Western Blot Analysis of Proacrosin/Acrosin in Frozen Dog Sperm During In Vitro Capacitation

Acrosin is an acrosomal protease synthesized as an inactive precursor, proacrosin, which is processed via autoproteolysis into active forms alpha- and beta-acrosin. In this paper, a comparative study on the immunoreactivity of acrosin during in vitro capacitation of frozen and fresh (control) canine sperm using Western blot analysis is reported. Semen samples were obtained by digital stimulation and ejaculates processed as fresh and frozen samples and then capacitated for 0, 30, 60 and 90 min. At each time period, samples were analyzed with monoclonal antibody C5F10 by Western blot. The antibody specifically recognized, in fresh and frozen/thawed spermatozoa, a 40-, 32- and 27-kDa bands corresponding to proacrosin, alpha- and beta-acrosin, respectively, during capacitation. Western immunoblots showed that the beta-acrosin reactivity in fresh sperm was directly proportional to the time of capacitation, whereas a decreased reactivity of active form of acrosin was observed with frozen-thawed sperm (p   < 0.05). These results suggest that proacrosin is activated to beta-acrosin earlier in frozen/thawed dog spermatozoa than in fresh dog spermatozoa.     

The antihistaminic chlorpheniramine inhibitsin vitro growth of several fungi isolated from harvested fruits

Chlorpheniramine (CPA) is an antihistaminic that changes the conformation of DNA and inhibits polyamine biosynthesis in mammalian cells. In the present work, we tested the effect of CPA on four genera of fungi species (Altemaria alternata, Botrytis cinerea, Cladosporium cladosporioides and threePenicillium spp.) grownin vitro. Similar growth inhibitions of these genera were produced by 0.5 raM iprodione, CPA and histidinol, but CPA was the most effective. The CPA sensitivities of the twoB. cinerea strains were different. Putrescine did not restore the fungal growth inhibited by CPA.     

Proteins and Carbohydrates from Red Seaweeds: Evidence for Beneficial Effects on Gut Function and Microbiota

Based on their composition, marine algae, and namely red seaweeds, are good potential functional foods. Intestinal mucosal barrier function refers to the capacity of the intestine to provide adequate containment of luminal microorganisms and molecules. Here, we will first outline the component of seaweeds and will summarize the effects of these on the regulation of mucosal barrier function. Special attention will be paid to unique components of red seaweeds: proteins and derived peptides (e.g., phycobiliproteins, glycoproteins that contain “cellulose binding domains”, phycolectins and the related mycosporine-like amino acids) together with polysaccharides (e.g., floridean starch and sulfated galactans, such as carrageenans, agarans and “dl-hybrid”) and minerals. These compounds have been shown to exert prebiotic effects, to regulate intestinal epithelial cell, macrophage and lymphocyte proliferation and differentiation and to modulate the immune response. Molecular mechanisms of action of peptides and polysaccharides are starting to be elucidated, and evidence indicating the involvement of epidermal growth factor receptor (EGFR), insulin-like growth factor receptor (IGFR), Toll-like receptors (TLR) and signal transduction pathways mediated by protein kinase B (PKB or AKT), nuclear factor-κB (NF-κB) and mitogen activated protein kinases (MAPK) will also be summarized. The need for further research is clear, but in vivo experiments point to an overall antiinflammatory effect of these algae, indicating that they can reinforce membrane barrier function.     

The genome of the diatom Thalassiosira pseudonana: ecology, evolution, and metabolism

Diatoms are unicellular algae with plastids acquired by secondary endosymbiosis. They are responsible for approximately 20% of global carbon fixation. We report the 34 million-base pair draft nuclear genome of the marine diatom Thalassiosira pseudonana and its 129 thousand-base pair plastid and 44 thousand-base pair mitochondrial genomes. Sequence and optical restriction mapping revealed 24 diploid nuclear chromosomes. We identified novel genes for silicic acid transport and formation of silica-based cell walls, high-affinity iron uptake, biosynthetic enzymes for several types of polyunsaturated fatty acids, use of a range of nitrogenous compounds, and a complete urea cycle, all attributes that allow diatoms to prosper in aquatic environments.     

Bioaccumulation of Copper by Zea mays: Impact on Root, Shoot and Leaf Growth

In the present study, the growth and the Cu²⁺accumulation by roots, shoots and leaves of Zea mays were examined using copper sulphate in the range of 10^?4 to 10^?2 M. Plants of Z. mays did not show inhibition of growth in the presence of 10^?4 to 10^?2 M Cu²⁺; however, it was observed growth effects on root when different Cu²⁺ solution concentrations were used. Only the seedlings exposed to 10^?2 M exhibited substantial root growth reduction, yielding only 56% of length with respect to the control. Seedlings exposed to 10^?4 M Cu²⁺ exhibited 16% and 42% growth increase in shoots and leaves, respectively, when compared with the controls. The seedlings treated with 10^?3 and 10^?2 M Cu²⁺ were inhibited in shoot and leaf growth. The fresh weights in roots, shoots and leaves significantly decreased at 10^?2 M Cu²⁺. The tolerance index, based on root length, was not significantly different for the three different treatments with copper. However, the total accumulation rate was very low at 10^?4 and 10^?3 M compared to 10^?2 Cu treatments. The capacity of copper accumulation by roots, shoots and leaves of Z. mays plants increased concomitant to the copper concentration, arriving to 382 times more in roots, 157 in shoots and only 16 in leaves, all compared to the controls. Cu could be accumulated by roots, shoots and leaves when the initial concentrations were 10^?3 and 10^?4 M. However, when it was 10^?2 M, the metal could not be accumulated by leaf and shoot levels; the roots could increase their copper accumulation capacity three times compared to the control. Z. mays has potential ability to accumulate Cu without being overly sensitive to Cu toxicity.     

Validation of a Commercial Enzyme Immunoassay for Detection of Clostridium difficile Toxins in Feces of Horses with Acute Diarrhea

Background: Clostridium difficile infection (CDI) is a recognized cause of colitis in the horse. Identification of its toxins is important for management of individual cases and for prevention of transmission and zoonosis. In humans, CDI diagnosis is performed with enzyme immunoassays, none of which have been validated for horses. Hypothesis/Objectives: (1) Establish which test for CDI diagnosis was more frequently used by diagnostic laboratories, (2) determine the identified test's performance, sensitivity, and specificity, and (3) validate its use in diarrheic horses. Animals: Samples were obtained from 72 horses presented with acute diarrhea and hospitalized at the Ontario Veterinary College, University of Guelph. Methods: A survey was conducted to establish which of the tests for CDI diagnosis in horses is most commonly used throughout North America. A questionnaire was sent to all laboratories registered in the Veterinary Infection Control Society and the American Association of Veterinary Laboratory Diagnosticians. The performance of the test was evaluated by comparison to a cell cytotoxicity assay (CTA), the accepted Gold Standard for C. difficile toxin detection. Results: The Techlab C. difficile Tox A/B II ELISA was the most frequently used test. Compared with the CTA, no significant difference was observed, and a good level of agreement (93%) was obtained. The diagnostic performance of the ELISA test was adequate (84% sensitivity and 96% specificity). Conclusions and Clinical Importance: Results demonstrate that the Techlab C. difficile Tox A/B II ELISA is a reliable, adequate, and practical tool for identification of C. difficile toxins in horse feces.     

Field trial measuring the compatibility of methoxyfenozide and flonicamid with Orius laevigatus Fieber (Hemiptera: Anthocoridae) and Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) in a commercial pepper greenhouse

BACKGROUND: Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) and Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) are among the most serious pests of sweet peppers in greenhouses. Chemical control is difficult because of their high reproductive rates and insecticide resistance, and seasonal inoculative releases of Orius laevigatus (Fieber) and Amblyseius swirskii (Athias‐Henriot) are commonly used to reduce their populations. As chemical treatments are often needed in the crop against other pests, the side effects of methoxyfenozide (an insect growth regulator against lepidopteran pests) and flonicamid (a selective feeding inhibitor against sucking insects) were studied in both beneficial organisms in a commercial greenhouse. RESULTS: Orius laevigatus and A. swirskii were released at commercial rates (4–5 and 100 m^?2), and a strong establishment and a very homogeneous distribution were reached. One pesticide treatment with the maximum field recommended concentration of methoxyfenozide and flonicamid (96 and 100 mg AI L^?1) was done when they were well established, and their population levels were not affected either immediately or up to 30 days after treatment. CONCLUSION: The results are indicative of no impact of methoxyfenozide and flonicamid on the two natural enemies in the field, and both can be considered as potential alternatives to be included in IPM programmes in sweet pepper. Copyright © 2011 Society of Chemical Industry