Cloning and expression of an insect Ca2+-ATPase from Heliothis virescens

AbstractA complementary DNA for the Tobacco Budworm, Heliothis virescens, sarco(endo)plasmic reticulum-type Ca2+-ATPase (HVSERCA) has been cloned and sequenced. cDNA fragments of adult rabbit fast-twitch muscle Ca2+-ATPase (SERCA1a) were used as heterologous probes to isolate a partial cDNA clone coding for a protein with high homology to the Ca2+-ATPase from Drosophila melanogaster (DRSERCA) and vertebrate ER/SR Ca2+ pumps. The entire cDNA clone contains an ORF encoding a protein of 1000 amino acids which shares the characteristic motifs of a P-type ATPase. HVSERCA shares 89% identity with DRSERCA, 80% identity with the Artemia Ca2+-ATPase and 72% identity with avian and mammalian SERCAs. An insect Ca2+-ATPase-specific polyclonal antiserum has been raised against a fusion protein containing sequence from the cytoplasmic domain of HVSERCA. Heterologous expression of the insect pump in COS-7 cells has been demonstrated by immunocytochemistry and the reticular pattern of staining is consistent with an ER localisation. However, the expressed enzyme from COS-7 cells does not appear to be active

CAPRI and RASAL impose different modes of information processing on Ras due to contrasting temporal filtering of Ca(2+)

The versatility of Ca(2+) as a second messenger lies in the complex manner in which Ca(2+) signals are generated. How information contained within the Ca(2+) code is interpreted underlies cell function. Recently, we identified CAPRI and RASAL as related Ca(2+)-triggered Ras GTPase-activating proteins. RASAL tracks agonist-stimulated Ca(2+) oscillations by repetitively associating with the plasma membrane, yet CAPRI displays a long-lasting Ca(2+)-triggered translocation that is refractory to cytosolic Ca(2+) oscillations. CAPRI behavior is Ca(2+)- and C2 domain–dependent but sustained recruitment is predominantly Ca(2+) independent, necessitating integration of Ca(2+) by the C2 domains with agonist-evoked plasma membrane interaction sites for the pleckstrin homology domain. Using an assay to monitor Ras activity in real time, we correlate the spatial and temporal translocation of CAPRI with the deactivation of H-Ras. CAPRI seems to low-pass filter the Ca(2+) signal, converting different intensities of stimulation into different durations of Ras activity in contrast to the preservation of Ca(2+) frequency information by RASAL, suggesting sophisticated modes of Ca(2+)-regulated Ras deactivation

Validation of the Dutch translation of the quality of recovery-15 scale

BACKGROUND: The 15-item Quality of Recovery-15 (QoR-15) scale is strongly recommended as a standard patient-reported outcome measure assessing the quality of recovery after surgery and anesthesia in the postoperative period. This study aimed to validate the Dutch translation of the questionnaire (QoR-15NL). MATERIALS AND METHODS: An observational, prospective, single-centre cohort study was conducted. Patients who underwent surgery under general anesthesia completed the QoR-15NL (preoperatively (t1) and twice postoperatively (t2 and t3)) and a visual analogue scale (VAS) for general recovery at t2. A psychometric evaluation was performed to assess the QoR-15NL's validity, reliability, responsiveness, reproducibility and feasibility. RESULTS: Two hundred and eleven patients agreed to participate (recruitment rate 94%), and 165 patients were included (completion rate 78%). The QoR-15NL score correlated with the VAS for general recovery (rs = 0.59). Construct validity was further demonstrated by confirmation of expected negative associations between the QoR-15NL and duration of surgery (rs = -0.25), duration of Post Anesthesia Care Unit stay (rs = -0.31), and duration of hospital stay (rs = -0.27). The QoR-15NL score decreased significantly according to the extent of surgery. Cronbach's alpha was 0.87, split-half reliability was 0.8, and the test-retest intra-class coefficient was 0.93. No significant floor- or ceiling effect was observed. CONCLUSION: The QoR-15NL scale is a valid, easy-to-use, and reliable outcome assessment tool with high responsiveness for patient-reported quality of recovery after surgery and general anesthesia in the Dutch-speaking population. The QoR-15NL's measurement properties are comparable to the original questionnaire and other translated versions. TRIAL REGISTRATION: not applicable

The nuclear receptors of Biomphalaria glabrata and Lottia gigantea: Implications for developing new model organisms

© 2015 Kaur et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedNuclear receptors (NRs) are transcription regulators involved in an array of diverse physiological functions including key roles in endocrine and metabolic function. The aim of this study was to identify nuclear receptors in the fully sequenced genome of the gastropod snail, Biomphalaria glabrata, intermediate host for Schistosoma mansoni and compare these to known vertebrate NRs, with a view to assessing the snail's potential as a invertebrate model organism for endocrine function, both as a prospective new test organism and to elucidate the fundamental genetic and mechanistic causes of disease. For comparative purposes, the genome of a second gastropod, the owl limpet, Lottia gigantea was also investigated for nuclear receptors. Thirty-nine and thirty-three putative NRs were identified from the B. glabrata and L. gigantea genomes respectively, based on the presence of a conserved DNA-binding domain and/or ligand-binding domain. Nuclear receptor transcript expression was confirmed and sequences were subjected to a comparative phylogenetic analysis, which demonstrated that these molluscs have representatives of all the major NR subfamilies (1-6). Many of the identified NRs are conserved between vertebrates and invertebrates, however differences exist, most notably, the absence of receptors of Group 3C, which includes some of the vertebrate endocrine hormone targets. The mollusc genomes also contain NR homologues that are present in insects and nematodes but not in vertebrates, such as Group 1J (HR48/DAF12/HR96). The identification of many shared receptors between humans and molluscs indicates the potential for molluscs as model organisms; however the absence of several steroid hormone receptors indicates snail endocrine systems are fundamentally different.The National Centre for the Replacement, Refinement and Reduction of Animals in Research, Grant Ref:G0900802 to CSJ, LRN, SJ & EJR [www.nc3rs.org.uk]

The gender specific frequency of risk factor and CHD diagnoses prior to incident MI: A community study

BACKGROUND: CHD is a chronic disease often present years prior to incident AMI. Earlier recognition of CHD may be associated with higher levels of recognition and treatment of CHD risk factors that may delay incident AMI. To assess timing of CHD and CHD risk factor diagnoses prior to incident AMI. METHODS: This is a 10-year population based medical record review study that included all medical care providers in Olmsted County, Minnesota for all women and a sample of men residing in Olmsted County, MN with confirmed incident AMI between 1995 and 2000. RESULTS: All medical care for the 10 years prior to incident AMI was reviewed for 150 women and 148 men (38% sample) in Olmsted County, MN. On average, women were older than men at the time of incident AMI (74.7 versus 65.9 years, p < 0.0001). 30.4% of the men and 52.0% of the women received diagnoses of CHD prior to incident AMI (p = 0.0002). Unrecognized and untreated CHD risk factors were present in both men (45% of men 5 years prior to AMI) and women (22% of women 5 years prior to first AMI), more common in men and those without a diagnosis of CHD prior to incident AMI (p < 0.0001). CONCLUSION: A CHD diagnosis prior to incident AMI is associated with higher rates of recognition and treatment of CHD risk factors suggesting that diagnosing CHD prior to AMI enhances opportunities to lower the risk of future CHD events

Lymnaea schirazensis, an Overlooked Snail Distorting Fascioliasis Data: Genotype, Phenotype, Ecology, Worldwide Spread, Susceptibility, Applicability

BACKGROUND: Lymnaeid snails transmit medical and veterinary important trematodiases, mainly fascioliasis. Vector specificity of fasciolid parasites defines disease distribution and characteristics. Different lymnaeid species appear linked to different transmission and epidemiological patterns. Pronounced susceptibility differences to absolute resistance have been described among lymnaeid populations. When assessing disease characteristics in different endemic areas, unexpected results were obtained in studies on lymnaeid susceptibility to Fasciola. We undertook studies to understand this disease transmission heterogeneity. METHODOLOGY/PRINCIPAL FINDINGS: A ten-year study in Iran, Egypt, Spain, the Dominican Republic, Mexico, Venezuela, Ecuador and Peru, demonstrated that such heterogeneity is not due to susceptibility differences, but to a hitherto overlooked cryptic species, Lymnaea schirazensis, confused with the main vector Galba truncatula and/or other Galba/Fossaria vectors. Nuclear rDNA and mtDNA sequences and phylogenetic reconstruction highlighted an old evolutionary divergence from other Galba/Fossaria species, and a low intraspecific variability suggesting a recent spread from one geographical source. Morphometry, anatomy and egg cluster analyses allowed for phenotypic differentiation. Selfing, egg laying, and habitat characteristics indicated a migration capacity by passive transport. Studies showed that it is not a vector species (n = 8572 field collected, 20 populations): snail finding and penetration by F. hepatica miracidium occur but never lead to cercarial production (n = 338 experimentally infected). CONCLUSIONS/SIGNIFICANCE: This species has been distorting fasciolid specificity/susceptibility and fascioliasis geographical distribution data. Hence, a large body of literature on G. truncatula should be revised. Its existence has henceforth to be considered in research. Genetic data on livestock, archeology and history along the 10,000-year post-domestication period explain its wide spread from the Neolithic Fertile Crescent. It is an efficient biomarker for the follow-up of livestock movements, a crucial aspect in fascioliasis emergence. It offers an outstanding laboratory model for genetic studies on susceptibility/resistance in F. hepatica/lymnaeid interaction, a field of applied research with disease control perspectives