Protective CD8+ T-cell responses to cytomegalovirus driven by rAAV/GFP/IE1 loading of dendritic cells

Background: Recent studies demonstrate that recombinant adeno-associated virus (rAAV)-based antigen loading of dendritic cells (DCs) generates in vitro, significant and rapid cytotoxic T-lymphocyte (CTL) responses against viral antigens. Methods: We used the rAAV system to induce specific CTLs against CVM antigens for the development of cytomegalovirus HCMV) gene therapy. As an extension of the versatility of the rAAV system, we incorporated immediate-early 1 (IE1), expressed in HCMV. Our rAAV vector induced a strong stimulation of CTLs directed against the HCMV antigen IE1. We then investigated the efficiency of the CTLs in killing IE1 targeted cells. Results: A significant MHC Class I-restricted, anti-IE1-specificCTL killing was demonstrated against IE1 positive peripheral blood mononuclear cells (PBMC) after one, in vitro, stimulation. Conclusion: In summary, single PBMC stimulation with rAAV/IE1 pulsed DCs induces strong antigen specific-CTL generation. CTLs were capable to lyse low doses of peptides pulsed into target cells. These data suggest that AAV-based antigen loading of DCs is highly effective for generating human CTL responses against HCMV antigens

Neonatal Ventilator Associated Pneumonia: A Quality Improvement Initiative Focusing on Antimicrobial Stewardship

Background and Aims: Neonatal ventilator associated pneumonia (VAP) is a common nosocomial infection and a frequent reason for empirical antibiotic therapy in NICUs. Nonetheless, there is no international consensus regarding diagnostic criteria and management. In a first step, we analyzed the used diagnostic criteria, risk factors and therapeutic management of neonatal VAP by a literature review. In a second step, we aimed to compare suspected vs. confirmed neonatal VAP episodes in our unit according to different published criteria and to analyze interrater-reliability of chest x-rays. Additionally, we aimed to evaluate the development of VAP incidence and antibiotic use after implementation of multifaceted quality improvement changes regarding antimicrobial stewardship and infection control (VAP-prevention-bundle, early-extubation policy, antimicrobial stewardship rounds).Methods: Neonates until 44 weeks of gestation with suspected VAP, hospitalized at our level-III NICU in Lucerne from September 2014 to December 2017 were enrolled. VAP episodes were analyzed according to 4 diagnostic frameworks. Agreement regarding chest x-ray interpretation done by 10 senior physicians was assessed. Annual incidence of suspected and confirmed neonatal VAP episodes and antibiotic days were calculated and compared for the years 2015, 2016, and 2017.Results: 17 studies were identified in our literature review. Overall, CDC-guidelines or similar criteria, requesting radiographic changes as main criteria, are mostly used. Comparison of suspected vs. confirmed neonatal VAP episodes showed a great variance (20.4 vs. 4.5/1,000 ventilator-days). The interrater-reliability of x-ray interpretation was poor (intra-class correlation 0.25). Implemented changes resulted in a gradual decline in annual VAP incidence and antibiotic days from 2015 compared with 2017 (28.8 vs. 7.4 suspected episodes/1,000 ventilator-days, 5.5 vs. 0 confirmed episodes/1,000 ventilator-days and 211 vs. 34.7 antibiotic days/1,000 ventilation-days, respectively).Conclusion: The incidence of suspected VAP and concomitant antibiotic use is much higher than for confirmed VAP, therefore inclusion of suspected episodes should be considered for accurate evaluation. There is a high diagnostic inconsistency and a low reliability of interpretation of chest x-rays regarding VAP. Implementation of combined antimicrobial stewardship and infection control measures may lead to an effective decrease in VAP incidence and antibiotic use

Admixture and reproductive skew shape the conservation value of ex situ populations of the Critically Endangered eastern black rhino

Small populations of endangered species risk losing already eroded genetic diversity, important for adaptive potential, through the effects of genetic drift. The magnitude of drift can be mitigated by maximising the effective population size, as is the goal of genetic management strategies. Different mating systems, specifically those leading to reproductive skew, exacerbate genetic drift by distorting contributions. In the absence of an active management strategy, reproductive skew will have long-term effects on the genetic composition of a population, particularly where admixture is present. Here we examine the contrasting effects of conservation management strategies in two ex situ populations of the Critically Endangered eastern black rhino (Diceros bicornis michaeli), one managed as a semi-wild population in South Africa (SAx), and one managed under a mean-kinship breeding strategy in European zoos. We use molecular data to reconstruct pedigrees for both populations and validate the method using the zoo studbook. Using the reconstructed pedigree and studbook we show there is male sex-specific skew in both populations. However, the zoo’s mean-kinship breeding strategy effectively reduces reproductive skew in comparison to a semi-wild population with little genetic management. We also show that strong male reproductive skew in SAx has resulted in extensive admixture, which may require a re-evaluation of the population’s original intended role in the black rhino meta-population. With a high potential for admixture in many ex situ populations of endangered species, molecular and pedigree data remain vital tools for populations needing to balance drift and selection

The Monarch Initiative in 2019: an integrative data and analytic platform connecting phenotypes to genotypes across species.

In biology and biomedicine, relating phenotypic outcomes with genetic variation and environmental factors remains a challenge: patient phenotypes may not match known diseases, candidate variants may be in genes that haven\u27t been characterized, research organisms may not recapitulate human or veterinary diseases, environmental factors affecting disease outcomes are unknown or undocumented, and many resources must be queried to find potentially significant phenotypic associations. The Monarch Initiative (https://monarchinitiative.org) integrates information on genes, variants, genotypes, phenotypes and diseases in a variety of species, and allows powerful ontology-based search. We develop many widely adopted ontologies that together enable sophisticated computational analysis, mechanistic discovery and diagnostics of Mendelian diseases. Our algorithms and tools are widely used to identify animal models of human disease through phenotypic similarity, for differential diagnostics and to facilitate translational research. Launched in 2015, Monarch has grown with regards to data (new organisms, more sources, better modeling); new API and standards; ontologies (new Mondo unified disease ontology, improvements to ontologies such as HPO and uPheno); user interface (a redesigned website); and community development. Monarch data, algorithms and tools are being used and extended by resources such as GA4GH and NCATS Translator, among others, to aid mechanistic discovery and diagnostics

Prescribing indicators at primary health care centers within the WHO African region: a systematic analysis (1995-2015)

Abstract Background Rational medicine use is essential to optimize quality of healthcare delivery and resource utilization. We aim to conduct a systematic review of changes in prescribing patterns in the WHO African region and comparison with WHO indicators in two time periods 1995–2005 and 2006–2015. Methods Systematic searches were conducted in PubMed, Scopus, Web of science, Africa-Wide Nipad, Africa Journals Online (AJOL), Google scholar and International Network for Rational Use of Drugs (INRUD) Bibliography databases to identify primary studies reporting prescribing indicators at primary healthcare centres (PHCs) in Africa. This was supplemented by a manual search of retrieved references. We assessed the quality of studies using a 14-point scoring system modified from the Downs and Black checklist with inclusions of recommendations in the WHO guidelines. Results Forty-three studies conducted in 11 African countries were included in the overall analysis. These studies presented prescribing indicators based on a total 141,323 patient encounters across 572 primary care facilities. The results of prescribing indicators were determined as follows; average number of medicines prescribed per patient encounter = 3.1 (IQR 2.3–4.8), percentage of medicines prescribed by generic name =68.0 % (IQR 55.4–80.3), Percentage of encounters with antibiotic prescribed =46.8 % (IQR 33.7–62.8), percentage of encounters with injection prescribed =25.0 % (IQR 18.7–39.5) and the percentage of medicines prescribed from essential medicines list =88.0 % (IQR 76.3–94.1). Prescribing indicators were generally worse in private compared with public facilities. Analysis of prescribing across two time points 1995–2005 and 2006–2015 showed no consistent trends. Conclusions Prescribing indicators for the African region deviate significantly from the WHO reference targets. Increased collaborative efforts are urgently needed to improve medicine prescribing practices in Africa with the aim of enhancing the optimal utilization of scarce resources and averting negative health consequences

REQUITE: A prospective multicentre cohort study of patients undergoing radiotherapy for breast, lung or prostate cancer

Purpose: REQUITE aimed to establish a resource for multi-national validation of models and biomarkers that predict risk of late toxicity following radiotherapy. The purpose of this article is to provide summary descriptive data. Methods: An international, prospective cohort study recruited cancer patients in 26 hospitals in eight countries between April 2014 and March 2017. Target recruitment was 5300 patients. Eligible patients had breast, prostate or lung cancer and planned potentially curable radiotherapy. Radiotherapy was prescribed according to local regimens, but centres used standardised data collection forms. Pre-treatment blood samples were collected. Patients were followed for a minimum of 12 (lung) or 24 (breast/prostate) months and summary descriptive statistics were generated. Results: The study recruited 2069 breast (99% of target), 1808 prostate (86%) and 561 lung (51%) cancer patients. The centralised, accessible database includes: physician-(47,025 forms) and patient-(54,901) reported outcomes; 11,563 breast photos; 17,107 DICOMs and 12,684 DVHs. Imputed genotype data are available for 4223 patients with European ancestry (1948 breast, 1728 prostate, 547 lung). Radiation-induced lymphocyte apoptosis (RILA) assay data are available for 1319 patients. DNA (n = 4409) and PAXgene tubes (n = 3039) are stored in the centralised biobank. Example prevalences of 2-year (1-year for lung) grade >= 2 CTCAE toxicities are 13% atrophy (breast), 3% rectal bleeding (prostate) and 27% dyspnoea (lung). Conclusion: The comprehensive centralised database and linked biobank is a valuable resource for the radiotherapy community for validating predictive models and biomarkers. Patient summary: Up to half of cancer patients undergo radiation therapy and irradiation of surrounding healthy tissue is unavoidable. Damage to healthy tissue can affect short-and long-term quality-of-life. Not all patients are equally sensitive to radiation "damage" but it is not possible at the moment to identify those who are. REQUITE was established with the aim of trying to understand more about how we could predict radiation sensitivity. The purpose of this paper is to provide an overview and summary of the data and material available. In the REQUITE study 4400 breast, prostate and lung cancer patients filled out questionnaires and donated blood. A large amount of data was collected in the same way. With all these data and samples a database and biobank were created that showed it is possible to collect this kind of information in a standardised way across countries. In the future, our database and linked biobank will be a resource for research and validation of clinical predictors and models of radiation sensitivity. REQUITE will also enable a better understanding of how many people suffer with radiotherapy toxicity

Stick-tight fleas in the nostrils and below the tongue: evolution of an extraordinary infestation site in Hectopsylla (Siphonaptera: Pulicidae)

BLANK STEPHANM, KUTZSCHER CHRISTIAN, Masello J, PILGRIM ROBERTLC, QUILLFELDT PETRA. Stick-tight fleas in the nostrils and below the tongue: evolution of an extraordinary infestation site in Hectopsylla (Siphonaptera: Pulicidae). Zoological Journal of the Linnean Society. 2007;149(1):117-137.Association with terrestrial mammals as hosts and a sessile mode of life in females are ground plan traits of the stick-tight fleas associated with Hectopsylla+Tunga. Hectopsylla comprises the lineages H. pulex+[H. psittaci-group +H. broscus-group]. The stem species of the H. psittaci-group has switched to birds. Hectopsylla narium sp. nov. infests the nestlings of the burrowing parrot (Cyanoliseus patagonus patagonus; Psittacidae). The infestation sites inside the nasal cavity, and later during the breeding season also below the tongue, are unique among fleas. These sites provide a concealed habitat for the sessile, immobile females, where they are safe against cleaning activities of their host and the host's parents. The fleas are able to disperse actively within the bird colony. Their dispersal over long distances can only be assumed to happen accidentally, as fleas have never been found on adult parrots during field studies in Río Negro, Patagonia, Argentina. Data on the species bionomics, morphological descriptions and illustrations, and an identification key for H. narium and related taxa are presented. A lectotype is designated for H. psittaci. The family group name Tunginae is proposed as a new synonym of Hectopsyllinae

Hectopsylla narium Blank & Kutzscher & Masello & Pilgrim & Quillfeldt 2007

BIONOMICS OF <i> <i>HECTOPSYLLA</i> NARIUM</i> <p> The fleas have been found on 204 of 380 studied nestlings of the burrowing parrot. An additional 100 adult parrots checked were completely free from fleas. Various larval stages and several teneral moult adults of both sexes have been collected from the sandy bottom of nine nest chambers. Within the studied colony flea larvae have also been found in tunnels of the parrot occupied by a barn owl [<i>Tyto alba tuidara</i> (J.E. Gray, 1829), Tytonidae; one nest with five larvae studied] and southern martins [<i>Progne elegans</i> (Baird, 1865), Hirundinidae; two nests studied, one with one larva]. Nestlings of the latter two species remained free from adult fleas.</p> <p> Legend: 0, 1, 2 – character states; p – 0/1 polymorphism; q – 0/2 polymorphism; r – 1/2/3 polymorphism; s – 0/1/2 polymorphism; x – state not scored;? – state unknown or uncertain. See text for definitions of character states. In row 17 the character states for the <i>H. broscus</i> -group (rows 8–16) are summarized using polymorphic character states where necessary.</p> <p>Freshly emerged adults of both sexes vivaciously crawl and can jump up to 25 cm in horizontal distance. Females become sessile by anchoring with their heavily denticulate lacinia in the host skin, remaining fully exposed. The uptake of blood distends their abdomen strongly by stretching the intersegmental membrane between the sclerotized terga and sterna (Fig. 2).</p> <p> Fig. 3. <i>H</i>. knighti (♀, holotype). Fig. 4. <i>H</i>. psittaci (♀). Fig. 5. <i>H</i>. pulex (♂). Fig. 6. <i>H</i>. pulex (♀).</p> <p> EXTRAORDINARY INFESTATION SITE IN <i>HECTOPSYLLA</i> 121</p> <p>The female/male sex ratio for the total number of specimens was 5.9: 1. Females ranged from four to 13 times as abundant as males on individual nestlings. Males were observed mating with females inside the nasal cavities, but they were never found feeding on the nestlings.</p> <p>Feeding females have usually been found in the nasal cavity (Fig. 27) and on the comparatively dry area under the tongue. Up to 11 live fleas or up to ten live and seven dead fleas were collected from the nostrils of individual nestlings (16 December 1999 to 3 January 2000). The respiration of such heavily infested nestlings was clearly impeded. An infestation under the tongue seems to occur mainly towards the end of the breeding season, when the nostrils are already largely occupied. On 26 December 2004 we found a nestling with 42 live fleas, 32 females and two males below the tongue and seven females and one male in the nostrils. We also found three nestlings with an infestation of the foot with one, one, 18 fleas, respectively. In the third case the fleas formed a crowded assemblage at the base of two toes.</p> <p> Nestlings become infested at 7–14 days, when the diameter of the nostrils is large enough for fleas to pass (Fig. 27). Females remain on the nestlings until they die <i>in situ</i>. Dead fleas are discernible by their darker colour and shrivelled body.</p> <p>Continuous observations using a miniature security camera inside nest tunnels revealed that adult parrots groom their nestlings approximately every 4 h but not the nestlings’ nostrils or oral cavities. Adult parrots have been observed to groom their own nostrils with their claws.</p>Published as part of <i>Blank, Stephan M., Kutzscher, Christian, Masello, Juan F., Pilgrim, Robert L. C. & Quillfeldt, Petra, 2007, Stick-tight fleas in the nostrils and below the tongue: evolution of an extraordinary infestation site in Hectopsylla (Siphonaptera: Pulicidae), pp. 117-137 in Zoological Journal of the Linnean Society 149 (1)</i> on pages 119-121, DOI: 10.1111/j.1096-3642.2006.00239.x, <a href="http://zenodo.org/record/5427621">http://zenodo.org/record/5427621</a&gt