Impacts of Fungal Stalk Rot Pathogens on Physicochemical Properties of Sorghum Grain.

Stalk rot diseases are among the most ubiquitous and damaging fungal diseases of sorghum (Sorghum bicolor (L.) Moench) worldwide. Although reports of quantitative yield losses to stalk rots are available, the impact of stalk rot on grain quality attributes is unknown. This study was conducted to test whether stalk rot diseases could affect grain mineral (N, P, K; Ca, Mg, Cu, Fe, Mn, and Zn) and macronutrient (protein, fat, and starch) content, ash content, and physical traits (unit grain weight, hardness, and diameter). A field experiment was conducted in 2013 and 2014 with four sorghum genotypes (two hybrids and two lines). Plants from each genotype were inoculated with four stalk rot pathogens (Fusarium andiyazi, F. proliferatum, F. thapsinum, and Macrophomina phaseolina) and mock-inoculated with phosphate-buffered saline (control). Grains collected from infected and control plants were analyzed for macronutrient and ash content using near-infrared reflectance spectroscopy, grain hardness and diameter using the single-kernel characterization system, and mineral content using the Rapid Flow Analyzer (Model RFA-300 for N) and inductively coupled plasma spectrometer (for P, K, Ca, Mg, Cu, Fe, Mn, and Zn). Although stalk rot pathogens significantly reduced unit grain weight, they did not significantly affect grain hardness and diameter and, therefore, may not affect milling quality. Pathogens significantly reduced all macronutrient and most mineral contents across genotypes and environments on a per-unit-grain basis, except N and Mg, which were affected in a genotype- and environment-specific manner, and Fe, which was not significantly affected. Most minerals tested were significantly and negatively correlated with disease severity (lesion length) and total grain weight per panicle. The hybrid tested (Pioneer 84G62) exhibited reduced mineral and macronutritional changes after stalk rot infection, providing insights into the possibility of producing high-yielding, nutritionally stable hybrids under stalk rot disease pressure through dedicated breeding efforts.

Bioethics consultation practices and procedures: a survey of a large Canadian community of practice.

The literature fails to reflect general agreement over the nature of the services and procedures provided by bioethicists, and the training and core competencies this work requires. If bioethicists are to define their activities in a consistent way, it makes sense to look for common ground in shared communities of practice. We report results of a survey of the services and procedures among bioethicists affiliated with the University of Toronto Joint Centre for Bioethics (JCB). This is the largest group of bioethicists working in healthcare organizations in Canada. The results suggest there are many common services and procedures of JCB bioethicists. This survey can serve as a baseline for further exploration of the work of JCB bioethicists. Common practices exist with respect to the domains of practice, individual reporting relationships, service availability within business hours and the education and training of the bioethicist.

Auditory function in Pelizaeus-Merzbacher disease.

Pelizaeus-Merzbacher disease (PMD; MIM 312080), an inherited defect of central nervous system myelin formation, affects individuals in many ways, including their hearing and language abilities. The aim of this study was to assess the auditory abilities in 18 patients with PMD by examining the functional processes along the central auditory pathways using auditory brainstem responses (ABR) and cortical auditory evoked potentials (CAEP) in response to speech sounds. The significant ABR anomalies confirm the existence of dyssynchrony previously described at the level of the brainstem in patients with PMD. Despite the significant auditory dyssynchrony observed at the level of the brainstem, CAEPs were present in most patients, albeit somehow abnormal in terms of morphology and latency, resembling a type of auditory neuropathy spectrum disorder.

Characterization of polymeric proteins from vitreous and floury sorghum endosperm.

Differences in protein content and composition between vitreous and floury endosperm were investigated using a number of different techniques. Differences in protein cross-linking between vitreous and floury endosperm were investigated using differential solubility, size exclusion chromatography (SEC), and analysis of sulfhydryl content and composition. Vitreous endosperm was found to have higher levels of total protein and kafirins, but floury endosperm had a higher proportion of gamma-kafirins than the vitreous. Floury endosperm was found to have higher levels of SDS-soluble proteins than SDS-insoluble proteins extracted using sonication than vitreous endosperm. Conversely, vitreous endosperm had a greater proportion of the insoluble proteins. SEC analysis of the polymeric proteins revealed that the insoluble proteins had more polymeric proteins than did the soluble proteins, indicating greater cross-linking and a larger Mw distribution. Vitreous endosperm was also found to have a greater percentage (i.e., a higher ratio of disulfide to total sulfhydryls) of disulfide bonds than floury endosperm. These results show that the proteins in vitreous endosperm have a higher degree of cross-linking and a greater Mw distribution than those found in floury endosperm.

Development of a 96-well plate iodine binding assay for amylose content determination.

Cereal starch amylose/amylopectin (AM/AP) is critical in functional properties for food and industrial applications. Conventional methods of AM/AP are time consuming and labor intensive making it difficult to screen the large sample sets necessary for evaluating breeding samples and investigating environmental impact on starch development. The objective was to adapt and optimize the iodine binding assay in a 96-well plate format for measurement at both λ 620 nm and λ 510 nm. The standard curve for amylose content was scaled to a 96-well plate format and demonstrated R(2) values of 0.999 and 0.993 for single and dual wavelengths, respectively. The plate methods were applicable over large ranges of amylose contents: high amylose maize starch at 61.7±2.3%, normal wheat starch at 29.0±0.74%, and a waxy maize starch at 1.2±0.9%. The method exhibited slightly greater amylose content values than the Concanavalin A method for normal type starches; but is consistent with cuvette scale iodine binding assays.

Immunopathology of cicatricial pemphigoid: studies of complement deposition.

Immunopathologic investigations were conducted on the sera and oral mucosal tissue specimens of 23 patients with cicatricial pemphigoid. A linear, continuous basement membrane zone pattern was noted in 83% of oral mucosal biopsy specimens studied. This pattern is indistinguishable from the pattern noted in immunofluorescence studies of bullous pemphigoid, herpes gestationis, and some cases of desquamative gingivitis. Complement studies provided data supportive of classical pathway activation in cicatricial pemphigoid tissue. Deposition of IgA with Factor B, properdin, and C3 raised the possibility of alternative pathway activation, a question requiring further study. Circulating antibasement membrane zone antibodies were noted in the sera of two patients with cicatricial pemphigoid.

Acquired reversible autistic syndrome in acute encephalopathic illness in children.

In seeking the neurologic substrate of the autistic syndrome of childhood, previous studies have implicated the medial temporal lobe or the ring of mesolimbic cortex located in the mesial frontal and temporal lobes. During an acute encephalopathic illness, a clinical picture developed in three children that was consistent with infantile autism. This development was reversible. It was differentiated from acquired epileptic aphasia, and the language disorder was differentiated aphasia. One child has rises in serum herpes simplex titers, and a computerized tomographic (CT) scan revealed an extensive lesion of the temporal lobes, predominantly on the left. The other two, with similar clinical syndromes, had normal CT scans, and no etiologic agent was defined. These cases are examples of an acquired and reversible autistic syndrome in childhood, emphasizing the clinical similarities to bilateral medial temporal lobe disease as described in man, including the Klüver-Bucy syndrome seen in postencephalitic as well as postsurgical states.

Pemphigus involving the eyes.

Two persons with pemphigus vulgaris involving the eyes were studied. In one case, in vivo-fixed intercellular deposition of IgG was demonstrated in a specimen from the conjunctiva.

Pemphigus vegetans of Hallopeau: immunofluorescent studies.

Immunofluorescent techniques were used in the study of tissue from a patient with pemphigus vegetans of Hallopeau. The findings were typical of pemphigus, confirming the propriety of classifying this entity as a benign variant of pemphigus, rather than a disease of purely microbial etiology. The question is raised whether other types of vegetating pyodermas actually are separate diseases or represent a spectrum of pemphigus with the clinical appearance dependent on the patient's immunity to his disease. Immunofluorescent studies are proposed as a basis for classification.

Cicatricial pemphigoid of Brunsting-Perry. Immunofluorescent studies.

Six patients with a vesiculobullous eruption of the type described by Brunsting and Perry as benign pemphigoid were studied by direct and indirect immunofluorescence. All six showed linear deposits of IgG but not IgM or IgA at the epidermal-dermal junction. One case also showed C3 deposition and one patient had circulating antibasement membrane zone antibodies in a titer of 1280. These data provide strong evidence that this condition belongs to the cicatricial pemphigoid-bullous pemphigoid spectrum of disease.

Vesicular pemphigoid.

Seven patients had a chronic eruption of small vesicles that was initially considered to be atypical dermatitis herpetiformis. Histopathologic studies disclosed a subepidermal bulla that was compatible with either bullous pemphigoid or dermatitis herpetiformis. Immunofluorescence studies demonstrated deposition of IgG or C3, Or both, at the basement-membrane zone (BMZ) in six of seven patients. In four patients, circulating BMZ antibodies were detected also. We belive that these cases represent a vesicular form of bullous pemphigoid.

Systemic lupus erythematosus presenting as a bullous eruption in a child.

An 8-year-old girl presented with a generalized bullous eruption clinically resembling bullous pemphigoid or chronic bullous disease of childhood. Further study revealed immunopathologic findings seen in patients with epidermolysis bullosa acquisita or bullous systemic lupus erythematosus (SLE). Although she did not fulfill the American Rheumatism Association (Atlanta) criteria for SLE at her presentation, one year later she went on to do so. As well as being the youngest patient reported with bullous SLE, our patient is noteworthy because the bullous eruption was the initial manifestation of her SLE. Bullous SLE should be considered in the differential diagnosis of children presenting with generalized bullous eruptions.

Optic atrophy following prophylactic chemotherapy and cranial radiation for acute lymphocytic leukemia.

Two patients with acute lymphocytic leukemia developed progressive optic nerve and chiasmal lesions eight to nine months after the initiation of identical chemotherapy protocols that included intrathecal medication and prophylactic radiation of only 2,400 rads to the central nervous system. Both patients eventually lost all vision despite additional radiotherapy, and there was no evidence of leukemia involving the central nervous system after acute lymphocytic leukemia was diagnosed. Optic nerve biopsy in one case showed changes consistent with radiation necrosis.

Immunofluorescent studies of ocular cicatricial pemphigoid.

Using direct and indirect immunofluorescent techniques, we studied ten patients with cicatricial pemphigoid (chronic cicatricial conjunctivitis). Serologic and histologic evaluation of 13 conjunctival, 12 mucous membrane, and 11 skin specimens from the ten patients showed that three had immunoglobulin deposition at the basement membrane zone of the conjunctiva. One patient had immunoglobulin deposited in the affected skin and oral mucosa. Two patients had circulating basement membrane zone antibodies that bound to their own tissue but not to guinea pig lip or human skin.

Electrophoresis of cereal storage proteins.

Cereal proteins have been studied by a number of analytical techniques over the years. One of the major methodologies utilized by cereal chemists has been electrophoresis. Starting with moving boundary electrophoresis and progressing to slab gels and high-performance capillary electrophoresis, innovative methods have been developed to provide high resolution separations of difficult to separate proteins. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), acid-PAGE, isoelectric focusing, free zone CE, and even high-resolution two-dimensional HPLC-HPCE methods have been developed to separate cereal proteins. This review focuses on electrophoretic methods for separating and characterizing cereal storage proteins.

High-performance capillary electrophoresis of cereal proteins.

Cereal grains are widely used of human foods and animal feed throughout the world. Cereals provide dietary protein, which also often has a functional role, as wheat gluten does in bread. Cereal proteins are unique in many ways: they are highly complex and heterogeneous, are often difficult to extract, and aggregate readily, making them difficult to characterize. Because of the economic importance and widespread use of cereal proteins, however, many techniques have been used for their analysis. High-performance capillary electrophoresis (HPCE) is one of the newest techniques to be so used. This review describes the development of charge- and size-based HPCE methods for analysis of cereal grain proteins, and the use of these methods for cultivar identification, classification, and prediction of quality. HPCE is versatile, rapid, easily automated, readily quantified, and provides high-resolution separations. Clearly, HPCE is a valuable addition to other methods of cereal protein analysis and should, in time, be applicable to all protein classes from all cereals.

Diagnosis and management of chronic oral mucosal bullous diseases.

Chronic blistering diseases involving the mouth are frequently difficult to diagnose correctly and can present a problem with management. Pemphigus vulgaris, cicatricial pemphigoid, and recurrent oral erythema multiforme may be difficult to separate from each other and from other erosive diseases such as recurrent aphthous stomatitis and erosive lichen planus. Essential to establishing a correct diagnosis is a pertinent history, a physical examination, and histopathologic and direct immunofluorescence studies. Oral pemphigus vulgaris can usually be managed successfully with prednisone alone but occasionally another immunosuppressive medication in concurrence with prednisone is necessary. Dapsone is the most efficacious treatment for oral cicatricial pemphigoid, and brief courses of prednisone usually control individual episodes of severe recurrent oral erythema multiforme. It is important that physicians who undertake treatment of these diseases be fully aware of the untoward effects of the medications that are being utilized.

A simple system for pea transformation.

The lateral cotyledonary meristems of germinatingPisum sativum cv. Puget seeds were used to develop a reproducibleAgrobacterium tumefaciens-mediated transformation system. This procedure exhibits distinct advantages over those previously reported, in that it uses dry seed as starting material, and the highly regenerable cotyledonary meristems rapidly produce transgenic shoots without an intermediate callus phase. This transformation regime facilitates the rapid generation of phenotypically normal, self-fertile plants containing functional transgenes inherited in a Mendelian fashion.

Sodium dodecyl sulfate capillary electrophoresis of wheat proteins. 1. Uncoated capillaries.

Four different polymer/buffer systems (a commercial polymer from Bio-Rad, dextran, poly(ethylene oxide) (PEO), and non-crosslinked poly(acrylamide)) were evaluated for use in sodium dodecyl sulfate capillary electrophoresis (SDS-CE) separations of wheat proteins. These polymers were chosen on the basis of published reports of their use in uncoated or dynamically coated capillaries. Each polymer was optimized (where possible) by manipulating the polymer concentration and buffer concentration, and through the use of organic modifiers such as methanol and ethylene glycol. The addition of ethylene glycol to the separation buffer was found to improve the resolution of the separations, despite dilution of the sieving polymers. When PEO was used as the sieving polymer, however, no improvement was seen when ethylene glycol was added. Despite producing similar separations of molecular mass markers, the polymers did not all produce similar wheat protein separations. The commercial reagent and dextran produced similar separations, while the poly(acrylamide) produced faster separations than either. The poly(acrylamide) displayed much lower resolution in the 40-60 kDa range than the other polymers, though this polymer was able to separate the high molecular mass glutenin subunits (HMM-GS) without the use of added organic solvent. PEO produced much different wheat protein separations than the other polymers, despite similar separations of the molecular weight markers. This may have been due to interaction between the wheat proteins and PEO. Each polymer system also predicted different molecular masses of the various wheat protein fractions separated, with the PEO and poly(acrylamide) grossly overestimating the masses for all protein classes. This could have been due to protein-polymer interactions. Further work was done with the Bio-Rad buffer modified by the addition of ethylene glycol. Several different wheat protein fractions as well as proteins extracted from several different cultivars were separated with this buffer and compared. SDS-CE separations were also compared to SDS-poly(acrylamide) gel electrophoresis (PAGE) and several differences in the migration pattern of HMM-GS were noted.