Dynamic movement of the calcium sensor STIM1 and the calcium channel Orai1 in activated T-cells: puncta and distal caps

The proteins STIM1 and Orai1 are the long sought components of the store-operated channels required in T-cell activation. However, little is known about the interaction of these proteins in T-cells after engagement of the T-cell receptor. We found that T-cell receptor engagement caused STIM1 and Orai1 to colocalize in puncta near the site of stimulation and accumulate in a dense structure on the opposite side of the T-cell. FRET measurements showed a close interaction between STIM1 and Orai1 both in the puncta and in the dense cap-like structure. The formation of cap-like structures did not entail rearrangement of the entire endoplasmic reticulum. Cap formation depended on TCR engagement and tyrosine phosphorylation, but not on channel activity or Ca(2+) influx. These caps were very dynamic in T-cells activated by contact with superantigen pulsed B-cells and could move from the distal pole to an existing or a newly forming immunological synapse. One function of this cap may be to provide preassembled Ca(2+) channel components to existing and newly forming immunological synapses.     

Lessons Learned about Research and Management: A Case Study from a Midwest Lowland Savanna,U.S.A.

Integrating research and practice is a fundamental challenge for restoration and conservation. Scientists recognize restoration as an opportunity to test basic ecological principles, and that core scientific practices such as establishing control plots, replication, monitoring, and data management are fundamental to learning from restoration and management. However, practical constraints inherent to many projects often work against fully integrating research and management. In 2005, we had the opportunity to evaluate a lowland savanna that had been undergoing restoration by The Nature Conservancy since 1997. We used the data generated as a case study to evaluate how the restoration process might have changed had scientists been involved from the outset. Prescribed fire is the primary tool used to restore upland savanna and was chosen by The Nature Conservancy to manage the lowland savanna. In open areas dominated by Reed canary grass (Phalaris arundinacea), fire was associated with reduced cover of this highly invasive species and an increase in native species richness. However, other results suggested that management options beyond the use of fire are needed to restore this lowland savanna. In retrospect, an approach that integrated the skills of researchers and managers at the outset of the study would have altered many of the restoration goals and processes. Pretreatment data, greater replication, and systematically varying treatments could have provided a more robust assessment of treatment effects. We recognize, however, that impediments in staffing, funding, and time were barriers to following this course of action.     

Seasonal expressed sequence tags of rainbow smelt (Osmerus mordax) revealed by subtractive hybridization and the identification of two genes up-regulated during winter

The rainbow smelt (Osmerus mordax) is freeze-resistant and maintains swimming and feeding activity during winter. In order to identify genes differentially expressed in smelt liver response to winter water temperatures, a large-scale analysis of gene expression using suppression subtractive hybridization was carried out using samples obtained in fall and winter. Forward and reverse subtractions were performed, subtraction-enriched products were cloned, and clones were sequenced from both of the resulting libraries. When 27 of these genes were screened by semi-quantitative RT-PCR to identify candidates for differential expression based generally on 2-fold changes in expression, one encoding FK506-binding protein 5 was classified as up-regulated in response to seasonal change, another encoding the mitochondrial solute carrier 25 member 25 (ATP-Mg/Pi carrier) was similarly classified with seasonal change and low temperature shift, and the one encoding the 78 kDa glucose-regulated protein was provisionally classified as down-regulated with low temperature shift. Analysis of fall (warm) and winter (cold) seasonal samples by quantitative PCR (qPCR) revealed significant up-regulation of genes encoding FK506-binding protein 51 and the mitochondrial solute carrier, whereas the gene encoding the glucose-regulated protein showed no significant change in expression. The mitochondrial solute carrier and FK506-binding protein results may relate to changes in cortisol action, as both are regulated by cortisol in other species.     

An improved multi-component sex attractant for trapping male western yellowstriped armyworm, Spodoptera praefica (Grote) (Lepidoptera: Noctuidae)

Abstract 1 Chemical analyses of solvent extracts of pheromone glands of female western yellowstriped armyworm moths Spodoptera praefica (Grote) indicated the presence of (Z)‐7‐dodecenol (Z)‐7‐dodecenyl acetate (Z)‐9‐dodecenyl acetate (Z)‐9‐tetradecenyl acetate and (Z)‐11‐hexadecenyl acetate. 2 In field tests of combinations of these chemicals, small numbers of male S. praefica were captured in traps baited with (Z)‐7‐dodecenyl acetate. Numbers of males captured in traps were greatly increased in response to blends that included both (Z)‐7‐dodecenyl acetate with either (Z)‐9‐tetradecenyl acetate (Z)‐9‐dodecenyl acetate. The combination of (Z)‐7‐dodecenyl acetate and (Z)‐9‐tetradecenyl acetate provided the strongest sex attractant for use in trapping male S. praefica. 3 Males of the cabbage looper Trichoplusia ni (Hübner) were captured in traps baited with blends possessing (Z)‐7‐dodecenyl acetate, and were greatly reduced in traps baited with blends that included (Z)‐7‐dodecenol. 4 Multi‐component blends that included (Z)‐7‐dodecenol attracted males of the alfalfa looper Autographa californica (Speyer). 5 Males of Peridroma saucia (Hübner) and Mamestra configurata Walker were captured in traps that included (Z)‐9‐tetradecenyl acetate with (Z)‐11‐hexadecenyl acetate. 6 These responses by other species of moths to S. praefica pheromone components and blends may still complicate the use of any lure for S. praefica.     

Force platform for rats measures fore and hind forces concurrently

Animal models are commonly used to test the efficacy of impact loading regimens on bone strength. We designed an inexpensive force platform to concurrently measure the separate peak vertical impact forces produced by the fore and hindfeet of immature F-344 rats when dropped onto the platform. The force platform consisted of three load cells placed in a triangular pattern under a flat plate. Rats were dropped from heights of 30, 45 and 60 cm onto the platform so that they landed on all four feet concurrently. The peak vertical impact forces produced by the feet of the rats were measured using a sampling frequency of 100 kHz. The location of each foot at landing relative to the load cells, and the force received by each load cell were combined in a series of static equations to solve for the vertical impact forces produced by the fore and hindfeet. The forces produced by feet when rats stood on the single platform were similarly determined. The forces exerted separately by the fore and hindfeet of young rats when landing on the plate as a ratio to standing forces were then calculated. Rats when standing bore more weight on their hindfeet but landed with more weight on their forefeet, which provides rationale for the greater response to landing forces of bones in the forelimbs than those in the hindlimbs. This system provided a useful method to simultaneously measure peak vertical impact forces in fore and hindfeet in rats.     

Prediction of outcome of non-small cell lung cancer patients treated with chemotherapy and bortezomib by time-course MALDI-TOF-MS serum peptide profiling

Background  

Only a minority of patients with advanced non-small cell lung cancer (NSCLC) benefit from chemotherapy. Serum peptide profiling of NSCLC patients was performed to investigate patterns associated with treatment outcome.     

Proximate mechanisms driving circadian control of neuroendocrine function: Lessons from the young and old

Circadian rhythms impact a variety of behavioral and physiological functions contributing to longevity and successful reproduction. In their natural environments, individuals of a species are faced with a multitude of challenges and the coordination of internal processes and behavior with external pressures has been hypothesized to be an important target of natural selection. Several lines of evidence from cyanobacteria, Drosophila, and plants provide strong support for an important role of the circadian clock in survival and reproductive success. Similarly in mammals, disruptions in circadian function markedly impact reproduction and lifespan. The present review discusses research outlining the proximate and ultimate mechanisms responsible for the central and peripheral control of the reproductive axis. Because precise temporal coordination of the endocrine system is particularly crucial for reproduction by females, the present overview focuses on the role of circadian timing in this sex.     

Interaction between fidgetin and protein kinase A-anchoring protein AKAP95 is critical for palatogenesis in the mouse

The gene defective in fidget mice encodes fidgetin, a member of the AAA (ATPases associated with diverse cellular activities) family of ATPases. Using a yeast two-hybrid screen, we identified cAMP-dependent protein kinase A anchoring protein 95 kDa (AKAP95) as a potential fidgetin-binding protein. Epitope-tagged fidgetin co-localized with endogenous AKAP95 in the nuclear matrix, and the physical interaction between fidgetin and AKAP95 was further confirmed by reciprocal immunoprecipitation. To evaluate the biological significance of the fidgetin-AKAP95 binding, we created AKAP95 mutant mice through a gene trap strategy. Akap95 mutant mice are surprisingly viable with no overt phenotype. However, a significant number of mice carrying both Akap95 and fidget mutations die soon after birth due to cleft palate, consistent with the overlapping expression of AKAP95 and fidgetin in the branchial arches during mouse embryogenesis. These results expand the spectrum of the pleiotropic phenotypes of fidget mice and provide new leads on the in vivo function of AKAP95.