Potential protein markers in children with Autistic Spectrum Disorder (ASD) revealed by salivary proteomics.

The lack of specific pharmacological therapy for Autistic Spectrum Disorder (ASD) and its clinical heterogeneity demand efforts directed toward the identification of biomarkers to aid in diagnosis. Proteomics offers a new perspective for studying the altered proteins associated with autism spectrum disorders (ASD) and we have saliva as an easy-to-collect biological fluid with important biomolecules for investigating biomarkers in various diseases. In this sense, saliva could be used to identify potential biomarkers of ASD. In the current work, saliva samples were collected from children with different degrees of ASD and healthy children and proteomics approaches were applied to generate data on differentially expressed proteins between groups which will serve as a basis for future validation studies as protein markers. Data are available via ProteomeXchange with identifier PXD030065. As results, 132 proteins were present in 80% of the saliva pools of all analyzed groups. Twenty-five proteins were identified as overexpressed in the group of severe and mild/moderate ASD carriers, among which, eight were identified as potential biomarkers for ASD.

Analysis of daunorubicin and its metabolite daunorubicinol in plasma and urine with application in the evaluation of total, renal and metabolic formation clearances in patients with acute myeloid leukemia.

This report presents improved analysis methods of daunorubicin (DAUN) and its metabolite daunorubicinol (DAUNOL) in small volumes of plasma, as total and unbound concentrations, as well as in urine. This study also presents the pharmacokinetics of DAUN and DAUNOL in patients (n = 12) diagnosed with acute myeloid leukemia treated with intravenous DAUN (60 mg/m2/day, for three days). Serial blood and urine samples were collected up to 144 h after the beginning of the first infusion. The analytical methods presented no significant matrix effect. The linear ranges were 0.1-1000 ng/mL in plasma, 0.05-40 ng/mL in ultrafiltrate and 0.5-3000 ng/ml in urine. The precision and accuracy presented coefficients of variation and standard errors lower than 15 % in the three matrices. The methods allowed for the quantification of samples up to 144 h after the beginning of the first infusion. Unbound fractions for DAUN and DAUNOL were 23.91 % (17.33-32.99) and 29.23 % (25.84-33.07), respectively. The fraction recovered in urine was 4.40 % (3.87-5.03) for DAUN and 7.91 % (6.86-9.19) for DAUNOL. Total 292.96 L/h (261.74-327.90), renal 13.01 L/h (11.44-14.88), and hepatic 280.26 L/h (248.40-317.91) clearances of DAUN, as well as the DAUNOL formation clearance 23.41 L/h (19.09-28.97), were evaluated.

Cellulose cone tip as a sorbent material for multiphase electrical field-assisted extraction of cocaine from saliva and determination by LC-MS/MS.

A porous and hydrophilic sorbent material was used in an extraction system, assisted by electric fields, for the extraction of cocaine in saliva and subsequent determination by ultra-high-performance liquid chromatography associated with sequential triple quadrupole mass spectrometry (UHPLC-MS/MS). The cellulose-based material was characterized by scanning electron microscopy, infrared spectroscopy, thermogravimetric analysis, and X-ray diffraction. The time and voltage variables applied in the extraction process were investigated through a Doehlert experimental design, and with the best conditions found (35min and 300 V) some validation parameters were evaluated. The established working range was 1-100 µg L-1 (R2 > 0.99), and the detection and quantification limits determined were 0.3 and 0.8 µg L-1, respectively. Recoveries from 80 to 115% and coefficient of variation ≤15 and 16% for intra-day and inter-day assays, respectively, were obtained for sample concentrations of LOQ, 5, 25, and 75 µg L-1, indicating satisfactory accuracy and precision for the proposed method. In addition, the method presented no matrix effect, and the extraction efficiency was between 56 and 70%. The results showed that the material used has adequate physicochemical characteristics and can be applied as a sorbent and electrolyte support in multiphase extractions using electric fields.

Direct Analysis of Amphetamine Stimulants in a Whole Urine Sample by Atmospheric Solids Analysis Probe Tandem Mass Spectrometry.

Amphetamine-type stimulants (ATS) are among illicit stimulant drugs that are most often used worldwide. A major challenge is to develop a fast and efficient methodology involving minimal sample preparation to analyze ATS in biological fluids. In this study, a urine pool solution containing amphetamine, methamphetamine, ephedrine, sibutramine, and fenfluramine at concentrations ranging from 0.5 pg/mL to 100 ng/mL was prepared and analyzed by atmospheric solids analysis probe tandem mass spectrometry (ASAP-MS/MS) and multiple reaction monitoring (MRM). A urine sample and saliva collected from a volunteer contributor (V1) were also analyzed. The limit of detection of the tested compounds ranged between 0.002 and 0.4 ng/mL in urine samples; the signal-to-noise ratio was 5. These results demonstrated that the ASAP-MS/MS methodology is applicable for the fast detection of ATS in urine samples with great sensitivity and specificity, without the need for cleanup, preconcentration, or chromatographic separation. Thus ASAP-MS/MS could potentially be used in clinical and forensic toxicology applications.

Dissolved ozone in biological fluid monitored by optical device operating in the red-infrared region

INTRODUCTION: When a gas is used for therapy, often the kinetic behavior and their distribution in biological systems is not known, leading to unsatisfactory results for clinical application. The use of ozone in living organisms has been scientifically released worldwide under the name of ozone therapy. The efficacy of this technique is determined primarily by the diffusion of gas within the tissues or fluids and which determines their action in the entire target region. We propose the development of technique to monitoring the O3 dissolved in the biological fluid using an optical device operating in the red-infrared region. METHODS: The recombination of O3 in O2 enables the monitoring of the latter by the measurement of SpO2, and, based on this phenomenon, we propose to use an optical device operating in the red-infrared region to monitoring indirectly the diffusion of O3 in fluids. The system was based on optomechanical arrangement using a capsule containing fluid that was ozonated or oxygenated during the process. A pulse oximeter is a noninvasive device used for continuously measure of SpO2 resulting from the recombination of ozone. RESULTS: The measurements of SpO2 when subjected to ozone and oxygen, showed an increased rate of SpO2 function of time for both cases reaching its peak in 80s and 160s, respectively. The experimental data concerning the SpO2 saturation as a function of time can be fitted by the theoretical model, showing a good correlation between them. CONCLUSION: A technique was developed using an optical device operating in the red-infrared region to monitoring ozone dissolved in biological fluid, showing a simple and effective way to indirectly monitoring the presence of ozone in fluids.

Determinación de la bioactividad de capas de alginato de sodio en discos de hidroxiapatita / Determination of bioactivity in sodium alginate layers of hydroxyapatite disks

OBJETIVO: el propósito central de este trabajo es evaluar la bioactividad in vitro de capas de alginato de sodio en discos de hidroxiapatita. MÉTODOS: los discos de hidroxiapatita fueron elaborados mediante procesos sucesivos de prensado y de sinterizado en un horno eléctrico. Las capas de alginato de sodio se obtuvieron empleando el método de sobrepresión y una disolución acuosa de alginato de sodio al 5 %. En el ensayo de bioactividad las muestras a estudiar fueron sumergidas en fluido biológico simulado. La caracterización de las muestras se realizó empleando microscopia electrónica de barrido y energía dispersiva de rayos X. RESULTADOS: en las muestras de hidroxiapatita sometidas al ensayo de bioactividad, con y sin capas de alginato de sodio, se observó la formación de precipitados ricos en calcio y fósforo. Además, se determinó que con el aumento del tiempo de inmersión en el fluido biológico simulado se incrementan las dimensiones de los aglomerados formados por partículas apatíticas. CONCLUSIONES: los resultados experimentales corroboran que la hidroxiapatita es bioactiva y demuestran que las capas estudiadas de alginato de sodio en discos de hidroxiapatita poseen un comportamiento bioactivo.

OBJECTIVE: the main purpose of the study is to evaluate in vitro bioactivity in sodium alginate layers of hydroxyapatite disks. METHODS: the hydroxyapatite disks were manufactured by successive pressing and sintering in an electric furnace. The sodium alginate layers were obtained by overpressure and a 5% sodium alginate aqueous solution. For the bioactivity assay, the study samples were soaked in simulated biological fluid. Characterization of the samples was conducted by scanning electron microscopy and energy dispersive X rays. RESULTS: the bioactivity assay of hydroxyapatite samples with and without sodium alginate layers revealed the formation of precipitates rich in calcium and phosphorus. It was also found that an increase in the time of immersion in the simulated biological fluid brought about an increase in the size of agglomerates of apatite particles. CONCLUSIONS: experimental results show that hydroxyapatite is indeed bioactive, and that the sodium alginate layers of hydroxyapatite disks which were studied behave bioactively.

Bomba de perfusión programable para uso biomédico

Este trabajo presenta el funcionamiento de un instrumento biomédico, diseñado y construido para cubrir los requerimientos de un grupo de investigadores en el área de bioquímica. El instrumento desarrollado tiene como propósito inyectar un fluido biológico a un sistema específico en una secuencia definida y programable, cuyo compuesto químico lo define el investigador. Dicha secuencia permite establecer diferentes velocidades de inyección del fluido en función del tiempo; todo ello, en un mismo ciclo de trabajo. Para cumplir con las características de funcionamiento se diseñó un sistema electrónico con microcontroladores y un sistema mecánico de precisión, que puede manejar una o dos inyectadoras de 3 centímetros cúbicos (cc) de uso comercial. La acción del sistema mecánico sobre las inyectadoras se encarga de inyectar el líquido en el medio o cuerpo de experimentación. El prototipo de este instrumento se está utilizando en el estudio del metabolismo de los atletas durante el ejercicio, en el laboratorio de investigación del Departamento de Bioquímica de la Facultad de Medicina de la Universidad de Los Andes.

This work presents the operation of a biomedical instrument, designed and built to cover the requirements of investigators group at the biochemistry area. The developed instrument has as purpose to inject a biological fluid, to a specific system in a defined and programmable sequence whose chemical compound the investigator defines. This sequence allows to establish different speeds of injection of the fluid in function of the time; everything it, in oneself work cycle. To fulfill the operation characteristics an electronic system it was designed with microcontrolers and a mechanical system of precision that it can manage an or two syringes of 3 cubic centimeters (cc) of commercial use. The action of the mechanical system on the syringes takes charge of inject the liquid in the means or experimentation body. The prototype of this instrument is used in the study of the metabolism of the athletes during the exercise, at the laboratory of investigation of the department of Biochemistry of the Ability of Medicine at the University of The Andes.