Relationships of Quality Characteristics with Size‐Exclusion HPLC Chromatogram of Protein Extract in Soft White Winter Wheats

This study investigated relationships between molecular weight distributions of unreduced grain proteins and grain, flour, and end‐use quality characteristics of soft white winter wheats grown in Oregon. Absorbance area and area percentage values of protein fractions separated by size‐exclusion HPLC (SE‐HPLC) showed significant correlations with quality characteristics, indicating associations of molecular weight distributions of proteins with quality characteristics. Specifically, high molecular weight polymeric protein fractions appeared to have a detrimental effect on soft wheat quality. This was shown by significant positive correlations with single kernel hardness index, and mixograph water absorption and tolerance, and negative correlations with break flour yield, cookie diameter, and cake volume. Higher proportions of soluble monomeric protein fraction eluted after the main gliadin peak, were associated with soft wheat quality due to negative associations with single kernel hardness index and mixograph water absorption and tolerance, and positive associations with break flour yield, cookie diameter, and cake volume. Calibration models were developed by the application of multivariate analyses to the SE‐HPLC data. These models explained >90% of the variation in mixograph water absorption and cookie diameter and thickness.     

Effects of Impact Dehuller Rotor Speed on Dehulling Characteristics of Diverse Oat Genotypes Grown in Different Environments

Commercial processing of oats for human consumption generally requires impact dehulling to isolate groats from the hull. Impact dehulling involves feeding oat grain into the center of a spinning rotor that expels the grain against the walls of the dehuller. The force of the impact breaks the hull from the groat. We have tested the effect of rotor speed on dehulling efficiency, groat breakage, and unbroken groat yield on 18 oat genotypes from replicated plots in six different environments. Dehulling efficiency and groat breakage increased with rotor speed with all genotypes and environments, but there were significant genotypic and environmental effects as well. In general, genotypes with higher test weight and kernel density dehulled more efficiently at slower rotor speeds. Oat genotypes with higher oil and protein concentrations in their groats tended to break less during dehulling. Oats from hotter, drier environments suffered greater groat breakage. Maximal unbroken groat yield represented a balance between dehulling efficiency and groat breakage, but groat proportion and dehulling efficiency appeared to be the most important factors contributing to groat yield.     

Transduction and Transplantation of Spermatogonia into the Testis of Ram Lambs through the Extra-testicular Rete

Spermatogonial transplantation will provide a new way to study spermatogenesis in domestic animals, disseminate male genetics and produce transgenic animals, if efficiency can be improved. We evaluated a 'surgical' method for transplanting donor cells into testes of ram lambs, where the head of the epididymis is reflected, and a catheter introduced into the extra-testicular rete testis. We also tested transduction of ram spermatogonia with a lentiviral (LV) vector as a means to identify permanent colonization, and introduce genes into donor cells. Eight ram lambs, 11- to 13-week olds, were the recipients: in five, spermatogonia were injected into one testis, and the contralateral testis was an un-manipulated control: in two, spermatogonia were injected into one testis and the contralateral was sham-injected: in one, both testes were injected. Six lambs received spermatogonia labelled with a cell-tracking dye and these were collected 1 or 2 weeks after transplantation; three lambs received spermatogonia transduced with a LV vector driving the expression of enhanced Green Fluorescence Protein and these were collected after 2 months. Donor cells were detected by immunohistochemistry in tubules of seven of nine recipient testes. Approximately 22% of tubule cross-sections contained donor cells immediately after transplantation, and 0.2% contained virally transduced cells 2 months after transplantation. The onset of spermatogenesis was delayed, and there were lesions in both injected and sham-injected testes. Despite the effects of the surgery, elongated spermatids were present in one recipient testis 2 months after surgery. The results suggest that, after modifying the surgical and transduction techniques, this approach will be a means to produce good colonization by donor spermatogonia in sheep testes.     

Isolation of Canine Mammary Cells With Stem Cell Properties and Tumour-Initiating Potential

Recent data suggest that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self-renewal or by progenitor cells that have acquired the capacity for cell self-renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long-term non-adherent culture conditions. We showed that cells obtained from spheres that display self-renewing properties, have multi-lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour-initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue.     

Relationships of High Molecular Weight Glutenin Subunit Composition and Molecular Weight Distribution of Wheat Flour Protein with Water Absorption and Color Characteristics of Noodle Dough

Colors of noodle doughs made from hard white winter wheat flours from Oregon were measured at optimum noodle water absorptions (NWA). Partial correlations, removing effect of protein concentration, indicated that NWA had negative relationships with 0 hr L* and 24 hr b*, and positive relationships with 0 and 24 hr a*. Kernel hardness index had positive simple and partial correlations with NWA without any significant (P < 0.05) correlation with color parameters. High molecular weight glutentin subunits (HMW‐GS) significantly (P < 0.05) affected all measured noodle parameters except for 0 hr L*. Covariance analysis, using protein concentration as a covariate, indicated that HMW‐GS significantly affected NWA and a* (P < 0.01). Wheat cultivars with HMW‐GS 17+18 showed significantly higher mean NWA and a* values than those with alternative Glu‐B1 subunits. Protein molecular weight distributions affected noodles, as shown by significant correlations with absorbance areas and % areas of protein size exclusion (SE) HPLC chromatograms. Protein fractions that had positive correlations with redness had negative correlations with yellowness. Applying multivariate analyses to SE‐HPLC data to derive calibration models to predict fresh noodle dough a* and b* values had R² > 0.91 and cross validations values of R² > 0.75.     

Variation in kernel characteristics and protein molecular weight distribution of Langdon durum-wild emmer wheat chromosome substitution lines

Triticum turgidum L. var. dicoccoides (DIC) provides a useful source of genes to improve agronomic and quality characteristics of durum wheat. Research was performed to identify DIC chromosomes that carry useful genes for quality improvement. Langdon-T. dicoccoides substitution lines consisting of 13 lines based on DIC accession PI 481521, 10 lines on PI 478742, and 2 lines on Israel A were evaluated for kernel characteristics and protein molecular weight distribution. DIC chromosomes 3A from PI 481521, and 1A and 7A from PI 478742 increased kernel hardness. Chromosome 2A from PI 481521 increased kernel weight, which resulted in increased semolina yield. Some substituted DIC chromosomes also affected grain protein concentration and protein molecular weight distribution. For example, chromosomes 2A, 5B, and 7B from PI 481521, and 6B from Israel A increased total protein concentration, which was primarily attributed to an increase of SDS soluble gliadins. Chromosome 6B from PI 478742 was unique in that it led to an increase in SDS insoluble high molecular weight polymeric proteins, which contributed to increased dough mixing strength. Results from this research indicated that chromosome 6B from PI 478742 is a potential gene source to improve dough characteristics of durum wheat by increasing insoluble high molecular weight polymeric protein concentration.     

Experimental studies of the role of the little raven (Corvus mellori) in surveillance for West Nile virus in Australia

Objective To study the potential role of an Australian corvid, the little raven (Corvus mellori), in the surveillance for exotic West Nile virus (WNV) in Australia. Method In a series of trials, little ravens were infected with WNV (strain 4132 New York 1999) and Kunjin virus (strain K42886) by the intramuscular route. They were observed for 20 days during which blood and swab samples were taken for virus isolation. Tissue samples were taken from ravens humanely killed during the acute infection period, and at the termination of the trials, for virus isolation, histopathology and immunohistochemistry. Results Ravens infected with WNV became mildly ill, but all recovered and seroconverted. Blood virus titres peaked around 3 to 4 days after inoculation at levels between 10^3.0 to 10^7.5 plaque forming units/mL. Virus or viral antigen was detected in spleen, liver, lung, kidney, intestine, testis and ovary by virus isolation and/or immunohistochemistry. WNV was detected in oral and cloacal swabs from 2 to 7 days post inoculation. The molecular and pathogenic characteristics of the inocula were consistent with them being of high virulence, as expected for this isolate. Ravens infected with Kunjin virus developed viraemia and seroconverted, although they did not develop disease. Conclusions Little ravens do not develop severe disease in response to virulent WNV infection and for this reason may not be important sentinel hosts in the event of an outbreak of WNV, as in North America. However, as they have relatively high viraemias, they may be able to support virus cycles.     

EFFECTS OF ANESTHETIC PROTOCOL ON NORMAL CANINE BRAIN UPTAKE OF 18F‐FDG ASSESSED BY PET/CT

The purpose of this study was to assess the effects of four anesthetic protocols on normal canine brain uptake of 2‐deoxy‐2‐[¹⁸F]fluoro‐d ‐glucose (FDG) using positron emission tomography/computed tomography (PET/CT). Five clinically normal beagle dogs were anesthetized with (1) propofol/isoflurane, (2) medetomidine/pentobarbital, (3) xylazine/ketamine, and (4) medetomidine/tiletamine–zolazepam in a randomized cross‐over design. The standard uptake value (SUV) of FDG was obtained in the frontal, parietal, temporal and occipital lobes, cerebellum, brainstem and whole brain, and compared within and between anesthetic protocols using the Friedman test with significance set at P<0.05. Significant differences in SUVs were observed in various part of the brain associated with each anesthetic protocol. The SUV for the frontal and occipital lobes was significantly higher than in the brainstem in all dogs. Dogs receiving medetomidine/tiletamine–zolazepam also had significantly higher whole brain SUVs than the propofol/isoflurane group. We concluded that each anesthetic protocol exerted a different regional brain glucose uptake pattern. As a result, when comparing brain glucose uptake using PET/CT, one should consider the effects of anesthetic protocols on different regions of the glucose uptake in the dog's brain.     

IMAGING DIAGNOSIS—FDG‐PET/CT OF A CANINE SPLENIC PLASMA CELL TUMOR

An 8‐year‐old Shih Tzu developed abdominal pain and hyperglobulinemia. A round splenic mass was noted radiographically and sonographically. The patient was evaluated by fluorodeoxyglucose positron emission tomography coupled with computed tomography (FDG‐PET/CT). There was no evidence of metastasis or bone marrow involvement on PET/CT images. The standardized uptake value (SUV) of the splenic mass was increased over the reference range (SUV=4.83). The patient was diagnosed as splenic extramedullary plasmacytoma through immunohistopathologic study. After the splenectomy, the globulin level normalized and the patient is alive without complications.     

Deleterious Effects of Progestagen Treatment in VEGF Expression in Corpora Lutea of Pregnant Ewes

The aim of the current study was to determine the possible effects of progestagen oestrous synchronization on vascular endothelial growth factor (VEGF) expression during sheep luteogenesis and the peri‐implantation period and the relationship with luteal function. At days 9, 11, 13, 15, 17 and 21 of pregnancy, the ovaries from 30 progestagen treated and 30 ewes cycling after cloprostenol injection were evaluated by ultrasonography and, thereafter, collected and processed for immunohistochemical evaluation of VEGF; blood samples were drawn for evaluating plasma progesterone. The progestagen‐treated group showed smaller corpora lutea than cloprostenol‐treated and lower progesterone secretion. The expression of VEGF in the luteal cells increased with time in the cloprostenol group, but not in the progestagen‐treated group, which even showed a decrease between days 11 and 13. In progestagen‐treated sheep, VEGF expression in granulosa‐derived parenchymal lobule capillaries was correlated with the size of the luteal tissue, larger corpora lutea had higher expression, and tended to have a higher progesterone secretion. In conclusion, the current study indicates the existence of deleterious effects from exogenous progestagen treatments on progesterone secretion from induced corpora lutea, which correlate with alterations in the expression of VEGF in the luteal tissue and, this, presumably in the processes of neoangiogenesis and luteogenesis.