Chronic exposure to xenobiotic pollution leads to significantly higher total glutathione and lower reduced to oxidized glutathione ratio in red blood cells of children with autism.

Analyses of reduced glutathione (GSH), oxidized glutathione (GSSG), and total glutathione (tGSH) in red blood cell samples from 30 children diagnosed with autism and 30 age, gender, and socioeconomic status matched controls were undertaken. The children's ages ranged from 2 to 9. Samples were obtained from subjects residing in Western Pennsylvania, an area of the United States greatly affected by high levels of mercury deposition and airborne PM 2.5 particulates. Liquid chromatography - mass spectrometry was utilized by following EPA Method 6800 for sample analyses. The children with autism had a significantly lower mean red blood cell (RBC) reduced to oxidized glutathione ratio (GSH/GSSG) compared to the control children (p = 0.025). In addition, compared to the controls, the children with autism had significantly higher RBC tGSH values (p = 0.0076) and GSH values (p = 0.022). These results suggest that exposure to toxic elements may prompt compensatory increases in production of GSH in children with autism in environments higher in toxins. The compensation did not fully correct the anti-oxidant properties of exposure to xenobiotics as demonstrated by the significantly lower GSH/GSSG in children with autism compared to controls. Out of a set of glutathione biomarkers, GSH/GSSG may best determine the degree of compensation for oxidative stress in children with autism.

A cleanroom sleeping environment's impact on markers of oxidative stress, immune dysregulation, and behavior in children with autism spectrum disorders.

BACKGROUND: An emerging paradigm suggests children with autism display a unique pattern of environmental, genetic, and epigenetic triggers that make them susceptible to developing dysfunctional heavy metal and chemical detoxification systems. These abnormalities could be caused by alterations in the methylation, sulfation, and metalloprotein pathways. This study sought to evaluate the physiological and behavioral effects of children with autism sleeping in an International Organization for Standardization Class 5 cleanroom. METHODS: Ten children with autism, ages 3-12, slept in a cleanroom for two weeks to evaluate changes in toxin levels, oxidative stress, immune dysregulation, and behavior. Before and after the children slept in the cleanroom, samples of blood and hair and rating scale scores were obtained to assess these changes. RESULTS: Five children significantly lowered their concentration of oxidized glutathione, a biomarker of oxidative stress. The younger cohort, age 5 and under, showed significantly greater mean decreases in two markers of immune dysregulation, CD3% and CD4%, than the older cohort. Changes in serum magnesium, influencing neuronal regulation, correlated negatively while changes in serum iron, affecting oxygenation of tissues, correlated positively with age. Changes in serum benzene and PCB 28 concentrations showed significant negative correlations with age. The younger children demonstrated significant improvements on behavioral rating scales compared to the older children. In a younger pair of identical twins, one twin showed significantly greater improvements in 4 out of 5 markers of oxidative stress, which corresponded with better overall behavioral rating scale scores than the other twin. CONCLUSIONS: Younger children who slept in the cleanroom altered elemental levels, decreased immune dysregulation, and improved behavioral rating scales, suggesting that their detoxification metabolism was briefly enhanced. The older children displayed a worsening in behavioral rating scale performance, which may have been caused by the mobilization of toxins from their tissues. The interpretation of this exploratory study is limited by lack of a control group and small sample size. The changes in physiology and behavior noted suggest that performance of larger, prospective controlled studies of exposure to nighttime or 24 hour cleanroom conditions for longer time periods may be useful for understanding detoxification in children with autism. TRIAL REGISTRATION: Clinical Trial Registration Number NCT02195401 (Obtained July 18, 2014).

Molecular speciated isotope dilution mass spectrometric methods for accurate, reproducible and direct quantification of reduced, oxidized and total glutathione in biological samples.

Novel protocols were developed to accurately quantify reduced (GSH), oxidized (GSSG) and total (tGSH) glutathione in biological samples using molecular speciated isotope dilution mass spectrometry (SIDMS). For GSH and GSSG measurement, the sample was spiked with isotopically enriched analogues of the analytes ((310)GSH and (616)GSSG), along with N-ethylmaleimide (NEM), and treated with acetonitrile to solubilize the endogenous analytes via protein precipitation and equilibrate them with the spikes. The supernatant was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the analytes were quantified with simultaneous tracking and correction for auto-oxidation of GSH to GSSG. For tGSH assay, a (310)GSH-spiked sample was treated with dithiothreitol (DTT) to convert disulfide-bonded glutathione to GSH. After removing the protein, the supernatant was analyzed by LC-MS/MS and the analyte was quantified by single-spiking isotope dilution mass spectrometry (IDMS). The mathematical relationships in IDMS and SIDMS quantifications are based on isotopic ratios and do not involve calibration curves. The protocols were validated using spike recovery tests and by analyzing synthetic standard solutions. Red blood cell (RBC) and saliva samples obtained from healthy subjects, and whole blood samples collected and shipped from a remote location were analyzed. The concentrations of tGSH in the RBC and whole blood samples were 2 orders of magnitude higher than those found in saliva. The fractions of GSSG were 0.2-2.2% (RBC and blood) and 15-47% (saliva) of the free glutathione (GSH + 2xGSSG) in the corresponding samples. Up to 3% GSH was auto-oxidized to GSSG during sample workup; the highest oxidations (>1%) were in the saliva samples.

Measuring intrabladder pressure with the head of the bed elevated 30°: evidence to support a change in practice.

BACKGROUND: A 30° head-of-bed elevation is recommended for most critically ill patients. Measuring intrabladder pressure with the patient in this position is controversial. OBJECTIVE: To assess the feasibility of measuring intrabladder pressure with a 30° head-of-bed elevation. METHODS: A prospective, randomized, and experimental study. Patients had intrabladder pressure measured first while positioned supine with a 30° head-of-bed elevation and 25 mL of saline instilled into the bladder and again after the patients were randomly repositioned to supine without any head-of-bed elevation (flat) or with a 30° head-of-bed elevation while supine or in right lateral or left lateral position with either 25, 50, or 200 mL of saline instilled into the patient's bladder. RESULTS: Intrabladder pressures measured with the patient in all 3 head-of-bed elevated positions were higher than pressures measured with patients supine and flat after instillation of 25 mL of saline into the bladder, but intrabladder pressure did not differ between the 30° head-of-bed elevated positions and the supine and flat positions when 50 or 200 mL of saline was instilled into the bladder. Two-way analysis of variance showed a significant interaction between volume of saline instilled (P = .05), patient's position (P = .007), and bladder instill volume and position interaction (P = .004). CONCLUSION: It is feasible to measure intrabladder pressure with a 30° head-of-bed elevation, and that position could be an alternative to supine positioning of patients for measurement of intrabladder pressure.

The plasma zinc/serum copper ratio as a biomarker in children with autism spectrum disorders.

The frequency of zinc deficiency, copper toxicity and low zinc/copper in children with autism spectrum disorders (ASDs) may indicate decrement in metallothionein system functioning. A retrospective review of plasma zinc, serum copper and zinc/copper was performed on data from 230 children with autistic disorder, pervasive developmental disorder-NOS and Asperger's syndrome. The entire cohort's mean zinc level was 77.2 microg dl(-1), mean copper level was 131.5 microg dl(-1), and mean Zn/Cu was 0.608, which was below the 0.7 cut-off of the lowest 2.5% of healthy children. The plasma zinc/serum copper ratio may be a biomarker of heavy metal, particularly mercury, toxicity in children with ASDs.

Inhibition of alveolar osteitis in mandibular tooth extraction sites using platelet-rich plasma.

Alveolar osteitis (AO), also known as dry socket, continues to be a complication of tooth removal. Platelet-rich plasma (PRP) can be used to accelerate both soft and hard tissue healing. This paper is a retrospective review of the benefits of PRP in AO prevention. PRP was obtained from patients for use in the postremoval alveolar sockets of mandibular molar extraction sites. Statistical analysis of 904 extraction sites with and without PRP use was examined. PRP significantly reduced the incidence of AO by 62.1%, from 9.57% in patients not receiving PRP to 3.63% in patients who received PRP (P = .00043). PRP use had benefits in all subpopulations. The odds of AO occurring in patients not receiving PRP treatment following tooth extraction was 2.81 times greater than in patients receiving PRP treatment immediately following tooth extraction. Four statistically significant risk factors for AO were identified: complete impaction, oral contraceptive use, bruxism, and failure to administer PRP. The application of PRP can significantly reduce the incidence of AO even in patients with risk factors for AO, such as removal of impacted teeth, bruxism, and oral contraceptive use. PRP may be of benefit because it helps initiate clot formation, provides growth factors to facilitate the healing process, and contains concentrated white blood cells to inhibit infection. The use of PRP following tooth extraction is a simple, cost-effective technique that can be used to decrease the incidence of AO and therefore decrease postoperative pain.