Evaluation of cell wall preparations for proteomics: a new procedure for purifying cell walls from Arabidopsis hypocotyls

Background  

The ultimate goal of proteomic analysis of a cell compartment should be the exhaustive identification of resident proteins; excluding proteins from other cell compartments. Reaching such a goal closely depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific difficulties: (i) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure, (ii) polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins. Several reported procedures to isolate cell walls for proteomic analyses led to the isolation of a high proportion (more than 50%) of predicted intracellular proteins. Since isolated cell walls should hold secreted proteins, one can imagine alternative procedures to prepare cell walls containing a lower proportion of contaminant proteins.     

Hypolipidemic effect in cholesterol-fed rats of a soluble fiber-rich product obtained from cocoa husks

A new soluble cocoa fiber product (SCFP), obtained after enzymatic treatment of cocoa husks, was characterized and its potential health effects studied in an animal model of dietary-induced hypercholesterolemia. The SCFP was rich in soluble dietary fiber (DF) and antioxidant polyphenols. Consumption of a cholesterol-rich diet containing the SCFP as a source of DF resulted in lower food intake and body weight gain in comparison with control groups consuming cholesterol-free or cholesterol-rich diets with cellulose as DF. The cholesterol-rich diet caused remarkable hypercholesterolemia. However, the SCFP diminished the negative impact of the cholesterol-rich diet, buffering the decrease of high density lipoprotein-cholesterol, and the increase of total and low density lipoprotein-cholesterol levels, and lipid peroxidation (malondialdehyde levels) induced by the fatty diet. The SCFP also decreased triglyceride levels to values lower than those in the group fed the cholesterol-free diet. These results put forward the potential application of the SCFP as a dietary supplement or functional food ingredient.     

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Background: Pseudomonas aeruginosa is one of the opportunistic pathogens causing frequent hospital-acquired life-threatening infections in mechanically ventilated patients. The most significant virulence factor of P. aeruginosa is T3SS. PcrV is an important structural protein of the T3SS. Methods:In the current investigation, a recombinant scFv mAb against the PcrV protein was expressed in EnBase® (fed-batch) cultivation mode. The pETiteTM N-His SUMO Kan vector, including anti-PcrV scFv gene, was transformed into Escherichia coli (BL21) cells. The expression and solubility of anti-PcrV scFv protein were investigated at two different temperatures (25 °C and 30 °C) and at different induction times (4, 6, 8, 12, and 24 hours). Results:Increased efficiency was achieved by EnBase® compared to LB broth; owing to the slow release of glucose, the maximum level of solubility and total protein expression was observed in EnBase® cultivation system at 30 °C and 24 h post induction. Furthermore, IC₅₀ for anti-PcrV scFv protein was determined to be approximately 7 μg/mL. Conclusion:Anti-PcrV scFv produced in this study showed promising in vitro results, protecting RBC from lysis by P. aeruginosa (exoU+). Key Words: Fed Batch, recombinant protein, Pseudomonas aeruginosa, scFv     

Quantitative Mapping and Assessment of Environmentally Sensitive Areas to Desertification in Central Iran

Desertification is one of the main environmental and also social and economic problems facing Iran. Seventeen out of 31 Iranian provinces, which are home to approximately 70% of the total population, are affected by desertification. This study aimed to use geographic information system (GIS) and fuzzy logic for mapping environmentally sensitive areas to desertification based on Mediterranean Desertification and Land Use approach in Isfahan province, central Iran. Six desertification indicators including climate, soil, vegetation, soil erosion, groundwater, and management and policy quality were used to determine various types of environmentally sensitive areas to desertification. Seventeen desertification indices affecting the quality of each indicator were spatially mapped and assigned a value between 0 and 1 using a fuzzy logic option of ArcSDM3 software in GIS environment. Results showed that a 21·7% of the study area was classified as critical, 70% as fragile and 5·5% as potential, and 2·9% of the area was not affected by desertification. In the town of Borkhar, 64·2% of the area was classified as critical, followed by the towns of Isfahan and Nayin with 40·2% and 31·8%, respectively. Results at provincial scale indicated that the climate indicator and humidity index with a weighting mean of 0·71 and 0·77 were the most affective factors in the desertification of the study area. The developed model in this study can be used for mapping desertification status in other 16 provinces that contain desert areas. These assessments provide a GIS‐based desertification database that Iran as a member of the United Nation Convention to Combat Desertification can use to report the condition of desertification at national scale. Copyright © 2013 John Wiley & Sons, Ltd.     

Role of GABAB Receptor and L-Arg in GABA-Induced Vasorelaxation in Non-diabetic and Streptozotocin-Induced Diabetic Rat Vessels

Background:

Hypertension is considered an independent risk factor for cardiovascular mortality in diabetic patients. The present study was designed to determine the role of gamma amino butyric acid B (GABA_B) receptor and L-arginine (L-Arg) in GABA-induced vasorelaxation in normal and streptozotocin-induced diabetic rat vessels.

Methods:

Diabetes was induced by a single i.p. injection of streptozotocin (STZ, 60 mg/kg). Eight weeks later, superior mesenteric arteries of all groups were isolated and perfused according to the McGregor method.

Results:

Baseline perfusion pressure of STZ diabetic rats was significantly higher than non-diabetic rats in both intact and denuded endothelium. In the presence of faclofen, a selective GABA_B receptor blocker, GABA-induced relaxation in intact and denuded endothelium mesenteric beds of STZ diabetic rats was suppressed, but this response in non-diabetic rats was not suppressed. Our results showed that in the presence of L-Arg, a nitric oxide precursor, GABA induced vasorelaxation in both diabetic and non-diabetic vessels.

Conclusion:

From the results of this study, it may be concluded that the vasorelaxatory effect of GABA in diabetic vessel is mediated by the GABA_B receptor and nitric oxide, but it seems that in non-diabetic vessel GABA_B receptor does not play any role in GABA-induced vasorelaxation, but nitric oxide induced GABA relaxation in non-diabetic vessel. Key Words: Gamma amino butyric acid (GABA), Diabetes, GABA_B receptor     

Study of structure-activity relationship in Aurein 1.2 analogs

Two new analogs of Aurein 1.2 antimicrobial peptide were synthesized and the antimicrobial activities were investigated. The results showed that the activity of G1R/F3W analog was higher than the native peptide and the F3W analog. Circular dichroism studies also showed that the secondary structure of the F3W was concentration-dependent, whereas, there was no such relationship seen in the case of G1R/F3W analog. It has been proposed that G1R/F3W activity was based on a single mechanism (snorkeling), while Aurein 1.2 and F3W utilized the snorkeling mechanism at low concentrations (0-0.01 mM) and the carpet mechanism at higher concentrations (0.01-0.1 mM). This study suggests that one pay attention to the concentration of biomolecules in peptide-based drug design.     

ULTRASONOGRAPHIC EVALUATION OF THE FETUS AND INTRAUTERINE ENVIRONMENT IN HEALTHY MARES DURING LATE GESTATION

Mares with uncomplicated pregnancies (n = 33) underwent transabdominal sonography to evaluate fetal well-being, obtain measurements of fetal size and characterize the intrauterine environment. Data from the last examination obtained prior to parturition were used for statistical analysis. All mares had one active fetus with good fetal tone. The maximal vertical depth of amniotic fluid (7.9 ± 3.5 cm) was less than allantoic (13.4 ± 4.4 cm) and fewer echogenic particles were detected in amniotic fluid. The maximal uteroplacental thickness was 1.38 ± 0.23 cm (retrospective) and 1.15 ±± 0.24 cm (propspective). In 3 mares small anechoic spaces were imaged between the uterus and placenta. Fetuses had a regular cardiac rhythm with a mean heart rate of 75 ± 7 beats/minute and breathing movements. The diameter of the fetal aorta (mean = 22.8 ± 2.15 mm) was significantly correlated with neonatal foal weight (P<0.0008, r = 0.72) and maternal prepartum weight (P<0.002, r = 0.86). This information of the normal intrauterine environment and fetal well-being can be used to develop a biophysical profile specific for the equine fetus.     

Serologic survey for Leptospira spp. in captive neotropical felids in Foz do Igua?u, Paraná, Brazil

Leptospirosis is a bacterial zoonosis of worldwide distribution and is endemic in tropical countries, where rodents and other wild mammals are abundant and may act as reservoirs. Leptospirosis has become a concern in captive wild animals, due mostly to their exposure to contaminated urine or environment. Although domestic cats (Felis catus) have been reported refractory to leptospirosis, serology and disease in captive wild felids is still unclear. In this study 57 adult, clinically healthy felids, including 1 Geoffroy's cat (Leopardus geoffroyi), 3 jaguarundis (Puma yagouaroundi), 17 margays (Leopardus wiedii), 22 little spotted cats (Leopardus tigrinus), and 14 ocelots (Leopardus pardalis) kept in captivity at the Sanctuary at the Itaipu Binacional hydroelectric power plant (Bela Vista Biological Sanctuary), Foz do Iguacu City, Paraná State, Brazil, were serologically surveyed for the presence of antibodies against 28 serovars of Leptospira spp. by microagglutination test (MAT). Two animals (3.5%) were seropositive: one male ocelot to the serovar Cynopteri (titer 100) and one female margay to Autumnalis (100) and Butembo (200). The captive-born, 5-yr-old ocelot had been solitary housed in an individual cage. The approximately 21-yr-old wild-caught margay was also kept individually. None of the tested animals showed signs ofleptospirosis. During a study conducted 4 yr previously in the same facility, this particular margay also tested positive for the same two serovars, among others. The present study indicates that the felids tested for Leptospira spp. by MAT were exposed to serovars, but did not demonstrate clinical signs of disease. Comparison with a previous study suggests that serovar titers may vary over time and that leptospirosis dynamics remains unclear in wild felids.     

Ozone Formation Potentials of Volatile Organic Compounds and Ozone Sensitivity to Their Emission in the Megacity of São Paulo, Brazil

In the present study, a three-dimensional Eulerian photochemical model was employed to estimate the impact that organic compounds have on tropospheric ozone formation in the Metropolitan Area of São Paulo (MASP). In the year 2000, base case simulations were conducted in two periods: August 22–24 and March 13–15. Based on the pollutant concentrations calculated by the model, the correlation coefficient relative to observations for ozone ranged from 0.91 to 0.93 in both periods. In the simulations employed to evaluate the ozone potential of individual VOCs, as well as the sensitivity of ozone to the VOC/NO _x emission ratio, the variation in anthropogenic emissions was estimated at 15% (according to tests performed previously variations of 15% were stable). Although there were significant differences between the two periods, ozone concentrations were found to be much more sensitive to VOCs than to NO _x in both periods and throughout the study domain. In addition, considering their individual rates of emission from vehicles, the species/classes that were most important for ozone formation were as follows: aromatics with a kOH?>?2?×?10⁴ ppm^?1 min^?1; olefins with a kOH?<?7?×?10⁴ ppm^?1 min^?1; olefins with a kOH?>?7?×?10⁴ ppm^?1 min^?1; ethene; and formaldehyde, which are the principal species related to the production, transport, storage and combustion of fossil fuels.