Cas9-mediated gene-editing in the malaria mosquito Anopheles stephensi by ReMOT Control

Innovative tools are essential for advancing malaria control and depend on an understanding of molecular mechanisms governing transmission of malaria parasites by Anopheles mosquitoes. CRISPR/Cas9-based gene disruption is a powerful method to uncover underlying biology of vector-pathogen interactions and can itself form the basis of mosquito control strategies. However, embryo injection methods used to genetically manipulate mosquitoes (especially Anopheles) are difficult and inefficient, particularly for non-specialist laboratories. Here, we adapted the ReMOT Control (Receptor-mediated Ovary Transduction of Cargo) technique to deliver Cas9 ribonucleoprotein complex to adult mosquito ovaries, generating targeted and heritable mutations in the malaria vector Anopheles stephensi without injecting embryos. In Anopheles, ReMOT Control gene editing was as efficient as standard embryo injections. The application of ReMOT Control to Anopheles opens the power of CRISPR/Cas9 methods to malaria laboratories that lack the equipment or expertise to perform embryo injections and establishes the flexibility of ReMOT Control for diverse mosquito species

A perspective on the impacts of microplastics on mosquito biology and their vectorial capacity

Microplastics (plastic particles <5 mm) permeate aquatic and terrestrial ecosystems and constitute a hazard to animal life. Although much research has been conducted on the effects of microplastics on marine and benthic organisms, less consideration has been given to insects, especially those adapted to urban environments. Here, we provide a perspective on the potential consequences of exposure to microplastics within typical larval habitat on mosquito biology. Mosquitoes represent an ideal organism in which to explore the biological effects of microplastics on terrestrial insects, not least because of their importance as an infectious disease vector. Drawing on evidence from other organisms and knowledge of the mosquito life cycle, we summarise some of the more plausible impacts of microplastics including physiological, ecotoxicological and immunological responses. We conclude that although there remains little experimental evidence demonstrating any adverse effect on mosquito biology or pathogen transmission, significant knowledge gaps remain, and there is now a need to quantify the effects that microplastic pollution could have on such an important disease vector

Limited influence of the microbiome on the transcriptional profile of female Aedes aegypti mosquitoes

The microbiome is an assemblage of microorganisms living in association with a multicellular host. Numerous studies have identified a role for the microbiome in host physiology, development, immunity, and behaviour. The generation of axenic (germ-free) and gnotobiotic model systems has been vital to dissecting the role of the microbiome in host biology. We have previously reported the generation of axenic Aedes aegypti mosquitoes, the primary vector of several human pathogenic viruses, including dengue virus and Zika virus. In order to better understand the influence of the microbiome on mosquitoes, we examined the transcriptomes of axenic and conventionally reared Ae. aegypti before and after a blood meal. Our results suggest that the microbiome has a much lower effect on the mosquito's gene expression than previously thought with only 170 genes influenced by the axenic state, while in contrast, blood meal status influenced 809 genes. The pattern of expression influenced by the microbiome is consistent with transient changes similar to infection rather than sweeping physiological changes. While the microbiome does seem to affect some pathways such as immune function and metabolism, our data suggest the microbiome is primarily serving a nutritional role in development with only minor effects in the adult

The microbiome and mosquito vectorial capacity: rich potential for discovery and translation

Microbiome research has gained considerable interest due to the emerging evidence of its impact on human and animal health. As in other animals, the gut-associated microbiota of mosquitoes affect host fitness and other phenotypes. It is now well established that microbes can alter pathogen transmission in mosquitoes, either positively or negatively, and avenues are being explored to exploit microbes for vector control. However, less attention has been paid to how microbiota affect phenotypes that impact vectorial capacity. Several mosquito and pathogen components, such as vector density, biting rate, survival, vector competence, and the pathogen extrinsic incubation period all influence pathogen transmission. Recent studies also indicate that mosquito gut-associated microbes can impact each of these components, and therefore ultimately modulate vectorial capacity. Promisingly, this expands the options available to exploit microbes for vector control by also targeting parameters that affect vectorial capacity. However, there are still many knowledge gaps regarding mosquito–microbe interactions that need to be addressed in order to exploit them efficiently. Here, we review current evidence of impacts of the microbiome on aspects of vectorial capacity, and we highlight likely opportunities for novel vector control strategies and areas where further studies are required

Wolbachia endosymbionts in two Anopheles species indicates independent acquisitions and lack of prophage elements.

Wolbachia is a genus of obligate bacterial endosymbionts that infect a diverse range of arthropod species as well as filarial nematodes, with its single described species, Wolbachia pipientis , divided into several ‘supergroups’ based on multilocus sequence typing. Wolbachia strains in mosquitoes have been shown to inhibit the transmission of human pathogens, including Plasmodium malaria parasites and arboviruses. Despite their large host range, Wolbachia strains within the major malaria vectors of the Anopheles gambiae and Anopheles funestus complexes appear at low density, established solely on PCR-based methods. Questions have been raised as to whether this represents a true endosymbiotic relationship. However, recent definitive evidence for two distinct, high-density strains of supergroup B Wolbachia within Anopheles demeilloni and Anopheles moucheti has opened exciting possibilities to explore naturally occurring Wolbachia endosymbionts in Anopheles for biocontrol strategies to block Plasmodium transmission. Here, we utilize genomic analyses to demonstrate that both Wolbachia strains have retained all key metabolic and transport pathways despite their smaller genome size, with this reduction potentially attributable to degenerated prophage regions. Even with this reduction, we confirmed the presence of cytoplasmic incompatibility (CI) factor genes within both strains, with wAnD maintaining intact copies of these genes while the cifB gene was interrupted in wAnM, so functional analysis is required to determine whether wAnM can induce CI. Additionally, phylogenetic analysis indicates that these Wolbachia strains may have been introduced into these two Anopheles species via horizontal transmission events, rather than by ancestral acquisition and subsequent loss events in the Anopheles gambiae species complex. These are the first Wolbachia genomes, to our knowledge, that enable us to study the relationship between natural strain Plasmodium malaria parasites and their anopheline hosts.</jats:p

RZ Piscium Hosts a Compact and Highly Perturbed Debris Disk

RZ Piscium (RZ Psc) is well-known in the variable star field because of its numerous, irregular optical dips in the past five decades, but the nature of the system is heavily debated in the literature. We present multiyear infrared monitoring data from Spitzer and WISE to track the activities of the inner debris production, revealing stochastic infrared variability as short as weekly timescales that is consistent with destroying a 90-km-size asteroid every year. ALMA 1.3 mm data combined with spectral energy distribution modeling show that the disk is compact ($\sim$0.1--13 au radially) and lacks cold gas. The disk is found to be highly inclined and has a significant vertical scale height. These observations confirm that RZ Psc hosts a close to edge-on, highly perturbed debris disk possibly due to migration of recently formed giant planets which might be triggered by the low-mass companion RZ Psc B if the planets formed well beyond the snowlines.Comment: 16 pages, 5 figures, accepted for publication in Ap

Origins of the extragalactic background at 1mm from a combined analysis of the AzTEC and MAMBO data in GOODS-N

We present a study of the cosmic infrared background, which is a measure of the dust obscured activity in all galaxies in the Universe. We venture to isolate the galaxies responsible for the background at 1mm; with spectroscopic and photometric redshifts we constrain the redshift distribution of these galaxies. We create a deep 1.16mm map (sigma ~ 0.5mJy) by combining the AzTEC 1.1mm and MAMBO 1.2mm datasets in GOODS-N. This combined map contains 41 secure detections, 13 of which are new. By averaging the 1.16mm flux densities of individually undetected galaxies with 24um flux densities > 25uJy, we resolve 31--45 per cent of the 1.16mm background. Repeating our analysis on the SCUBA 850um map, we resolve a higher percentage (40--64 per cent) of the 850um background. A majority of the background resolved (attributed to individual galaxies) at both wavelengths comes from galaxies at z > 1.3. If the ratio of the resolved submillimeter to millimeter background is applied to a reasonable scenario for the origins of the unresolved submillimeter background, 60--88 per cent of the total 1.16mm background comes from galaxies at z > 1.3.Comment: 12 pages, 10 figures. Accepted by MNRAS. The combined map is publicly available at http://www.astro.umass.edu/~pope/goodsn_mm

Clubbing masculinities: Gender shifts in gay men's dance floor choreographies

This is an Author's Accepted Manuscript of an article published in Journal of Homosexuality, 58(5), 608-625, 2011 [copyright Taylor & Francis], available online at: http://www.tandfonline.com/10.1080/00918369.2011.563660This article adopts an interdisciplinary approach to understanding the intersections of gender, sexuality, and dance. It examines the expressions of sexuality among gay males through culturally popular forms of club dancing. Drawing on political and musical history, I outline an account of how gay men's gendered choreographies changed throughout the 1970s, 80s, and 90s. Through a notion of “technologies of the body,” I situate these developments in relation to cultural levels of homophobia, exploring how masculine expressions are entangled with and regulated by musical structures. My driving hypothesis is that as perceptions of cultural homophobia decrease, popular choreographies of gay men's dance have become more feminine in expression. Exploring this idea in the context of the first decade of the new millennium, I present a case study of TigerHeat, one of the largest weekly gay dance club events in the United States

The association of plasma biomarkers with computed tomography-assessed emphysema phenotypes

Rationale: Chronic obstructive pulmonary disease (COPD) is a phenotypically heterogeneous disease. In COPD, the presence of emphysema is associated with increased mortality and risk of lung cancer. High resolution computed tomography (HRCT) scans are useful in quantifying emphysema but are associated with radiation exposure and high incidence of false positive findings (i.e., nodules). Using a comprehensive biomarker panel, we sought to determine if there was a peripheral blood biomarker signature of emphysema. Methods: 114 plasma biomarkers were measured using a custom assay in 588 individuals enrolled in the COPDGene study. Quantitative emphysema measurements included percent low lung attenuation (%LAA) ≤ −950 HU, ≤ − 910 HU and mean lung attenuation at the 15th percentile on lung attenuation curve (LP15A). Multiple regression analysis was performed to determine plasma biomarkers associated with emphysema independent of covariates age, gender, smoking status, body mass index and FEV1. The findings were subsequently validated using baseline blood samples from a separate cohort of 388 subjects enrolled in the Treatment of Emphysema with a Selective Retinoid Agonist (TESRA) study. Results: Regression analysis identified multiple biomarkers associated with CT-assessed emphysema in COPDGene, including advanced glycosylation end-products receptor (AGER or RAGE, p < 0.001), intercellular adhesion molecule 1 (ICAM, p < 0.001), and chemokine ligand 20 (CCL20, p < 0.001). Validation in the TESRA cohort revealed significant associations with RAGE, ICAM1, and CCL20 with radiologic emphysema (p < 0.001 after meta-analysis). Other biomarkers that were associated with emphysema include CDH1, CDH 13 and SERPINA7, but were not available for validation in the TESRA study. Receiver operating characteristics analysis demonstrated a benefit of adding a biomarker panel to clinical covariates for detecting emphysema, especially in those without severe airflow limitation (AUC 0.85). Conclusions: Our findings, suggest that a panel of blood biomarkers including sRAGE, ICAM1 and CCL20 may serve as a useful surrogate measure of emphysema, and when combined with clinical covariates, may be useful clinically in predicting the presence of emphysema compared to just using covariates alone, especially in those with less severe COPD. Ultimately biomarkers may shed light on disease pathogenesis, providing targets for new treatments. Electronic supplementary material The online version of this article (doi:10.1186/s12931-014-0127-9) contains supplementary material, which is available to authorized users