Selective Estrogen Receptor Modulators Suppress Hif1α Protein Accumulation in Mouse Osteoclasts

Anti-bone resorptive drugs such as bisphosphonates, the anti-RANKL antibody (denosumab), or selective estrogen receptor modulators (SERMs) have been developed to treat osteoporosis. Mechanisms underlying activity of bisphosphonates or denosumab in this context are understood, while it is less clear how SERMs like tamoxifen, raloxifene, or bazedoxifene inhibit bone resorption. Recently, accumulation of hypoxia inducible factor 1 alpha (Hif1α) in osteoclasts was shown to be suppressed by estrogen in normal cells. In addition, osteoclast activation and decreased bone mass seen in estrogen-deficient conditions was found to require Hif1α. Here, we used western blot analysis of cultured osteoclast precursor cells to show that tamoxifen, raloxifene, or bazedoxifene all suppress Hif1α protein accumulation. The effects of each SERM on osteoclast differentiation differed in vitro. Our results suggest that interventions such as the SERMs evaluated here could be useful to inhibit Hif1α and osteoclast activity under estrogen-deficient conditions.     

Hierarchical neighbor effects on mycorrhizal community structure and function

Theory predicts that neighboring communities can shape one another's composition and function, for example, through the exchange of member species. However, empirical tests of the directionality and strength of these effects are rare. We determined the effects of neighboring communities on one another through experimental manipulation of a plant‐fungal model system. We first established distinct ectomycorrhizal fungal communities on Douglas‐fir seedlings that were initially grown in three soil environments. We then transplanted seedlings and mycorrhizal communities in a fully factorial experiment designed to quantify the direction and strength of neighbor effects by focusing on changes in fungal community species composition and implications for seedling growth (a proxy for community function). We found that neighbor effects on the composition and function of adjacent communities follow a dominance hierarchy. Specifically, mycorrhizal communities established from soils collected in Douglas‐fir plantations were both the least sensitive to neighbor effects, and exerted the strongest influence on their neighbors by driving convergence in neighbor community composition and increasing neighbor seedling vigor. These results demonstrate that asymmetric neighbor effects mediated by ecological history can determine both community composition and function.     

Evidence for a presynaptic action of chlordimeform at the insect neuromuscular junction

The mechanism of action of chlordimeform on the mealworm nerve-muscle preparation was studied with microelectrodes. Chlordimeform affected neither the mean amplitude nor the frequency of spontaneous miniature excitatory postsynaptic potentials. Extracellular focal recordings show that in the presence of 0.8 mM chlordimeform the presynaptic spike is almost unchanged, but the quantal content for evoked transmitter release is reduced. It is suggested that chlordimeform decreases the influx of calcium at the presynaptic terminal during the active phase of the nerve terminal action potential, thereby inhibiting evoked transmitter release.     

Proton NMR studies of nucleotide and amine storage in the dense granules of pig platelets

1H NMR measurements have been conducted at 360 MHz on isolated pig platelet dense granules. Resonances of the H8, H2 protons of the adenine ring, H1' protons of the ribose moiety, and the aromatic hydrogens of 5-hydroxytryptamine (5HT) have been identified in spectra of intact dense granules. Like the 31P resonances of the nucleotides contained in the dense granules (U?urbil et al., 1984), the line widths and the intensities of these resonances were sensitive to sample temperature and osmolarity of the suspension medium. Their chemical shifts indicate that 5HT in the granule interior is predominantly bound to the nucleotides through ring-stacking interactions. Association of 5HT with the nucleotides was also confirmed by the presence of intermolecular nuclear Overhauser effect (NOE) between 5HT and nucleotide protons. Large and negative intermolecular NOE's observed among the nucleotide H8, H2 and H1' protons, together with upfield shifts undergone by these protons within the dense granules, demonstrate that the nucleotides form a complex where they are in close proximity of each other. The formation of this complex apparently does not require the presence of amines since removal of 5HT and histamine did not change the chemical shifts of the nucleotide protons. From T1 and T2 data, rotational correlation time of 4 ns was calculated for the nucleotides in the dense granule interior at 35 degrees C. A resonance tentatively identified as H2 of histamine was found to shift upon manipulation of the intragranular pH; it was used as an indicator of pH changes within the granule interior during 5HT uptake and showed that 5HT accumulation increases the intragranular pH. These results demonstrate that 5HT is first taken up in response to the inside acidic pH gradient across the granule membrane and is subsequently sequestered in a matrix formed by the divalent cations and the nucleotides.     

Live Imaging-Based Model Selection Reveals Periodic Regulation of the Stochastic G1/S Phase Transition in Vertebrate Axial Development

In multicellular organism development, a stochastic cellular response is observed, even when a population of cells is exposed to the same environmental conditions. Retrieving the spatiotemporal regulatory mode hidden in the heterogeneous cellular behavior is a challenging task. The G1/S transition observed in cell cycle progression is a highly stochastic process. By taking advantage of a fluorescence cell cycle indicator, Fucci technology, we aimed to unveil a hidden regulatory mode of cell cycle progression in developing zebrafish. Fluorescence live imaging of Cecyil, a zebrafish line genetically expressing Fucci, demonstrated that newly formed notochordal cells from the posterior tip of the embryonic mesoderm exhibited the red (G1) fluorescence signal in the developing notochord. Prior to their initial vacuolation, these cells showed a fluorescence color switch from red to green, indicating G1/S transitions. This G1/S transition did not occur in a synchronous manner, but rather exhibited a stochastic process, since a mixed population of red and green cells was always inserted between newly formed red (G1) notochordal cells and vacuolating green cells. We termed this mixed population of notochordal cells, the G1/S transition window. We first performed quantitative analyses of live imaging data and a numerical estimation of the probability of the G1/S transition, which demonstrated the existence of a posteriorly traveling regulatory wave of the G1/S transition window. To obtain a better understanding of this regulatory mode, we constructed a mathematical model and performed a model selection by comparing the results obtained from the models with those from the experimental data. Our analyses demonstrated that the stochastic G1/S transition window in the notochord travels posteriorly in a periodic fashion, with doubled the periodicity of the neighboring paraxial mesoderm segmentation. This approach may have implications for the characterization of the pathophysiological tissue growth mode.     

Syntheses of Female Sex Pheromone Analogues of the German Cockroach and Their Biological Activity

Several analogues of 3,1l-dimethyl-2-nonacosanone, one component of the sex pheromone of the German cockroach, were synthesized. Their activity for the male to raise his wings was assayed and summerized in Tables I and II.     

Streamside assessment of sperm performance for wild charr

Streamside assessment of sperm quality was conducted using wild charr dwelling in mountain stream habitats. Through a portable combination of microscope and video camera, male conspecifics were observed to differ in the parameters of temporal changes in the frequency of active sperm, actual point-to-point distance that a sperm moved, and content of abnormal sperm cells. Frequency of active sperm, measured at 30 s after activation, could be a representative parameter of sperm motility because of its highly significant relatedness with other variables. To allow more rapid assessment, this parameter is proved to be appropriate for the estimation of sperm quality in the field.