Visualizing signals moving in cells

Cells display a highly complex spatiotemporal organization, required to exert a wide variety of different functions, for example, detection, processing, and propagation of nerve impulses by neurons; contraction and relaxation by muscle cells; movement by leukocytes; and adsorption and secretion of nutrients and metabolites by epithelial cells lining the gut. Successful execution of these complex processes requires highly dynamic information transfer between different regions and compartments within cells. Through the development of fluorescent sensors for intracellular signaling molecules coupled with improved microscopic imaging techniques, it has now become possible to investigate signal propagation in cells with high spatial and temporal resolution.     

Electromyographic activity of the hyoepiglotticus muscle and control of epiglottis position in horses

OBJECTIVE: To determine whether the hyoepiglotticus muscle has respiratory-related electromyographic activity and whether electrical stimulation of this muscle changes the position and conformation of the epiglottis, thereby altering dimensions of the aditus laryngis. ANIMAL: 6 Standardbred horses. PROCEDURE: Horses were anesthetized, and a bipolar fine-wire electrode was placed in the hyoepiglotticus muscle of each horse. Endoscopic images of the nasopharynx and larynx were recorded during electrical stimulation of the hyoepiglotticus muscle in standing, unsedated horses. Dorsoventral length and area of the aditus laryngis were measured on images obtained before and during electrical stimulation. Electromyographic activity of the hyoepiglotticus muscle and nasopharyngeal pressures were measured while horses exercised on a treadmill at 50, 75, 90, and 100% of the speed that produced maximum heart rate. RESULTS: Electrical stimulation of the hyoepiglotticus muscle changed the shape of the epiglottis, displaced it ventrally, and significantly increased the dorsoventral length and area of the aditus laryngis. The hyoepiglotticus muscle had inspiratory activity that increased significantly with treadmill speed as a result of an increase in phasic and tonic activity. Expiratory activity of the hyoepiglotticus muscle did not change with treadmill speed in 4 of 6 horses. CONCLUSIONS AND CLINICAL RELEVANCE: Findings reported here suggest that contraction of the hyoepiglotticus muscle increases dimensions of the airway in horses by depressing the epiglottis ventrally during intense breathing efforts. The hyoepiglotticus muscle may be an important muscle for dilating the airway in horses, and contraction of the hyoepiglotticus muscle may induce conformational changes in the epiglottis.     

Unilateral shunt formation with thoracic aortic dissection in a whippet

A three‐year‐old neutered male whippet was presented with intermittent, exercise‐induced paraparesis. Femoral pulses were bilaterally absent. Neurologic examination was suggestive of a thoracolumbar myelopathy. Blood pressure measurements revealed hypotension in both pelvic limbs, hypertension in the right thoracic limb and it was immeasurable in the left thoracic limb. Echocardiography was within reference limits. A clear vascular pulsation was palpable on the right ventral abdominal wall. Computed tomographic angiography revealed a dissection of the aortic wall between the left subclavian artery and the brachiocephalic trunk with subsequent thrombus formation. A shunt between the right internal thoracic, cranial and caudal epigastric arteries to preserve blood flow to the pelvic limbs was visualized. Necropsy was declined by the owner. This is the first case report describing the formation of a unilateral vascular shunt following a thoracic aortic occlusion, which presented as exercise‐induced paraparesis .     

Classification of Ilex species based on metabolomic fingerprinting using nuclear magnetic resonance and multivariate data analysis

The metabolomic analysis of 11 Ilex species, I. argentina, I. brasiliensis, I. brevicuspis, I. dumosavar. dumosa, I. dumosa var. guaranina, I. integerrima, I. microdonta, I. paraguariensis var. paraguariensis, I. pseudobuxus, I. taubertiana, and I. theezans, was carried out by NMR spectroscopy and multivariate data analysis. The analysis using principal component analysis and classification of the (1)H NMR spectra showed a clear discrimination of those samples based on the metabolites present in the organic and aqueous fractions. The major metabolites that contribute to the discrimination are arbutin, caffeine, phenylpropanoids, and theobromine. Among those metabolites, arbutin, which has not been reported yet as a constituent of Ilex species, was found to be a biomarker for I. argentina,I. brasiliensis, I. brevicuspis, I. integerrima, I. microdonta, I. pseudobuxus, I. taubertiana, and I. theezans. This reliable method based on the determination of a large number of metabolites makes the chemotaxonomical analysis of Ilex species possible.     

The effect of Lumbricus rubellus and Lumbricus terrestris on zinc distribution and availability in artificial soil columns

This study investigated the impact of epigeic and (epi)anecic earthworms on the distribution and availability of zinc in the soil profile. Experiments were carried out with Lumbricus rubellus and Lumbricus terrestris in perspex columns (Ø 10 cm), filled with 20 to 23 cm non-polluted soil [organic matter 2%, clay 2.9%, pH 6.4 (0.01 M CaCl₂)], that was covered by a 3- to 5-cm layer of aged zinc-spiked soil (500 mg Zn/kg dry soil) and another 2 cm non-polluted soil on top. After 80 days, columns were sacrificed and sampled in a depth profile. Earthworm casts, deposited on top of the soil, were collected. Each sample was analyzed for total and 0.01 M CaCl₂-exchangeable zinc concentrations. L. rubellus did not go deeper than 3 cm into the soil and therefore no effect on zinc distribution in the soil could be detected. For L. terrestris, total zinc concentrations in the non-polluted layers were slightly but significantly higher in columns with earthworms, and so were the CaCl₂-exchangeable zinc concentrations in the polluted layers of these columns. Casts of L. terrestris collected from the soil surface showed higher total zinc concentrations than those from non-polluted soil. Casts were mainly placed on top of the soil. This study showed that these epigeic and (epi)anecic species have only a slight effect on zinc availability, and that deep burrowing species, like L. terrestris, are able to transport polluted soil from deeper layers to the soil surface.     

Degradation of starchy endosperm cell walls in nongerminating sterilized barley by fungi

Strains of fungi from different origins, including isolates of the natural microflora of barley, were screened for their ability to modify barley starchy endosperm cell walls in situ. In an initial step, fungi were selected that degrade the major component of the cell walls, that is, (1-->3),(1-->4)-beta-D-glucan, in vitro on artificial media. Nongerminating, sterilized barley, obtained by gamma-irradiation, was inoculated with such fungi and subjected to solid state fermentation under conditions resembling those of a traditional malting process. For some strains of fungi, a clear correlation between the production of endo-beta-glucanase and the friability of the treated kernels was found. Image analysis of Calcofluor stained longitudinal sections of barley kernels fermented with the endo-beta-glucanase producing strains showed that starchy endosperm cell walls were modified. As malt quality is inversely related to its (1-->3),(1-->4)-beta-D-glucan content, the selected strains have high potential to be used as starter cultures during malt production, contributing to the processing quality of the final product.     

The effect of two endogeic earthworm species on zinc distribution and availability in artificial soil columns

The objective of this study was to determine the impact of earthworm bioturbation on the distribution and availability of zinc in the soil profile.Experiments were carried out with Allolobophora chlorotica and Aporrectodea caliginosa in 24 perspex columns (∅ 10 cm), filled with 20-23 cm non-polluted soil (OM 2%, clay 2.9%, pH 0.01 M CaCl₂ 6.4), that was covered by a 3-5 cm layer of aged zinc spiked soil (500 mg Zn/kg dry soil) and another 2 cm non-polluted soil on top. After 80 and 175 days, columns were sacrificed and each cm from the top down to a depth of 15 cm was sampled. Earthworm casts, placed on top of the soil, were collected. Each sample was analyzed for total and CaCl₂-exchangeable zinc concentrations.Effects of earthworm bioturbation were most pronounced after 175 days. For A. chlorotica, total and CaCl₂-exchangeable zinc concentrations in the polluted layers were lower with than without earthworms. Total zinc concentrations in the non-polluted layers were higher in columns with earthworms. Casts of A. chlorotica collected on the soil surface showed slightly higher total zinc concentrations than non-polluted soil. Casts were found throughout the whole column. For A. caliginosa there were no differences in total zinc concentration between columns with and without earthworms. CaCl₂-exchangeable zinc concentrations in the polluted layers were lower for columns with earthworms. Casts were mainly placed on top of the soil and contained total zinc concentrations intermediate between those in non-polluted and polluted soil layers.This study shows that different endogeic earthworm species have different effects on zinc distribution and availability in soils. A. chlorotica transfers soil throughout the whole column, effectively mixing it, while A. caliginosa decreases metal availability and transfers polluted soil to the soil surface.     

The V-antigen of Yersinia forms a distinct structure at the tip of injectisome needles

Many pathogenic bacteria use injectisomes to deliver effector proteins into host cells through type III secretion. Injectisomes consist of a basal body embedded in the bacterial membranes and a needle. In Yersinia, translocation of effectors requires the YopB and YopD proteins, which form a pore in the target cell membrane, and the LcrV protein, which assists the assembly of the pore. Here we report that LcrV forms a distinct structure at the tip of the needle, the tip complex. This unique localization of LcrV may explain its crucial role in the translocation process and its efficacy as the main protective antigen against plague.