Putative ligand binding sites of two functionally characterized bark beetle odorant receptors

Background: Bark beetles are major pests of conifer forests, and their behavior is primarily mediated via olfaction. Targeting the odorant receptors (ORs) may thus provide avenues towards improved pest control. Such an approach requires information on the function of ORs and their interactions with ligands, which is also essential for understanding the functional evolution of these receptors. Hence, we aimed to identify a high-quality complement of ORs from the destructive spruce bark beetle Ips typographus (Coleoptera, Curculionidae, Scolytinae) and analyze their antennal expression and phylogenetic relationships with ORs from other beetles. Using 68 biologically relevant test compounds, we next aimed to functionally characterize ecologically important ORs, using two systems for heterologous expression. Our final aim was to gain insight into the ligand-OR interaction of the functionally characterized ORs, using a combination of computational and experimental methods. Results: We annotated 73 ORs from an antennal transcriptome of I. typographus and report the functional characterization of two ORs (ItypOR46 and ItypOR49), which are responsive to single enantiomers of the common bark beetle pheromone compounds ipsenol and ipsdienol, respectively. Their responses and antennal expression correlate with the specificities, localizations, and/or abundances of olfactory sensory neurons detecting these enantiomers. We use homology modeling and molecular docking to predict their binding sites. Our models reveal a likely binding cleft lined with residues that previously have been shown to affect the responses of insect ORs. Within this cleft, the active ligands are predicted to specifically interact with residues Tyr84 and Thr205 in ItypOR46. The suggested importance of these residues in the activation by ipsenol is experimentally supported through site-directed mutagenesis and functional testing, and hydrogen bonding appears key in pheromone binding. Conclusions: The emerging insight into ligand binding in the two characterized ItypORs has a general importance for our understanding of the molecular and functional evolution of the insect OR gene family. Due to the ecological importance of the characterized receptors and widespread use of ipsenol and ipsdienol in bark beetle chemical communication, these ORs should be evaluated for their potential use in pest control and biosensors to detect bark beetle infestations

The evolutionary history of Trichoptera (Insecta): A case of successful adaptation to life in freshwater

The insect order Trichoptera (caddisflies) forms the second most species-rich monophyletic group of animals in freshwater. So far, several attempts have been made to elucidate its evolutionary history with both morphological and molecular data. However, none have attempted to analyse the time frame for its diversification. The order is divided into three suborders - Annulipalpia, Integripalpia and Spicipalpia'. Historically, the most problematic taxon to place within the order is Spicipalpia', whose larvae do not build traditional cases or filtering nets like the majority of the caddisflies. They have previously been proposed to be the sister group of all other Trichoptera or more advanced within the order, with equivocal monophyly and with different interordinal placements among various studies. In order to resolve the evolutionary history of the caddisflies as well as timing their diversification, we utilized fragments of three nuclear (carbamoylphosphate synthethase, isocitrate dehydrogenase and RNA polymerase II) and one mitochondrial (cytochrome oxidase I) protein coding genes, with 16 fossil trichopteran taxa used for time calibration. The spicipalpian' families are recovered as ancestral to all other caddisflies, though paraphyletic. We recover stable relationships among most families and superfamilies, resolving many previously unrecognized phylogenetic affinities amongst extant families. The origin of Trichoptera is estimated to be around 234Ma, i.e. Middle - Late Triassic

Limnología inicial de Laguna Pozo Verde, Costa Rica: Batimetría, agua, sedimentos y diatomeas

Introducción : Costa Rica tiene cientos de lagos, muchos de los cuales nunca han sido estudiados científicamente.  Objetivo: Conocer limnológia  de la Laguna Pozo Verde en el Parque Nacional Juan Castro Blanco, Costa Rica (~1935 m de elevación), para proporcionar datos de referencia para estudiar cambios futuros. Métodos: Medimos la profundidad y temperatura del agua y la profundidad de Secchi; sedimentos superficiales analizados; y examinó mapas e imágenes de satélite. Resultados: Aunque algunos la describen como formada por procesos volcánicos, la Laguna Pozo Verde probablemente se formó en un deslizamiento de tierra, que ocurre con frecuencia en esta zona lluviosa en la empinada ladera sur del inactivo volcán Porvenir. Nuestros sondeos mostraron una profundidad máxima de 9,25 m cerca del centro del lago. El agua era moderadamente transparente (profundidad de Secchi 2,6 m), con pH circunneutro y temperaturas de 15,9 a 18,1 °C, con estratificación débil a 0,5 m. Los sedimentos superficiales contenían 27% de materia orgánica y tenían relaciones C/N y valores de isótopos de carbono estables consistentes con algas lacustres y plantas C 3 ; Surirella angusta compuso más del 90% de las diatomeas. Conclusión: La laguna es significativamente más omera de lo informado y los sedimentos superficiales albergar una combinación de diatomeas única entre los 88 lagos examinados en Costa Rica.Introducción: Costa Rica tiene cientos de lagos, muchos nunca estudiados científicamente. Objetivo: Conocer la limnología de Laguna Pozo Verde en el Parque Nacional Juan Castro Blanco, Costa Rica (~1935m de elevación), como referencia para estudiar futuros cambios. Métodos: Medimos la profundidad y temperatura del agua y la profundidad de Secchi; analizamos sedimentos superficiales; y examinamos mapas e imágenes satelitales. Resultados: Aunque algunos la describen como formada por procesos volcánicos, es probable que Laguna Pozo Verde se haya formado en un deslizamiento de tierra, algo común en esta zona lluviosa de la empinada ladera sur del Volcán Porvenir (inactivo). Hallamos una profundidad máxima de 9,25m cerca del centro. El agua era moderadamente transparente (profundidad de Secchi 2,6m), con un pH cercano a neutro y temperaturas de 15,9-18,1°C, y una débil estratificación a 0,5m. Los sedimentos superficiales contenían un 27% de materia orgánica y tenían cocientes C/N y valores de isótopos de carbono estables, coherentes con algas lacustres y plantas C 3 ; más del 90% de las diatomeas eran Surirella angusta . Conclusión: La laguna es significativamente más somera de lo informado y los sedimentos superficiales albergan una combinación de diatomeas única entre los 88 lagos examinados en Costa Rica

Temporal course of cerebrospinal fluid dynamics and amyloid accumulation in the aging rat brain from three to thirty months

<p>Abstract</p> <p>Background</p> <p>Amyloid accumulation in the brain parenchyma is a hallmark of Alzheimer's disease (AD) and is seen in normal aging. Alterations in cerebrospinal fluid (CSF) dynamics are also associated with normal aging and AD. This study analyzed CSF volume, production and turnover rate in relation to amyloid-beta peptide (Aβ) accumulation in the aging rat brain.</p> <p>Methods</p> <p>Aging Fischer 344/Brown-Norway hybrid rats at 3, 12, 20, and 30 months were studied. CSF production was measured by ventriculo-cisternal perfusion with blue dextran in artificial CSF; CSF volume by MRI; and CSF turnover rate by dividing the CSF production rate by the volume of the CSF space. Aβ40 and Aβ42 concentrations in the cortex and hippocampus were measured by ELISA.</p> <p>Results</p> <p>There was a significant linear increase in total cranial CSF volume with age: 3-20 months (<it>p </it>< 0.01); 3-30 months (<it>p </it>< 0.001). CSF production rate increased from 3-12 months (<it>p </it>< 0.01) and decreased from 12-30 months (<it>p </it>< 0.05). CSF turnover showed an initial increase from 3 months (9.40 day^-1) to 12 months (11.30 day^-1) and then a decrease to 20 months (10.23 day^-1) and 30 months (6.62 day^-1). Aβ40 and Aβ42 concentrations in brain increased from 3-30 months (<it>p </it>< 0.001). Both Aβ42 and Aβ40 concentrations approached a steady state level by 30 months.</p> <p>Conclusions</p> <p>In young rats there is no correlation between CSF turnover and Aβ brain concentrations. After 12 months, CSF turnover decreases as brain Aβ continues to accumulate. This decrease in CSF turnover rate may be one of several clearance pathway alterations that influence age-related accumulation of brain amyloid.</p

Amyloid-beta transporter expression at the blood-CSF barrier is age-dependent

<p>Abstract</p> <p>Background</p> <p>Age is the major risk factor for many neurodegenerative diseases, including Alzheimer's disease (AD). There is an accumulation of amyloid-beta peptides (Aβ) in both the AD brain and the normal aging brain. Clearance of Aβ from the brain occurs via active transport at the blood-brain barrier (BBB) and blood-cerebrospinal fluid barrier (BCSFB). With increasing age, the expression of the Aβ efflux transporters is decreased and the Aβ influx transporter expression is increased at the BBB, adding to the amyloid burden in the brain. Expression of the Aβ transporters at the choroid plexus (CP) epithelium as a function of aging was the subject of this study.</p> <p>Methods</p> <p>This project investigated the changes in expression of the Aβ transporters, the low density lipoprotein receptor-related protein-1 (LRP-1), P-glycoprotein (P-gp), LRP-2 (megalin) and the receptor for advanced glycation end-products (RAGE) at the BCSFB in Brown-Norway/Fischer rats at ages 3, 6, 9, 12, 20, 30 and 36 months, using real time RT-PCR to measure transporter mRNA expression, and immunohistochemistry (IHC) to measure transporter protein in isolated rat CP.</p> <p>Results</p> <p>There was an increase in the transcription of the Aβ efflux transporters, LRP-1 and P-gp, no change in RAGE expression and a decrease in LRP-2, the CP epithelium influx transporter, at the BCSFB with aging. Decreased Aβ42 concentration in the CP, as measured by quantitative IHC, was associated with these Aβ transporter alterations.</p> <p>Conclusions</p> <p>Age-dependent alterations in the CP Aβ transporters are associated with a decrease in Aβ42 accumulation in the CP, and are reciprocal to the changes seen in these transporters at the BBB, suggesting a possible compensatory role for the BCSFB in Aβ clearance in aging.</p

The effectiveness of manual stretching in the treatment of plantar heel pain: a systematic review

Background: Plantar heel pain is a commonly occurring foot complaint. Stretching is frequently utilised as a treatment, yet a systematic review focusing only on its effectiveness has not been published. This review aimed to assess the effectiveness of stretching on pain and function in people with plantar heel pain. Methods: Medline, EMBASE, CINAHL, AMED, and The Cochrane Library were searched from inception to July 2010. Studies fulfilling the inclusion criteria were independently assessed, and their quality evaluated using the modified PEDro scale. Results: Six studies including 365 symptomatic participants were included. Two compared stretching with a control, one study compared stretching to an alternative intervention, one study compared stretching to both alternative and control interventions, and two compared different stretching techniques and durations. Quality rating on the modified Pedro scale varied from two to eight out of a maximum of ten points. The methodologies and interventions varied significantly between studies, making meta-analysis inappropriate. Most participants improved over the course of the studies, but when stretching was compared to alternative or control interventions, the changes only reached statistical significance in one study that used a combination of calf muscle stretches and plantar fascia stretches in their stretching programme. Another study comparing different stretching techniques, showed a statistically significant reduction in some aspects of pain in favour of plantar fascia stretching over calf stretches in the short term. Conclusions: There were too few studies to assess whether stretching is effective compared to control or other interventions, for either pain or function. However, there is some evidence that plantar fascia stretching may be more effective than Achilles tendon stretching alone in the short-term. Appropriately powered randomised controlled trials, utilizing validated outcome measures, blinded assessors and long-term follow up are needed to assess the efficacy of stretching

Acceleration and Deceleration in the Internationalization Process of the Firm

By adopting a processual and dynamic view on internationalization, we develop the concepts of acceleration and deceleration, providing analytical tools to enhance our understanding of the non-linearity and multidimensionality of internationalization. We argue that acceleration and deceleration are embedded in the internationalization process and are a consequence of the firm’s capability to absorb and integrate acquired knowledge, and to find and exploit opportunities. In addition, we advance the idea that changes in speed are further influenced by how the firm integrates and coordinates the resources it has deployed within and across various internationalization dimensions. Thus, it emerges that the overall evolution of commitment to internationalization is more complex than received theories tend to present; therefore, empirical studies should aim to include a wide set of international activities and processes embedded in time

Antecedents and consequences of effectuation and causation in the international new venture creation process

The selection of the entry mode in an international market is of key importance for the venture. A process-based perspective on entry mode selection can add to the International Business and International Entrepreneurship literature. Framing the international market entry as an entrepreneurial process, this paper analyzes the antecedents and consequences of causation and effectuation in the entry mode selection. For the analysis, regression-based techniques were used on a sample of 65 gazelles. The results indicate that experienced entrepreneurs tend to apply effectuation rather than causation, while uncertainty does not have a systematic influence. Entrepreneurs using causation-based international new venture creation processes tend to engage in export-type entry modes, while effectuation-based international new venture creation processes do not predetermine the entry mod

A new classification of the long-horned caddisflies (Trichoptera: Leptoceridae) based on molecular data

Background: Leptoceridae are among the three largest families of Trichoptera (caddisflies). The current classification is founded on a phylogenetic work from the 1980's, based on morphological characters from adult males, i.e. wing venation, tibial spur formula and genital morphology. In order to get a new opinion about the relationships within the family, we undertook a molecular study of the family based on sequences from five genes, mitochondrial COI and the four nuclear genes CAD, EF-1 alpha, IDH and POL. Results: The resulting phylogenetic hypotheses are more or less congruent with the morphologically based classification, with most genera and tribes recovered as monophyletic, but with some major differences. For monophyly of the two subfamilies Triplectidinae and Leptocerinae, one tribe of each was removed and elevated to subfamily status; however monophyly of some genera and tribes is in question. All clades except Leptocerinae, were stable across different analysis methods. Conclusions: We elevate the tribes Grumichellini and Leptorussini to subfamily status, Grumichellinae and Leptorussinae, respectively. We also propose the synonymies of Ptochoecetis with Oecetis and Condocerus with Hudsonema.authorCount :