PKCdelta modulates p21WAF1/CIP1 ability to bind to Cdk2 during TNFalpha-induced apoptosis

Cyclin-dependent kinase 2 (Cdk2) activity is thought to be involved in cell death-associated chromatin condensation and other manifestations of apoptotic death. Here we show that during TNFalpha-induced apoptosis, PKCdelta is activated in a caspase-3-dependent manner and phosphorylates p21(WAF1/CIP1), a specific cyclin-dependent kinase inhibitor, on (146)Ser. This residue is located near a cyclin-binding motif (Cy2) that plays an important role in the interaction between p21(WAF1/CIP1) and Cdk2, and its phosphorylation modulates the ability of p21(WAF1/CIP1) to associate with Cdk2. The phosphorylation of p21(WAF1/CIP1) is temporally related to the activation kinetics of Cdk2 activity during the apoptosis. We propose that during TNFalpha-induced apoptosis, PKCdelta-mediated phosphorylation of p21(WAF1/CIP1) at (146)Ser attenuates the Cdk2 binding of p21(WAF1/CIP1) and thereby upregulates Cdk2 activity.     

Community structure and food web based on stable isotopes (δN and δC) analysis of a North Eastern Atlantic maerl bed

Maerl beds are highly biodiverse biogenic substrata that have been receiving increasing attention in the last decade. Although maerl beds represent important nursery areas for commercial fishes and molluscs, little is known on the trophic web of their communities. Community structure parameters of maerl bed of the Bay of Brest (species richness, abundance, biomass and dominating species) were studied in parallel with the carbon and nitrogen isotopic composition of their main benthic species (macrofaunal, and megafaunal organisms) in order to assess the trophic levels and differences in the potential food sources of maerl inhabitants. The major potential sources of energy were identified to originate either from epiphytic macroalgae and microphytobenthos both growing on maerl thalli, together with sedimenting (sedimentary) particulate organic matter (POM) originating from the water column. The majority of the macro- and megafaunal organisms investigated were filter feeders, selective-deposit feeders and predators/scavengers. Filter feeders fall into three different groups representing different trophic pathways (i) sponges feeding directly on POM (water column filter feeders I), (ii) ascidians and holothurians feeding on POM and probably captured pelagic preys (water column filter feeders II), and (iii) filter feeding molluscs and crustaceans were hypothesised to feed on microphytobenthos or on decaying sedimented POM (Interface filter feeders). Selective deposit feeders were also divided into two subgroups. Carnivores were also distinguished between those with scavenging habits and true predators. Coupling of the trophic levels observed with the community biomass structure revealed that most of the benthic biomass derives its food from detritic sedimented POM and/or microphytobenthos, with interface filter feeders (23% of the biomass), selective deposit feeders (12%). Carnivores made up to 14% of the total biomass. Generally stable isotopes ratio mean values overlap and cover a large range within feeding types, indicating a strong overlap in food sources and a high degree of complexity of the food web presumably due to the diversity of the potential food sources.     

Mitochondrial F1Fo-ATP Synthase: The Small Subunits e and g Associate with Monomeric Complexes to Trigger Dimerization

Mitochondrial F₁F_o-ATP synthase catalyzes the formation of ATP from ADP and inorganic phosphate. The enzyme is found in monomeric, dimeric and higher oligomeric forms in the inner mitochondrial membrane. Dimerization of ATP synthase complexes is a prerequisite for the generation of larger oligomers that promote membrane bending and formation of tubular cristae membranes. Two small proteins of the membrane-embedded F_o-domain, subunit e (Su e; Atp21) and Su g (Atp20), were identified as dimer-specific subunits of yeast ATP synthase and shown to be required for stabilization of the dimers. We have identified two distinct monomeric forms of yeast ATP synthase. Su e and Su g are present not only in the dimer but also in one of the monomeric forms. We demonstrate that Su e and Su g sequentially assemble with monomeric ATP synthase to form a dimerization-competent primed monomer. We conclude that association of Su e and Su g with monomeric F₁F_o-ATP synthase represents an initial step of oligomer formation.     

Temporary drought can selectively suppress Schoenoplectus mucronatus in rice

Outdoor pot experiments were conducted in California to quantify differences in rice and Schoenoplectus mucronatus susceptibility to drought and to identify morphological and physiological traits that would favor rice over S. mucronatus under drought. Plants were grown in flooded soil for approximately 5 weeks, and then subjected to different drought periods after which pots were re-flooded. Chlorophyll fluorescence assays revealed that rice and S. mucronatus Fv/Fm first became <0.8 after leaf water potential (Ψ_leaf) had decreased to approximately −4 MPa and −2 MPa, respectively. Thus, by suffering less photosynthetic damage from drought, rice had better recovery after re-flooding than S. mucronatus. When drought reduced Ψ_leaf to −3 MPa, S. mucronatus re-growth was nearly suppressed but that of rice was unaffected. Rice plants depleted soil moisture 1.6 faster than S. mucronatus due to larger and deeper roots and a high water-spending strategy (when Ψ_leaf decreased from approximately −0.5 MPa to −2.5 MPa, ¹³δ increased from −27.8 to −27.4 and from −28.1 to −26.0 for rice and S. mucronatus, respectively). Rice under interspecific competition sustained its Ψ_leaf by extracting more water from greater depths, while causing severe moisture stress and photosynthetic damage to S. mucronatus. Thus temporary drought enhanced rice competitiveness over S. mucronatus, supporting the concept of using brief drought as a tool for S. mucronatus suppression in rice. The Ψ_leaf developed by the end of the drought period predicted rice yields (R² = 0.77, P < 0.0001) and the capacity of S. mucronatus to recover from drought upon irrigation resumption (R² = 0.62, P < 0.001). Brief (8-10 d) drought imposed on 5-week-old rice did not significantly depress late-season rice biomass growth or grain yields, while S. mucronatus never fully recovered from drought. Rice yields were only reduced after Ψ_leaf reached values below approximately −2.5 MPa. Longer drought (∼20 d) delayed maturity and reduced rice yields by approximately 60-80%. The dry-down approach could help suppress weeds similar to S. mucronatus in organic rice where premium prices can compensate for lower grain yield.     

Intracellular Ca2+ transients in delta-sarcoglycan knockout mouse skeletal muscle

Background

δ-Sarcoglycan (δ-SG) knockout (KO) mice develop skeletal muscle histopathological alterations similar to those in humans with limb muscular dystrophy. Membrane fragility and increased Ca²⁺ permeability have been linked to muscle degeneration. However, little is known about the mechanisms by which genetic defects lead to disease.

Methods

Isolated skeletal muscle fibers of wild-type and δ-SG KO mice were used to investigate whether the absence of δ-SG alters the increase in intracellular Ca²⁺ during single twitches and tetani or during repeated stimulation. Immunolabeling, electrical field stimulation and Ca²⁺ transient recording techniques with fluorescent indicators were used.

Results

Ca²⁺ transients during single twitches and tetani generated by muscle fibers of δ-SG KO mice are similar to those of wild-type mice, but their amplitude is greatly decreased during protracted stimulation in KO compared to wild-type fibers. This impairment is independent of extracellular Ca²⁺ and is mimicked in wild-type fibers by blocking store-operated calcium channels with 2-aminoethoxydiphenyl borate (2-APB). Also, immunolabeling indicates the localization of a δ-SG isoform in the sarcoplasmic reticulum of the isolated skeletal muscle fibers of wild-type animals, which may be related to the functional differences between wild-type and KO muscles.

Conclusions

δ-SG has a role in calcium homeostasis in skeletal muscle fibers.

General significance

These results support a possible role of δ-SG on calcium homeostasis. The alterations caused by the absence of δ-SG may be related to the pathogenesis of muscular dystrophy.     

Isotope analysis reveals proportional change and site-selection variation of river- and lake-produced eggs of a landlocked migratory fish

Changes in the proportions of river- and lake-produced eggs of a landlocked amphidromous fish, ayu (Plecoglossus altivelis altivelis) in the Lake Biwa water system, Japan, were monitored by stable isotope analysis, based on different δ¹⁵N and δ¹³C values of prey organisms between the lake and its tributaries. During the 3 month reproduction season, the δ¹⁵N values of spawned eggs decreased with time. This result implies that there was a shift from lake-produced eggs to river-produced eggs within a reproductive season, based on the observation that adult fish in the lake had previously been shown to have eggs with distinctly higher δ¹⁵N values in their ovaries than those in the tributaries. This explanation was also supported by the change in δ¹³C values of the spawned eggs. Furthermore, eggs with lower δ¹⁵N and higher δ¹³C values tended to be spawned at less variable depths, suggesting that females spawning river-produced eggs selected the spawning sites from a narrower range. We conclude that stable isotope ratios of spawned eggs can be indicators of the relative contributions of different food chains and can enable comparisons of reproductive characteristics between types of egg.     

Design,synthesis and structure-activity relationship study of piperazinone-containing thieno[3,2-d]pyrimidine derivatives as new PI3Kδ inhibitors

Two classes of piperazinone-containing thieno[3,2-d]pyrimidines were designed and synthesized as new PI3Kδ inhibitors in this study. Detailed SAR study with respect to the piperazinone substituents at the 6-position of thieno[3,2-d]pyrimidine core demonstrated that piperazinone-containing thieno[3,2-d]pyrimidines would be more potent and selective for PI3Kδ than their piperazine counterparts, which led to the discovery of several potent PI3Kδ inhibitors with comparable or better antiproliferative activity against a panel of non-Hodgkin lymphoma (NHL) cell lines as compared with idelalisib. Our study will promote the development of new PI3Kδ inhibitors based on piperazinone-containing thieno[3,2-d]pyrimidine scaffold.     

Effects of the early stage of decomposition on change in carbon and nitrogen isotopes in Sphagnum litter

Abstract

Isotope and elemental composition of carbon (C) and nitrogen (N) as well as its mass loss were measured for Sphagnum fuscum litter after one and two years of incubation in three different soil zones defined by the position of water table in a pristine Sphagnum-dominated peatland on the coast of western Canada. Mass losses were greater for the first year than for the second year, and the greatest loss was found in the oxic zone closest to the peatland surface. Early stage of decomposition clearly affected isotope signatures in Sphagnum litter. Litter δ¹³C values significantly decreased after the first year of incubation. The depletion of ¹³C content during the first year might be related to the loss of more isotopically enriched soluble constituents coupled with the large mass loss. Litter δ¹⁵N values significantly increased after the first year of incubation in spite of the large mass loss. Litters incubated in the oxic zone had the greatest mass loss and ¹⁵N enrichment, suggesting that the enrichment was the result of interactions with soil microbes and preferential loss of lighter N. Conversely, litters incubated in the anoxic zone had smaller mass loss and the amount of N significantly increased, suggesting that the incorporation of bacterial biomass might also contribute to the ¹⁵N enrichment. The ¹⁵N enrichment trend continued in the second year, but the change was not significant as the first year. Increases in the δ¹⁵N values with depth in the near surface Sphagnum peat core suggests that the enrichment trend of litter ¹⁵N abundance with age is likely to continue for much longer periods than observed over the two-year period of this study.     

Microbial Response to Organic Matter Enrichment in the Oligotrophic Levantine Basin (Eastern Mediterranean)

The response of a deep-water benthic microbial community to organic matter (OM) enrichment was studied in the unexplored region of the SW Cretan margin (E. Mediterranean). A food pulse of 0.5 g C m^?2 was simulated by adding ¹³C-labelled diatoms to sediment cores retrieved from 1079 m depth. The diatom addition resulted in a significant increase in the sediment community oxygen consumption (SCOC). After 6 days, ～50 mg C m^?2 of the added material was processed by the microbial community. The major carbon sink was respiration, which accounted for ～96% of the total processed material. The carbon uptake rate (12 mg C m^?2 d^?1) was considerably lower than previously published values in the E. Mediterranean at similar depths. The microbial community in our study site is distinct, as revealed by the unusually high presence of branched phospholipid fatty acids (PLFA). Previous studies have revealed that the slope under investigation may act as a conduit of OM from the shallow shelf to the deep basins, resulting in the prevalence of relatively refractory OM at mid-slope depths. We postulate that sedimentary processes affect the amount of bioavailable sedimentary OM and consequently the structure and physiological state of bacterial community in our study site. The distinct microbial community composition at our station compared to more stable adjacent slopes could explain the limited response of the microbial community to the addition of labile OM. Supplemental materials are available for this article. Go to the publisher's online edition of Geomicrobiology Journal to view the free supplemental file.