Scalable Production and Purification of Adeno-Associated Viral Vectors (AAV).

Here we describe methods for the production of adeno-associated viral (AAV) vectors by transient transfection of HEK293 cells grown in serum-free medium in orbital shaken bioreactors and the subsequent purification of vector particles. The protocol for expression of AAV components is based on polyethyleneimine (PEI) mediated transfection of a 2-plasmid system and is specified for production in milliliter to liter scales. After PEI and plasmid DNA (pDNA) complex formation the diluted cell culture is transfected without a prior concentration step or medium exchange. Following a 3-day batch process, cell cultures are further processed using different methods for lysis and recovery. Methods for the purification of viral particles are described, including iodixanol gradient purification, immunoaffinity chromatography, and ultrafiltration, as well as quantitative PCR to quantify vector titer

Integration of submersible transect data and high-resolution multibeam sonar imagery for a habitat-based groundfish assessment of Heceta Bank, Oregon

In the face of dramatic declines in groundfish populations and a lack of sufficient stock assessment information, a need has arisen for new methods of assessing groundfish populations. We describe the integration of seafloor transect data gathered by a manned submersible with high-resolution sonar imagery to produce a habitat-based stock assessment system for groundfish. The data sets used in this study were collected from Heceta Bank, Oregon, and were derived from 42 submersible dives (1988–90) and a multibeam sonar survey (1998). The submersible habitat survey investigated seafloor topography and groundfish abundance along 30-minute transects over six predetermined stations and found a statistical relationship between habitat variability and groundfish distribution and abundance. These transects were analyzed in a geographic information system (GIS) by using dynamic segmentation to display changes in habitat along the transects. We used the submersible data to extrapolate fish abundance within uniform habitat patches over broad areas of the bank by means of a habitat classification based on the sonar imagery. After applying a navigation correction to the submersible-based habitat segments, a good correlation with major boundaries on the backscatter and topographic boundaries on the imagery were apparent. Extrapolation of the extent of uniform habitats was made in the vicinity of the dive stations and a preliminary stock assessment of several species of demersal fish was calculated. Such a habitat-based approach will allow researchers to characterize marine communities over large areas of the seafloor

Epidemiology of Coxiella burnetii infection in Africa: a OneHealth systematic review

Background: Q fever is a common cause of febrile illness and community-acquired pneumonia in resource-limited settings. Coxiella burnetii, the causative pathogen, is transmitted among varied host species, but the epidemiology of the organism in Africa is poorly understood. We conducted a systematic review of C. burnetii epidemiology in Africa from a “One Health” perspective to synthesize the published data and identify knowledge gaps.<p></p> Methods/Principal Findings: We searched nine databases to identify articles relevant to four key aspects of C. burnetii epidemiology in human and animal populations in Africa: infection prevalence; disease incidence; transmission risk factors; and infection control efforts. We identified 929 unique articles, 100 of which remained after full-text review. Of these, 41 articles describing 51 studies qualified for data extraction. Animal seroprevalence studies revealed infection by C. burnetii (≤13%) among cattle except for studies in Western and Middle Africa (18–55%). Small ruminant seroprevalence ranged from 11–33%. Human seroprevalence was <8% with the exception of studies among children and in Egypt (10–32%). Close contact with camels and rural residence were associated with increased seropositivity among humans. C. burnetii infection has been associated with livestock abortion. In human cohort studies, Q fever accounted for 2–9% of febrile illness hospitalizations and 1–3% of infective endocarditis cases. We found no studies of disease incidence estimates or disease control efforts.<p></p> Conclusions/Significance: C. burnetii infection is detected in humans and in a wide range of animal species across Africa, but seroprevalence varies widely by species and location. Risk factors underlying this variability are poorly understood as is the role of C. burnetii in livestock abortion. Q fever consistently accounts for a notable proportion of undifferentiated human febrile illness and infective endocarditis in cohort studies, but incidence estimates are lacking. C. burnetii presents a real yet underappreciated threat to human and animal health throughout Africa.<p></p&gt

How U.S. Ocean Policy and Market Power Can Reform the Coral Reef Wildlife Trade

As the world’s largest importer of marine ornamental species for the aquaria, curio, home décor, and jewelry industries, the United States has an opportunity to leverage its considerable market power to promote more sustainable trade and reduce the effects of ornamental trade stress on coral reefs worldwide. Evidence indicates that collection of some coral reef animals for these trades has caused virtual elimination of local populations, major changes in age structure, and promotion of collection practices that destroy reef habitats. Management and enforcement of collection activities in major source countries such as Indonesia and the Philippines remain weak. Strengthening US trade laws and enforcement capabilities combined with increasing consumer and industry demand for responsible conservation can create strong incentives for improving management in source countries. This is particularly important in light of the March 2010 failure of the parties to the Convention on International Trade in Endangered Species (CITES) to take action on key groups of corals

Framing the work: A coparenting model for guiding infant mental health engagement with families

: When working with families of infants and toddlers, intentionally looking beyond dyadic child-parent relationship functioning to conceptualize the child's socioemotional adaptation within their broader family collective can enhance the likelihood that clinical gains will be supported and sustained. However, there has been little expert guidance regarding how best to frame infant-family mental health therapeutic encounters for the adults responsible for the child's care and upbringing in a manner that elevates their mindfulness about and their resolve to strengthen the impact of their coparenting collective. This article describes a new collaborative initiative organized by family-oriented infant mental health professionals across several different countries, all of whom bring expansive expertise assessing and working with coparenting and triangular family dynamics. The Collaborative's aims are to identify a means for framing initial infant mental health encounters and intakes with families with the goal of assessing and raising family consciousness about the relevance of coparenting. Initial points of convergence and growing points identified by the Collaborative for subsequent field study are addressed

Analysis of Temporal Expressions Annotated in Clinical Notes

Annotating the semantics of time in language is important. THYME is a recent temporal annotation standard for clinical texts. This paper examines temporal expressions in the first major corpus released under this standard. It investigates where the standard has proven difficult to apply, and gives a series of recommendations regarding temporal annotation in this important domain

Development of an HPV Educational Protocol for Adolescents

To develop an educational protocol about HPV and Pap tests for adolescents, to evaluate the protocol for understandability and clarity, and to evaluate the protocol for its effectiveness in increasing knowledge about HPV