Novel Microdialysis Technique Reveals a Dramatic Shift in Metabolite Secretion during the Early Stages of the Interaction between the Ectomycorrhizal Fungus Pisolithus microcarpus and Its Host Eucalyptus grandis.

The colonisation of tree roots by ectomycorrhizal (ECM) fungi is the result of numerous signalling exchanges between organisms, many of which occur before physical contact. However, information is lacking about these exchanges and the compounds that are secreted by each organism before contact. This is in part due to a lack of low disturbance sampling methods with sufficient temporal and spatial resolution to capture these exchanges. Using a novel in situ microdialysis approach, we sampled metabolites released from Eucalyptus grandis and Pisolithus microcarpus independently and during indirect contact over a 48-h time-course using UPLC-MS. A total of 560 and 1530 molecular features (MFs; ESI- and ESI+ respectively) were identified with significant differential abundance from control treatments. We observed that indirect contact between organisms altered the secretion of MFs to produce a distinct metabolomic profile compared to either organism independently. Many of these MFs were produced within the first hour of contact and included several phenylpropanoids, fatty acids and organic acids. These findings show that the secreted metabolome, particularly of the ECM fungus, can rapidly shift during the early stages of pre-symbiotic contact and highlight the importance of observing these early interactions in greater detail. We present microdialysis as a useful tool for examining plant-fungal signalling with high temporal resolution and with minimal experimental disturbance.

Rationale and design of the Baylor Infant Twin Study-A study assessing obesity-related risk factors from infancy.

BACKGROUND: Early childhood (0-3 years) is a critical period for obesity prevention, when tendencies in eating behaviors and physical activity are established. Yet, little is understood about how the environment shapes children's genetic predisposition for these behaviors during this time. The Baylor Infant Twin Study (BITS) is a two phase study, initiated to study obesity risk factors from infancy. Data collection has been completed for Phase 1 in which three sub-studies pilot central measures for Phase 2. A novel infant temperament assessment, based on observations made by trained researchers was piloted in Behavior Observation Pilot Protocol (BOPP) study, a new device for measuring infant feeding parameters (the "orometer") in the Baylor Infant Orometer (BIO), and methods for analyzing DNA methylation in twins of unknown chorionicity in EpiTwin. METHODS: EpiTwin was a cross-sectional study of neonatal twins, while up to three study visits occurred for the other studies, at 4- (BOPP, BIO), 6- (BOPP), and 12- (BOPP, BIO) of age. Measurements for BOPP and BIO included temperament observations, feeding observations, and body composition assessments while EpiTwin focused on collecting samples of hair, urine, nails, and blood for quantifying methylation levels at 10 metastable epialleles. Additional data collected include demographic information, zygosity, chorionicity, and questionnaire-based measures of infant behaviors. RESULTS: Recruitment for all three studies was completed in early 2020. EpiTwin recruited 80 twin pairs (50% monochorionic), 31 twin pairs completed the BOPP protocol, and 68 singleton infants participated in BIO. CONCLUSIONS: The psychometric properties of the data from all three studies are being analyzed currently. The resulting findings will inform the development of the full BITS protocol, with the goal of completing assessments at 4-, 6-, 12-, and 14-month of age for 400 twin pairs.

Quantification of nutritive sucking among preterm and full-term infants.

PURPOSE: We developed summaries of oral bottle-feeding skills among preterm (<37 gestational weeks) and full-term (≥37 gestational weeks) infants using a mechanical device (Orometer) to measure intraoral pressure changes, with accompanying automated software and analytics. We then compared the rates of change in feeding skills over several weeks (feeding trends) between preterm and full-term infants. We also compared group means at 40 weeks post menstrual age (PMA). PATIENTS AND METHODS: Healthy full-term and preterm infants capable of oral feeding were recruited from the Pediatric Outpatient Clinic at University of California San Francisco, Fresno, and from the Oregon Health & Science University Doernbecher Neonatal Critical Care Unit, respectively. Feeding skill was quantified using an Orometer and automated suck-analysis software. Factor analysis reduced the >40 metrics produced by the Orometer system to the following seven factors that accounted for >99% of the sample covariance: suck vigor, endurance, resting, irregularity, frequency, variability, and bursting. We proposed that these factors represent feeding skills and they served as the dependent variables in linear models estimating trends in feeding skills over time for full-term and preterm infants (maturation). At approximately 40 weeks PMA we compared mean feedings skills between infants born preterm and those born full-term using predictions from our models. RESULTS: Feeding skills for 117 full-term infants and 82 preterm infants were first captured at mean PMA of 42.3 and 36.0 weeks, respectively. For some feeding skills, preterm and full-term infants showed different trends over time. At 37-40 weeks PMA, preterm infants took approximately 15% fewer sucks than infants born full-term (p=0.06) and generally had weaker suck vigor, greater resting, and less endurance than full-term babies. Preterm infant-feeding skills appeared similar to those of full-term infants upon reaching ≥40 weeks PMA, although preterm infants showed greater variability for all factors. CONCLUSION: The Orometer device, accompanying software, and analytic methods provided a framework for describing trends in oral feeding, thereby allowing us to characterize differences in maturation of feeding between healthy preterm and full-term infants.

Quantification of intraoral pressures during nutritive sucking: methods with normal infants.

We report quantitative measurements of ten parameters of nutritive sucking behavior in 91 normal full-term infants obtained using a novel device (an Orometer) and a data collection/analytical system (Suck Editor). The sucking parameters assessed include the number of sucks, mean pressure amplitude of sucks, mean frequency of sucks per second, mean suck interval in seconds, sucking amplitude variability, suck interval variability, number of suck bursts, mean number of sucks per suck burst, mean suck burst duration, and mean interburst gap duration. For analyses, test sessions were divided into 4 × 2-min segments. In single-study tests, 36 of 60 possible comparisons of ten parameters over six pairs of 2-min time intervals showed a p value of 0.05 or less. In 15 paired tests in the same infants at different ages, 33 of 50 possible comparisons of ten parameters over five time intervals showed p values of 0.05 or less. Quantification of nutritive sucking is feasible, showing statistically valid results for ten parameters that change during a feed and with age. These findings suggest that further research, based on our approach, may show clinical value in feeding assessment, diagnosis, and clinical management.

Analysis and treatment of problem behavior evoked by music.

The present study investigated the effects of differential negative reinforcement of other behavior (DNRO) on problem behavior evoked by music in a 7-year-old child with pervasive developmental disorder. Following an auditory stimulus assessment, DNRO was used to reduce problem behavior to near-zero levels. Results are discussed in terms of identifying establishing operations to govern treatment options and the use of negative reinforcement when arbitrarily imposed positive reinforcers do not compete with escape from auditory stimulation.

An evaluation of simultaneous presentation and differential reinforcement with response cost to reduce packing.

We evaluated the effects of multiple treatment procedures, including simultaneous presentation of preferred foods, on the packing behavior of a 9-year-old girl with autism. A reversal design was used to assess the effects of differential reinforcement with response cost alone and with simultaneous presentation. In addition, simultaneous presentation was assessed independent of differential reinforcement with response cost. Results indicated that simultaneous presentation reduced packing and that differential reinforcement with response cost was not necessary to maintain these reductions. Results are discussed in terms of the use of simultaneous presentation for packing as an alternative to consequence manipulations.

Differential impact of response effort within a response chain on use of mands in a student with autism.

We investigated the effects of response effort on the use of mands during functional communication training (FCT) in a participant with autism. The number of links in a picture exchange response chain determined two levels of response effort. Each level was paired with a fixed ratio (FR3) schedule of reinforcement for aggression in a reversal design. Responding to either schedule produced access to a preferred item. The participant opted for the low effort mand while aggression decreased significantly. However, the high effort mand did not compete with the FR3 schedule for aggression. Results are discussed in terms of response effort within a response chain of a picture exchange system and competing ratio schedules for problem behavior during mand training.