Jeanne P. Ryan

A Pilot Proteomic Analysis of Salivary Biomarkers in Autism Spectrum Disorder

Autism spectrum disorder (ASD) prevalence is increasing, with current estimates at 1/68–1/50 individuals diagnosed with an ASD. Diagnosis is based on behavioral assessments. Early diagnosis and intervention is known to greatly improve functional outcomes in people with ASD. Diagnosis, treatment monitoring and prognosis of ASD symptoms could be facilitated with biomarkers to complement behavioral assessments. Mass spectrometry (MS) based proteomics may help reveal biomarkers for ASD. In this pilot study, we have analyzed the salivary proteome in individuals with ASD compared to neurotypical control subjects, using MS‐based proteomics. Our goal is to optimize methods for salivary proteomic biomarker discovery and to identify initial putative biomarkers in people with ASDs. The salivary proteome is virtually unstudied in ASD, and saliva could provide an easily accessible biomaterial for analysis. Using nano liquid chromatography‐tandem mass spectrometry, we found statistically significant differences in several salivary proteins, including elevated prolactin‐inducible protein, lactotransferrin, Ig kappa chain C region, Ig gamma‐1 chain C region, Ig lambda‐2 chain C regions, neutrophil elastase, polymeric immunoglobulin receptor and deleted in malignant brain tumors 1. Our results indicate that this is an effective method for identification of salivary protein biomarkers, support the concept that immune system and gastrointestinal disturbances may be present in individuals with ASDs and point toward the need for larger studies in behaviorally‐characterized individuals. Autism Res 2015, 8: 338–350.

Evidence of practice effects in variants of the Trail Making Test during serial assessment.

Practice effects often obscure detection of meaningful intraindividual cognitive change in serial assessment. The Trail Making Test and four of its variants (i.e., Trail Making Test of the Delis–Kaplan Executive Functioning System, Comprehensive Trail Making Test, Connections Task, and Planned Connections) were administered to college-aged participants over a 3-week period with 7 days separating each session. Linear growth analysis yielded statistically significant average change in slope across time periods at the p < .05 level for each of the five instruments. The results raise concern about clinical interpretations based upon repeated use of each instrument in serial assessment.

Sports-related and gender differences on neuropsychological measures of frontal lobe functioning.

ObjectiveTo determine similarities and differences in the performance of female and male athletes on neuropsychological measures of frontal lobe functioning. DesignA cross-sectional study of male and female college-aged athletes involved in one of the following sports: hockey, basketball, softball, lacrosse, soccer, swimming, and track. SettingDivision III college. ParticipantsA total of 262 athletes (male, n = 157; female, n = 105) participated in the study. Main Outcome MeasuresControlled Oral Word Association (letters F, A, S), Cognitive Assessment System (Planned Codes, Planned Connections, Number Detection), and WAIS-R-NI Vocabulary were administered to all athletes. ResultsMANCOVA was performed with gender and sport as fixed factors. Female athletes displayed faster and more accurate performance on perceptual-motor tasks (P < 0.01) and on one condition of a verbal fluency task (P < 0.01) compared with male athletes. Male hockey athletes showed superior perceptual-motor speed and accuracy (P < 0.01) compared with male athletes in the track/swimming group. Evaluators were naive to athletes’ gender and sport. ConclusionGender- and sport-specific performances on perceptual-motor and verbal fluency tasks were found. Adding cognitive components to base functions eliminates gender- and sports-related distinctions, suggesting that existing differences are related to basic, fundamental skills, which are inherent and practiced within the respective sport. Understanding the differences and similarities across sports and gender on various neurocognitive measures is relevant for determining group differences in studies examining the consequences of mild traumatic brain injury among athletes.

Mass spectrometry as a tool for studying autism spectrum disorder

Autism spectrum disorders (ASDs) are increasing in incidence but have an incompletely understood etiology. Tools for uncovering clues to the cause of ASDs and means for diagnoses are valuable to the field. Mass Spectrometry (MS) has been a useful method for evaluating differences between individuals with ASDs versus matched controls. Different biological substances can be evaluated using MS, including urine, blood, saliva, and hair. This technique has been used to evaluate relatively unsupported hypotheses based on introduction of exogenous factors, such as opiate and heavy metal excretion theories of ASDs. MS has also been used to support disturbances in serotonin-related molecules, which have been more consistently observed in ASDs. Serotonergic system markers, markers for oxidative stress, cholesterol system disturbances, peptide hypo-phosphorylation and methylation have been measured using MS in ASDs, although further analyses with larger numbers of subjects are needed (as well as consideration of behavioral data). Refinements in MS and data analysis are ongoing, allowing for the possibility that future studies examining body fluids and specimens from ASD subjects could continue to yield novel insights. This review summarizes MS investigations that have been conducted to study ASD to date and provides insight into future promising applications for this technique, with focus on proteomic studies.

Treating clients with Asperger’s syndrome and autism

Asperger’s syndrome (AS) is a form of autism spectrum disorder (ASD) affecting many individuals today. Although neurobiological correlates for AS have been identified, like many ASDs, AS is not completely understood. AS as a distinct disorder is also not universally accepted and in the DSM-5 AS is not considered a separate nosological entity. In contrast to some other ASDs, individuals with AS are commonly characterized by having standard or higher than average intelligence, yet difficulties in social skills and communication can present challenges for these individuals in everyday functioning. Counseling a person with AS or autism presents a unique challenge for the mental health care provider. We have compiled this review consisting of some recent ideas regarding counseling the client with AS with the goal of providing some clinical insights and practical clues. Although the focus of the present paper is largely on AS, many of these strategies could also apply to individuals with high-functioning autism (HFA).

Salivary proteomics and biomarkers in neurology and psychiatry

Biomarkers are greatly needed in the fields of neurology and psychiatry, to provide objective and earlier diagnoses of CNS conditions. Proteomics and other omics MS‐based technologies are tools currently being utilized in much recent CNS research. Saliva is an interesting alternative biomaterial for the proteomic study of CNS disorders, with several advantages. Collection is noninvasive and saliva has many proteins. It is easier to collect than blood and can be collected by professionals without formal medical training. For psychiatric and neurological patients, supplying a saliva sample is less anxiety‐provoking than providing a blood sample, and is less embarrassing than producing a urine specimen. The use of saliva as a biomaterial has been researched for the diagnosis of and greater understanding of several CNS conditions, including neurodegenerative diseases, autism, and depression. Salivary biomarkers could be used to rule out nonpsychiatric conditions that are often mistaken for psychiatric/neurological conditions, such as fibromyalgia, and potentially to assess cognitive ability in individuals with compromised brain function. As MS and omics technology advances, the sensitivity and utility of assessing CNS conditions using distal human biomaterials such as saliva is becoming increasingly possible.

Three trail making tests for use in neuropsychological assessments with brief intertest intervals

Practice effects in serial neuropsychological assessment have led to the use of alternate forms to measure change in cognitive functioning. The construct validity of three variants of the Trail Making Test was explored over a 3-week period in a sample of 158 undergraduate students. Using confirmatory factor analysis, a two-factor (sequencing-shifting) model was identified to best represent the data. Latent means structural analysis indicated the absence of order effects, lending support for the construct validity of the three tests. The study provides evidence that the three tests can be offered as potential alternative instruments in serial assessment.

Comparative two‐dimensional polyacrylamide gel electrophoresis of the salivary proteome of children with autism spectrum disorder

In the last decades, prevalence of autism spectrum disorder (ASD) has been on the rise. However, clear aetiology is still elusive and improvements in early diagnosis are needed. To uncover possible biomarkers present in ASD, we used two‐dimensional polyacrylamide gel electrophoresis and nanoliquid chromatography‐tandem mass spectrometry (nanoLC‐MS/MS), to compare salivary proteome profiling of children with ASD and controls. A total of 889 spots were compared and only those spots with a fold change ≥1.7 and a P‐value <0.05 or a fold change of ≥3.0 between ASD cases and controls were analysed by nanoLC‐MS/MS. Alpha‐amylase, CREB‐binding protein, p532, Transferrin, Zn alpha2 glycoprotein, Zymogen granule protein 16, cystatin D and plasminogen were down‐regulated in ASD. Increased expression of proto‐oncogene Frequently rearranged in advanced T‐cell lymphomas 1 (FRAT1), Kinesin family member 14, Integrin alpha6 subunit, growth hormone regulated TBC protein 1, parotid secretory protein, Prolactin‐inducible protein precursor, Mucin‐16, Ca binding protein migration inhibitory factor‐related protein 14 (MRP14) was observed in individuals with ASD. Many of the identified proteins have previously been linked to ASD or were proposed as risk factors of ASD at the genetic level. Some others are involved in pathological pathways implicated in ASD causality such as oxidative stress, lipid and cholesterol metabolism, immune system disturbances and inflammation. These data could contribute to protein signatures for ASD presence, risk and subtypes, and advance understanding of ASD cause as well as provide novel treatment targets for ASD.

Mass Spectrometry for the Study of Autism and Neurodevelopmental Disorders

Mass spectrometry (MS) has been increasingly used to study central nervous system disorders, including autism spectrum disorders (ASDs). The first studies of ASD using MS focused on the identification of external toxins, but current research is more directed at understanding endogenous protein changes that occur in ASD (ASD proteomics). This chapter focuses on how MS has been used to study ASDs, with particular focus on proteomic analysis. Other neurodevelopmental disorders have been investigated using this technique, including genetic syndromes associated with autism such as fragile X syndrome and Smith-Lemli-Opitz syndrome.

Proteomics and Cholesterol in Autism

Autism Spectrum Disorder (ASD) diagnosis is increasing worldwide. ASDs are characterized by impaired social function, stereotyped behaviors/interests and communication deficits. ASD causes are poorly understood and treatments are largely limited to behavioral interventions once problems have developed and been detected. Here we discuss the potential use of mass spectrometry and proteomics in early diagnosis of ASD. The potential link between at least some subtypes of ASD, the cholesterol system and proteins that interact with cholesterol is also discussed.