Solvent tuning of electrochemical potentials in the active sites of HiPIP versus ferredoxin

A persistent puzzle in the field of biological electron transfer is the conserved iron-sulfur cluster motif in both high potential iron-sulfur protein (HiPIP) and ferredoxin (Fd) active sites. Despite this structural similarity, HiPIPs react oxidatively at physiological potentials, whereas Fds are reduced. Sulfur K-edge x-ray absorption spectroscopy uncovers the substantial influence of hydration on this variation in reactivity. Fe-S covalency is much lower in natively hydrated Fd active sites than in HiPIPs but increases upon water removal; similarly, HiPIP covalency decreases when unfolding exposes an otherwise hydrophobically shielded active site to water. Studies on model compounds and accompanying density functional theory calculations support a correlation of Fe-S covalency with ease of oxidation and therefore suggest that hydration accounts for most of the difference between Fd and HiPIP reduction potentials.     

Activation of endogenous Cdc42 visualized in living cells

Signaling proteins are tightly regulated spatially and temporally to perform multiple functions. For Cdc42 and other guanosine triphosphatases, the subcellular location of activation is a critical determinant of cell behavior. However, current approaches are limited in their ability to examine the dynamics of Cdc42 activity in living cells. We report the development of a biosensor capable of visualizing the changing activation of endogenous, unlabeled Cdc42 in living cells. With the use of a dye that reports protein interactions, the biosensor revealed localized activation in the trans-Golgi apparatus, microtubule-dependent Cdc42 activation at the cell periphery, and activation kinetics precisely coordinated with cell extension and retraction.     

Laryngeal function in normal dogs administered isoflurane following partial clearance of alfaxalone or propofol

ObjectiveTo assess laryngeal function in normal dogs administered isoflurane following partial clearance of alfaxalone or propofol.Study designRandomized experimental crossover study.AnimalsA group of 12 purpose-bred, male Beagle dogs.MethodsDogs were randomly assigned to one of two treatments: alfaxalone–isoflurane (ALF-ISO) or propofol–isoflurane (PRO-ISO) and anesthetized for three video laryngoscopy examinations. The alternate treatment occurred after ≥ 14 days interval. Examinations were performed after induction of anesthesia (LS-A), after 20 minutes of breathing isoflurane via a facemask (LS-B) and after a further 20 minutes of isoflurane (LS-C). Parameters of objective laryngeal function included inspiratory rima glottidis surface area (RGSA-I), expiratory rima glottidis surface area (RGSA-E) and % RGSA increase, calculated from three consecutive respiratory cycles in the final 15 seconds of each video laryngoscopy examination. The % RGSA increase was calculated using [(RGSA-I – RGSA-E)/RGSA-E] × 100. Subjective laryngeal function was evaluated independently by two experienced surgeons blinded to treatment.ResultsThe % RGSA increase within each treatment was greater for LS-B and LS-C than for LS-A (ALF-ISO: p = 0.03, PRO-ISO: p = < 0.001). There was no difference within each treatment from LS-B compared with LS-C. RGSA-I increased within each treatment from LS-A to both LS-B and LS-C (ALF-ISO: p = 0.002) and to LS-C (PRO-ISO: p = 0.006). Subjective laryngeal function scores improved from LS-A to LS-C.Conclusions and clinical relevanceLaryngeal function improved from postinduction examination following either 20 or 40 minutes of anesthesia with isoflurane via facemask. This study demonstrates that isoflurane may have a lesser effect on arytenoid abduction activity compared with more commonly used intravenous induction anesthetics (alfaxalone and propofol).     

Field calibration of a neutron moisture meter in a cracking grey clay

Summary A neutron moisture meter was field calibrated in a cracking grey clay prepared for furrow irrigation at Narrabri, N.S.W. Neutron counts were taken in successive 0.1 m increments between the 0 and 1.5 m depths. Concomitant measurements using undisturbed soil cores provided independent estimates of volumetric water content. Separate linear calibrations were required for the 0–0.1 m, 0.1–0.2 m and 0.2–1.5 m depth increments. Correction for bias due to cracking and changes in bulk density slightly improved the calibrations. The accuracy of predicted soil water content was improved relative to previous calibrations. A precision of ±0.01 m³m^–3 required 3 samples per mean by the neutron method or 11 samples per mean by the core sampling method.     

Aroma release and delivery following the consumption of beverages

Processes controlling aroma release and delivery during and after the consumption of a beverage were studied using real-time physiological and aroma release measurements. The key processes were as follows. During swallowing, a portion of the buccal gas phase was transferred first to the throat and then to the nasal passages via the tidal breath flow. This mechanism accounted for the sharp pulse of aroma seen at the beginning of the swallow breath and on subsequent swallows. The persistence effect was due to liquid-air partition from beverage coated on the throat and was dependent on the concentration of volatile compounds in the beverage. Lipid in the beverage caused a decrease in the intensity of volatile compounds on the breath, but the presence of a thickening agent had no effect on persistence.     

Pharmacokinetics of pioglitazone after multiple oral dose administration in horses

Pioglitazone is a thiazolidinedione class of antidiabetic agent with proven efficacy in increasing insulin sensitivity in humans with noninsulin-dependent diabetes mellitus, a syndrome of insulin resistance sharing similarities with equine metabolic syndrome. The purpose of this study was to determine the pharmacokinetics of pioglitazone in adult horses following multiple oral dose administration. Pioglitazone hydrochloride (1 mg/kg) was administered orally for 11 doses at 24-h intervals, and plasma samples were collected. Initially, a pilot study was performed using one horse; and thereafter the drug was administered to six horses. Samples were analyzed by liquid chromatography with tandem mass spectrometry, and pharmacokinetic parameters were calculated using noncompartmental modeling. The maximum plasma concentration was 509.1 ± 413.5 ng/mL achieved at 1.88 ± 1.39 h following oral administration of the first dose, and 448.1 ± 303.5 ng/mL achieved at 2.83 ± 1.81 h (mean ± SD) following the eleventh dose. Apparent elimination half-life was 9.94 ± 4.57 and 9.63 ± 5.33 h after the first and eleventh dose, respectively. This study showed that in healthy horses, pioglitazone administered at a daily oral dose of 1 mg/kg results in plasma concentrations and total drug exposure approximating, but slightly below, those considered therapeutic in humans.     

Disentangling the identities and distribution patterns of the introduced beachfleas Orchestia gammarellus and Platorchestia platensis (Crustacea: Amphipoda: Talitridae) in South Africa

Two introduced beachfleas, Orchestia gammarellus and Platorchestia platensis, have been recorded from South Africa, but have been misidentified by some earlier researchers and, prior to this study, each was also known from just a few, scattered observations, such that their true distributions remained obscure. We illustrate both species to clarify the features that identify and distinguish them and determine their true distributions by re-examining historical samples and by collecting new material from estuaries and lagoons along the west and south coasts of South Africa. Adult males of O. gammarellus have slender antennae and characteristic expanded, flattened, oar-like distal segments on pereopod 7. Adult males of P. platensis have the peduncle of antenna 2 strongly swollen and have bulbous, but not flattened, distal segments on pereopod 7. Both species occur under wrack and other debris along the banks of estuaries and sheltered lagoons. Orchestia gammarellus is found only along the south-west coast, in the Berg River Estuary, Langebaan Lagoon and the Diep River Estuary. Previous records from Knysna were misidentifications of P. platensis. Platorchestia platensis is found at one west coast site, Langebaan Lagoon, but is widely distributed from Cape Point eastwards to at least Algoa Bay. Both species share the same habitats as several other native beachfleas and competitive interactions between native and introduced species merit investigation.     

Evaluation of an Immunochromatographic Test to the Diagnosis of Canine Brucellosis Caused by Brucella canis

This study evaluated the performance of an immunochromatographic test (ICT) for the diagnosis of canine brucellosis caused by Brucella canis, comparing its results with that of the rapid slide agglutination test with and without the use of 2‐mercaptoethanol and the agar gel immunodiffusion test (AGID). The microbiological culture, PCR and clinical examination were used as reference. According to the results obtained in clinical examination, blood culture, culture of semen and vaginal swab and PCR in blood, semen and vaginal swab, a total of 102 dogs were divided into three groups: B. canis‐infected dogs (Group 1), B. canis‐non‐infected dogs (Group 2) and dogs with suspected brucellosis (Group 3). The diagnostic sensitivity of RSAT, 2ME‐RSAT, AGID and ICT in Group 1 was, respectively, 75%, 37.5%, 27.8% and 89.58%. The diagnostic specificity of RSAT, 2ME‐RSAT, AGID and ICT in Group 2 was, respectively, 91%, 100%, 100%, and 100%. In dogs with suspected brucellosis, 9.67% were RSAT positive, none was positive by 2ME‐RSAT, 3.22% were AGID positive and 6.45% were ICT positive. The main drawback concerning canine brucellosis diagnosis is the lack of a highly sensitive serological assay to be used as a screening test to the rapid identification of infected animals. The ICT showed a high diagnostic specificity and a diagnostic sensitivity value greater than that observed in the RSAT, 2ME‐RSAT and AGID. However, 10.41% of infected dogs had negative results by ICT. These dogs were positive by microbiological culture and/or PCR, indicating active infection and consequently a higher potential of spreading Brucella. Although rapid and simple to perform, the ICT lacked sensitivity to be used as a screening test.