A Ca2-activated cation-selective channel in the basolateral membrane of the cortical thick ascending limb of Henle's loop of the mouse

The patch-clamp technique was used to investigate the properties of a cation-selective channel in the basolateral membrane of microdissected collagenase-treated fragments of cortical thick ascending limbs of Henle's loop from mouse kidney. The channel activity was seldom observed in cell-attached patches (2 out 15 studied cases). In inside-out excised patches immersed in symmetrical NaCl Ringer's solutions, the unit channel conductance was ohmic and ranged from 22 to 33 pS (mean, 26.8 +/- 0.6 pS, n = 24). When NaCl was replaced by KCl (n = 8) or sodium gluconate (n = 2) on the cytoplasmic side of the membrane, single-channel currents still reversed at 0 mV and the conductance was unchanged. The reversal potential was +28.8 +/- 0.4 mV (n = 8) when a NaCl concentration (140 vs. 42 mmol/l) gradient was applied, close to the expected value (approx. 30 mV) for a cation selective channel. The channel was found to discriminate poorly between Na+, K+, Cs+, and Li+ ions. The activity of the channel was not clearly voltage-dependent but was dependent upon the free Ca2+ concentration on the cytoplasmic side of the membrane. We conclude that the channel resembles the non-selective cation channel which has been previously described in several tissues.     

Triterpenoid glycosides from the bark of Mimosa tenuiflora.

Two new saponins were isolated from Mimosa tenuiflora and their structures established as 3-O-[alpha-L-rhamnopyranosyl(1----2)-beta-D-glucopyranosyl-(1----3]-(alp ha-L- arabinopyranosyl-(1----4]-beta-D-xylopyranosyl-(1----2)]-[beta-D- xylopyranosyl-(1----4)]-beta-D-glucopyranosyl)-28-O-alpha-L-rhamnopyrano syl oleanolic acid and 3-O-[alpha-L-rhamnopyranosyl-(1----2)-beta-D-glucopyranosyl-(1----3]-(al pha- L-arabinopyranosyl-(1----4]beta-D-xylopyranosyl-(1----2)]-[beta-D- xylopyranosyl-(1----4)]-beta-D-glucopyranosyl) oleanolic acid.     

Transport of chlorine in the proximal tubule. Its effects on water-electrolyte absorption

Several studies in rat kidney have established that an appreciable fraction of proximal absorption is passive in nature and occurs across the highly conductive paracellular pathway. Passive absorption is generally ascribed to the transepithelial Cl- distribution, luminal Cl- activity (alpha lCl) being higher than plasma Cl- activity (alpha pCl). The inequality alpha lCl greater than alpha pCl generates a transepithelial diffusion potential, lumen positive, which taken together with the chemical potential differences of Cl- and Na+ across the epithelium gives rise to transepithelial electrochemical potential differences for Cl- and Na+ favoring their absorption. The alpha lCl greater than alpha pCl distribution is traditionally ascribed to preferential bicarbonate absorption. We argue that HCO3- absorption alone cannot generate a non equilibrium transepithelial Cl- distribution. Other mechanisms are necessary. Our measurements in amphibian proximal tubule demonstrate that the intracellular Cl- activity, alpha cCl, is higher than the theoretical value predicted for equilibrium. This distribution is the result of two basolateral coupled transport processes (Cl-/HCO3- exchange and Cl-/Na+ cotransport). It contributes to the exit of Cl- from cell to lumen (by passive diffusion and K+/Cl- cotransport), yielding alpha lCl values higher than the theoretical value for equilibrium with regard to plasma. Thus, a small transcellular flux of Cl- (without solvent) proceeds from interstitium to lumen. It compensates the dissipative tendency of a much higher paracellular Cl- absorptive flux (in association with water) on the transepithelial Cl- gradient. The result is a steady-state luminal Cl- distribution above equilibrium, along the major part of the proximal tubule.     

Characterization of the mouse ClC-K1/Barttin chloride channel

Several Cl⁻ channels have been described in the native renal tubule, but their correspondence with ClC-K1 and ClC-K2 channels (orthologs of human ClC-Ka and ClC-Kb), which play a major role in transcellular Cl⁻ absorption in the kidney, has yet to be established. This is partly because investigation of heterologous expression has involved rat or human ClC-K models, whereas characterization of the native renal tubule has been done in mice. Here, we investigate the electrophysiological properties of mouse ClC-K1 channels heterologously expressed in Xenopus laevis oocytes and in HEK293 cells with or without their accessory Barttin subunit. Current amplitudes and plasma membrane insertion of mouse ClC-K1 were enhanced by Barttin. External basic pH or elevated calcium stimulated currents followed the anion permeability sequence Cl⁻ > Br⁻ > NO₃⁻ > I⁻. Single-channel recordings revealed a unit conductance of ~ 40 pS. Channel activity in cell-attached patches increased with membrane depolarization (voltage for half-maximal activation: ~ − 65 mV). Insertion of the V166E mutation, which introduces a glutamate in mouse ClC-K1, which is crucial for channel gating, reduced the unit conductance to ~ 20 pS. This mutation shifted the depolarizing voltage for half-maximal channel activation to ~ + 25 mV. The unit conductance and voltage dependence of wild-type and V166E ClC-K1 were not affected by Barttin. Owing to their strikingly similar properties, we propose that the ClC-K1/Barttin complex is the molecular substrate of a chloride channel previously detected in the mouse thick ascending limb (Paulais et al., J Membr. Biol, 1990, 113:253–260).     

On molecular recognition of an uranyl chelate by monoclonal antibodies

The energy landscape of the uranyl (UO2) chelate dissociated from a monoclonal antibody U08S was investigated using dynamic force spectroscopy (DFS). The uranyl ion (UO2(2+)) is chelated with the ligand dicarboxy-phenanthroline (DCP). The monoclonal antibody U08S was raised against UO2-DCP and does not cross-react with DCP alone. The results of plotting the most probable force against the logarithm of the loading rate show two distinguished values of slopes of multiple fitting lines, as observed in our previous study on that system with monoclonal antibody U04S (Odorico et al., 2007a. Biophys. J. 93: 645-654.). It indicates an unbinding process undergoing at least two activation states. We have generated the histogram of unbinding events with respect to the composite stiffness of the complex between the protein and the uranyl compound. Combining the model of Bell and Evans with that of Williams, we have estimated the number of parallel bonds involved in the unbinding process and determined the value of stiffness for individual bonds. We propose that the uranyl compound binds to the two antibodies U04S and U0c at structurally equivalent locations and forms the interaction with similar coordination modes. In addition, the unbinding process goes through two steps; the first weakens the bonding of the central metal with AspL50 of the antibody and the second breaks other non-bonded interactions of the compound with the antibody.     

Olfactory responses of western flower thrips (Frankliniella occidentalis) populations to a non‐pheromone lure

Western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), is a major pest of horticultural crops worldwide. The search for alternative pest management techniques has resulted in increasing interest in the use of kairomones and other behaviour‐modifying chemicals to mitigate the impact of this pest. In this study, we determined whether the origin of populations, feeding history, and/or genotype influence the response of WFT to the thrips kairomone lure methyl isonicotinate (MI) in a Y‐tube olfactometer study. Four New Zealand thrips populations were tested: (1) from a commercial glasshouse capsicum crop, (2) from a long‐established laboratory colony (>222 generations) kept on chrysanthemums, (3) from a laboratory colony (6–9 generations) kept on French dwarf beans, and (4) thought to be a separate cryptic non‐pest species from outdoor yellow tree lupins, Lupinus arboreus Sims (Fabaceae). In the laboratory tests, significantly more WFT from all four populations chose the MI‐laden arm of a Y‐tube olfactometer when it contained 1 μl methyl isonicotinate (61.3–73.2%) compared with the blank no‐odour arm. No differences in response to MI were found between the two laboratory and the one glasshouse WFT populations. Both laboratory populations and the greenhouse population belonged to the ‘glasshouse pest’ genotype of WFT. However, the cryptic non‐pest WFT genotype responded more strongly to MI than any of the other populations, although the response was only significantly stronger than that of the long‐established laboratory population. Significant differences were also found among populations in the average time taken for thrips to make a choice to enter either arm of the Y‐tube olfactometer, with the cryptic non‐pest lupin genotype taking the shortest time, followed by thrips from the capsicum glasshouse. The results are discussed with respect to the variability in olfactory perception and olfactory behaviour within a species and the relevance to the use of such a kairomone lure in pest management programmes.     

Host Age Preference Behavior in <Emphasis Type="Italic">Aphidius ervi</Emphasis> Haliday (Hymenoptera: Aphidiidae)

Many parasitoid species have preference for certain stages of hosts to parasitize but the underlying behavioral mechanisms of such preference are still poorly understood, making it difficult to evaluate host-parasitoid interactions and their effects on the success of biological control programs. Here, we report our work on a parasitoid Aphidius ervi Haliday on the pea aphid Acyrthosiphon pisum (Harris). We show that with the increase of host age, female parasitoids are more likely to encounter and to attack their hosts but the hosts develop increasingly greater defensive capabilities. Encounter almost always triggers attack attempt; however, increasing attack attempts do not proportionally lead to ovipositor probings and increasing ovipositor probings do not proportionally translate into ovipositions. These asymmetric responses may be interpreted as that A. ervi females prefer to parasitize older aphids for higher fitness return but those aphids can better defend themselves, and as a consequence, A. ervi females may achieve the highest gain by attacking aphids of intermediate ages. We suggest that A. ervi females forage in a manner consistent with the optimal foraging theory, trading off host handling time with fitness returns.     

Response of Female <Emphasis Type="Italic">Frankliniella occidentalis</Emphasis> (Pergande) to Visual Cues and <Emphasis Type="Italic">Para-</Emphasis>anisaldehyde in a Flight Chamber

The effect of visual cue color and size, volume of para-anisaldehyde (plant-derived semiochemical), and airflow on thrips behavior were examined in a flight chamber. After 5 min more female Frankliniella occidentalis (western flower thrips) landed on sticky traps containing yellow plastic squares (100 cm²) (55.2% of those that flew landed on the trap) than blue (21%), white (4.7%), or transparent traps (2%). The percentage of thrips caught on traps increased with increasing size of the visual cues (0 and 1 cm² (4%), 4 cm² (16%), 25 cm² (44–49%), 100 cm² (60%)). Using a yellow (100 cm²) square, fewer thrips flew in the presence of 1.0 ml (47%) or 2 ml (55%) of para-anisaldehyde than of 0.5 ml (78%). However, more thrips landed on a trap with a 100 cm² yellow square when 1 ml of para-anisaldehyde (81%) was added than when 0.5 ml (55%) or 2 ml (62%) were added. Airflow (0–0.3 m/s) did not affect the percentage of thrips that flew or landed on traps. Results suggest that thrips responded to a yellow cue in the absence of UV. Further, the volume of para-anisladehyde affected the percentage that flew or landed on a trap containing a yellow cue.     

An ensemble model for predicting Rhopalosiphum padi abundance

A novel modeling method is proposed to predict the abundance of the main vector of barley yellow dwarf virus in autumn sown cereal crops. An ensemble model based on artificial neural networks (ANN) was developed to predict the number of Rhopalosiphum padi (L.) (Homoptera: Aphididae) caught in traps during the autumn flight period at Lincoln, Canterbury, New Zealand, over the period 1982–2003. Artificial neural networks were trained using historical weather data and aphid data collected from a suction trap. Model results were compared with those obtained using multiple regression (MR) models using the same independent variables. Both ANN and MR models were validated by leave‐one‐out validation, in other words, by sequentially jackknifing each year out of the data set, fitting a model to the remaining data, then using that model to predict the number of aphids for each jackknifed year. A linear ensemble of ANN models further improved the predictions and represented the trends in the number of aphids over the 22‐year period very well. The r² between the predicted and observed numbers of aphids for the ANN models changed from 0.68 to 0.83 using the linear ensemble model, but the ensemble approach did not change the prediction for the MR models. The absolute mean error (ABSME) of prediction was much lower for the ANN ensemble predictions compared to that for the MR models. The ABMSE for the ANN models dropped from 109 to 86 aphids compared to an ABMSE reduction from 245 to 220 aphids for the MR models. We discuss the potential for ensemble models for predicting insect abundance when long‐term historical data are available.