Application of isotope dilution analysis to the determination of sulfate in glycoproteins and oligosaccharides

Isotope dilution analysis, using a probe of ³⁵S-labeled BaSO₄, is proposed for the determination of sulfate in hydrolysates of glycoproteins and other glycoconjugates. A scaled-down version of the method of Klockow is presented. A modified radioassay in strongly acidic conditions, effective in the range of 0.5 to 10 nmol sulfate per sample, has been developed.     

THE ROLES OF FLUCTUATING SELECTION AND LONG-TERM DIAPAUSE IN MICROEVOLUTION OF DIAPAUSE TIMING IN A FRESHWATER COPEPOD

Direct observations of selection response in natural, unmanipulated populations in the wild are rare. Those that exist have resulted from major changes in environment during an ongoing study. Selection response should be more common and more readily observable in short-lived organisms where the direction of selection changes from year to year. We examined how the interaction of fluctuating selection, and emergence from long-term diapause, caused ongoing microevolutionary change over eight years in an important life-history trait (diapause timing) in the freshwater calanoid copepod Diaptomus sanguineus. Emergence from long-term diapause releases into the population lineages that did not experience the most recent bout of selection, thereby promoting the maintenance of the heritable trait variation that allows continual selection response. A mechanistic selection model was created on the basis of field and laboratory studies to predict how interannual variations in predation intensity generate year-to-year changes in mean diapause timing and in net reproductive success for alternate trait values. The predicted selection response and the estimated effect of emergence from diapause were both significantly correlated with observed changes in trait mean. A linear model combining selection response and emergence from diapause explained 59% of the variance in year-to-year changes in trait mean. According to this model, strong selection occurred in about half of the years studied, and the average annual contributions to changes in trait mean from selection and emergence were roughly equal. Thus, both fluctuating natural selection and emergence from prolonged diapause affect the expression of diapause timing by D. sanguineus. Fluctuating selection is ubiquitous in nature and may provide opportunities in other populations to witness ongoing natural selection without directional trends in mean phenotype.     

Diapause dynamics of two diaptomid copepod species in a large lake

Skistodiaptomus oregonensis and Leptodiaptomus minutus produce diapausing eggs in a large lake, Oneida Lake, in New York State. The timing of the switch from production of subitaneous (immediately hatching) eggs to diapausing eggs for both species is in October. This timing is consistent with a pattern, reviewed here, for other populations of diaptomid copepods: populations living in large lakes tend to begin production of diapausing eggs later in the season than those living in small lakes. Populations living in temporary ponds tend to switch still earlier in the season. All populations reviewed here live in the north temperate zone. The sediments of Oneida Lake contain densities of diaptomid diapausing eggs on the order of 10⁵ m^–2 per cm below the sediment surface down to 5 cm. Below this sediment depth, egg densities decline. The highest egg densities were found in sediments under the deepest water. Diapausing eggs of L. minutus survive in the sediments at least two years, as shown by the recovery of the population after a year in which no new diapausing eggs were produced, and probably for two or more decades. Long-term dormancy can have the effect of ensuring the continuation of a population through periods of poor recruitment, and can help create conditions for the coexistence of competing species. Other investigators have suggested that S. oregonensis and L. minutus are competitors in other lakes when they co-occur.     

Environmental variability and the persistence of multiple emergence strategies

Studies of plant and animal populations have demonstrated the occurrence of multiple and mixed life history strategies such as polymodal timing of germination and emergence from dormancy. We present the results of a simulation model used to test whether between-year variance in mortality can lead to the persistence of multiple hatching strategies considered over an ecological time scale (50 years). The model is based on the general life history characteristics of a population of planktonic copepods (Diaptomus sanguineus) in Bullhead Pond, Rhode Island. Our model results demonstrate that, given a range of between-year variance in mortality, multiple strategies for timing of emergence can persist in a common environment for ecologically relevant periods of time. A qualitative test of the model comparing field estimates of mean and variance of mortality in Bullhead Pond with the region of persistence indicates that the model results are in approximate agreement with field estimates. The results suggest that variability in year-to-year selection pressures, such as predation or harsh winters, may play an important role in determining the evolution of life histories.     

Consumer movement dynamics as hidden drivers of stream habitat structure: suckers as ecosystem engineers on the night shift

Ecosystem engineering can control the spatial and temporal distribution of resources and movement by engineering organisms within an ecosystem can mobilize resources across boundaries and distribute engineering effects. Movement patterns of fishes can cause physical changes to aquatic habitats though nesting or feeding, both of which often vary in space and time. Here we present evidence of ecosystem engineering by the Sonora sucker Catostomus insignis, a dominant fish in streams of the southwestern United States, and show how cryptic nocturnal movement patterns and bioturbation activities control heterogeneity in benthic substrates, and in sediment and carbon export. Sonora suckers exhibit distinct diel movement patterns, spending daylight hours in refuge habitats (typically deep pools) while moving into shallow habitats at night to feed. Feeding by suckers creates substantial disturbance in soft sediments that are patchy in space and time. These disturbances moved up to 2.4 × 10⁴ cm³ of sediment per square meter per week in locations that are up to hundreds of meters away from sucker daytime refuges. The diel cycles in feeding activity (i.e. nocturnal digging in benthic substrates) caused nighttime pulses in suspended sediment that comprised up to 32% of the daily suspended load and organic matter transport of a stream reach. During the daytime, this particulate transport settles in habitats beyond the location of the initial disturbance, thus redistributing both sediment and organic matter. Our data indicate that cryptic movement by ecosystem engineers can distribute their effects in space and time generating heterogeneity in resources and suggest that habitat modifications restricting consumer movement may alter the impact of engineering activities.     

Does rapid evolution matter? Measuring the rate of contemporary evolution and its impacts on ecological dynamics

Rapid contemporary evolution due to natural selection is common in the wild, but it remains uncertain whether its effects are an essential component of community and ecosystem structure and function. Previously we showed how to partition change in a population, community or ecosystem property into contributions from environmental and trait change, when trait change is entirely caused by evolution (Hairston et al. 2005). However, when substantial non-heritable trait change occurs (e.g. due to phenotypic plasticity or change in population structure) that approach can mis-estimate both contributions. Here, we demonstrate how to disentangle ecological impacts of evolution vs. non-heritable trait change by combining our previous approach with the Price Equation. This yields a three-way partitioning into effects of evolution, non-heritable phenotypic change and environment. We extend the approach to cases where ecological consequences of trait change are mediated through interspecific interactions. We analyse empirical examples involving fish, birds and zooplankton, finding that the proportional contribution of rapid evolution varies widely (even among different ecological properties affected by the same trait), and that rapid evolution can be important when it acts to oppose and mitigate phenotypic effects of environmental change. Paradoxically, rapid evolution may be most important when it is least evident.     

Density‐ and trait‐mediated top–down effects modify bottom–up control of a highly endemic tropical aquatic food web

Benthic invertebrates mediate bottom–up and top–down influences in aquatic food webs, and changes in the abundance or traits of invertebrates can alter the strength of top–down effects. Studies assessing the role of invertebrate abundance and behavior as controls on food web structure are rare at the whole ecosystem scale. Here we use a comparative approach to investigate bottom–up and top–down influences on whole anchialine pond ecosystems in coastal Hawai‘i. In these ponds, a single species of endemic atyid shrimp (Halocaridina rubra) is believed to structure epilithon communities. Many Hawaiian anchialine ponds and their endemic fauna, however, have been greatly altered by bottom–up (increased nutrient enrichment) and top–down (introduced fish predators) disturbances from human development. We present the results of a survey of dissolved nutrient concentrations, epilithon biomass and composition, and H. rubra abundance and behavior in anchialine ponds with and without invasive predatory fish along a nutrient concentration gradient on the North Kona coast of Hawai‘i. We use linear models to assess 1) the effects of nutrient loading and fish introductions on pond food web structure and 2) the role of shrimp density and behavior in effecting that change. We find evidence for bottom–up food web control, in that nutrients were associated with increased epilithon biomass, autotrophy and nutrient content as well as increased abundance and size of H. rubra. We also find evidence for top–down control, as ponds with invasive predatory fish had higher epilithon biomass, productivity, and nutrient content. Top–down effects were transmitted by both altered H. rubra abundance, which changed the biomass of epilithon, and H. rubra behavior, which changed the composition of the epilithon. Our study extends experimental findings on bottom–up and top–down control to the whole ecosystem scale and finds evidence for qualitatively different effects of trait‐ and density‐mediated change in top–down influences.     

Somato-Dendritic Localization and Signaling by Leptin Receptors in Hypothalamic POMC and AgRP Neurons

Leptin acts via neuronal leptin receptors to control energy balance. Hypothalamic pro-opiomelanocortin (POMC) and agouti-related peptide (AgRP)/Neuropeptide Y (NPY)/GABA neurons produce anorexigenic and orexigenic neuropeptides and neurotransmitters, and express the long signaling form of the leptin receptor (LepRb). Despite progress in the understanding of LepRb signaling and function, the sub-cellular localization of LepRb in target neurons has not been determined, primarily due to lack of sensitive anti-LepRb antibodies. Here we applied light microscopy (LM), confocal-laser scanning microscopy (CLSM), and electron microscopy (EM) to investigate LepRb localization and signaling in mice expressing a HA-tagged LepRb selectively in POMC or AgRP/NPY/GABA neurons. We report that LepRb receptors exhibit a somato-dendritic expression pattern. We further show that LepRb activates STAT3 phosphorylation in neuronal fibers within several hypothalamic and hindbrain nuclei of wild-type mice and rats, and specifically in dendrites of arcuate POMC and AgRP/NPY/GABA neurons of Leprb ^+/+ mice and in Leprb ^db/db mice expressing HA-LepRb in a neuron specific manner. We did not find evidence of LepRb localization or STAT3-signaling in axon-fibers or nerve-terminals of POMC and AgRP/NPY/GABA neurons. Three-dimensional serial EM-reconstruction of dendritic segments from POMC and AgRP/NPY/GABA neurons indicates a high density of shaft synapses. In addition, we found that the leptin activates STAT3 signaling in proximity to synapses on POMC and AgRP/NPY/GABA dendritic shafts. Taken together, these data suggest that the signaling-form of the leptin receptor exhibits a somato-dendritic expression pattern in POMC and AgRP/NPY/GABA neurons. Dendritic LepRb signaling may therefore play an important role in leptin’s central effects on energy balance, possibly through modulation of synaptic activity via post-synaptic mechanisms.     

Side chain SAR of bicyclic β-lactamase inhibitors (BLIs). 2. N-Alkylated and open chain analogs of MK-8712

The bridged monobactam β-lactamase inhibitor MK-8712 (1) effectively inhibits class C β-lactamases. Side chain N-alkylated and ring-opened analogs of 1 were prepared and evaluated for combination with imipenem to overcome class C β-lactamase mediated resistance. Although some analogs were more potent inhibitors of AmpC, none exhibited better synergy with imipenem than 1.