Activity of a KATP+ channel in Arum spadix mitochondria during thermogenesis

This report demonstrates that mitochondria isolated from thermogenic Arum spadices possess an ATP-sensitive potassium channel--responsible for electrical potential (DeltaPsi) collapse and mitochondrial swelling--whose characteristics are similar to those previously described in pea and wheat mitochondria. In order to study the relationship between this K(ATP)(+) channel and the uncoupled respiration, linked to thermogenesis, K(+) transport activities were compared with those of mitochondria that were isolated from pea stems, soybean suspension cell cultures and Arum tubers. The channel from Arum spadices is highly active and its major features are (i) potassium flux is performed primarily in an inward-rectifying manner; (ii) the influx of K(+) is associated with a matrix volume increase in both energized and non-energized mitochondria; and (iii) its activity depends on the redox state of electron transport chain (ETC) and oxygen availability. In particular, this paper shows that the K(ATP)(+) channel is inwardly activated in parallel with the alternative oxidase (AO). The activation is linked to an ETC-oxidized state and to high oxygen consumption. The putative role of this K(ATP)(+) channel is discussed in relation to flowering of thermogenic Arum spadices.     

Glutamate-induced glioma cell proliferation is prevented by functional expression of the glutamate transporter GLT-1

A tetracycline-dependent inducible system was used to achieve controlled expression of the glutamate transporter 1 (GLT-1) in C6 glioma cells. Non-induced cells show modest glutamate uptake and, in the presence of L-cystine, these cells tend to release substantial amounts of glutamate. Overnight exposure to doxycycline increased D-[3H]-aspartate uptake, reaching similar capacity as observed in cultured astrocytes. Efficient clearance of exogenously applied glutamate was evidenced in these cells, even in the presence of l-cystine. The addition of glutamate (100 microM) to the medium of non-induced cells significantly increased their proliferation rate, an effect that was blocked when the expression of GLT-1 was induced. This suggests that impaired glutamate uptake capacity in glioma cells indirectly contributes to their proliferation.     

Epithelial Sodium Channel in a Human Trophoblast Cell Line (BeWo)

Atomic force microscopy was used to image Bacillus thuringiensis (Bt) toxins interacting with their natural targets, Manduca sexta midgut brush border membranes (BBMs), as well as with dipalmitoylphosphatidylcholine-dioleoylphosphatidylcholine (DPPC-DOPC) solid-supported lipid bilayers. In lipid bilayers, Cry1Aa formed structures 30-60 nm wide and 3-7 nm high, mostly at the interface of domains formed by the two different lipids or at the edge of DOPC-enriched domains. BBM vesicles, in the absence of toxin, formed flat membrane fragments of up to 25 microm(2) and 4.2 nm high, with irregular embedded structures. After incubation with Cry1Aa, Cry1Ac and Cry1C, which are active against M. sexta, new structures, 35 nm wide and 5.1-6.7 nm high, were observed in some membrane fragments, sometimes only in particular regions. Their density, which reached a plateau within 4 h, was toxin- and concentration-dependent. The structures formed by Cry1Ac were often grouped into dense, two-dimensional arrangements. No such specific interactions were observed with Cry1Ba, which is inactive against M. sexta. This study provides the first visual demonstration of specific interactions of Bt toxins with insect midgut BBMs at the nanometric scale. The observed structures likely represent the protein complexes forming functional Bt pores in target membranes.     

Protonation and hydrogen bonding of Ca2+ site residues in the E2P phosphoenzyme intermediate of sarcoplasmic reticulum Ca2+-ATPase studied by a combination of infrared spectroscopy and electrostatic calculations

Protonation of the Ca²⁺ ligands of the SR Ca²⁺-ATPase (SERCA1a) was studied by a combination of rapid scan FTIR spectroscopy and electrostatic calculations. With FTIR spectroscopy, we investigated the pH dependence of CO bands of the Ca²⁺-free phosphoenzyme (E2P) and obtained direct experimental evidence for the protonation of carboxyl groups upon Ca²⁺ release. At least three of the infrared signals from protonated carboxyl groups of E2P are pH dependent with pK_a values near 8.3: a band at 1758 cm⁻¹ characteristic of nonhydrogen-bonded carbonyl groups, a shoulder at 1720 cm⁻¹, and part of a band at 1710 cm⁻¹, both characteristic of hydrogen-bonded carbonyl groups. The bands are thus assigned to H⁺ binding residues, some of which are involved in H⁺ countertransport. At pH 9, bands at 1743 and 1710 cm⁻¹ remain which we do not attribute to Ca²⁺/H⁺ exchange. We also obtained evidence for a pH-dependent conformational change in β-sheet or turn structures of the ATPase. With MCCE on the E2P analog E2(), we assigned infrared bands to specific residues and analyzed whether or not the carbonyl groups of the acidic Ca²⁺ ligands are hydrogen bonded. The carbonyl groups of Glu⁷⁷¹, Asp⁸⁰⁰, and Glu⁹⁰⁸ were found to be hydrogen bonded and will thus contribute to the lower wave number bands. The carbonyl group of some side-chain conformations of Asp⁸⁰⁰ is left without a hydrogen-bonding partner; they will therefore contribute to the higher wave number band.     

Triple mutation of Cys26, Trp35, and Cys126 in stromal ascorbate peroxidase confers H2O2 tolerance comparable to that of the cytosolic isoform

Ascorbate peroxidase (APX) isoforms localized in the stroma and thylakoid of the chloroplast play a principle role in detoxifying hydrogen peroxide (H₂O₂) generated in photosystem I; however, once the ascorbate is depleted, the enzyme is attacked by H₂O₂ and rapidly loses its activity. Here, we report that radical transfer across the porphyrin moiety and amino acid residues in the reaction intermediate and H₂O₂-mediated enzyme inactivation involve cooperative interactions of the Cys26, Trp35, and Cys126 residues of stromal APX. The wild-type enzyme had a half-time of inactivation of <10 s, while the triple mutant of the three residues retained 50% of the initial activity after H₂O₂ treatment for 3 min. The H₂O₂ tolerance of this mutant was comparable to that of the H₂O₂-tolerant APX isoform localized in the cytosol.     

IKs通道新型阻断剂293B对犬离体右心房心率和收缩力的影响

Chromanol2 93B( 2 93B)是一种作用于缓慢激活的延迟整流钾电流 (IKs)的新型选择性阻断剂。本实验采用犬离体血液灌流右心房标本 ,观察了 2 93B对心房标本心率及收缩力的影响。结果表明 ,2 93B降低犬离体右心房血液灌流标本的心率和收缩力 ,2 93B的负性频率和肌力作用与M胆碱能受体无关 ,此外 ,2 93B对 β -受体激动剂的正性频率和肌力作用没有影响。上述结果提示 ,在犬右心房组织中 ,2 93B的负性频率和负性肌力作用与M胆碱能受体和 β-受体无关。     

Immediate Effect of Specific Nostril Manipulating Yoga Breathing Practices on Autonomic and Respiratory Variables

The effect of right, left, and alternate nostril yoga breathing (i.e., RNYB, LNYB, and ANYB, respectively) were compared with breath awareness (BAW) and normal breathing (CTL). Autonomic and respiratory variables were studied in 21 male volunteers with ages between 18 and 45 years and experience in the yoga breathing practices between 3 and 48 months. Subjects were assessed in five experimental sessions on five separate days. The sessions were in fixed possible sequences and subjects were assigned to a sequence randomly. Each session was for 40 min; 30 min for the breathing practice, preceded and followed by 5 min of quiet sitting. Assessments included heart rate variability, skin conductance, finger plethysmogram amplitude, breath rate, and blood pressure. Following RNYB there was a significant increase in systolic, diastolic and mean pressure. In contrast, the systolic and diastolic pressure decreased after ANYB and the systolic and mean pressure were lower after LNYB. Hence, unilateral nostril yoga breathing practices appear to influence the blood pressure in different ways. These effects suggest possible therapeutic applications.     

Drosophila RalA is essential for the maintenance of Jak/Stat signalling in ovarian follicles

Small GTPases of the Ras-like (Ral) family are crucial for signalling functions in both normal and cancer cells; however, their role in a developing organism is poorly understood. Here, we identify the Drosophila Ral homologue RalA as a new key regulator of polar-cell differentiation during oogenesis. Polar cells have a crucial role in patterning the egg chamber and in recruiting border cells, which undergo collective and guided migration. We show that RalA function is essential for the maintenance of anterior and posterior polar-cell fate and survival. RalA is required cell autonomously to control the expression of polar-cell-specific markers, including the Jak/Stat ligand Unpaired. The loss of RalA also causes a cell non-autonomous phenotype owing to reduced Jak/Stat signalling in neighbouring follicle cells. As a result, border-cell assembly and migration as well as the polarization of the oocyte are defective. Thus, RalA is required in organizing centres to control proper patterning and migration in vivo.     

Boundary coding in the rat subiculum

The spatial mapping function of the hippocampal formation is likely derived from two sets of information: one based on the external environment and the other based on self-motion. Here, we further characterize ‘boundary vector cells’ (BVCs) in the rat subiculum, which code space relative to one type of cue in the external environment: boundaries. We find that the majority of cells with fields near the perimeter of a walled environment exhibit an additional firing field when an upright barrier is inserted into the walled environment in a manner predicted by the BVC model. We use this property of field repetition as a heuristic measure to define BVCs, and characterize their spatial and temporal properties. In further tests, we find that subicular BVCs typically treat drop edges similarly to walls, including exhibiting field repetition when additional drop-type boundaries are added to the testing environment. In other words, BVCs treat both kinds of edge as environmental boundaries, despite their dissimilar sensory properties. Finally, we also report the existence of ‘boundary-off cells’, a new class of boundary-coding cells. These cells fire everywhere except where a given BVC might fire.