Selective one-step extraction of Arabidopsis thaliana seed oleosins using organic solvents

Oleosins are hydrophobic proteins from oleaginous seeds, surrounding and stabilizing oil bodies. They are known to display interesting interfacial properties. Specific sera were raised against four different A. thaliana oleosins and used in dot-blot assays for oleosin quantification. These assays were used to set up extraction of oleosins from A. thaliana seeds. One mixture of chloroform/methanol gave optimal oleosin extraction. Extracted proteins represented 9% of seed proteins and were identified by immunoblot and proteomic analyses. Oleosins accounted for 79% of the extracted proteins. This simple one-step procedure allows selective extraction and concentration of oleosins from seeds without tedious oil body purification. Oleosin extract was indeed used to demonstrate the presence of the rare oleosin S5 in mature seeds. Moreover, this method will be useful to investigate the potential use of oleosins as emulsifier and to question their possible allergenicity.     

Transcutaneous Oxygen and Carbon Dioxide Monitoring in Normal Cats

Feasibility of noninvasive oxygenation and ventilation monitoring using continuous transcutaneous oxygen (PO₂-_TC) and carbon dioxide (PCO₂-_TC) measurements was investigated in six healthy adult male cats anesthetized with isoflurane. Concurrent arterial blood gases, inspired oxygen concentration (FIO₂), end-tidal carbon dioxide (ETCO₂), PO₂-_TC were recorded during hyperoxia/normocapnia, normoxia/normocapnia, hypoxia/normocapnia, hypocapnia/hyperoxia, and hypercapnia/hyperoxia. Dorsolateral thorax and dorsal pelvis probe sites were evaluated. Probe site did not significantly affect the parameters. During normoxia and hypoxia, mean PO₂-_TC was insignificantly greater than mean PaO₂ (p > 0.154), but during hyperoxia PO₂-_TC was less than Pao₂ (p < 0.002). At each assessment (except hypercapnia for the dorsal pelvis probe site) PCO₂- _TC was greater than PaCO₂. Correlations between PO₂-_TC and FIO₂ (p < 0.05), and PaO₂ (p <0.001), and between PCO₂-_TCand ETCO₂ (p < 0.001), and PaCO₂ (p < 0.05) were good; however, the measured values of PO₂-_TC and PCO₂-_TC were not directly comparable to the measured values of PaO₂ and PaCO₂, respectively. Clinical utility of transcutaneous monitoring in cats will require development of the appropriate conversion equation for carbon dioxide and modifications for practical application.     

Antitrypanosomal activity of polycarpol from Piptostigma preussi (Annonaceae)

Polycarpol, sitosterol and sitosterol-3-O-β-D-glucoside isolated for the first time from Piptostigma preussi (Annonaceae) occur regularly in some Annonaceae such as Piptostigma genus. Polycarpol exhibits interesting antitrypanosomal activity with an ED₅₀ value of 5.11 µM on Trypanosoma brucei cells. Moreover, it inhibits T. brucei glycolytic enzymes GAPDH and PFK with IC₅₀ values of 650 and 180 µM respectively.     

Evaluation of the discriminatory power of variable number tandem repeat (VNTR) typing of Mycobacterium bovis strains

The discriminatory power of variable number tandem repeat (VNTR) typing based on 16 known loci (12 MIRUs, 3 ETRs and VNTR 3232) was assessed for Mycobacterium bovis strains collected sequentially at the slaughterhouse of N'Djaména, Chad. Of 67 M. bovis strains analyzed, 67% were clustered. In this study, VNTR typing was highly discriminative with an overall allelic diversity (h(oa)) of 0.922. We defined five loci (ETR A, B, C and MIRU 26, 27) as highly (h>0.25), two loci (MIRU 4, and VNTR 3232) as moderately (0.11相似文献   

Malignant hyperthermia in a horse anesthetized with halothane

Ahealthy, 565-kg, 14-year-old Quarter Horse gelding, negative for equine protozoal myelitis on serum Western blot and for hyperkalemic periodic paralysis (HYPP) by polymerase chain reaction, was selected for use in an electrophysiology study performed under inhalation anesthesia. The study was performed by following the guidelines of an Animal Use and Care Committee Protocol of the University of California at Davis (UCD 8578). Anesthesia was induced with halothane delivered via a face-mask. The horse was intubated 17 minutes later with an orotracheal tube and anesthesia was maintained with halo-thane by using a semiclosed, large-animal breathing circuit. Although muscle relaxation was otherwise good, the horse's ears remained tensed in a caudal direction throughout the study     

Spiroplasma citri Spiralin Acts In Vitro as a Lectin Binding to Glycoproteins from Its Insect Vector Circulifer haematoceps

ABSTRACT In order to understand the molecular mechanisms underlying transmission of Spiroplasma citri by the leafhopper Circulifer haematoceps, we screened leafhopper proteins as putative S. citri-binding molecules using a spiroplasma overlay assay of protein blots (Far-western assay). Insect proteins were separated by one- or two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, blotted, and probed with S. citri proteins. In this in vitro assay, we found that spiroplasma proteins exhibited affinity for seven leafhopper proteins. The interactions between S. citri proteins and insect proteins with molecular masses of 50 and 60 kDa were found to be sugar sensitive. These insect proteins were identified as high mannose N-glycoproteins, which support an interaction of glycoprotein-lectin type with S. citri proteins. Lectin detection in S. citri has revealed only one protein of 24 kDa. Using a leafhopper protein overlay assay on an S. citri protein blot, one spiroplasma protein with a similar molecular mass of 24 kDa was shown to display an insect protein-binding capacity. This protein was identified as the spiralin, which is the most abundant membrane protein of S. citri. Far-western experiments performed with purified spiralin and insect glycoproteins confirmed the binding of spiralin to the insect glycoproteins of 50 and 60 kDa. Thus, the spiralin could play a key role in the transmission of S. citri by mediating spiroplasma adherence to epithelial cells of insect vector gut or salivary gland.     

Comparison of different tube materials and use of Chinese finger trap or four friction suture technique for securing gastrostomy, jejunostomy, and thoracostomy tubes in dogs

Objective— To compare Chinese finger trap (CFT) and 4 friction suture (FFS) techniques to secure gastrostomy (GT), jejunostomy (JT), and thoracostomy (TT) tubes of different materials.
Study Design— Prospective experimental study.
Animals— Canine cadavers (n=20).
Methods— Randomly, GT (n=20), JT (20), and TT (20) were inserted using 2 different suture techniques (10 for each tube type) and either silicone or another material (10 for each type). Axial distraction was applied to each tube until failure. Force and displacement to failure and failure mode were recorded and compared between techniques and materials for GT, JT, and TT.
Results— CFT failed most commonly by suture breakage whereas FFS failed mainly by tube slippage ( P =.003). For GT, failure occurred more commonly by tube slippage (n=15; P <.001) whereas tube breakage was more common for JT (n=10; P <.001) and suture breakage for TT (n=14; P =.022). Silicone had higher force to failure than latex with GT, lower force to failure than red rubber with JT, and lower displacement to failure than polyvinylchloride with TT.
Conclusions— Different failure modes occurred for CFT (suture breakage) and FFS (tube slippage) and among different tube types (tube slippage with GT, tube breakage with JT, and suture breakage with TT). Based on study results, CFT is preferred to FFS for anchoring silicone GT and TT. Silicone GT, red rubber JT, and polyvinylchloride TT were more secure than latex GT, silicone JT, and silicone TT, respectively.
Clinical Relevance— CFT should be preferred over FFS based on force and displacement to failure, but tube type and tissue reaction could influence anchoring strength.     

Effects of temporal floral resource availability and non-crop habitats on broad bean pollination

Landscape Ecology - Flowering plants can enhance wild insect populations and their pollination services to crops in agricultural landscapes, especially when they flower before the focal crop....