites. We found that viral genomes from 7 swab specimens and 3 cultured specimens collected over a 4-month period from the same individual were 98.5% identical. We observed a >2-fold difference in the number of minority variants between swab specimens from lesions, swab specimens from nonlesion sites, and cultured specimens. This virus appeared distinct in its phylogenetic relationship to other strains, and it contained novel coding variations in 21 viral proteins. This included a truncation in the UL11 tegument protein, which is involved in viral egress and spread. Normal immune responses were identified, suggesting that unique viral genomic features may contribute to the recurrent genital infection that this participant experiences.

Adamma Aghaizu, Jennifer Tosswill, Daniela De Angelis, Helen Ward, Gwenda Hughes, Gary Murphy, Valerie Delpech

by Adamma Aghaizu, Jennifer Tosswill, Daniela De Angelis, Helen Ward, Gwenda Hughes, Gary Murphy, Valerie Delpech Introduction The HIV epidemic in England is largely concentrated among heterosexuals who are predominately black African and men who have sex with men (MSM). We present for the first time trends in annual HIV incidence for adults attending sexual health clinics, where 80% of all HIV diagnoses are made. Methods We identified newly diagnosed incident HIV using a recent infection testing algorithm (RITA) consisting of a biomarker (AxSYM assay, modified to determine antibody avidity), epidemiological and clinical information. We estimated HIV incidence using the WHO RITA formula for cross-sectional studies, with HIV testing data from sexual health clinics as the denominator. Results From 2009 to 2013, each year, between 9,700 and 26,000 black African heterosexuals (of between 161,000 and 231,000 heterosexuals overall) were included in analyses. For the same period, annually between 19,000 and 55,000 MSM were included. Estimates of HIV incidence among black Africans increased slightly (although non-significantly) from 0.15% (95% C.I.0.05%-0.26%) in 2009 to 0.19% (95% C.I.0.04%-0.34%) in 2013 and was 4-5-fold higher than among all heterosexuals among which it remained stable between 0.03% (95% C.I.0.02%-0.05%) and 0.05% (95% C.I.0.03%-0.07%) over the period. Among MSM incidence was highest and increased (non-significantly) from 1.24% (95%C.I 0.96–1.52%) to 1.46% (95% C.I 1.23%-1.70%) after a peak of 1.52% (95%C.I 1.30%-1.75%) in 2012. Conclusion These are the first nationwide estimates for trends in HIV incidence among black African and heterosexual populations in England which show black Africans, alongside MSM, remain disproportionately at risk of infection. Although people attending sexual health clinics may not be representative of the general population, nearly half of black Africans and MSM had attended in the previous 5 years. Timely and accurate incidence estimates will be critical in monitoring the impact of the reconfiguration of sexual health services in England, and any prevention programmes such as pre-exposure prophylaxis.

On the Wednesday before the summer solstice in the United States, President Donald Trump ended his administration’s policy of forced separation of immigrant children from their parents at the U.S.–Mexico border — a practice characterized by the president of the American Academy of Pediatrics (AAP)……

Tiffany M. Lowe-Power, Devanshi Khokhani, Caitilyn Allen

The plant wilt pathogen Ralstonia solanacearum thrives in the water-transporting xylem vessels of its host plants. Xylem is a relatively nutrient-poor, high-flow environment but R. solanacearum succeeds there by tuning its own metabolism and altering xylem sap biochemistry. Flow influences many traits that the bacterium requires for pathogenesis. Most notably, a quorum sensing system mediates the pathogen’s major transition from a rapidly dividing early phase that voraciously consumes diverse food sources and avidly adheres to plant surfaces to a slower-growing late phase that can use fewer nutrients but produces virulence factors and disperses effectively.

Carteron, Laurent; Solari, Daria; Patet, Camille; Quintard, Hervé; Miroz, John-Paul; Bloch, Jocelyne; Daniel, Roy T.; Hirt, Lorenz; Eckert, Philippe; Magistretti, Pierre J.; Oddo, Mauro

Objectives: Lactate promotes cerebral blood flow and is an efficient substrate for the brain, particularly at times of glucose shortage. Hypertonic lactate is neuroprotective after experimental brain injury; however, human data are limited. Design: Prospective study (clinicaltrials.gov NCT01573507). Setting: Academic ICU. Patients: Twenty-three brain-injured subjects (13 traumatic brain injury/10 subarachnoid hemorrhage; median age, 59 yr [41–65 yr]; median Glasgow Coma Scale, 6 [3–7]). Interventions: Three-hour IV infusion of hypertonic lactate (sodium lactate, 1,000 mmol/L; concentration, 30 µmol/kg/min) administered 39 hours (26–49 hr) from injury. Measurements and Main Results: We examined the effect of hypertonic lactate on cerebral perfusion (using transcranial Doppler) and brain energy metabolism (using cerebral microdialysis). The majority of subjects (13/23 = 57%) had reduced brain glucose availability (baseline pretreatment cerebral microdialysis glucose, < 1 mmol/L) despite normal baseline intracranial pressure (10 [7–15] mm Hg). Hypertonic lactate was associated with increased cerebral microdialysis lactate (+55% [31–80%]) that was paralleled by an increase in middle cerebral artery mean cerebral blood flow velocities (+36% [21–66%]) and a decrease in pulsatility index (–21% [13–26%]; all p < 0.001). Cerebral microdialysis glucose increased above normal range during hypertonic lactate (+42% [30–78%]; p < 0.05); reduced brain glucose availability correlated with a greater improvement of cerebral microdialysis glucose (Spearman r = –0.53; p = 0.009). No significant changes in cerebral perfusion pressure, mean arterial pressure, systemic carbon dioxide, and blood glucose were observed during hypertonic lactate (all p > 0.1). Conclusions: This is the first clinical demonstration that hypertonic lactate resuscitation improves both cerebral perfusion and brain glucose availability after brain injury. These cerebral vascular and metabolic effects appeared related to brain lactate supplementation rather than to systemic effects. Drs. Carteron and Solari are co-first authors and contributed equally to this work. Supported, in part, by grants from the Swiss National Science Foundation (320030_138191 and 32003B_155957 [to Dr. Oddo]), the Novartis Foundation for Biomedical Research (to Dr. Oddo), and the Société Française d’Anesthésie et de Réanimation and the Fondation des Gueules Cassées (both to Dr. Carteron). Dr. Hirt received other support from Boehringer Ingelheim (speaker fees and travel grants, not related to this work) and research grants not in relation with this work from the Swiss Science Foundation, the Biaggi Foundation, and the Juchum Foundation. Dr. Oddo received support for article research from the Swiss National Science Foundation. The remaining authors have disclosed that they do not have any potential conflicts of interest. For information regarding this article, E-mail: mauro.oddo@chuv.ch Copyright © by 2018 by the Society of Critical Care Medicine and Wolters Kluwer Health, Inc. All Rights Reserved.

Abstract Background Plasmodium falciparum is the most virulent parasite of the five Plasmodium species that cause human malaria, and biological analysis of the parasite is critical for the development of novel strategies for disease control. DNA endonucleases are important for maintaining the biological activity, gene stability of the parasite and interaction with host immune systems. In this study, ten sequences of DNA endonucleases were found in the genome of P. falciparum 3D7 clone, seven of them were predicted to contain an endonuclease/exonuclease/phosphatase (IPR005135) domain which plays an important role in DNA catalytic activity. The seven DNA endonucleases of P. falciparum were systematically investigated. Methods Plasmodium falciparum 3D7 clone was cultured in human O+ RBCs, RNA was extracted at 8, 16, 24, 32, 40, and 48 h post invasion and real-time quantitative PCR was carried out to analyse the transcription of the seven DNA endonuclease genes in asexual stages. Immunofluorescence assay was carried out to confirm the location of the encoded proteins expressed in the erythrocytic stages. Finally, the catalytic activity of the DNA nucleases were tested. Results Of the seven proteins analysed, two proteins were not soluble. Fragments derived from the rest five endonuclease sequences were successfully expressed as soluble proteins, and which were used to generate antisera for protein localization. The proteins were all located in the nucleus at ring and trophozoite stages. While at schizont stage, proteins encoded by PF3D7_1238600, PF3D7_0107200 and PF3D7_0319200 were in the punctuated forms in the parasite most likely around nuclei of the merozoites. But the proteins encoded by PF3D7_0305600 and PF3D7_1363500 were distributed around the infected erythrocyte membrane. The enzymatic activity of the recombinant GST-PF3D7_1238600 was very efficient without divalent iron, while the activity of the rest four enzymes was iron dependent. Further, divalent irons did not show any specific enhancement on the activity of GST-PF3D7_1238600, but the activity of GST-PF3D7_0107200, GST-PF3D7_1363500 and GST-PF3D7_0319200 were Cu2+ dependent. The activity of GST-PF3D7_0305600 was dependent on Mg2+ and Mn2+. Except GST-PF3D7_1363500, four of the GST tagged recombinant proteins hydrolysed the supercoiled circular plasmid DNA with or without divalent metal ions. The GST-PF3D7_1363500 protein only changed the supercoiled circular plasmid DNA into nicked plasmids, even with Cu2+. Conclusions Fragments derived from five of the endonuclease sequences of P. falciparum 3D7 clone were successfully expressed. The proteins displayed diverse cell distribution, biochemical and enzymatic activities, which indicated that they carried different biological function in the development of the parasite in the erythrocytes. The DNA repair and DNA degradation capacity of the DNA endonucleases in the biology of the parasite remained further studied.

Abstract Background Dengue and West Nile viruses are highly cross-reactive and have numerous parallels in geography, potential vector host (Aedes family of mosquitoes), and initial symptoms of infection. While the vast majority (> 80%) of both dengue and West Nile virus infections result in asymptomatic infections, a minority of individuals experience symptomatic infection and an even smaller proportion develop severe disease. The mechanisms by which these infections lead to severe disease in a subset of infected individuals is incompletely understood, but individual host differences including genetic factors and immune responses have been proposed. We sought to identify genetic risk factors that are associated with more severe disease outcomes for both viruses in order to shed light on possible shared mechanisms of resistance and potential therapeutic interventions. Methods We applied a search strategy using four major databases (Medline, PubMed, Embase, and Global Health) to find all known genetic associations identified to date with dengue or West Nile virus disease. Here we present a review of our findings and a meta-analysis of genetic variants identified. Results We found genetic variations that are significantly associated with infections of these viruses. In particular we found variation within the OAS1 (meta-OR = 0.83, 95% CI: 0.69–1.00) and CCR5 (meta-OR = 1.29, 95% CI: 1.08–1.53) genes is significantly associated with West Nile virus disease, while variation within MICB (meta-OR = 2.35, 95% CI: 1.68–3.29), PLCE1 (meta-OR = 0.55, 95% CI: 0.42–0.71), MBL2 (meta-OR = 1.54, 95% CI: 1.02–2.31), and IFN-γ (meta-OR = 2.48, 95% CI: 1.30–4.71), is associated with dengue disease. Conclusions Despite substantial heterogeneity in populations studied, genes examined, and methodology, significant associations with genetic variants were found across studies within both diseases. These gene associations suggest a key role for immune mechanisms in susceptibility to severe disease. Further research is needed to elucidate the role of these genes in disease pathogenesis and may reveal additional genetic factors associated with disease severity.

Mercedes Marín, Adrián Ruiz, Cristina Iglesias, Lidia Quiroga, Emilia Cercenado, Pablo Martín-Rabadán, Emilio Bouza, Belén Rodríguez-Sánchez

The identification of Nocardia isolates still represents a challenge for MALDI-TOF MS despite its acceptance for most bacterial and fungal isolates. In this study we evaluate the identification of Nocardia isolates using direct spotting and an updated database.

Chong, S., Dugast-Darzacq, C., Liu, Z., Dong, P., Dailey, G. M., Cattoglio, C., Heckert, A., Banala, S., Lavis, L., Darzacq, X., Tjian, R.

Many eukaryotic transcription factors (TFs) contain intrinsically disordered low-complexity domains (LCDs), but how they drive transactivation remains unclear. Here, live-cell single-molecule imaging reveals that TF-LCDs form local high-concentration interaction hubs at synthetic and endogenous genomic loci. TF-LCD hubs stabilize DNA binding, recruit RNA polymerase II (Pol II), and activate transcription. LCD-LCD interactions within hubs are highly dynamic, display selectivity with binding partners, and are differentially sensitive to disruption by hexanediols. Under physiological conditions, rapid and reversible LCD-LCD interactions occur between TFs and the Pol II machinery without detectable phase separation. Our findings reveal fundamental mechanisms underpinning transcriptional control and suggest a framework for developing single-molecule imaging screens for novel drugs targeting gene regulatory interactions implicated in disease.

Alemnesh H. Mirkuzie

by Alemnesh H. Mirkuzie About 40% of the new HIV infections in Ethiopia are among children < 15 years of age. The great majority of these infections occur through Mother-to-child HIV transmission (MTCT). For prevention of MTCT, the national guidelines has been revised to incorporate scientific advances in HIV prevention, treatment and care. Since 2005, the country has been implementing a peer mentor programme called Mother Support Group (MSG), which provides psychosocial and adherence support for HIV positive mothers. This study examined implementation of PMTCT guidelines revisions and outcomes of HIV exposed babies in the MSG in Addis Ababa. Retrospective routine data were collected between 2005 and August 2013 from seven randomly selected primary health facilities. Odds ratios and 95% confidence intervals were calculated using logistic regression models. Several guidelines revisions were made between 2001 and 2013 in HIV testing approaches, prophylactic antiretroviral options, infant feeding recommendations and infant HIV testing algorithms. Revisions on the CD4 thresholds were associated with a significant increase in the proportion of women initiating antiretroviral treatment from 0 in 2005 to 62% in 2013. Revisions in infant feeding recommendations led to a 92.3% reported practice of exclusive breastfeeding in 2013 compared to 60.9% in 2005. Two and four percent of the HIV exposed babies were HIV positive by six and 18 months respectively. Not receiving prophylactic ART and receiving mixed feeding were independent predictors for babies having an HIV positive antibody test at 18 months. The rate of HIV status disclosure increased significantly year by year. Over the years, the PMTCT recommendations have moved from having a solo focus on PMTCT to holistic and inclusive approaches emphasizing survival beyond HIV prevention. The data reflect favourable outcomes of HIV exposed babies in terms of averted MTCT though serious gaps in data quality remain. For successful implementation of Option-B plus, the identified gaps in the MSG need to be addressed.

Abstract Background Five species of Plasmodium are known to infect humans. For proper treatment of malaria, accurate identification of the parasite species is crucial. The current gold standard for malaria diagnosis is microscopic examination of Giemsa-stained blood smears. Since the parasite species are identified by microscopists who manually search for the parasite-infected red blood cells (RBCs), misdiagnosis due to human error tends to occur in case of low parasitaemia or mixed infection. Then, molecular methods, such as polymerase chain reaction or loop-mediated isothermal amplification (LAMP), are required for conclusive identification of the parasite species. However, since molecular methods are highly sensitive, false-positive results tend to occur due to contamination (carry over) or the target gene products may be detected even after clearance of the parasites from the patient’s blood. Therefore, accurate detection of parasites themselves by microscopic examination is essential for the definitive diagnosis. Thus, the method of in situ LAMP for the parasites was developed. Results Red blood cell suspensions, including cultured Plasmodium falciparum, strain 3D7, infected-RBCs, were dispersed on cyclic olefin copolymer (COC) plate surfaces rendered hydrophilic by reactive ion-etching treatment using a SAMCO RIE system (hydrophilic-treated), followed by standing for 10 min to allow the RBCs to settle down on the plate surface. By rinsing the plate with RPMI 1640 medium, monolayers of RBCs formed on almost the entire plate surface. The plate was then dried with a hair drier. The RBCs were fixed with formalin, followed by permeabilization with Triton X-100. Then, amplification of the P. falciparum 18S rRNA gene by the LAMP reaction with digoxigenin (DIG)-labelled dUTP and a specific primer set was performed. Infected RBCs as fluorescence-positive cells with anti-DIG antibodies conjugated with fluorescein using fluorescent microscopy could be detected. Conclusions The present work shows that the potential of in situ LAMP for the identification of Plasmodium species at the single cell level on hydrophilic-treated COC palates, allowing highly sensitive and accurate malaria diagnosis. The findings will improve the efficacy of the gold standard method for malaria diagnosis.

Robin, F., Auzou, M., Bonnet, R., Lebreuilly, R., Isnard, C., Cattoir, V., Guerin, F.

The study evaluated the in vitro activity of ceftolozane-tazobactam (C/T) against 94 unique clinical isolates of Enterobacter cloacae complex (ECC). No difference was observed according to the ECC cluster. The in vitro activity greatly varied depending on the β-lactamase-producing profile: 100%, 67% and 19% of wild-type, ESBL-producing, and AmpC-overproducing strains were susceptible to C/T, respectively. The use of C/T could be of interest for the treatment of some infections caused by ESBL-producing AmpC-non-overexpressing ECC isolates.

Van Dalem, A., Herpol, M., Echahidi, F., Peeters, C., Wybo, I., De Wachter, E., Vandamme, P., Pierard, D.

We tested the in vitro susceptibility of ceftazidime-avibactam and ceftolozane-tazobactam and 13 other antibiotics against 91 Burkholderia cepacia complex (BCC) strains isolated from cystic fibrosis patients since 2012. Highest susceptibility (82%) was found for trimethoprim-sulfamethoxazole. Respectively, 81% and 63% of all BCC strains were susceptible to ceftazidime-avibactam and ceftolozane-tazobactam. For temocillin, ceftazidime, piperacillin-tazobactam and meropenem, at least 50% of the strains were susceptible. B. stabilis seems to be more resistant than other BCC species.

Autumn Zuckerman, Andrew Douglas, Sam Nwosu, Leena Choi, Cody Chastain

by Autumn Zuckerman, Andrew Douglas, Sam Nwosu, Leena Choi, Cody Chastain Barriers remain in the hepatitis C virus (HCV) cascade of care (CoC), limiting the overall impact of direct acting antivirals. This study examines movement between the stages of the HCV CoC and identifies reasons why patients and specific patient populations fail to advance through care in a real world population. We performed a single-center, ambispective cohort study of patients receiving care in an outpatient infectious diseases clinic between October 2015 and September 2016. Patients were followed from treatment referral through sustained virologic response. Univariate and multivariate analyses were performed to identify factors related to completion of each step of the CoC. Of 187 patients meeting inclusion criteria, 120 (64%) completed an evaluation for HCV treatment, 119 (64%) were prescribed treatment, 114 (61%) were approved for treatment, 113 (60%) initiated treatment, 107 (57%) completed treatment, and 100 (53%) achieved a sustained virologic response. In univariate and multivariate analyses, patients with Medicaid insurance were less likely to complete an evaluation and were less likely to be approved for treatment. Treatment completion and SVR rates are much improved from historical CoC reports. However, linkage to care following referral continues to be a formidable challenge for the HCV CoC in the DAA era. Ongoing efforts should focus on linkage to care to capitalize on DAA treatment advances and improving access for patients with Medicaid insurance.

Lai Lai San, Khin Saw Aye, Nan Aye Thida Oo, Mu Mu Shwe, Yukari Fukushima, Stephen V Gordon, Yasuhiko Suzuki, Chie Nakajima

Myanmar is a WHO high tuberculosis (TB) burden country with a high multidrug-resistant (MDR)-TB burden. Significantly a high prevalence of the Beijing genotype of Mycobacterium tuberculosis (MTB) among MDR-MTB has been reported previously. To explore whether an association exists between the prevalence of the Beijing MTB genotype and MDR-TB in Myanmar, we performed detailed genetic characterization of TB clinical isolates.

Hasan, M. M., Teixeira, J. E., Huston, C. D.

Entamoeba histolytica is a protozoan parasite that causes invasive amoebiasis when it invades the human colon. Tissue invasion requires a shift from an adhesive lifestyle in the colonic lumen to a motile and extracellular matrix (ECM) degradative lifestyle in the colonic tissue layers. How the parasite regulates between these two lifestyles is largely unknown. Previously, we showed that silencing the E. histolytica surface metalloprotease EhMSP-1 results in parasites that are hyperadherent and less motile. Trying to better understand the molecular mechanism of this phenotype, we now show that the EhMSP-1 silenced parasites cannot efficiently form specialized dot-like polymerized actin (F-actin) structures upon interaction with the human ECM component fibronectin. We characterized these F-actin structures and found that they are very short-lived structures that are the sites of fibronectin degradation. Motile mammalian cells form F-actin structures called invadosomes that are similar in stability and function to these amebic actin dots. Therefore, we propose here that E. histolytica forms amebic invadosomes to facilitate colonic tissue invasion.

Sonia Andersson, Karen Belkić, Miriam Mints, Ellinor Östensson

by Sonia Andersson, Karen Belkić, Miriam Mints, Ellinor Östensson Objective Self-sampling to test for high risk human papilloma virus (HPV) is becoming an increasingly important component of cervical cancer screening. The aim of this observational study is to examine how women treated for high-grade cervical intraepithelial neoplasia (CIN) view HPV self-sampling. Methods Invited to participate in the present study were patients who had undergone treatment of high-grade CIN (grade 2 or higher) and were followed-up at 6-months at the Karolinska University Hospital, Stockholm. The participants were instructed as to how to perform HPV self-sampling. Thereafter, the participants completed a questionnaire about HPV self-sampling and other cervical cancer screening methods, as well as about self-perceived risk of cervical cancer without regular gynecologic follow-up and about specific knowledge regarding HPV, CIN and cervical cancer. Results Altogether 479 women enrolled in this study. The participation rate was 96.6%. Nearly 75% of the participants stated they would consider performing the HPV self-sampling prior to their next gynecologic follow-up. Confidence in HPV self-sampling was a significant independent predictor of willingness to perform HPV self-sampling. However, confidence in HPV self-sampling was significantly lower than confidence in Papanicolaou smears and in HPV testing with samples collected by health professionals. Higher specific knowledge about HPV, CIN and cervical cancer was also a significant independent predictor of willingness to perform HPV self-sampling, as was having travelled longer distance to attend gynecologic follow-up. Participants with lower income and without completed university education expressed significantly higher confidence in HPV self-sampling and lower confidence in Papanicolaou smears than the other women. Conclusions To the best of our knowledge, this is the first study to examine the views of women treated for high-grade CIN vis-à-vis HPV self-sampling. The latter is an acceptable option for the vast majority of this cohort of women.

Daniel Slunge, Anders Boman

by Daniel Slunge, Anders Boman The purpose of this study is to analyse the role of risk perceptions and exposure for protective behaviour against tick bites and the related diseases Lyme borreliosis (LB) and tick-borne encephalitis (TBE), both of which are growing health concerns. We use data from a national survey in Sweden with respondents in geographical areas with substantial differences in both abundance of ticks and incidence of LB and TBE. We find that the share of respondents who frequently use protective clothing (64%), perform tick checks (63%) or avoid tall grass while in areas with ticks (48%) is relatively high. However, the use of protective measures is uneven and a considerably lower share tuck their trousers into their socks (18%), use repellent against ticks (16%) or use a combination of protective measures. Thirty-one per cent of the respondents report one or more tick bites in the last year and 68% report one or more lifetime tick bites, indicating that it is difficult to protect oneself from tick bites. There is a strong positive association between exposure and checking the skin for ticks, but exposure is only weakly associated with other protective measures. Tick bites are perceived as a serious health risk by as many as 43% of the respondents. The perception that a single tick bite is serious is negatively associated with actual exposure to ticks, while the opposite is true for the perception that tick bites constitute a serious lifetime health risk. This indicates a learning effect in relation to risk perceptions and the performance of tick checks, but not in relation to other protective measures. Recommendations include informing people of the risks associated with tick bites, the efficacy of various protective measures and the importance of combining multiple types of protection. Given the high exposure to tick bites, the growing incidence of TBE and LB, and the difficulties in preventing tick bites, other preventive measures should be further discussed, including vaccination programmes.

Soucy A, Hurteau G, Metzger D.

AbstractDisease tolerance can preserve host homeostasis and limit the negative impact of infections. We report that vaccinated mice survived pulmonary challenge with the extremely virulent SchuS4 strain of F. tularensis, despite the persistence of large numbers (~104) of organisms. Transfer of 100 of these resident bacteria to naïve animals caused 100% lethality, demonstrating that virulence was maintained. Tissue damage in the lung was limited over the course of infection, and was associated with increased levels of amphiregulin. Mice depleted of CD4+ cells had reduced amphiregulin and succumbed to infection. In addition, neutralization of IFNγ or depletion of CD8+ cells resulted in increased pathogen loads, bacteremia, and death of the host. Conversely, depletion of Ly6G+ neutrophils had no effect on survival and actually resulted in reduced bacterial levels. Understanding the interplay between host resistance and disease tolerance will provide new insights into the understanding of chronic infectious diseases.

Abstract Background Malaria parasites and related haemosporidian parasites are widespread and may cause severe diseases in birds. These pathogens should be considered in projects aiming breeding of birds for purposes of sustained ex situ conservation. Cranes are the ‘flagship species’ for health assessment of wetland ecosystems, and the majority of species are endangered. Malaria parasites and other haemosporidians have been reported in cranes, but the host-parasite relationships remain insufficiently understood. Morbidity of cranes due to malaria has been reported in Beijing Zoo. This study report prevalence, diversity and distribution of malaria parasites and related haemosporidians in cranes in Beijing Zoo and suggest simple measures to protect vulnerable individuals. Methods In all, 123 cranes (62 adults and 61 juveniles) belonging to 10 species were examined using PCR-based testing and microscopic examination of blood samples collected in 2007–2014. All birds were maintained in open-air aviaries, except for 19 chicks that were raised in a greenhouse with the aim to protect them from bites of blood-sucking insects. Bayesian phylogenetic analysis was used to identify the closely related avian haemosporidian parasites. Results Species of Plasmodium (5 lineages), Haemoproteus (1) and Leucocytozoon (2) were reported. Malaria parasites predominated (83% of all reported infections). The overall prevalence of haemosporidians in juveniles was approximately seven-fold higher than in adults, indicating high susceptibility of chicks and local transmission. Juvenile and adult birds hosted different lineages of Plasmodium, indicating that chicks got infection from non-parent birds. Plasmodium relictum (pSGS1) was the most prevalent malaria parasite. Mortality was not reported in adults, but 53% of infected chicks died, with reports of co-infection with Plasmodium and Leucocytozoon species. All chicks maintained in the greenhouse were non-infected and survived. Species of Leucocytozoon were undetectable by commonly used PCR protocol, but readily visible in blood films. Conclusion Crane chicks often die due to malaria and Leucocytozoon infections, which they likely gain from wild free-living birds in Beijing Zoo. Molecular diagnostics of crane Leucocytozoon parasites needs improvement. Because the reported infections are mainly chick diseases, the authors recommend maintaining of juvenile birds in vector-free facilities until the age of approximately 6 months before they are placed in open-air aviaries.

Christy A. Braham, Peter J. White, Nimalan Arinaminpathy

by Christy A. Braham, Peter J. White, Nimalan Arinaminpathy Objective To assess the quality of tuberculosis (TB) care in Pakistan, through determining comparison of healthcare practitioners’ knowledge and practices to national and international TB care guidelines. Methods Studies reporting on knowledge, attitudes and practices of public and private practitioners with TB patients were selected through searching electronic databases and grey literature. Findings Of 1458 reports, 20 full-texts were assessed, of which 11 met the eligibility and quality criteria; all studies focused on private sector care. Heterogeneity precluded meta-analysis. In 3 of 4 studies, over 50% of practitioners correctly identified a cough as the main TB symptom. However, 4 out of 6 studies showed practitioners’ compliance to be low (under 50%) for the use of sputum microscopy in diagnosis. The poorest quality care occurred in the later stages of treatment, with low compliance in prescribing practices for continuation-phase care and in monitoring and recording treatment progress, the latter of which is particularly critical for treatment success. Conclusion TB care was variable and generally inadequate, with both a lack of knowledge and a small ‘know-do’ gap evident—practitioners did not use methods that they know they should use. A lack of recent evidence found suggests that the quality of current practices may not be fully captured and further research is needed, especially on non-allopathic, rural and public-sector contexts. Improved training of practitioners, greater availability of recommended diagnostic tools and expansion of public-private partnerships are suggestions for improving the quality of TB care in Pakistan.

Cho, W.-K., Spille, J.-H., Hecht, M., Lee, C., Li, C., Grube, V., Cisse, I. I.

Models of gene control have emerged from genetic and biochemical studies, with limited consideration of the spatial organization and dynamics of key components in living cells. Here we used live cell super-resolution and light sheet imaging to study the organization and dynamics of the Mediator coactivator and RNA polymerase II (Pol II) directly. Mediator and Pol II each form small transient and large stable clusters in living embryonic stem cells. Mediator and Pol II are colocalized in the stable clusters, which associate with chromatin, have properties of phase-separated condensates, and are sensitive to transcriptional inhibitors. We suggest that large clusters of Mediator, recruited by transcription factors at large or clustered enhancer elements, interact with large Pol II clusters in transcriptional condensates in vivo.

Bruce Barrett, Mary S. Hayney, Daniel Muller, David Rakel, Roger Brown, Aleksandra E. Zgierska, Shari Barlow, Supriya Hayer, Jodi H. Barnet, Elisa R. Torres, Christopher L. Coe

by Bruce Barrett, Mary S. Hayney, Daniel Muller, David Rakel, Roger Brown, Aleksandra E. Zgierska, Shari Barlow, Supriya Hayer, Jodi H. Barnet, Elisa R. Torres, Christopher L. Coe Background Practice of meditation or exercise may enhance health to protect against acute infectious illness. Objective To assess preventive effects of meditation and exercise on acute respiratory infection (ARI) illness. Design Randomized controlled prevention trial with three parallel groups. Setting Madison, Wisconsin, USA. Participants Community-recruited adults who did not regularly exercise or meditate. Methods 1) 8-week behavioral training in mindfulness-based stress reduction (MBSR); 2) matched 8-week training in moderate intensity sustained exercise (EX); or 3) observational waitlist control. Training classes occurred in September and October, with weekly ARI surveillance through May. Incidence, duration, and area-under-curve ARI global severity were measured using daily reports on the WURSS-24 during ARI illness. Viruses were identified multiplex PCR. Absenteeism, health care utilization, and psychosocial health self-report assessments were also employed. Results Of 413 participants randomized, 390 completed the trial. In the MBSR group, 74 experienced 112 ARI episodes with 1045 days of ARI illness. Among exercisers, 84 had 120 episodes totaling 1010 illness days. Eighty-two of the controls had 134 episodes with 1210 days of ARI illness. Mean global severity was 315 for MBSR (95% confidence interval 244, 386), 256 (193, 318) for EX, and 336 (268, 403) for controls. A prespecified multivariate zero-inflated regression model suggested reduced incidence for MBSR (p = 0.036) and lower global severity for EX (p = 0.042), compared to control, not quite attaining the p

Wong S, Au T, Chan S, et al.

AbstractBackgroundHuman papillomavirus (HPV) is a known causative factor in the etiology of cervical cancer.MethodsHPV DNA genotyping was performed in menstrual blood (MB) collected in napkins from patients with cervical intraepithelial neoplasia (CIN), HPV infection and sexually active apparently normal subjects. In the same patient cohort, MB TAP1 I333V and TAP1 D637G gene polymorphisms were examined.ResultsThe sensitivity, specificity, positive and negative predictive values of HPV DNA to detect CIN or HPV infection was 83% (223/268), 98% (131/134), 99% (223/226) and (74%) 131/176, respectively. Moreover, HPV DNA was found in 24% (28/118) patients who had loop electrosurgical excision procedure treatment and 0% (0/76) HPV infected or CIN1 patient with proven recovery. On the other hand, the risk to develop high grade cervical intraepithelial neoplasia was significantly reduced for AG and GG genotypes when compared to AA genotype and for carriers with a G allele when compared to those with an A allele for both polymorphisms.ConclusionsMB HPV DNA is a potential non-invasive marker for screening and monitoring of squamous intraepithelial lesion. Together with TAP1 I333V and TAP1 D637G gene polymorphisms, the combined test may be useful to stratify high risk patients for better follow-up strategies.Clinical TrailTrial number: 0378261…

Felipe Hernandes Coutinho, Gustavo Bueno Gregoracci, Juline Marta Walter, Cristiane Carneiro Thompson, Fabiano L. Thompson

Advances brought about by omics-based approaches have revolutionized our understanding of the diversity and ecological processes involving marine archaea, bacteria, and their viruses. This broad review discusses recent examples of how genomics, metagenomics, and ecogenomics have been applied to reveal the ecology of these biological entities. Three major topics are covered in this revision: (i) the novel roles of microorganisms in ecosystem processes; (ii) virus–host associations; and (iii) ecological associations of microeukaryotes and other microbes.

Fangfang Zeng, Pingxi Xu, Kaiming Tan, Paulo H. G. Zarbin, Walter S. Leal

by Fangfang Zeng, Pingxi Xu, Kaiming Tan, Paulo H. G. Zarbin, Walter S. Leal Insect repellents are widely used to fend off nuisance mosquitoes and, more importantly, to reduce or eliminate mosquito bites in areas where viruses and other vector-borne diseases are circulating. Synthesized more than six decades ago, DEET is the most widely used insect repellent. Plant-derived compounds are used in a plethora of commercial formulations and natural recipes to repel mosquitoes. They are also used as fragrances. We analysed Bombshell® to identify the constituent(s) eliciting a previously reported “off- label” repellence activity. The two major fragrance ingredients in Bombshell, i.e., methyl dihydrojasmonate and lilial, demonstrated strong repellence against the southern house mosquito, Culex quinquefasciatus, in laboratory assays. Both compounds activated a previously identified DEET-sensitive odorant receptor, CquiOR136. These compounds were also major constituents of Ivanka Trump eau de parfum. The methyl dihydrojasmonate content was higher in the Ivanka Trump perfume than in Bombshell, the reverse being true for lilial. Both Bombshell and Ivanka Trump eaux de parfums retained activity for as long as 6 hours in laboratory assays. Although wearing these perfumes may repel nuisance mosquitoes, their use as “off-label” repellents against infected mosquitoes is not recommended.

Matthew J. Saunders , Tom Wingfield , Marco A. Tovar , Niamh Herlihy , Claudio Rocha , Karine Zevallos , Rosario Montoya , Eric Ramos , Sumona Datta , Carlton A. Evans

Abstract Objectives Mobile phone interventions have been advocated for tuberculosis care, but little is known about access of target populations to mobile phones. We studied mobile phone access among patients with tuberculosis, focusing on vulnerable patients and patients who later had adverse treatment outcomes. Methods In a prospective cohort study in Callao, Peru, we recruited and interviewed 2584 patients with tuberculosis between 2007 and 2013 and followed them until 2016 for adverse treatment outcomes using national treatment registers. Subsequently, we recruited a further 622 patients between 2016 and 2017. Data were analysed using logistic regression and by calculating relative risks (RR). Results Between 2007 and 2013, the proportion of the general population of Peru without mobile phone access averaged 7.8% but for patients with tuberculosis was 18% (P < 0.001). Patients without access were more likely to hold a lower socioeconomic position, suffer from food insecurity and be older than 50 years (all P < 0.01). Compared to patients with mobile phone access, patients without access at recruitment were more likely to subsequently have incomplete treatment (20% vs. 13%, RR = 1.5; P = 0.001) or an adverse treatment outcome (29% vs. 23% RR = 1.3; P = 0.006). Between 2016 and 2017, the proportion of patients without access dropped to 8.9% overall, but remained the same (18%) as in 2012 among the poorest third. Conclusion Access to mobile phones among patients with tuberculosis is insufficient, and rarest in patients who are poorer and later have adverse treatment outcomes. Thus, mobile phone interventions to improve tuberculosis care may be least accessed by the priority populations for whom they are intended. Such interventions should ensure access to mobile phones to enhance equity.

Poovelikunnel, T. T., Budri, P. E., Shore, A. C., Coleman, D. C., Humphreys, H., Fitzgerald-Hughes, D.

Sequential methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients following attempted mupirocin nasal decolonisation showed an increase in mupirocin resistance (MR) from 6.6% to 20%. MR isolates from patients who failed decolonization yielded indistinguishable spa types and carried multiple antimicrobial- and antiseptic-resistance genes, which may guide infection control and prevention.

Kim, S.-Y., Jhun, B. W., Moon, S. M., Shin, S. H., Jeon, K., Kwon, O. J., Yoo, I. Y., Huh, H. J., Ki, C.-S., Lee, N. Y., Shin, S. J., Daley, C. L., Suh, G. Y., Koh, W.-J.

Data are limited on the frequency of gyrA and gyrB mutations in fluoroquinolone-resistant isolates of the Mycobacterium avium complex (MAC) and M. abscessus complex (MABC). In our analysis, we did not find any resistance-associated mutations in gyrA or gyrB in 105 clinical isolates of MAC or MABC, including 72 moxifloxacin-resistant isolates. Our findings suggest that mechanisms other than gyrA and gyrB mutations contribute to moxifloxacin resistance in these organisms.

Rattiporn Kosuwin , Meng Feng , Takashi Makiuchi , Chaturong Putaporntip , Hiroshi Tachibana , Somchai Jongwutiwes

Abstract Background Thrombospondin‐related anonymous protein (TRAP) is a prime candidate for a malaria vaccine. Antibodies to Plasmodium vivax TRAP (PvTRAP) occur upon natural infection while specific antigenic domains remain to be addressed. Methods The PvTRAP sequences were determined from 73 P. vivax isolates from Tak and Ubon Ratchathani provinces collected in 2013. The recombinant proteins representing four variants each for domain II (A domain) and domain IV (thrombospondin repeat region) of PvTRAP circulating in these areas were used as antigens in enzyme‐linked immunosorbent assay against 246 serum samples from P. vivax‐infected patients in both provinces collected during 2013 and 2014. Results The prevalence of total IgG antibodies to at least one variant antigen of domain II and domain IV was 63.8% and 71.5%, respectively. Differential IgG antibody responses to these variant antigens of each domain were observed. Total IgG antibody responses to the variant antigens of each domain upon pairwise comparisons were highly correlated, suggesting immunological cross‐reactivity in the majority of serum samples. A smaller proportion of serum samples contained non‐cross‐reactive antibodies to variants of each domain; particularly domain II in which amino acid differences significantly influenced antibody recognition. Previous malaria exposure positively affected antibody responses to domain IV. Positive seroconversion and rising antibody titres occurred within a few weeks after resolution of infections. Conclusions Both domains II and IV are targets of naturally acquired IgG antibodies. Despite sequence variation in these domains, most antibody responses were cross‐reactive. A cross‐sectional evaluation of antibodies to PvTRAP during acute infection could underestimate the seroprevalence.

Vacca, P., Fazio, C., Neri, A., Ambrosio, L., Palmieri, A., Stefanelli, P.

The aim of the study was to evaluate the antimicrobial susceptibility of 866 Neisseria meningitidis invasive strains during 11-years of surveillance in Italy. Two and six strains were resistant to ciprofloxacin and rifampin, respectively. Forty-five percent were penicillin intermediate (PenI) associated with hypervirulent serogroup C clonal complex 11. All the strains were susceptible to cephalosporins.

Liz Paul

Nipah virus is a rare but dreaded zoonotic pathogen causing infections with alarmingly high mortality rates of 40-75 %. The ongoing outbreak in Kerala has witnessed 16 cases so far out of which 13 succumbed .The situation has created total panic and false perceptions among the public. As a Clinical Microbiologist belonging to the affected area, my contribution is giving health education and implementing strict infection control practices in the laboratory and in the hospital . ( As a part of Roles and responsibilities as Infection control officer of my Hospital)…

Lea Bayer, Jessica Gümpel, Gerd Hause, Martin Müller, Thomas Grunwald

by Lea Bayer, Jessica Gümpel, Gerd Hause, Martin Müller, Thomas Grunwald Papillomavirus capsids are known to have the ability to package DNA plasmids and deliver them both in vitro and in vivo. Of all known papillomavirus types, human papillomaviruses (HPVs) are by far the most intensely studied. Although HPVs work well as gene transfer vectors, their use is limited as most individuals are exposed to this virus either through a HPV vaccination or natural infection. To circumvent these constraints, we produced pseudovirions (PsVs) of ten non-human papillomavirus types and tested their transduction efficiencies in vitro. PsVs based on Macaca fascicularis papillomavirus-11 and Puma concolor papillomavirus-1 were further tested in vivo. Intramuscular transduction by PsVs led to months-long expression of a reporter plasmid, indicating that PsVs have potential as gene delivery vectors.