Startle-evoked changes in diaphragmatic activity during wakefulness and sleep

Tonic inhibition of some respiratory muscles occurs as part of the generalized muscle atonia of rapid-eye-movement sleep (REMS). A second type of inhibition of the diaphragm during REMS, fractionations, consists of brief pauses in the diaphragmatic electromyogram (DIA EMG) in association with phasic events. Because motor inhibition can occur as part of the startle response, and the brain is highly activated during REMS, we hypothesized that the neural basis of the fractionations might be activation of a startle network. To test this hypothesis, tone bursts (100 dB, 20-ms duration at 15-s intervals) were applied to cats at a fixed inspiratory level in the DIA moving average during REMS, non-rapid-eye-movement sleep (NREMS), and wakefulness. Parallel sham studies (no tone applied) were obtained for each state. The response of the DIA EMG was averaged over 100 ms by using the tone pulse as a trigger, and the following parameters of the DIA EMG were measured: latency to peak and/or nadir, increment or decrement in activity, and duration of peak and/or nadir. After a tone, all five animals studied displayed a profound suppression of DIA activity during REMS (latency to nadir 42.4 +/- 10.0 ms, duration of suppression 35.9 +/- 17.6 ms). Similarly, DIA activity was suppressed in all cats during NREMS (latency to nadir 40.9 +/- 13.3 ms, duration 23.9 +/- 13.4 ms). An excitatory response was observed in only two cats during NREMS and wakefulness. The similarity of startle-induced DIA EMG pauses to spontaneous fractionations of DIA activity during REMS suggests that the latter result from activation of a central startle system.     

The effect of long-distance running on plasma immunoreactive glucagon levels

Twelve highly conditioned long-distance runners were studied to determine the effects of marathon (42 km) and 10,000 m running on plasma immunoreactive glucagon (IRG), serum immunoreactive insulin (IRI), and serum glucose (G) levels. Blood samples were drawn just prior to and immediately upon completion of the run. Marathon running resulted in no significant change in G, IRI, or IRG levels. After running 10,000 m, plasma IRG levels did not change significantly, while IRI and G increased significantly. In evaluating the pooled data from both runs, a significant inverse correlation was observed between delta G and delta IRG. This relationship between delta G and delta IRG suggests that glucagon plays a role in maintaining normal blood glucose levels during strenuous exercise.     

The other ocean acidification problem: CO2 as a resource among competitors for ecosystem dominance

Predictions concerning the consequences of the oceanic uptake of increasing atmospheric carbon dioxide (CO₂) have been primarily occupied with the effects of ocean acidification on calcifying organisms, particularly those critical to the formation of habitats (e.g. coral reefs) or their maintenance (e.g. grazing echinoderms). This focus overlooks direct and indirect effects of CO₂ on non-calcareous taxa that play critical roles in ecosystem shifts (e.g. competitors). We present the model that future atmospheric [CO₂] may act as a resource for mat-forming algae, a diverse and widespread group known to reduce the resilience of kelp forests and coral reefs. We test this hypothesis by combining laboratory and field CO₂ experiments and data from ‘natural’ volcanic CO₂ vents. We show that mats have enhanced productivity in experiments and more expansive covers in situ under projected near-future CO₂ conditions both in temperate and tropical conditions. The benefits of CO₂ are likely to vary among species of producers, potentially leading to shifts in species dominance in a high CO₂ world. We explore how ocean acidification combines with other environmental changes across a number of scales, and raise awareness of CO₂ as a resource whose change in availability could have wide-ranging community consequences beyond its direct effects.     

The initiation of B cell clonal expansion occurs independently of pre-B cell receptor formation.

Current models of B cell development posit that clonal expansion occurs as a direct result of Ig H chain expression. To test this hypothesis, we isolated a population of early B cells in which H chain recombination is initiated and assessed V(H)DJ(H) rearrangements in both cycling and noncycling cells. We found that actively dividing cells within this population are enriched for H chain rearrangements that are productive when compared with their counterparts in G(0)/G(1), apparently supporting a role for H chain expression in initiating early B cell division; entrance into the cell cycle was accompanied by V(H) gene-dependent H chain selection. However, we also identified a phenotypically identical population of actively cycling early B cells in the absence of H chain expression in recombination activating gene knockout mice. In addition, actively cycling early B cells could be detected in pre-B cell receptor (pBCR)-negative lambda5 knockout mice, but we found no evidence for V(H)-dependent H chain selection in this population. Given these results, we suggest that the initiation of clonal expansion, at this early stage in B cell development, occurs independently of H chain expression. Although the cycling cell pool is enriched for pBCR-positive cells in mice expressing surrogate L chain, pBCR formation is not required for the initiation of cell division.     

Mutation and identification of the F plasmid locus determining resistance to acridine orange curing

A region in plasmid F responsible for resistance to acridine orange curing has been identified. Insertion of the transposable element for ampicillin resistance Tn3 at the 45.8- or 46.35-kilobase coordinate in mini-F plasmids results in resistance to curing by acridine orange and also results in increased plasmid copy numbers. In contrast, copy number mutants of mini-F induced by chemical mutagenesis are sensitive to acridine-induced loss of plasmid. Hence, the Tn3-induced acridine orange resistance does not represent a suppression of sensitivity because of elevated plasmid copy numbers. General hypotheses to explain our results are presented.     

Ankyrin-B syndrome: enhanced cardiac function balanced by risk of cardiac death and premature senescence

Here we report the unexpected finding that specific human ANK2 variants represent a new example of balanced human variants. The prevalence of certain ANK2 (encodes ankyrin-B) variants range from 2 percent of European individuals to 8 percent in individuals from West Africa. Ankyrin-B variants associated with severe human arrhythmia phenotypes (eg E1425G, V1516D, R1788W) were rare in the general population. Variants associated with less severe clinical and in vitro phenotypes were unexpectedly common. Studies with the ankyrin-B(+/-) mouse reveal both benefits of enhanced cardiac contractility, as well as costs in earlier senescence and reduced lifespan. Together these findings suggest a constellation of traits that we term "ankyrin-B syndrome", which may contribute to both aging-related disorders and enhanced cardiac function.     

Improving biological indicators to better assess the condition of streams

Biological indicators of stream condition are in use by water resource managers worldwide. The State of Maryland and many other organizations that use Indices of Biotic Integrity (IBIs) must determine when and how to refine their IBIs so that better stream condition information is provided. With completion of the second statewide round in 2004, the Maryland Biological Stream Survey (MBSS) had collected data from 2500 stream sites, more than doubling the number of sites that were available for the original IBI development. This larger dataset provided an opportunity for the MBSS to address the following shortcomings in the original IBIs: (1) substantial disturbance apparent in some reference sites, (2) fish IBIs could not be applied to very small streams, (3) natural variability within IBIs (based on regions) resulted in some stream types (e.g., coldwater and blackwater streams) receiving lower IBI scores and (4) one IBI was not able to discriminate degradation as desired (i.e., Coastal Plain fish IBI). Therefore, development of new fish and benthic macroinvertebrate IBIs was undertaken to achieve the goals of: (1) increased confidence that the reference conditions are minimally disturbed, (2) including more natural variation (such as stream size) across the geographic regions and stream types of Maryland and (3) increased sensitivity of IBIs by using more classes (strata) and different metric combinations. New fish IBIs were developed for four geographical and stream type strata: Coastal Plain, Eastern Piedmont, warmwater Highlands and coldwater Highlands streams; new benthic macroinvertebrate IBIs were developed for three geographical strata: Coastal Plain, Eastern Piedmont and Highlands streams. The addition of one new fish IBI and one new benthic macroinvertebrate IBI partitioned natural variability into more homogeneous strata. At the same time, smaller streams (i.e., those draining catchments <300 ac), which constituted a greater proportion of streams (25%) sampled in Round Two (2000–2004) than Round One (1995–1997), because of the finer map scale, were included in the reference conditions used to develop the new IBIs. The resulting new IBIs have high classification efficiencies of 83–96% and are well balanced between Type I and Type II errors. By scoring coldwater streams, smaller streams and to some extent blackwater streams higher, the new IBIs improve on the original IBIs. Overall, the new IBIs provide better assessments of stream condition to support sound management decisions, without requiring substantial changes by cooperating stream assessment programs.     

The human Mis12 complex is required for kinetochore assembly and proper chromosome segregation

During cell division, kinetochores form the primary chromosomal attachment sites for spindle microtubules. We previously identified a network of 10 interacting kinetochore proteins conserved between Caenorhabditis elegans and humans. In this study, we investigate three proteins in the human network (hDsn1Q9H410, hNnf1PMF1, and hNsl1DC31). Using coexpression in bacteria and fractionation of mitotic extracts, we demonstrate that these proteins form a stable complex with the conserved kinetochore component hMis12. Human or chicken cells depleted of Mis12 complex subunits are delayed in mitosis with misaligned chromosomes and defects in chromosome biorientation. Aligned chromosomes exhibited reduced centromere stretch and diminished kinetochore microtubule bundles. Consistent with this, localization of the outer plate constituent Ndc80HEC1 was severely reduced. The checkpoint protein BubR1, the fibrous corona component centromere protein (CENP) E, and the inner kinetochore proteins CENP-A and CENP-H also failed to accumulate to wild-type levels in depleted cells. These results indicate that a four-subunit Mis12 complex plays an essential role in chromosome segregation in vertebrates and contributes to mitotic kinetochore assembly.