Effectiveness of Telehealth Direct Therapy for Individuals with Autism Spectrum Disorder.

The field of applied behavior analysis (ABA) has utilized telehealth for clinical supervision and caregiver guidance with research supporting the use of both modalities. Research demonstrating effectiveness is crucial, as behavior analysts must ensure the services they provide are effective in order to be ethical. With the increased need for patients to access more services via telehealth, due to the novel coronavirus (COVID-19) pandemic, the current study evaluated the efficacy of telehealth direct therapy to teach new skills to individuals with autism spectrum disorder (ASD). This study examined the utility of natural environment teaching and discrete trial training strategies provided over a videoconferencing platform to teach new skills directly to seven individuals with varying ASD severity levels. The targeted skills were taught solely through telehealth direct therapy with varying levels of caregiver support across participants and included skills in the language, adaptive, and social domains. In a multiple baseline design, all seven participants demonstrated mastery and maintenance for all targets; in addition, generalization to family members was assessed for some targets. The evidence suggests that telehealth is a modality that is effective and can be considered for all patients when assessing the appropriate location of treatment.

A Measure for Supporting Implementation of Telehealth Direct Therapy With Treatment Integrity.

As telehealth direct therapy sessions are being increasingly provided for safety reasons during the COVID-19 pandemic, it is critical to ensure that sessions are implemented with integrity by direct service providers. Although existing research addresses the efficacy of the telehealth model, there is no literature on integrity measures tied to this service model. Without a framework or point of reference, clinicians new to the field of telehealth direct therapy may not be able or willing to attempt to implement it. The Telehealth Therapy Treatment Integrity Measure is designed to delineate components of effective telehealth direct therapy, including aspects of both instruction that should be generalized from in-person sessions to telehealth direct therapy sessions and new elements that are unique to telehealth direct therapy. A description of how the measure can be used to support the training, both initial and ongoing, of direct service providers is included. This measure can support clinicians in ensuring that direct service providers are working within their scope of competence when providing telehealth direct therapy.

Citrulline, a novel compatible solute in drought-tolerant wild watermelon leaves, is an efficient hydroxyl radical scavenger.

Drought-tolerant wild watermelon accumulates high levels of citrulline in the leaves in response to drought conditions. In this work, the hydroxyl radical-scavenging activity of citrulline was investigated in vitro. The second-order rate constant for the reaction between citrulline and hydroxyl radicals was found to be 3.9x10(9) M(-1) s(-1), demonstrating that citrulline is one of the most efficient scavengers among compatible solutes examined so far. Moreover, citrulline effectively protected DNA and an enzyme from oxidative injuries. Liquid chromatography-mass spectrometry analysis revealed that at least four major products were formed by the reaction between citrulline and hydroxyl radicals. Activities of metabolic enzymes were not inhibited by up to 600 mM citrulline, indicating that citrulline does not interfere with cellular metabolism. We reasoned, from these results, that citrulline contributes to oxidative stress tolerance under drought conditions as a novel hydroxyl radical scavenger.

Crystallization and preliminary X-ray diffraction analysis of glutathione-dependent dehydroascorbate reductase from spinach chloroplasts.

Glutathione-dependent dehydroascorbate reductase (GSH-DHAR) catalyzes the reduction of dehydroascorbate to ascorbate using reduced glutathione as the electron donor. GSH-DHAR from spinach chloroplasts produced in Escherichia coli was crystallized by the hanging-drop vapour-diffusion method. The crystals were monoclinic, space group C2, with unit-cell parameters a = 98.25, b = 39.96, c = 106.86 A, beta = 110.46 degrees. The asymmetric unit contained two molecules, giving a crystal volume per enzyme mass (V(M)) of 2.06 A(3) Da(-1) and a solvent content of 40.3%. A full set of X-ray diffraction data were collected to 2.2 A Bragg spacing from three native crystals with an overall R(merge) of 6.5% and a completeness of 93.4%.

Cyclic flow of electrons within PSII in thylakoid membranes.

In photosynthesis, the electrons released from PSII are considered to be shared mainly by carbon metabolism and the water-water cycle. We demonstrated previously that some electrons are utilized in a CO2- and O2-independent manner in leaves of wild watermelon [Miyake and Yokota (2000) Plant Cell Physiol: 41: 335]. In the present study, we examined the mechanism of this alternative flow of electrons in thylakoid membranes, isolated from fresh spinach leaves, by simultaneously measuring the quantum yield of PSII and the flux of the linear flow of electrons. In the presence of the protonophore nigericin, which eliminates the pH gradient across thylakoid membranes, the quantum yield and the flux of the linear flow of electrons were directly proportional to one another. The quantum yield at a given linear flux of electrons was much higher in the absence of nigericin than in its presence, indicating that an additional or alternative flow of electrons can occur independently of the linear flow in the absence of nigericin. In the presence of nigericin, the alternative flux of electrons increased with decreasing pH and with increasing reduction of the plastoquinone pool. Cyclic flow of electrons in PSII appears to be the most plausible candidate for the alternative flow of electrons. The flux reached 280 micromol x e(-) (mg Chl)(-1) x h(-1) and was similar to that of the CO2- and O2-independent alternative flow of electrons that we found in leaves of wild watermelon. The cyclic, alternative flow of electrons in PSII provides a possible explanation for the alternative flow of electrons observed in vivo.

Characterization of ascorbate peroxidases from unicellular red alga Galdieria partita.

Galdieria partita, a unicellular red alga isolated from acidic hot springs and tolerant to sulfur dioxide, has at least two ascorbate peroxidase (APX) isozymes. This was the first report to demonstrate that two isozymes of APX are found in algal cells. Two isozymes were separated from each other at the hydrophobic chromatography step of purification and named APX-A and APX-B after the elution order in the chromatography. APX-B accounted for 85% of the total activity. Both isozymes were purified. APXs from Galdieria were monomers whose molecular weights were about 28,000, similar to stromal APX of higher plants. APX-A cross-reacted with monoclonal antibody raised against APX of Euglena gracilis in immunoblotting, but APX-B did not, although the antibody can recognize all other APXs tested. The amino-terminal sequences of APX-A and -B from Galdieria had some homology with each other but little homology with those from other sources. Their Km values for ascorbate and hydrogen peroxide were comparable with those of APX from higher plants. Unlike the green algal enzymes, the donor specificities of Galdieria APXs were as high as those of plant chloroplastic APX. On the contrary, these APXs reduced tertiary-butyl hydroperoxide as an electron acceptor as APXs from Euglena and freshwater Chlamydomonas do. The inhibition of APX-A and -B by cyanide and azide, and characteristics of their light absorbance spectra indicated that they were heme peroxidases.

Aromatic monoamine-induced immediate oxidative burst leading to an increase in cytosolic Ca2+ concentration in tobacco suspension culture.

Aromatic monoamines may contribute to both chemical and physical protection of plants. Addition of phenylethylamine (PEA) and benzylamine to tobacco suspension culture (cell line BY-2) induced a very rapid and transient generation of two active oxygen species (AOS), H2O2 and superoxide anion, both detected with chemiluminescence. Electron spin resonance spectroscopy revealed that hydroxy radicals are also produced. With laser-scanning confocal microscopy, fluorescence spectroscopy and microplate fluorescence reading, intracellular H2O2 production was detected using dichlorofluorescin diacetate as a fluorescent probe. Following AOS production, cytosolic Ca2+ concentration ([Ca2+]c) of the tobacco cells, monitored with luminescence of transgenic aequorin, increased and attained to a peak level 12 s after PEA addition. The PEA-induced increase in [Ca2+]c was inhibited by a Ca2+ chelator, Ca2+ antagonists and AOS scavengers, suggesting that PEA-induced AOS triggered a Ca2+ influx across the plasma membrane.

Phenylethylamine-induced generation of reactive oxygen species and ascorbate free radicals in tobacco suspension culture: mechanism for oxidative burst mediating Ca2+ influx.

In the previous paper [Kawano et al. (2000a) Plant Cell Physiol. 41: 1251], we demonstrated that addition of phenylethylamine (PEA) and benzylamine can induce an immediate and transient burst of active oxygen species (AOS) in tobacco suspension culture. Detected AOS include H2O2, superoxide anion and hydroxyl radicals. Use of several inhibitors suggested the presence of monoamine oxidase-like H2O2-generating activity in the cellular soluble fraction. It was also suggested that peroxidase(s) or copper amine oxidase(s) are involved in the extracellular superoxide production as a consequence of H2O2 production. Since more than 85% of the PEA-dependent AOS generating activity was localized in the extracellular space (extracellular fluid + cell wall), extracellularly secreted enzymes, probably peroxidases, may largely contribute to the oxidative burst induced by PEA. The PEA-induced AOS generation was also observed in the horseradish peroxidase (HRP) reaction mixture, supporting the hypothesis that peroxidases catalyze the oxidation of PEA leading to AOS generation. In addition to AOS production, we observed that PEA induced an increase in monodehydroascorbate radicals (MDA) in the cell suspension culture and in HRP reaction mixture using electron spin resonance spectroscopy and the newly invented MDA reductase-coupled method. Here we report that MDA production is an indicator of peroxidase-mediated generation of PEA radical species in tobacco suspension culture.

Chloroplast development in Arabidopsis thaliana requires the nuclear-encoded transcription factor sigma B.

Development of plastids into chloroplasts, the organelles of photosynthesis, is triggered by light. However, little is known of the factors involved in the complex coordination of light-induced plastid gene expression, which must be directed by both nuclear and plastid genomes. We have isolated an Arabidopsis mutant, abc1, with impaired chloroplast development, which results in a pale green leaf phenotype. The mutated nuclear gene encodes a sigma factor, SigB, presumably for the eubacterial-like plastid RNA polymerase. Our results provide direct evidence that a nuclear-derived prokaryotic-like SigB protein, plays a critical role in the coordination of the two genomes for chloroplast development.

Responses of wild watermelon to drought stress: accumulation of an ArgE homologue and citrulline in leaves during water deficits.

Wild watermelon from the Botswana desert had an ability to survive under severe drought conditions by maintaining its water status (water content and water potential). In the analysis by two-dimensional electrophoresis of leaf proteins, seven spots were newly induced after watering stopped. One with the molecular mass of 40 kilodaltons of the spots was accumulated abundantly. The cDNA encoding for the protein was cloned based on its amino-terminal sequence and the amino acid sequence deduced from the determined nucleotide sequences of the cDNA exhibited homology to the enzymes belong to the ArgE/DapE/Acy1/Cpg2/YscS protein family (including acetylornithine deacetylase, carboxypeptidase and aminoacylase-1). This suggests that the protein is involved in the release of free amino acid by hydrolyzing a peptidic bond. As the drought stress progressed, citrulline became one of the major components in the total free amino acids. Eight days after withholding watering, although the lower leaves wilted significantly, the upper leaves still maintained their water status and the content of citrulline reached about 50% in the total free amino acids. The accumulation of citrulline during the drought stress in wild watermelon is an unique phenomenon in C3-plants. These results suggest that the drought tolerance of wild watermelon is related to (1) the maintenance of the water status and (2) a metabolic change to accumulate citrulline.