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Ingvarsson and Hollobaugh (2011) investigated tact- or echoic-to-intraverbal transfer of stimulus control to “wh” questions for three preschool-aged boys with autism. The current study was a systematic replication of this study with an adolescent girl with Down syndrome. A multielement design was used to compare the effectiveness and efficiency of picture or echoic prompts presented on an iPad or in vivo to teach “wh” questions. All prompt conditions were effective. Conclusions and recommendations for practice are presented.

Flours from five spelt cultivars grown over three years were evaluated as to their breadbaking quality and isolated starch properties. The starch properties included amylose contents, gelatinization temperatures (differential scanning calorimetry), granule size distributions, and pasting properties. Milled flour showed highly variable protein content and was higher than hard winter wheat, with short dough-mix times indicating weak gluten. High protein cultivars gave good crumb scores, some of which surpassed the HRW baking control. Loaf volume was correlated to protein and all spelt cultivars were at least 9–51% lower than the HRW control. Isolated starch properties revealed an increase in amylose in the spelt starches of 2–21% over the hard red winter wheat (HRW) control. Negative correlations were observed for the large A-type granules to bread crumb score, amylose level, and final pasting viscosity for cultivars grown in year 1999 and to pasting temperature in 1998 samples. Positive correlations were found for the small B- and C-type granules relative to crumb score, loaf volume, amylose, and RVA final pasting viscosity for cultivars grown in 1999, and to RVA pasting temperature for samples grown in 1998. The environmental impact on spelt properties seemed to have a greater effect than genetic control.

Starch was isolated from flour of four wheats representing hard red winter (Karl), hard red spring (Gunner), durum (Belfield 3), and spelt (WK 86035-8) wheat classes. Digital image analysis (IA) coupled with light microscopy was used to determine starch size distributions where the volume of granules was calculated as spherical particles or oblate spheroids. Starch granules were classified into three size ranges: A-type granules (> 15 µm), B-type granules (5–15 µm), and C-type granules (<5 µm). An error was noted in using digital image analysis because the perimeter of some granules touch the edge (PTE) of the field being analyzed. To correct for this error, the PTE granules were manually replaced into the field by measuring their diameters and entering them into the database. The results showed differences in the starch size distributions between the classes of wheat evaluated, as well as the method of analysis. Four laser diffraction sizing (LDS) instruments were used to measure granule distributions of the four classes of wheat. LDS compared with IA resulted in a ˜40% underestimation of the A-type granule diameter and a ˜50% underestimation of the B-type granule diameter. A correction factor (adjustment) was developed from IA data to correct LDS analysis. LDS data correlations before adjustments to IA data were R2= 0.02nsto 0.55***. After adjustment, these correlations improved to R2= 0.81*** to 0.93*** depending on the class of wheat starch evaluated.

A transmission electron microscopic (TEM) study was conducted on maturing wheat to determine the sequences of events involved during the drying of preripe starchy endosperm. Field-grown hard red winter wheat Karl and soft red winter wheat Clark were harvested at 21 days after flowering (DAF) and air-dried in the spike at 20°C for 2, 4, 8, 12, 24, 48, and 96 hr and until completely air-dried (7 days). Fresh samples of Karl and Clark were also harvested and prepared immediately for microscopy. Both wheat cultivars underwent similar changes during drying. Few changes were observed during the first 8 hr of drying. By 12 hr after the start of drying, the rough endoplasmic reticulum (RER) was distended and circularized. Protein bodies were irregular in shape and small autophagic vacuoles were found in the cytoplasm. An amorphous material was first observed in areas of the cytoplasm after 24 hr of drying. RER was typically associated with those regions. Endosperm tissue looked nearly mature after 48 hr of drying. Artificially induced senescence caused by harvesting wheat spikes prematurely caused the endosperm tissue to undergo a number of changes that resulted in the tissue looking normal when compared with wheat that was not prematurely harvested. This suggests that the wheat plant has great capacity to develop normally when subjected to environmental stresses. The methods used in this study can be used to investigate endosperm structural changes caused by adverse environmental stress.

Starches were isolated from 12 soft wheat (Triticum aestivumL.) cultivars and were characterized for waxy (Wx) allelic expression, thermal pasting characteristics, and starch granule size. Gels were produced from the thermally degraded starches and were evaluated using large deformation rheological measurements. Data were compared with cultivar kernel texture, milling characteristics, starch chemical analyses, and flour pasting characteristics. Larger flour yields were produced from cultivars that had larger starch granules. Flour yield also was correlated with lower amylose content and greater starch content. Harder starch gels were correlated with higher levels of amylose content and softer kernel texture. The cultivar Fillmore, which had a partial waxy mutation at the B locus, produced the highest peak pasting viscosity and the lowest gel hardness. Softer textured wheats had greater lipid-complexed amylose and starch phosphorus contents and had less total starch content. Among these wheats of the soft market class, softer textured wheats had larger starch granules and harder textured wheats had smaller starch granules. In part, this may explain why soft wheats vary in texture. The smaller granules have larger surface area available for noncovalent bonding with the endosperm protein matrix and they also may pack more efficiently, producing harder endosperm.

A technique was developed and evaluated whereby differentiating wheat endosperm tissue could be processed for the freeze‐etch, freeze‐fracture technique without the use of chemical fixatives. Field grown, developing hard red winter wheat (cv. Newton) caryopses were infiltrated with glycerol prior to freezing in liquid nitrogen‐cooled Freon‐22. Frozen samples were placed in cryogenic vials and stored in liquid nitrogen until needed. Freeze‐fracture was carried out under standard conditions. Evaluation of replicas from glycerol‐imbibed endosperm was made by comparing them to replicas obtained from freshly frozen untreated endosperm, and endosperm that had been prefixed in glutaraldehyde and paraformaldehyde. Other evaluations were made by comparing replicas of glycerol‐treated endosperm to thin sections obtained from wheat that had been imbibed with glycerol, fixed, dehydrated, and infiltrated and embedded in epoxy resin for routine electron microscopy. The results indicate that if the unfixed glycerol‐treated wheat endosperm is handled carefully, replicas can be obtained which show few artifacts due to glycerol and freezing. Typical artifacts such as vesiculation of RER, ice crystal damage, production of fusion intermediates, and membrane particle segregation can be nearly eliminated when this technique is applied to developing wheat endosperm.

The mobility of 13C specifically labeled branched chain end groups of iso-even fatty acids in intact, live Bacillus thuringiensis cells was studied by 13C nuclear magnetic resonance spectroscopy. This study apparently represents the first direct observation of branched chain carbon atoms in living cells. End groups were labeled using DL-[beta, delta, delta‘-13C]valine as a precursor chain initiator for iso-even fatty acid synthesis after using L-[delta, delta‘-14C]L-valine to determine optimal conditions for labeling of the membrane fatty acid end groups. Cell survival in the NMR was determined for various lengths of time at 28 and 39 degrees C. Subsequently, 13C-labeled vegetative cells, sporulating cells (three stages of development), and purified mature spores were analyzed by 13C NMR using corresponding unlabeled cells as controls. Spin lattice relaxation times (T1) were obtained for the enriched iso-branched region at 23.3 ppm and for the natural abundance peak for the glycerol backbone (carbons 1 and 3) of the membrane lipids at 61.7 ppm. The T1 of the glycerol carbons (0.08 s) did not change significantly with stage of development or temperature. The T1 of the iso-even enriched end group changed dramatically from vegetative cells (0.70s) to sporulating cells (0.28 s) at 28 degrees C. A decrease in the T1 was also observed at 39 degrees C from 0.91 s for vegetative cells to 0.54 s for sporulating cells. Accompanying the reduced mobility indicated by the T1 values, there was a general decline in the signal-to-noise ratios of identically acquired spectra as sporulation continued which culminated in the lack of discernible plasma membrane lipid resonances in purified mature spores. The progressive loss of signal appeared to have resulted from a continuous decline in the fraction of plasma membrane fatty acids with sufficient mobility to give signals above background.

Abstract: Calcium excretion and deposition were studied during sporogenesis in the myxomycete,Physarella oblonga. Prior to fruiting, the migrating vegetative plasmodium contains numerous, large channels connected to the external environment. During early sporangial development, these channels become smaller and are isolated as channel remnants. Plasmodial microfilament bundles also disappear at this time. Internally, two types of spheres, each about 0.5–1.5 μm in diameter, are formed in the channel remnants. A dense sphere appears to originate from small particles initially secreted from the mitochondria. These particles then enter the channels and channel remnants by reverse pinocytosis and fuse to form the dense spheres. A second, less dense sphere has no detectable cytoplasmic origin but also appears to form in the channels and channel remnants. Both types of spheres are concentrated by the fusion of channel remnants to form enlarged regions (knots and spikes) of the developing capillitium. Some of the channel remnants also fuse with the outer surface of the sporangium, depositing both spheres to the outside. Both spheres have been studied by various light and electron microscopic techniques and with specific histochemical and analytical procedures. The less dense spheres appear to contain a major portion of calcium in the form of calcium carbonate. These results are compared to other recent studies on calcium deposition in the Myxomycetes and a different mechanism for calcium deposition is proposed.

A transmission electron microscopic study was conducted on air- and freeze-dried developing wheat to determine the effects of drying on the structure of the starchy endosperm. Field-grown hard red winter wheat (Karl) and soft red winter wheat (Clark) were harvested at 15, 18, 21, 23, 25, 28, and 35 days after flowering (DAF). Wheat was dried by either air-drying in the spike at 28°C or freeze-drying following freezing in liquid nitrogen. Dried wheat was prepared for microscopy. Fresh samples of Karl and Clark were also harvested on the same days and prepared immediately for microscopy. The method of drying greatly affected cellular ultrastructure. The most pronounced change upon air-drying of developing samples was disappearance of individual protein bodies and conversion of the cytoplasm into a matrix-like material similar in appearance to storage protein matrix found in mature wheat endosperm. Freeze-dried wheats maintained nearly natural ultrastructure but exhibited various amounts of freeze damage. Conversion of protein bodies to a matrix was not observed in freeze-dried samples. The results suggest that hardness develops as a result of endosperm senescence rather than accumulation of particular grain components. Senescence may cause changes in the starch granule surface such that surrounding components bind tightly in hard wheats, whereas the binding is weaker in soft wheats. Therefore, the surface of starch granules might be more important than components the starch granules bind to in determining hardness.