Production of gluten-free wheat starch by peptidase treatment

The potential of peptidase-containing bran extracts from germinated cereals (wheat, emmer, barley) and a peptidase preparation from Aspergillus niger (AN-PEP) to degrade gluten in wheat starch below the threshold for gluten-free foods of 20 mg/kg was compared. The gluten-specific peptidase activity of the peptidases was determined by using gliadin as a protein-based substrate as well as the two celiac-active peptides PQPQLPYPQPQLPY (α-gliadin) and SQQQFPQPQQPFPQQP (γ-hordein). The peptidase activity of AN-PEP exceeded the activities of bran from germinated cereals by a factor up to 690,000. Three wheat starches with initial gluten contents of 110, 1679, and 2070 mg/kg, respectively, were incubated with bran extracts and AN-PEP, lyophilized, and residual gluten was quantitated by a competitive ELISA. Unlike peptidases from bran extracts, AN-PEP was capable of degrading gluten below 20 mg/kg in all starches. The absence of gluten in AN-PEP-treated starches was confirmed by liquid-chromatography-mass spectrometry. The properties of gluten-free starches were comparable to the native starches with the exception of a reduced viscosity after AN-PEP treatment. This problem could be overcome by using higher enzyme concentrations and shorter incubation times or by optimizing AN-PEP production for lower residual α-amylase activity.     

Modulation of the immunogenicity of the Trypanosoma congolense cysteine protease, congopain, through complexation with ��2-macroglobulin

The protozoan parasite Trypanosoma congolense is the main causative agent of livestock trypanosomosis. Congopain, the major lysosomal cysteine proteinase of T. congolense, contributes to disease pathogenesis, and antibody-mediated inhibition of this enzyme may contribute to mechanisms of trypanotolerance. The potential of different adjuvants to facilitate the production of antibodies that would inhibit congopain activity was evaluated in the present study. Rabbits were immunised with the recombinant catalytic domain of congopain (C2), either without adjuvant, with Freund’s adjuvant or complexed with bovine or rabbit α₂-macroglobulin (α₂M). The antibodies were assessed for inhibition of congopain activity. Rabbits immunised with C2 alone produced barely detectable anti-C2 antibody levels and these antibodies had no effect on recombinant C2 or native congopain activity. Rabbits immunised with C2 and Freund’s adjuvant produced the highest levels of anti-C2 antibodies. These antibodies either inhibited C2 and native congopain activity to a small degree, or enhanced their activity, depending on time of production after initial immunisation. Rabbits receiving C2-α₂M complexes produced moderate levels of anti-C2 antibodies and these antibodies consistently showed the best inhibition of C2 and native congopain activity of all the antibodies, with maximum inhibition of 65%. Results of this study suggest that antibodies inhibiting congopain activity could be raised in livestock with a congopain catalytic domain-α₂M complex. This approach improves the effectiveness of the antigen as an anti-disease vaccine candidate for African trypanosomosis.     

Food-purchasing patterns for home: a grocery store-intercept survey

OBJECTIVES: To identify the most common frequency of food-purchasing patterns and relate this pattern to characteristics of individuals and families.DESIGN: A customer-intercept survey was conducted in the greater Houston area, Texas, USA, in 2002. The frequency of food shopping at supermarkets, convenience stores and restaurants to buy food for eating at home was assessed.SUBJECTS: A total of 823 adults (78.5% female; mean age 37.4 years) who went to any of several grocery or convenience stores, including European, Hispanic and African Americans, and Asian/Pacific Islanders.RESULTS: Major food-shopping patterns were a weekly big trip with a few small trips (34.9%), biweekly big trips with a few small trips (21.9%), no big shopping trips (15.4%), a weekly big trip without small trips (13.9%), a monthly big trip (8.3%), and biweekly big trips without small trips (6.4%). While 61.1% of participants never went to convenience stores to buy fruit and vegetables (F&V) for eating at home, 67% went to restaurants for F&V. African American families shopped for food least frequently, while Asian American families shopped for food most frequently. Educational level was negatively associated with the use of convenience stores and positively associated with take-away from restaurants.CONCLUSIONS: There is substantial variability in the frequency of food shopping. Future research on food shopping should incorporate this variable.     

Use of multigeneration-family molecular dog leukocyte antigen typing to select a hematopoietic cell transplant donor for a dog with T-cell lymphoma

CASE DESCRIPTION: A 7-year-old Golden Retriever was examined because of anorexia, lethargy, vomiting, and gradual weight loss. CLINICAL FINDINGS: Splenomegaly, pancytopenia, high serum calcium concentration, and high alkaline phosphatase activity were detected. Magnetic resonance imaging revealed an enlarged mesenteric lymph node and increased signals from the bone marrow of the ilium and vertebral bodies. Histologic examination and immunophenotyping of biopsy specimens confirmed a stage V (b) T-cell malignant lymphoma. TREATMENT AND OUTCOME: Clinical remission was attained by use of 2 chemotherapy cycles, followed by an allogeneic hematopoietic cell transplant performed at 18 weeks after diagnosis. A donor was identified by molecular dog leukocyte antigen typing methods. The patient was conditioned with 2 fractions of 4 Gy total body irradiation delivered 3 hours apart at 7 cGy/min, followed by an IV infusion of recombinant canine granulocyte colony-stimulating factor mobilized leukapheresis product and postgrafting immunosuppression with cyclosporine. Chimerism analyses revealed full donor engraftment that has been maintained for at least 58 weeks after transplant. Remission has been confirmed by normal results of serum thymidine kinase assays and the absence of peripheral blood clonal T-cell receptor gene rearrangements. CLINICAL RELEVANCE: Systemic chemotherapy induces remissions; however, most dogs succumb to disease recurrence because of multidrug resistance. Outcome of allogeneic hematopoietic cell transplantation in dogs can be excellent because of improved donor-recipient selection by use of molecular dog leukocyte antigen typing, compared with early attempts, and better prevention of graft versus host disease, better supportive care, and substitution of peripheral blood mononuclear cells for bone marrow.     

Drug delivery systems: entering the mainstream

Drug delivery systems (DDS) such as lipid- or polymer-based nanoparticles can be designed to improve the pharmacological and therapeutic properties of drugs administered parenterally. Many of the early problems that hindered the clinical applications of particulate DDS have been overcome, with several DDS formulations of anticancer and antifungal drugs now approved for clinical use. Furthermore, there is considerable interest in exploiting the advantages of DDS for in vivo delivery of new drugs derived from proteomics or genomics research and for their use in ligand-targeted therapeutics.     

Framing and reframing the environmental risks and economic benefits of ethanol production in Iowa

Recent research exposing environmental and social externalities of biofuels has undermined the earlier national consensus that they would provide climate mitigation and rural development benefits, but support for ethanol remains strong in Iowa. The objective of this paper is to understand how stakeholder groups in Iowa have framed the benefits and risks associated with ethanol’s impact on the local economy and environment. Our case study draws on in-depth, semi-structured interviews conducted with key informants from agricultural organizations, environmental organizations, and government departments in the state. We find that in Iowa, widespread support for ethanol production exists among government, energy, and farm groups, and that they frame ethanol production as economically beneficial to rural communities and agriculture, while minimizing the possibility of associated environmental risks. Although participants from environmental organizations in Iowa express apprehension about the environmental impacts of expanded corn ethanol production, their unease is muted in relation to economic benefits and in relation to other environmental issues, and few have publicly voiced their concerns. To understand these findings, we draw from the environmental sociology literature that examines the role of powerful natural resource interests in framing the importance of resource extraction and commodity production to community identity and economy and in delegitimizing and naturalizing associated environmental issues and problems. We argue that powerful natural resource interests in Iowa both naturalize environmental problems related to ethanol production and engage in diversionary reframing to emphasize the economic benefits while minimizing or rejecting the potential environmental risks.