Serum proteomic analysis identifies sex-specific differences in lipid metabolism and inflammation profiles in adults diagnosed with Asperger syndrome.

BACKGROUND: The higher prevalence of Asperger Syndrome (AS) and other autism spectrum conditions in males has been known for many years. However, recent multiplex immunoassay profiling studies have shown that males and females with AS have distinct proteomic changes in serum. METHODS: Here, we analysed sera from adults diagnosed with AS (males = 14, females = 16) and controls (males = 13, females = 16) not on medication at the time of sample collection, using a combination of multiplex immunoassay and shotgun label-free liquid chromatography mass spectrometry (LC-MSE). The main objective was to identify sex-specific serum protein changes associated with AS. RESULTS: Multiplex immunoassay profiling led to identification of 16 proteins that were significantly altered in AS individuals in a sex-specific manner. Three of these proteins were altered in females (ADIPO, IgA, APOA1), seven were changed in males (BMP6, CTGF, ICAM1, IL-12p70, IL-16, TF, TNF-alpha) and six were changed in both sexes but in opposite directions (CHGA, EPO, IL-3, TENA, PAP, SHBG). Shotgun LC-MSE profiling led to identification of 13 serum proteins which had significant sex-specific changes in the AS group and, of these, 12 were altered in females (APOC2, APOE, ARMC3, CLC4K, FETUB, GLCE, MRRP1, PTPA, RN149, TLE1, TRIPB, ZC3HE) and one protein was altered in males (RGPD4). The free androgen index in females with AS showed an increased ratio of 1.63 compared to controls. CONCLUSION: Taken together, the serum multiplex immunoassay and shotgun LC-MSE profiling results indicate that adult females with AS had alterations in proteins involved mostly in lipid transport and metabolism pathways, while adult males with AS showed changes predominantly in inflammation signalling. These results provide further evidence that the search for biomarkers or novel drug targets in AS may require stratification into male and female subgroups, and could lead to the development of novel targeted treatment approaches.

Increased serum androstenedione in adults with autism spectrum conditions.

Molecular and behavioural evidence points to an association between sex-steroid hormones and autism spectrum conditions (ASC) and/or autistic traits. Prenatal androgen levels are associated with autistic traits, and several genes involved in steroidogenesis are associated with autism, Asperger Syndrome and/or autistic traits. Furthermore, higher rates of androgen-related conditions (such as Polycystic Ovary Syndrome, hirsutism, acne and hormone-related cancers) are reported in women with autism spectrum conditions. A key question therefore is if serum levels of gonadal and adrenal sex-steroids (particularly testosterone, estradiol, dehydroepiandrosterone sulfate and androstenedione) are elevated in individuals with ASC. This was tested in a total sample of n=166 participants. The final eligible sample for hormone analysis comprised n=128 participants, n=58 of whom had a diagnosis of Asperger Syndrome or high functioning autism (33 males and 25 females) and n=70 of whom were age- and IQ-matched typical controls (39 males and 31 females). ASC diagnosis (without any interaction with sex) strongly predicted androstenedione levels (p<0.01), and serum androstenedione levels were significantly elevated in the ASC group (Mann-Whitney W=2677, p=0.002), a result confirmed by permutation testing in females (permutation-corrected p=0.02). This result is discussed in terms of androstenedione being the immediate precursor of, and being converted into, testosterone, dihydrotestosterone, or estrogens in hormone-sensitive tissues and organs.

Real-time evaluation of experimental variation in large-scale LC-MS/MS-based quantitative proteomics of complex samples.

Quantitative proteomic profiling is becoming a widely used approach in systems biology and biomarker discovery. There is a growing realization that quantitative studies require high numbers of unpooled samples for increased statistical power. Large-scale quantitative analyses require an added degree of stringency due to the lengthy study periods and reliance on stability of analytical instrumentation. We present the inclusion of quality control samples alongside clinical samples in the preparation and nanoLC-MS analysis pipelines. These serve the purpose of monitoring, evaluating and reporting experimental variation measured in real-time. This concept is shown for two types of complex biological samples: serum samples and fibroblast samples. In both studies QC samples were added among dozens of clinical ones and analyzed using a label-free quantitative proteomic platform.

Elevated rates of testosterone-related disorders in women with autism spectrum conditions.

The androgen theory of autism proposes that autism spectrum conditions (ASC) are in part due to elevated fetal testosterone (FT) levels, which are positively correlated with a number of autistic traits and inversely correlated with social development and empathy. A medical questionnaire was completed by n=54 women with ASC, n=74 mothers of children with ASC, and n=183 mothers of typically developing children to test whether women with ASC have an increased rate of testosterone-related medical conditions, and to see whether mothers of children with ASC show similar abnormalities, as part of the 'broader autism phenotype'. Compared to controls, significantly more women with ASC reported (a) hirsutism, (b) bisexuality or asexuality, (c) irregular menstrual cycle, (d) dysmenorrhea, (e) polycystic ovary syndrome, (f) severe acne, (g) epilepsy, (h) tomboyism, and (i) family history of ovarian, uterine, and prostate cancers, tumors, or growths. Compared to controls, significantly more mothers of ASC children reported (a) severe acne, (b) breast and uterine cancers, tumors, or growths, and (c) family history of ovarian and uterine cancers, tumors, or growths. These results suggest current hormone abnormalities in women with ASC and their mothers. Direct investigations of serum testosterone levels and genetic susceptibility to high testosterone production or sensitivity in women with ASC would illuminate the origin of these conditions. The relationship between FT and current testosterone levels also needs to be clarified. The present results may be relevant to understanding the increased male risk to developing autism.

Continuity of care practices and substance use disorder patients' engagement in continuing care.

BACKGROUND: Substance use disorder (SUD) patients who engage in more continuing care have better outcomes, but information on practices associated with greater patient engagement and retention in continuing care remains elusive. OBJECTIVES: The objectives of this study were to determine if staff's continuity of care practices predict patients' engagement in continuing care in the 6 months after discharge from intensive SUD treatment and to determine if the impact of continuity of care practices on patients' engagement in continuing care differs for patients treated in inpatient/residential versus outpatient programs. RESEARCH DESIGN: Staff in 28 Veterans Affairs (VA) intensive SUD treatment programs with varying continuity of care practices provided data on 878 patients' alcohol and drug problems at treatment entry. At discharge, staff provided data on patients' motivation, treatment intensity, and on the continuity of care practices they used with each patient. VA administrative databases supplied data on patients' subsequent engagement in continuing care. Mixed-effects modeling was used to examine predictors of patients' engagement in care. RESULTS: Patients in outpatient programs who received more continuity of care engaged in continuing care significantly longer. More highly motivated outpatients, those with fewer alcohol problems at treatment entry, and patients who used VA services in the year before treatment also remained in continuing care longer. These findings did not hold for patients treated in inpatient/residential programs. CONCLUSIONS: Continuity of care practices predicted engagement in continuing care only for patients treated in outpatient SUD programs. More research is needed to identify effective continuity of care practices for patients treated in inpatient/residential programs.

Assessing continuity of care practices in substance use disorder treatment programs.

OBJECTIVE: The purpose of this article is to describe the development and psychometric properties of parallel program-level and individual-level versions of the Continuity of Care Practices Survey (CCPS-P and CCPS-I), a measure that assesses four dimensions of continuity of care practices in substance use disorder (SUD) treatment programs. CCPS subscales assess staff efforts to ensure provider continuity, maintain contact with patients, coordinate care among providers and connect patients to community resources. METHOD: Program-level CCPS data were obtained from directors/coordinators of 129 intensive inpatient/residential and outpatient Department of Veterans Affairs SUD programs. These data were used to examine the internal consistency and discriminant validity of the CCPS-P. A parallel individual-level CCPS-I completed by counselors for 835 patients in a subsample of 28 SUD programs, assessed the continuity of care services that staff provided to individual patients. These data were used to examine the predictive validity of the CCPS-P. RESULTS: CCPS-P and CCPS-I subscales demonstrated acceptable psychometric properties. Lack of significant correlations between CCPS-P subscales and SUD program characteristics (e.g., size, staffing) provided preliminary evidence for discriminant validity. CCPS-P subscales and the overall CCPS-P score predicted corresponding continuity of care services that staff provided to patients within programs, offering support for predictive validity. CONCLUSIONS: Managers can use the CCPS to monitor and improve SUD programs' continuity of care practices. The CCPS also enables researchers to determine the impact of continuity of care practices on the engagement of patients in continuing care and outcomes.