Leg coordination during turning on an extremely narrow substrate in a bug, Mesocerus marginatus (Heteroptera, Coreidae)

The turning movement of a bug, Mesocerus marginatus, is observed when it walks upside-down below a horizontal beam and, at the end of the beam, performs a sharp turn by 180 degrees . The turn at the end of the beam is accomplished in three to five steps, without strong temporal coordination among legs. During the stance, leg endpoints (tarsi) run through rounded trajectories, rotating to the same side in all legs. During certain phases of the turn, a leg is strongly depressed and the tarsus crosses the midline. Swing movements rotate to the same side as do leg endpoints in stance, in strong contrast to the typical swing movements found in turns or straight walk on a flat surface. Terminal location is found after the search through a trajectory that first moves away from the body and then loops back to find substrate. When a leg during stance has crossed the midline, in the following swing movement the leg may move even stronger on the contralateral side, i.e. is stronger depressed, in contrast to swing movements in normal walking, where the leg is elevated. These results suggest that the animals apply a different control strategy compared to walking and turning on a flat surface.     

Intracellular recordings from nonspiking interneurons in a semiintact,tethered walking insect

Nonspiking interneurons were investigated in a tethered, walking insect, Carausius morosus, that was able to freely perform walking movements. Experiments were carried out with animals walking on a lightweight, double-wheel treadmill. Although the animal was opened dorsally, the walking system was left intact. Intracellular recordings were obtained from the dorsal posterior neuropil of the mesothoracic ganglion. Nonspiking inter-neurons, in which modulations of the membrane potential were correlated with the walking rhythm, were described physiologically and stained with Lucifer Yellow. Interneurons are demonstrated in which membrane potential oscillations mirror the leg position or show correlation with the motoneuronal activity of the protractor and retractor coxae muscles during walking. Other interneurons showed distinct hyperpolarizations at certain important trigger points in the step cycle, for example, at the extreme posterior position. Through electrical stimulation of single, nonspiking interneurons during walking, the motoneuronal activity in two antagonistic muscles—protractor and retractor coxae—could be reversed and even the movement of the ipsilateral leg could be influenced. The nonspiking interneurons described appear to be important premotor elements involved in walking. They receive, integrate, and process information from different leg proprioceptors and drive groups of leg motoneurons during walking.     

Effects of temperature on properties of flight neurons in the locust

High ambient temperatures increase the wing-beat frequency in flying locusts, Locusta migratoria. We investigated parameters of circuit and cellular properties of flight motoneurons at temperatures permissive for flight (20–40 °C). As the thoracic temperature increased motoneuronal conduction velocity increased from an average of 4.40 m/s at 25 °C to 6.73 m/s at 35 °C, and the membrane time constant decreased from 11.45 ms to 7.52 ms. These property changes may increase locust wing-beat frequency by affecting the temporal summation of inputs to flight neurons in the central circuitry. Increases in thoracic temperature from 25–35 °C also resulted in a hyperpolarization of the resting membrane potentials of flight motoneurons from an average of-41.1 mV to -47.5 mV, and a decrease of input resistances from an average of 3.45 M to 2.00 M. Temperature affected the measured input resistance both by affecting membrane properties, and by altering synaptic input. We suggest that the increase in conduction velocity Q₁₀=1.53) and the decrease of membrane time constant (Q₁₀=0.62) would more than account for the wing-beat frequency increase (Q₁₀=1.15). Hyperpolarization of the resting membrane potential (Q₁₀=1.18) and reduction in input resistance (Q₁₀=0.54) may be involved in automatic compensation of temperature effects.Abbreviations ANOVA analysis of variance - CPG central pattern generator - DL dorsal longitudinal muscles - EMG electromyographic - MN motoneuron - PSP post synaptic potential - Q₁₀ temperature coefficient - RMP resting membrane potential - S.D. standard deviation - SR stretch receptor     

Detection of intracellular cytokines during antioxidant supplementation in corticoid-dependent asthmatics and modulation of adhesion molecule expression on cultured endothelial cells

In this study, we report on the interferon-γ (IFN-γ) and interleukin-4 (IL-4) cytokine responses to phorbol myristate acetate (PMA)+ionomycin-stimulated CD3+ lymphocytes in asthmatic subjects when compared with normal donors. There was a significantly lower production of intracellular IFN-γ in asthmatic patients. No difference was found for IL-4 production between these two groups. After administration of a multivitamin-mineral supplement containing selenium, zinc, vitamin A, vitamin B₆, vitamin C, and vitamin E for 6 mo, a significant increase in the percentage of CD3+/IL-4 positive cells (p<0.05) was found. The induction of endothelial cell adhesion molecule (CAM) expression in cultured human umbilical vein endothelial cells (HUVEC) and whole-blood mixture was studied using flow cytometry. The ICAM-1 and VCAM-1 expressions were higher in the patients than in control donors (p<0.05). There is a correlation between the increased percentage of CD3+/IFN-γ positive cells and reduced endothelial ICAM-1 and VCAM-1 expression after 6 mo of intervention period. No apparent effect of supplementation on CAM expression was found, suggesting that these changes do not arise from an antioxidant mechanism. This newly developed whole-blood technique for the assessment of CAM expression can be of use for monitoring therapy in inflammatory diseases.     

Silent ovulation, based on walking activity and milk progesterone concentrations, in Holstein cows housed in a free-stall barn

The objectives of the current study were to determine the incidence of silent ovulation (based on walking activity and milk progesterone profiles), identify risk factors for silent ovulation, and investigate its impact on reproductive performance in high-yielding dairy cows in free-stall housing. Overall, 277 lactations in 161 Holstein Friesian cows from a commercial dairy herd in northern Japan were studied. Walking activity (measured with pedometers) >80% above the mean for the preceding 2 d was defined as estrus, whereas day of ovulation was estimated using milk progesterone concentrations. Ovulation not preceded by increased walking activity was considered silent ovulation; the incidence was 55.2%, 23.8%, 21.3%, and 10.5% at the first, second, third, and fourth ovulations postpartum, respectively. Moderate and high milk yield significantly increased the risk of silent ovulation at second (odds ratio [OR] = 2.7 and 1.2; P = 0.04) and third and/or fourth ovulations (OR = 6.7 and 12.9; P = 0.03). Based on survival analysis, silent ovulations at the first, second, third, and/or fourth ovulations were associated with 28% (hazard ratio [HR] = 0.72), 55% (HR = 0.45), and 47% (HR = 0.53) reductions in pregnancy rate, respectively, and 41% (HR = 0.59), 66% (HR = 0.34), and 65% (HR = 0.35) reductions in artificial insemination (AI) submission rate. Cows with at least one silent ovulation (with the exception of the first ovulation) had a longer interval from calving to first AI (72 vs. 54 d, P < 0.001) and to achievement of pregnancy (133 vs. 80 d, P < 0.001). In conclusion, approximately one third of the ovulations (based on milk progesterone concentrations) in Holstein cows within 90 d postpartum were silent. Silent ovulations at the second to fourth ovulations were associated with high milk yields and at all ovulations were associated with impaired reproductive performance.     

Quantifying intracellular metabolites in yeast using a matrix with minimal interference from naturally occurring analytes

For quantification of intracellular metabolites, mass spectrometry combined with liquid chromatography, capillary electrophoresis, or gas chromatography is currently the method of choice, especially when combined with stable isotope-labeled internal standards (SIL-ISs). Due to the difficulties in finding a biological matrix free of intracellular metabolites, a standard addition based validation is needed. Here, we present an alternative by producing a matrix with minimal signal interferences on both the analytes and their SIL-ISs. The matrix was obtained by cultivating Saccharomyces cerevisiae in [¹³C₆]glucose/nonlabeled glucose (50:50, w/w) growth medium. The areas of both ¹²C₆ and ¹³C₆ fractions of ATP in the matrix were measured to be 2% of the sum of the areas of all ATP isotopes detected. The matrix allowed for spiking of both the nonlabeled and SIL-ISs and more straightforward validation. The intra- and inter-day accuracy and precision were ?80% and ?20%, respectively. The methodology was used for quantification of nucleotides, coenzymes, and redox compounds from Saccharomyces cerevisiae. The determined energy charge ratio was 0.9, whereas the Mal-CoA/Ac-CoA ratio was 0.04. The analysis of the redox compounds was challenging due to the oxidation of NADH and NADPH, when dissolved in water or tributylamine. The oxidation was reduced by dissolving them in ammonium acetate solution (pH 8.0).     

Insights into different dependence of dNTP triphosphohydrolase on metal ion species from intracellular ion concentrations in Thermus thermophilus

Deoxyribonucleoside triphosphate (dNTP) triphosphohydrolase (dNTPase) from Thermus thermophilus HB8 (TTHB8) hydrolyzes wide variety of dNTPs to deoxyribonucleoside and inorganic triphosphate in magnesium-dependent manner. In this paper, we assess the specificity for various metal ions and of the dNTP triphosphohydrolase activity of the dNTPase from TTHB8. Manganese and cobalt ions more effectively induced the activity for dNTPs than magnesium and, unexpectedly, brought about the degradation of single kind of dNTP. Manganese and cobalt concentrations of 10 nM were enough to induce the activity, while magnesium of about 1 mM was required for the induction of the activity. To further evaluate metal ions inherent to dNTPase in TTHB8 cells, we measured intracellular concentrations of major metal ions in TTHB8 cells by inductively coupled plasma emission spectroscopy and compared them with the dependence of metal ion concentration on dNTPase activity. Though cobalt ion was below detectable level, magnesium and manganese ions were detected at sufficient level to induce dNTPase activity. These results suggest that both manganese and magnesium ions are likely to be functional under intracellular condition. In addition, the proposed model of dNTPase activity induced by magnesium and multiple dNTPs was discussed based on the results obtained in this study.     

International Biodiversity Observation Year in Western-Pacific and Asian regions (DIWPA-IBOY): a case report on species rarity and spatio-temporal variability of species composition in Lepidoptera and Coleoptera communities from a temperate forest of northern Japan

An international project, DIWPA-IBOY, took place for simultaneously observing biodiversity throughout the Western-Pacific and Asian regions in 2001–2003, as one of the core projects for International Biodiversity Observation Year, a crosscutting network activity of DIVERSITAS (an international programme of biodiversity science). DIWPA-IBOY provides extensive data on species diversity obtained by the standardized method. Under this project, 51,742 individuals of Lepidoptera and 11,633 of Coleoptera were collected by light traps from the Tomakomai Experimental Forest of Hokkaido University, one of the core DIWPA-IBOY sites, in the cool-temperate region of northern Japan. Based on these data, this study examined the relative abundance distribution (RAD) to evaluate the amount of rare species in the Lepidoptera and Coleoptera communities. The beta diversities between sampling seasons, forest strata, and trap sites were also assessed to evaluate the spatio-temporal variability of species composition in these communities. In the analysis of the RAD, the best-fit model was selected from the log-Normal, Zipf–Mandelbrot, and Zipf models differing in the tail length of the RAD, i.e., the proportion of rare species. To explore the beta diversity between samples, the abundance-based Jaccard index with an unseen species estimator was calculated, and then a hierarchical clustering analysis was conducted. As a result of RAD analysis, the Coleoptera community was regarded as containing a larger proportion of rare species than the Lepidoptera community. The seasonal compartmentalization of the community, deduced from the beta-diversity analysis, was finer in Lepidoptera (seven assemblages recognized) than in Coleoptera (three assemblages). The spatial (vertical and horizontal) compartmentalization was negligible in both communities. The coincidence of the larger proportion of rare species and the lower beta diversity between seasons in the Coleoptera community was explained by the longer life spans of beetles compared to moths, based on the assumption that the length of life span acts as a temporal agent for mass effect on the analogy of the migration rate as a spatial agent for mass effect.     

Machine learning algorithms based on signals from a single wearable inertial sensor can detect surface- and age-related differences in walking

The aim of this study was to investigate if a machine learning algorithm utilizing triaxial accelerometer, gyroscope, and magnetometer data from an inertial motion unit (IMU) could detect surface- and age-related differences in walking. Seventeen older (71.5 ± 4.2 years) and eighteen young (27.0 ± 4.7 years) healthy adults walked over flat and uneven brick surfaces wearing an inertial measurement unit (IMU) over the L5 vertebra. IMU data were binned into smaller data segments using 4-s sliding windows with 1-s step lengths. Ninety percent of the data were used as training inputs and the remaining ten percent were saved for testing. A deep learning network with long short-term memory units was used for training (fully supervised), prediction, and implementation. Four models were trained using the following inputs: all nine channels from every sensor in the IMU (fully trained model), accelerometer signals alone, gyroscope signals alone, and magnetometer signals alone. The fully trained models for surface and age outperformed all other models (area under the receiver operator curve, AUC = 0.97 and 0.96, respectively; p ≤ .045). The fully trained models for surface and age had high accuracy (96.3, 94.7%), precision (96.4, 95.2%), recall (96.3, 94.7%), and f1-score (96.3, 94.6%). These results demonstrate that processing the signals of a single IMU device with machine-learning algorithms enables the detection of surface conditions and age-group status from an individual’s walking behavior which, with further learning, may be utilized to facilitate identifying and intervening on fall risk.     

Effect of caulerpenyne,a toxin extracted from Caulerpa taxifolia on mechanisms regulating intracellular pH in sea urchin eggs and sea bream hepatocytes

The proliferation of the green marine alga Caulerpa taxifolia in the Mediterranean led us to investigate the toxic effects on marine organisms of caulerpenyne (Cyn), the major secondary metabolite synthesized by the alga. This study was performed on sea urchin eggs (Paracentrotus lividus) and isolated hepatocytes from the sea bream (Sparus aurata), in which accumulation of the toxins by metabolic processes may be of significance. Cyn provoked an acidification of seawater containing both unfertilized and fertilized eggs, as revealed by a titrable efflux of protons. The pHi in unfertilized eggs continuously increased in the presence of Cyn, whereas there was a biphasic response in both fertilized eggs and isolated hepatocytes, with a decrease of the pHi followed by recovery to the initial value. Cyn inhibited the accumulation of ¹⁴C-methylamine in acidic granules present in the cortical zone of sea urchin eggs. Dicyclocarbodiimide (DCCD), a well-known H⁺-ATPase inhibitor, provoked a similar inhibition. Both molecules increased pH in the acidic compartments of isolated bream hepatocytes. These results suggest that Cyn inhibits intracellular sequestration of protons and thus liberates protons into the cell cytoplasm from which they leak toward the extracellular medium. © 1997 John Wiley & Sons, Inc.     

Effect of S20787, a novel Cl--HCO3- exchange inhibitor, on intracellular pH regulation in guinea pig ventricular myocytes

S20787 has recently been proposed to be a selective Cl--HCO3- anion exchange (AE) inhibitor in rat cardiomyocytes. The AE transporter mediates sarcolemmal acid influx but is only one part of the cardiac cell's dual acid loading mechanism, the other part being a sarcolemmal Cl--OH- exchanger (CHE). We have therefore (1) investigated the differential effects of S20787 on the AE and CHE transporters in isolated guinea pig ventricular myocytes and (2) re-examined the influence of the drug on other sarcolemmal acid transporters by monitoring its effect on intracellular pH (pH(i)) recovery from alkali or acid loads. The pH(i) was measured using microspectrofluorimetry (carboxy-SNARF-1). The results indicate that CHE activity was unaffected by the drug (1-20 microM), whereas up to 78% of AE activity was blocked (K(i) = 3.9 microM). Thus, S20787 targets only the AE component of the dual acid influx system. Activities of other acid-transporting carriers, such as Na+-H+ exchange, Na+-HCO3- co-transport and the monocarboxylic acid transporter, were unaffected by the drug. The inhibitory efficacy of S20787 for AE in guinea pig cardiomyocytes appears to be considerably higher (approximately 78%) than proposed previously for rat cardiomyocytes (50%). This is most likely because, in both cells, a significant fraction (20-30%) of acid influx is mediated through the S20787-insensitive CHE transporter. Previous studies made no allowance for the CHE component, which would result in an underestimation. S20787 is thus a highly selective AE inhibitor which may be useful as an experimental tool and a potential cardiac protective agent in the heart.     

The effect of endotoxin lipopolysaccharide from different bacterial species on the generation of intracellular inositol triphosphate and superoxide in a human phagocytic cell line

Abstract The acute effects of endotoxins and lipid A on two intracellular responses, inositol phosphate generation and superoxide production were analysed in the DMSO differentiated premyelocytic leukamic HL-60 cell line. Short-term incubation (1–10 min) with Escheria coli -type LPS, Salmonella -type LPS and Lipid A caused significant increases in cellular InsP₁ and InsP₃, compared with control cells ( P < 0.5 − P < 0.001). The Escheria coli -type B LPS released approximately twice the quantity of InsP₃ compared with Salmonella -type LPS ( P < 0.001). Lipid A-dependent stimulation of InsP₃ production was also detected. The rate of superoxide production increased 1–10 min after addition of both Escheria coli - and Salmonella -type LPS and Lipid A. Endotoxins and Lipid A caused a dose-dependent increase in intracellular oxidative activity. The superoxide response showed less species dependence and a higher response to particulate lipid A compared with the inositol phosphate response.     

Effects of a time varying strong magnetic field on release of cytosolic free Ca2+ from intracellular stores in cultured bovine adrenal chromaffin cells

This study was made to explain the mechanisms for the effects of exposure to a time varying 1.51 T magnetic field on the intracellular Ca(2+) signaling pathway. The exposure inhibited an increase in intracellular Ca(2+) concentration ([Ca(2+)](i)) in bovine chromaffin cells induced by addition of bradykinin (BK) to a Ca(2+) free medium. The exposure did not change BK induced production of inositol 1,4,5-trisphosphate (IP(3)). [Ca(2+)](i) was markedly increased in IP(3) loaded cells, and this increase was inhibited by the magnetic field exposure. A similar increase in [Ca(2+)](i) by other drugs, which stimulated Ca(2+) release from intracellular Ca(2+) stores, was again inhibited by the same exposure. However, transmembrane Ca(2+) fluxes caused in the presence of thapsigargin were not inhibited by the magnetic field exposure in a Ca(2+) containing medium. Inhibition of the BK induced increase in [Ca(2+)](i) by the exposure for 30 min was mostly recovered 1 h after exposure ended. Our results reveal that the magnetic field exposure inhibits Ca(2+) release from intracellular Ca(2+) stores, but that BK bindings to BK receptors of the cell membrane and intracellular inositol IP(3) production are not influenced.