Surveillance of molluscan (gastropod) intermediate hosts for the emerging infectious disease Angiostrongyliasis (Angiostrongylus cantonensis) in Oklahoma.

Angiostrongylus cantonensis, the rat lungworm, is the etiologic agent of an emerging infectious zoonosis, angiostrongyliasis. To date there is relatively little known about this parasite from a public health perspective. Because snails play a critical role in the rat lungworm's life cycle, as intermediate hosts, it is paramount that snail species able to harbor this disease are identified to increase awareness of the geographic spread of this disease locally and globally. Our research aimed to identify the parasite within endemic Oklahoman molluscan intermediate hosts and to provide current geographic distribution within these intermediate hosts. The most abundant aquatic snails and bivalves were collected by hand in southeast Oklahoma between the months of September 2016 and September 2017 (N=351). The sampling resulted in the collection of three species including Physa spp., Planorbella trivolvis and Corbicula fluminea, the Asian clam. Total digestion of tissue was performed before extraction of DNA for each sample. Quantitative polymerase chain reaction (qPCR) was utilized for amplification of the internal transcribed spacer 1 (ITS1) region of A. cantonensis. Melt-curve analysis was implemented with qPCR for secondary confirmation. Further confirmation was performed by resolving all amplified products on an agarose gel via electrophoresis and identifying the presence or absence of a band at the target DNA size belonging to A. cantonensis (267bp fragment). Further optimization was conducted to ensure highest quality of results including temperature gradients, primer concentration gradients, different primer sets (including self-created sets), varying cycle conditions, different modes of PCR such as touchdown-PCR, utilizing intercalating dyes as well as probe-based qPCR, and inclusion of more refined assay components such as a black hole quencher (BHQ). After performing the optimized assay on all 351 collected gastropod samples none yielded positive confirmation of A. cantonensis DNA. These results indicate that Oklahoma creates bottlenecks in many forms such as geographic, climatic and biologic that inhibit the ability of this parasite to gain a foothold beyond southeast Oklahoma. The absence of rat lungworm DNA within the most abundant aquatic intermediate hosts in this region confirm the unsuitability of these species as intermediate hosts for this parasite

The Use of Edge-Betweenness Clustering to Investigate Biological Function in Protein Interaction Networks

RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are.Abstract Background This paper describes an automated method for finding clusters of interconnected proteins in protein interaction networks and retrieving protein annotations associated with these clusters. Results Protein interaction graphs were separated into subgraphs of interconnected proteins, using the JUNG implementation of Girvan and Newman's Edge-Betweenness algorithm. Functions were sought for these subgraphs by detecting significant correlations with the distribution of Gene Ontology terms which had been used to annotate the proteins within each cluster. The method was implemented using freely available software (JUNG and the R statistical package). Protein clusters with significant correlations to functional annotations could be identified and included groups of proteins know to cooperate in cell metabolism. The method appears to be resilient against the presence of false positive interactions. Conclusion This method provides a useful tool for rapid screening of small to medium size protein interaction datasets.Published versio

Label-free enrichment of adrenal cortical progenitor cells using inertial microfluidics.

Passive and label-free isolation of viable target cells based on intrinsic biophysical cellular properties would allow for cost savings in applications where molecular biomarkers are known as well as potentially enable the separation of cells with little-to-no known molecular biomarkers. We have demonstrated the purification of adrenal cortical progenitor cells from digestions of murine adrenal glands utilizing hydrodynamic inertial lift forces that single cells and multicellular clusters differentially experience as they flow through a microchannel. Fluorescence staining, along with gene expression measurements, confirmed that populations of cells collected in different outlets were distinct from one another. Furthermore, primary murine cells processed through the device remained highly viable and could be cultured for 10 days in vitro. The proposed target cell isolation technique can provide a practical means to collect significant quantities of viable intact cells required to translate stem cell biology to regenerative medicine in a simple label-free manner

An Existential-Humanistic View of Personality Change: Co-Occurring Changes with Psychological Well-Being in a 10 Year Cohort Study

Increasingly, psychological research has indicated that an individual’s personality changes across the lifespan. We aim to better understand personality change by examining if personality change is linked to striving towards fulfilment, as suggested by existential–humanistic theories of personality dynamics. Using the Wisconsin Longitudinal Study, a cohort of 4,733 mid-life individuals across 10years, we show that personality change was significantly associated with change in existential well-being, represented by psychological well-being (PWB). Moreover, personality change was more strongly related to change in PWB than changes in other well-being indicators such as depression, hostility and life satisfaction. Personality changed to a similar degree and explained greater variation in our well-being measures than changes in socioeconomic variables. The findings indicate personality change is necessary for the holistic development of an individual, supporting a greater need to understand personality change and increasing room for use of personality measures as indicators of well-being and policy making

Effects of two contrasting canopy manipulations on growth and water use of London plane (Platanus x acerifolia) trees

Aims: Two contrasting canopy manipulations were compared to unpruned controls on London plane trees, to determine the effects on canopy regrowth, soil and leaf water relations. Methods: ‘Canopy reduction’, was achieved by removing the outer 30 % length of all major branches and ‘canopy thinning’, by removing 30 % of lateral branches arising from major branches. Results: Total canopy leaf areas recovered within two and three years of pruning for the canopy-thinned and reduced trees respectively. Canopy reduction increased mean leaf size, nitrogen concentration, canopy leaf area density and conserved soil moisture for up to 3 years, whereas canopy thinning had no effects. Another experiment compared more severe canopy reduction to unpruned trees. This produced a similar growth response to the previous experiment, but soil moisture was conserved nearer to the trunk. Analysis of 13C and 18O signals along with leaf water relations and soil moisture data suggested that lower boundary layer conductance within the canopy-reduced trees restricted tree water use, whereas for the canopy-thinned trees the opposite occurred. Conclusions: Only canopy reduction conserved soil moisture and this was due to a combination of reduced total canopy leaf area and structural changes in canopy architecture

A Versatile, Portable Intravital Microscopy Platform for Studying Beta-cell Biology In Vivo

The pancreatic islet is a complex micro-organ containing numerous cell types, including endocrine, immune, and endothelial cells. The communication of these systems is lost upon isolation of the islets, and therefore the pathogenesis of diabetes can only be fully understood by studying this organized, multicellular environment in vivo. We have developed several adaptable tools to create a versatile platform to interrogate β-cell function in vivo. Specifically, we developed β-cell-selective virally-encoded fluorescent protein biosensors that can be rapidly and easily introduced into any mouse. We then coupled the use of these biosensors with intravital microscopy, a powerful tool that can be used to collect cellular and subcellular data from living tissues. Together, these approaches allowed the observation of in vivo β-cell-specific ROS dynamics using the Grx1-roGFP2 biosensor and calcium signaling using the GcAMP6s biosensor. Next, we utilized abdominal imaging windows (AIW) to extend our in vivo observations beyond single-point terminal measurements to collect longitudinal physiological and biosensor data through repeated imaging of the same mice over time. This platform represents a significant advancement in our ability to study β-cell structure and signaling in vivo, and its portability for use in virtually any mouse model will enable meaningful studies of β-cell physiology in the endogenous islet niche

mTORC1 activity is essential for erythropoiesis and B cell lineage commitment

Mechanistic target of rapamycin (mTOR) is a serine/threonine protein kinase that mediates phosphoinositide-3-kinase (PI3K)/AKT signalling. This pathway is involved in a plethora of cellular functions including protein and lipid synthesis, cell migration, cell proliferation and apoptosis. In this study, we proposed to delineate the role of mTORC1 in haemopoietic lineage commitment using knock out (KO) mouse and cell line models. Mx1-cre and Vav-cre expression systems were used to specifically target Raptorfl/fl (mTORC1), either in all tissues upon poly(I:C) inoculation, or specifically in haemopoietic stem cells, respectively. Assessment of the role of mTORC1 during the early stages of development in Vav-cre+Raptorfl/fl mice, revealed that these mice do not survive post birth due to aberrations in erythropoiesis resulting from an arrest in development at the megakaryocyte-erythrocyte progenitor stage. Furthermore, Raptor-deficient mice exhibited a block in B cell lineage commitment. The essential role of Raptor (mTORC1) in erythrocyte and B lineage commitment was confirmed in adult Mx1-cre+Raptorfl/fl mice upon cre-recombinase induction. These studies were supported by results showing that the expression of key lineage commitment regulators, GATA1, GATA2 and PAX5 were dysregulated in the absence of mTORC1-mediated signals. The regulatory role of mTOR during erythropoiesis was confirmed in vitro by demonstrating a reduction of K562 cell differentiation towards RBCs in the presence of established mTOR inhibitors. While mTORC1 plays a fundamental role in promoting RBC development, we showed that mTORC2 has an opposing role, as Rictor-deficient progenitor cells exhibited an elevation in RBC colony formation ex vivo. Collectively, our data demonstrate a critical role played by mTORC1 in regulating the haemopoietic cell lineage commitment

The effect of gaze angle on visual acuity in infantile nystagmus

Purpose: Most individuals with infantile nystagmus (IN) have an idiosyncratic gaze angle at which their nystagmus intensity is minimized. Some adopt an abnormal head posture to use this “null zone,” and it has therefore long been assumed that this provides people with nystagmus with improved visual acuity (VA). However, recent studies suggest that improving the nystagmus waveform could have little, if any, influence on VA; that is, VA is fundamentally limited in IN. Here, we examined the impact of the null zone on VA. Methods: Visual acuity was measured in eight adults with IN using a psychophysical staircase procedure with reversals at three horizontal gaze angles, including the null zone. Results: As expected, changes in gaze angle affected nystagmus amplitude, frequency, foveation duration, and variability of intercycle foveation position. Across participants, each parameter (except frequency) was significantly correlated with VA. Within any given individual, there was a small but significant improvement in VA (0.08 logMAR) at the null zone as compared with the other gaze angles tested. Despite this, no change in any of the nystagmus waveform parameters was significantly associated with changes in VA within individuals. Conclusions: A strong relationship between VA and nystagmus characteristics exists between individuals with IN. Although significant, the improvement in VA observed within individuals at the null zone is much smaller than might be expected from the occasionally large variations in intensity and foveation dynamics (and anecdotal patient reports of improved vision), suggesting that improvement of other aspects of visual performance may also encourage use of the null zone