Evidence for absence of latch-bridge formation in muscular saphenous arteries

Large-diameter elastic arteries can produce strong contractions indefinitely at a high-energy economy by the formation of latch bridges. Whether downstream blood vessels also use latch bridges remains unknown. The zero-pressure medial thickness and lumen diameter of rabbit saphenous artery (SA), a muscular branch of the elastic femoral artery (FA), were, respectively, approximately twofold and half-fold that of the FA. In isolated FA and SA rings, KCl rapidly (< 16 s) caused strong increases in isometric stress (1.2 x 10(5) N/m2) and intracellular Ca2+ concentration ([Ca2+]i; 250 nM). By 10 min, [Ca2+]i declined to approximately 175 nM in both tissues, but stress was sustained in FA (1.3 x 10(5) N/m2) and reduced by 40% in SA (0.8 x 10(5) N/m2). Reduced tonic stress correlated with reduced myosin light chain (MLC) phosphorylation in SA (28 vs. 42% in FA), and simulations with the use of the four-state kinetic latch-bridge model supported the hypothesis that latch-bridge formation in FA, but not SA, permitted maintenance of high stress values at steady state. SA expressed more MLC phosphatase than FA, and permeabilized SA relaxed more rapidly than FA, suggesting that MLC phosphatase activity was greater in SA than in FA. The ratio of fast-to-slow myosin isoforms was greater for SA than FA, and on quick release, SA redeveloped isometric force faster than FA. These data support the hypothesis that maintained isometric force was 40% less in SA than in FA because expressed motor proteins in SA do not support latch-bridge formation.     

Effects of prefeeding, intercomponent-interval food, and extinction on temporal discrimination and pacemaker rate

This experiment investigated the effects of nonpharmacological disruption on temporal discrimination. Pigeons responded on a multiple schedule composed of fixed interval, color-matching, and temporal-discrimination components. The effects of three different disruptors (prefeeding, intercomponent-interval food, and extinction) were assessed. All disruptors decreased response rates during the fixed interval. Prefeeding and intercomponent-interval food had unsystematic effects on response patterning during the fixed interval, whereas extinction increased the relative response rate in the initial portions of the fixed interval. Accuracy of color matching was decreased by prefeeding and was not systematically affected by intercomponent-interval food and extinction. In the temporal-discrimination component, all disruptors flattened the psychophysical functions relating proportion long responses to sample duration. This result indicates a general disruption of temporal discrimination. In addition, parameter estimates derived from the behavioral theory of timing indicated all disruptors decreased pacemaker rate, a result consistent with the predictions of the theory. These results highlight the similarities between disruption of temporal discrimination by pharmacological and nonpharmacological manipulations.     

Olfactory discrimination among sex pheromone stereoisomers: chirality recognition by pink hibiscus mealybug males

Our previous field studies suggested that the two chiral centers in the sex pheromone of pink hibiscus mealybug, Maconellicoccus hirsutus, could elicit different male responses. The chiral center in the acid moiety of the pheromone seemed to be more critical than the alcohol portion of the pheromone molecule for attractiveness. The objective of the current study was to test this hypothesis by deploying stereoisomeric blends in pheromone traps. Captures of male M. hirsutus showed that pheromone with the naturally occurring (R)-maconelliyl (S)-2-methylbutanoate and (R)-lavandulyl (S)-2-methylbutanoate [R-S configuration] was most attractive and that pheromone with the unnatural S-S configuration was less attractive. In addition, the RS-R blend (containing R-R and S-R stereoisomers) yielded captures of male M. hirsutus that were comparable to blank controls, and an inhibitory effect was observed when R-R and S-R were combined with naturally occurring R-S blend. These results suggest a unique chirality recognition mechanism; olfactory discrimination among different pheromone stereoisomers depends upon both asymmetric centers. The S configuration on the acid moiety elicits attraction, whereas the R configuration induces inhibition. However, the attractive activity shows some degree of tolerance toward chirality change in the alcohol portion of the pheromone molecules.     

Gradual translocation of spatial correlates of neuronal firing in the hippocampus toward prospective reward locations

In a continuous T-maze alternation task, CA1 complex-spike neurons in the hippocampus differentially fire as the rat traverses overlapping segments of the maze (i.e., the stem) repeatedly via alternate routes. The temporal dynamics of this phenomenon were further investigated in the current study. Rats learned the alternation task from the first day of acquisition and the differential firing pattern in the stem was observed accordingly. More importantly, we report a phenomenon in which spatial correlates of CA1 neuronal ensembles gradually changed from their original firing locations, shifting toward prospective goal locations in the continuous T-maze alternation task. The relative locations of simultaneously recorded firing fields, however, were preserved within the ensemble spatial representation during this shifting. The within-session shifts in preferred firing locations in the absence of any changes in the environment suggest that certain cognitive factors can significantly alter the location-bound coding scheme of hippocampal neurons.     

In vivo imaging reveals dendritic targeting of laminated afferents by zebrafish retinal ganglion cells

Targeting of axons and dendrites to particular synaptic laminae is an important mechanism by which precise patterns of neuronal connectivity are established. Although axons target specific laminae during development, dendritic lamination has been thought to occur largely by pruning of inappropriately placed arbors. We discovered by in vivo time-lapse imaging that retinal ganglion cell (RGC) dendrites in zebrafish show growth patterns implicating dendritic targeting as a mechanism for contacting appropriate synaptic partners. Populations of RGCs labeled in transgenic animals establish distinct dendritic strata sequentially, predominantly from the inner to outer retina. Imaging individual cells over successive days confirmed that multistratified RGCs generate strata sequentially, each arbor elaborating within a specific lamina. Simultaneous imaging of RGCs and subpopulations of presynaptic amacrine interneurons revealed that RGC dendrites appear to target amacrine plexuses that had already laminated. Dendritic targeting of prepatterned afferents may thus be a novel mechanism for establishing proper synaptic connectivity.     

Bio-optical modeling of photosynthetic pigments in corals

The spectral reflectance of coral is inherently related to the amounts of photosynthetic pigments present in the zooxanthellae. There are no studies, however, showing that the suite of major photosynthetic pigments can be predicted from optical reflectance spectra. In this study, we measured cm-scale in vivo and in situ spectral reflectance for several colonies of the massive corals Porites lobata and Porites lutea, two colonies of the branching coral Porites compressa, and one colony of the encrusting coral Montipora flabellata in Kaneohe Bay, Oahu, Hawaii. For each reflectance spectrum, we collected a tissue sample and utilized high-performance liquid chromatography to quantify six major photosynthetic pigments, located in the zooxanthellae. We used multivariate multiple regression analysis with cross-validation to build and test an empirical linear model for predicting pigment concentrations from optical reflectance spectra. The model accurately predicted concentrations of chlorophyll a, chlorophyll c ₂, peridinin, diadinoxanthin, diatoxanthin and β-carotene, with correlation coefficients of 0.997, 0.941, 0.995, 0.996, 0.980 and 0.984, respectively. The relationship between predicted and actual concentrations was 1:1 for each pigment, except chlorophyll c ₂. This simple empirical model demonstrates the potential for routine, rapid, non-invasive monitoring of coral-zooxanthellae status, and ultimately for remote sensing of reef biogeochemical processes.     

Glenohumeral elevation-dependent influence of anterior glenohumeral capsular lesions on passive axial humeral rotation

The purpose of this study was to assess the effect of standardized anterior glenohumeral capsular lesions on axial humeral rotation in a full arc of glenohumeral elevation. Using a testing apparatus, the range of internal and external humeral rotation was assessed in an arc of glenohumeral elevation in the scapular plane with steps of 15 degrees in six isolated shoulder joint specimens. Cutting of the glenohumeral joint capsule 1 cm laterally from, and parallel to the glenoid rim was performed in seven steps of 1 cm till the anterior capsule was cut. Capsular lesions were made in three ways: from inferior, from superior and from the middle of the capsule. Anterior capsular lesions resulted in significant increase of external humeral rotation. This occurred particularly at 15-60 degrees glenohumeral elevation. Lesions of the inferior part of the capsule mainly increased external rotation at 30-60 degrees glenohumeral elevation, lesions of the superior part mainly in lower elevation angles and lesions of the middle more gradually in the range till 60 degrees of glenohumeral elevation. Cutting of the anterior glenohumeral capsule barely increased passive axial humeral rotation at elevation angles over 60 degrees. Above 60 degrees glenohumeral elevation, tightening of the inferior posterior glenohumeral joint capsule prevented both internal and, increasingly, external humeral rotation. From these observations it is concluded that increased external rotation correlates with progressive anterior capsular lesions, mainly below 60 degrees glenohumeral elevation. To assess anterior glenohumeral capsular lesions in patients, axial humeral rotation tests should probably not exceed 60 degrees glenohumeral elevation, i.e. 90 degrees thoracohumeral elevation.     

Structural insights into the design of nonpeptidic isothiazolidinone-containing inhibitors of protein-tyrosine phosphatase 1B

Structural analyses of the protein-tyrosine phosphatase 1B (PTP1B) active site and inhibitor complexes have aided in optimization of a peptide inhibitor containing the novel (S)-isothiazolidinone (IZD) phosphonate mimetic. Potency and permeability were simultaneously improved by replacing the polar peptidic backbone of the inhibitor with nonpeptidic moieties. The C-terminal primary amide was replaced with a benzimidazole ring, which hydrogen bonds to the carboxylate of Asp(48), and the N terminus of the peptide was replaced with an aryl sulfonamide, which hydrogen bonds to Asp(48) and the backbone NH of Arg(47) via a water molecule. Although both substituents retain the favorable hydrogen bonding network of the peptide scaffold, their aryl rings interact weakly with the protein. The aryl ring of benzimidazole is partially solvent exposed and only participates in van der Waals interactions with Phe(182) of the flap. The aryl ring of aryl sulfonamide adopts an unexpected conformation and only participates in intramolecular pi-stacking interactions with the benzimidazole ring. These results explain the flat SAR for substitutions on both rings and the reason why unsubstituted moieties were selected as candidates. Finally, substituents ortho to the IZD heterocycle on the aryl ring of the IZD-phenyl moiety bind in a small narrow site adjacent to the primary phosphate binding pocket. The crystal structure of an o-chloro derivative reveals that chlorine interacts extensively with residues in the small site. The structural insights that have led to the discovery of potent benzimidazole aryl sulfonamide o-substituted derivatives are discussed in detail.     

Effect of insecticides on mealybug destroyer (Coleoptera: Coccinellidae) and parasitoid Leptomastix dactylopii (Hymenoptera: Encyrtidae), natural enemies of citrus mealybug (Homoptera: Pseudococcidae)

In this study, we measured, under laboratory conditions, the direct and indirect effects of insecticides on mealybug destroyer, Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae), and parasitoid Leptomastix dactylopii Howard (Hymenoptera: Encyrtidae), natural enemies of citrus mealybug, Planococcus citri (Risso) (Homoptera: Pseudococcidae). The adult stages of both natural enemies were exposed to sprays of the insecticides buprofezin, pyriproxyfen, flonicamid, acetamiprid, dinotefuran, and clothianidin at label-recommended rates to assess direct mortality after 24, 48, and 72 h, respectively. The effects of the insecticides on L. dactylopii parasitization rate and percentage of parasitoid emergence also were monitored using the label and 4x the recommended label rate. Dinotefuran was extremely detrimental to the adult parasitoid at the label rate with 100% mortality after 24 h. Buprofezin, pyriproxyfen, and flonicamid were not harmful to L. dactylopii when applied at the label rate. At 4x the recommended label rate, dinotefuran, acetamiprid, and clothianidin were all harmful to the parasitoid with 100% mortality 72 h after application. Both buprofezin and flonicamid were not toxic to L. dactylopii with 100% adult survival after 72 h. Pyriproxyfen and flonicamid, at both the label and 4x the recommended label rate, did not negatively affect L. dactylopii parasitization rate or percentage of parasitoid emergence. Acetamiprid, dinotefuran, and clothianidin were toxic to C. montrouzieri adults with 100% mortality after 48 h, whereas buprofezin, pyriproxyfen, and flonicamid demonstrated minimal (10-20% mortality after 48 h) harmful effects to the predator. Based on the results from our study, the indirect effects of the insect growth regulator (IGR) buprofezin were not decisive; however, the IGR pyriproxyfen and the insecticide flonicamid were not directly or indirectly harmful to the predator C. montrouzieri and parastioid L. dactylopii, indicating that these insecticides are compatible with both natural enemies when used together for control of citrus mealybug in greenhouses and conservatories.     

Cutting edge: impaired transitional B cell production and selection in the nonobese diabetic mouse

Developing B cells undergo selection at multiple checkpoints to eliminate autoreactive clones. We analyzed B cell kinetics in the NOD mouse to establish whether these checkpoints are intact. Our results show that although bone marrow production is normal in NOD mice, transitional (TR) B cell production collapses at 3 wk of age, reflecting a lack of successful immature B cell migration to the periphery. This yields delayed establishment of the follicular pool and a lack of selection at the TR checkpoint, such that virtually all immature B cells that exit the bone marrow mature without further selection. These findings suggest that compromised TR B cell generation in NOD mice yields relaxed TR selection, affording autoreactive specificities access to mature pools.