Machine learning-based detection of acute psychosocial stress from body posture and movements.

Investigating acute stress responses is crucial to understanding the underlying mechanisms of stress. Current stress assessment methods include self-reports that can be biased and biomarkers that are often based on complex laboratory procedures. A promising additional modality for stress assessment might be the observation of body movements, which are affected by negative emotions and threatening situations. In this paper, we investigated the relationship between acute psychosocial stress induction and body posture and movements. We collected motion data from N = 59 individuals over two studies (Pilot Study: N = 20, Main Study: N = 39) using inertial measurement unit (IMU)-based motion capture suits. In both studies, individuals underwent the Trier Social Stress Test (TSST) and a stress-free control condition (friendly-TSST; f-TSST) in randomized order. Our results show that acute stress induction leads to a reproducible freezing behavior, characterized by less overall motion as well as more and longer periods of no movement. Based on these data, we trained machine learning pipelines to detect acute stress solely from movement information, achieving an accuracy of 75.0 ± 17.7 % (Pilot Study) and 73.4 ± 7.7 % (Main Study). This, for the first time, suggests that body posture and movements can be used to detect whether individuals are exposed to acute psychosocial stress. While more studies are needed to further validate our approach, we are convinced that motion information can be a valuable extension to the existing biomarkers and can help to obtain a more holistic picture of the human stress response. Our work is the first to systematically explore the use of full-body body posture and movement to gain novel insights into the human stress response and its effects on the body and mind.

Taming the Biological Activity of Pd(II) and Pt(II) Complexes with Triazolato "Protective" Groups: 1H, 77Se Nuclear Magnetic Resonance and X-ray Crystallographic Model Studies with Selenocysteine to Elucidate Differential Thioredoxin Reductase Inhibition.

The biological activity of Pd(II) and Pt(II) complexes toward three different cancer cell lines as well as inhibition of selenoenzyme thioredoxin reductase (TrxR) was modulated in an unexpected way by the introduction of triazolate as a "protective group" to the inner metal coordination sphere using the iClick reaction of [M(N3)(terpy)]PF6 [M = Pd(II) or Pt(II) and terpy = 2,2':6',2″-terpyridine] with an electron-poor alkyne. In a cell proliferation assay using A549, HT-29, and MDA-MB-231 human cancer cell lines, the palladium compound was significantly more potent than the isostructural platinum analogue and exhibited submicromolar activity on the most responsive cell line. This difference was also reflected in the inhibitory efficiency toward TrxR with IC50 values of 0.1 versus 5.4 µM for the Pd(II) and Pt(II) complexes, respectively. UV/Vis kinetic studies revealed that the Pt compound binds to selenocysteine faster than to cysteine [k = (22.9 ± 0.2)·10-3 vs (7.1 ± 0.2)·10-3 s-1]. Selective triazolato ligand exchange of the title compounds with cysteine (Hcys) and selenocysteine (Hsec)─but not histidine (His) and 9-ethylguanine (9EtG)─was confirmed by 1H, 77Se, and 195Pt NMR spectroscopy. Crystal structures of three of the four ligand exchange products were obtained, including [Pt(sec)(terpy)]PF6 as the first metal complex of selenocysteine to be structurally characterized.

Isostructural Series of Ni(II), Pd(II), Pt(II), and Au(III) Azido Complexes with a N

An isoelectronic and isostructural series of cyclometalated azido complexes [M(N3)(dpb)] with M = Ni(II), Pd(II), Pt(II), and Au(III) based on the N^C^N pincer ligand 1,3-di(2-pyridyl)phenide (dpb) was characterized by X-ray diffraction analysis and investigated for reactivity in the iClick reaction with a wide range of internal and terminal alkynes by using 1H and 19F NMR spectroscopy. Reaction rate constants were found to increase with greater charge density in the order Ni(II) > Pd(II) > Pt(II) > Au(III). Terminal alkynes R-C≡C-R' with strongly electron-withdrawing groups R and R' exhibited faster kinetics than those with electron-donating substituents in the order CF3 > ketone > ester > H > phenyl ≫ amide, while R = CH3 resulted in complete loss of reactivity. Four symmetrical triazolato complexes [M(triazolatoCOOCH3,COOCH3)(dpb)] with M = Ni(II), Pd(II), Pt(II), and Au(III) as well as four nonsymmetrically substituted triazolato complexes [Pt(triazolatoR,R')(dpb)] originating from terminal and internal alkynes were shown by X-ray crystal structure analysis to exclusively feature N2-coordination of the five-membered ring ligand. However, the Pt(II) triazolato complexes exist as a mixture of N1- and N2-coordinated species in solution. Torsion angles between the mean planes of the N^C^N pincer and the triazolato ligand increase from a nearly coplanar to a perpendicular arrangement when going from Au(III)/Pt(II)/Pd(II) to Ni(II), while different substituents R and R' on the alkyne have no influence on the torsion angle and were rationalized by DFT calculations. Finally, a carbohydrate derivative obtained by glucuronic acid conjugation to methyl propiolate demonstrates the facile biofunctionalization of metal complexes via the iClick reaction.

Sulfinyl-aminotroponiminates: alkali- (Li, Na, K) and heavy-metal (Bi) complexes.

The installation of electron-withdrawing functional groups at the carbocyclic backbone of aminotroponiminate (ATI) ligands is a versatile method for influencing the electronic properties of the resulting ATI complexes. We report here Li, Na, and K salts of an ATI ligand with a phenylsulfinyl substituent in the backbone. It is demonstrated that the sulfinyl group actively contributes to the coordination chemistry of these complexes, effectively competing with neutral donor ligands such as thf or pyridine in the solid state (XRD), in solution (DOSY NMR spectroscopy), and in the gas phase (DFT). The impact of the phenylsulfinyl group on the redox properties of the complexes have been investigated and access to sodium sodiate species through ligand-induced disproportionation has been studied. Transfer of the ATI ligand to the heavy p-block element bismuth has been demonstrated. Analytical techniques applied in this work include multinuclear and DOSY NMR spectroscopy, cyclic voltammetry, DFT calculations, and single-crystal X-ray diffraction analysis.

Sensitive determination of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol and complementary cannabinoids in hair using alkaline digestion and mixed-mode solid phase extraction followed by liquid-chromatography-tandem mass spectrometry.

Hair drug testing can be used for the evaluation of cannabis use with a large detection window, and is required for professional driving license granting in Brazil. A positive hair result for cannabis use requires quantification of the metabolite THC-COOH above the cutoff value of 0.2 ng/g. The achievement of such lower limit of quantification is challenging, particularly with the use of liquid chromatography coupled to triple quadrupole mass spectrometers (LC-MS/MS). In this study, a very sensitive LCMS/ MS assay for the simultaneous quantification of THC-COOH along with THC, CBD, and CBN was developed and validated. Sample preparation was based on hair hydrolysis, followed by selective ion-exchange solid-phase extraction. The extraction yield was 101.5-101.6% for THC-COOH, 92.3-97.4% for THC, 89.7-95.2% for CBN, and 104.9-121.1% for CBD. Internal standard corrected matrix effects were - 2.7 to - 1,1 for THCCOOH and - 11.5 to - 0.1% for the other analytes. The lower limit of quantification was 01 ng/g for THC-COOH and 25 ng/g for THC, CBD, and CBN. The assay fulfilled validation guidelines acceptance criteria. The measurement uncertainties were determined and the assay was ISO17025 accredited, being currently used in routine testing.

Simple extraction of toxicologically relevant psychotropic compounds and metabolites from whole blood using mini-QuEChERS followed by UPLC-MS/MS analysis.

The determination of psychotropic drugs and metabolites in blood is relevant in the context of both therapeutic drug monitoring and clinical and forensic toxicology. LC-MS/MS is the preferred method for these assays. However, LC-MS/MS is particularly susceptible to matrix ionization effects and appropriate sample preparation is required to minimize these effects. In this study, a simple, single-step, mini-QuEchERS extraction procedure, coupled to UPLC-MS/MS, was developed and validated for the determination of 15 toxicologically relevant compounds in whole blood, including psychoactive drugs and some metabolites. The assay was linear in the range of 25-1,000 ng ml-1 , fulfilling criteria for accuracy and precision. Extraction yields (71.9-87.7%) and matrix effects (-3.3 to +4.4%, with the exception of codeine, which had matrix effects of -35.36 to -28.14%) were acceptable for the majority of the evaluated compounds, using a single internal standard. The assay was applied to 238 clinical specimens from patients admitted to an emergency service, with 22 samples presenting quantifiable concentrations of 11 different compounds. The developed assay is a simple and efficient strategy for determination of target psychotropic drugs and metabolites in forensic and clinical toxicology.

Blood phosphatidyl ethanol levels as a tool to detect alcohol misuse in trauma patients.

INTRODUCTION: There is a strong need for a reliable marker of harmful alcohol consumption to identify injured patients that can benefit from alcohol interventions, and blood phosphatidyl ethanol (PEth) has not previously been tested on this population. This study aims to compare the performance of blood PEth concentration, blood alcohol concentration (BAC) and the Alcohol Use Disorders Identification Test Consumption (AUDIT-C) for the screening of alcohol misuse in trauma patients. METHODS: Prospective cross-sectional study of 238 adult patients presenting in the emergency department with any type of trauma. PEth concentration was determined in whole blood by high-performance liquid chromatography with tandem mass spectrometry. Consent, AUDIT-C score and demographic data were obtained. RESULTS: The sample consisted of majority male (67.6%), single (46.2%) and employed (66%) patients. The most common type of trauma was traffic collision (63.9%). The mean age was 41.7 years. We found a significant correlation between PEth levels with AUDIT-C score (Spearman's r = 0.654; p < .0001). PEth had an area under the ROC curve of 0.885 to detect hazardous alcohol consumption (AUDIT-C score ≥ 6) and PEth ≥23.9 ng/mL cutoff point provided 91.2% of sensitivity and 78.4% of specificity. Twelve patients reported alcohol abstinence, but had quantifiable levels of PEth. CONCLUSIONS: PEth levels and AUDIT-C score had a moderate correlation in our population. PEth was useful to identify 12 cases of underreporting of alcohol consumption habits. PEth shows promising results, but more research is needed to identify the best screening tool for alcohol misuse in trauma patients.

Validation of an analytical method for the simultaneous determination of 16 drugs and metabolites in hair in the context of driving license granting.

The use of psychoactive substances has been associated with increased risk for traffic accidents. Hair testing has become a routine practice in clinical and forensic toxicological laboratories, with a unique perspective in the investigation of drug consumption. The study aimed to develop and validate a UHPLC-MS/MS method for the determination of multiple drugs in hair, to be used for toxicological examination in driving license granting. Sample preparation was a one-step liquid extraction of milled hair with methanol, which was incubated for 15h at 50°C. The chromatographic separation was performed in a reversed phase column, with a run time of 2.2min. Measured compounds were cocaine, benzoylecgonine, norcocaine, anhydroecgonine methyl ester, cocaethylene, amphetamine, methamphetamine, methylenedioxyamphetamine, methylenedioxymethamphetamine, fenproporex, amfepramone, mazindol, codeine, morphine, 6-monoacetylmorphine, and tetrahydrocannabinol. The assay was linear for all substances (r>0.99), accurate (86.63-105.87 %), and precise, with a cv ranging from 1.9-13.5 % for intra-assay and 3.3-14.3 % for inter-assay. There was no significant carry over effect and the internal standard corrected matrix effect was minimal. The relative uncertainty percentages were below 9% for all the substances at cut-off values. The method was successfully applied to 50 hair samples from injured drivers, with 12% of positivity, including cocaine, MDMA and THC.

Determination of cannabinoids in plasma using salting-out-assisted liquid-liquid extraction followed by LC-MS/MS analysis.

The detection of the markers of Cannabis consumption in biological specimens is an important task for drug testing laboratories in varous contexts. A simple assay combining salting-out assisted liquid-liquid extraction sample preparation and LC-MS/MS analysis was applied to the measurement of Δ9 -tetrahydrocannabinol, 11-nor-9-carboxy-Δ9 -tetrahydrocannabinol (THC-COOH), 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in 100 µl plasma specimens. The assay had linearity of 1-100 ng ml-1 for THC-COOH and 0.5-50 ng ml-1 for the other tested cannabinoids. Assay validation criteria were fulfilled. Extraction yields (88.7-97.3%) and internal-standard correct matrix effects (-9.6 to +5.4%) were acceptable. The assay was applied to 238 clinical specimens from trauma patients, with 19 samples presenting quantifiable concentrations of at least one of the target compounds. The developed assay is a simple and efficient strategy for simultaneous measurement of Δ9 -tetrahydrocannabinol, THC-COOH, 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in plasma specimens.

Determination of Endogenous Concentrations of Uracil and Dihydrouracil in Dried Saliva Spots by LC-MS/MS: Method Development, Validation, and Clinical Application.

BACKGROUND: The aim of this study was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the measurement of uracil (U) and dihydrouracil (UH2) concentrations in dried saliva spots (DSSs), for the evaluation of dihydropyrimidine dehydrogenase (DPD) enzyme activity. RESULTS: Nine 18-mm diameter DSS discs were extracted with acetate:isopropyl alcohol (85:15, vol/vol) and analyzed by LC-MS/MS. The assay was linear in the range of 10-1000 ng·mL, with accuracy between 89% and 112% and precision between 5.7% and 13%. The metabolic ratio [UH2]/[U] was stable in DSS for up to 9 days at 45°C. Concentrations of U and UH2, as well as the metabolic ratio, were highly concordant between matrices. Using a metabolic ratio classification cutoff of 1.16 for the identification of slow DPD metabolizers, 98.7% concordance was achieved between SS and saliva. CONCLUSIONS: DSS samples could be a useful alternative for DPD activity screening, particularly in locations with limited access to highly equipped laboratories.

DPD functional tests in plasma, fresh saliva and dried saliva samples as predictors of 5-fluorouracil exposure and occurrence of drug-related severe toxicity.

OBJECTIVE: to evaluate plasma and salivary uracil (U) to dihydrouracil (UH2) ratios as tools for predicting 5-fluorouracil systemic exposure and drug-related severe toxicity, and clinically validate the use of dried saliva spots (DSS) as an alternative sampling strategy for dihydropyrimidine dehydrogenase (DPD) deficiency assessment. METHODS: Pre-chemotherapy plasma, fresh saliva and DSS samples were obtained from gastrointestinal patients (N = 40) for measurement of endogenous U and UH2 concentrations by LC-MS/MS. A second plasma sample collected during 5FU infusion was used for 5FU area under the curve (AUC) determination by HPLC-DAD. Data on toxicity was reported according to CTCAE. RESULTS: 15% of the patients developed severe 5FU-related toxicity, with neutropenia accounting for 67% of the cases. U, UH2 and [UH2,]/[U] were highly correlated between fresh and dried saliva samples (rs = 0.960; rs = 0.828; rs = 0.910, respectively). 5FU AUC ranged from 11.3 to 37.31 mg h L-1, with 46.2% of under-dosed and 10.3% over-dosed patients. The [UH2]/[U] ratios in plasma, fresh saliva and dried saliva samples were moderately correlated with 5FU AUC and adverse events grade, indicating a partial contribution of the variables to drug exposure (r = -0.412, rs = -0.373, rs = 0.377) and toxicity (r = -0.363, rs = -0.523, rs = 0.542). Metabolic ratios were lower in patients with severe toxicity (P < .01 salivary ratios, and P < .5 plasma ratios), and 5FU AUC were in average 47% higher in this group than in moderate toxicity. The diagnostic performance of [UH2]/[U] ratios in fresh saliva and DSS for the identification of patients with severe toxicity were comparable. CONCLUSIONS: The [UH2]/[U] metabolic ratios in plasma, fresh saliva and DSS were significantly associated with 5FU systemic exposure and toxicity degree. This study also demonstrated the applicability of DSS as alternative sampling for evaluating DPD activity.

Relación entre las maloclusiones y la respiración bucal en pacientes que asistieron al Servicio de Otorrinolaringología del Hospital Pediátrico San Juan de Dios; junio de 2005 / Relationship between malocclusions and mouth breathing in patients who attended the ORL Service of the Juan de Dios Pediatric Hospital, June 2005

La Insuficiencia Respiratoria Nasal es una condición patológica que se presenta con relativa frecuencia en los pacientes que acuden a consulta odontológica y de otorrinolaringología. Estos pacientes adquieren una postura adaptativa de las estructuras de la cabeza y la región del cuello, que afectan la relación de los maxilares y el desarrollo normal de la oclusión. El propósito del presente estudio fue demostrar si existe relación entre los pacientes con Insuficiencia Respiratoria Nasal y un tipo específico de maloclusión dentaria utilizando la clasificación de Angle. Así mismo se planteó como propósito interrelacionar el área de odontología con la de otorrinolaringología, con el fin de establecer un sistema de referencias de un área a otra para brindar una atención integral. El estudio se realizó, en 39 niños que asistieron al servicio de otorrinolaringología del Hospital San Juan de Dios en el mes de Junio de 2005, de edades comprendidas entre 6 y 12 años, de ambos sexos y que presentaban problemas otorrinos que causaran respiración bucal. A estos pacientes se les realizó el examen clínico bucal para determinar las maloclusiones presentes. Los resultados no evidenciaron relación entre la Insuficiencia Respiratoria Nasal y una maloclusión específica. Hubo una marcada presencia de apiñamiento anteroinferior, de mordidas abiertas y de características propias de la fascie adenoidea. El tipo de maloclusión predominante fue la Clase I y el principal problema de otorrinolaringología causante de insuficiencia respiratoria nasal fue la Hipertrofia Adenotonsilar.

Nasal Respiratory Insufficiency is a frequent disease in patients suffering from odontological and ORL conditions. These patients acquire an adaptive posture of the head and neck structures affecting the relationship between the maxilla, the mandible and the occlusion normal development. The purpose of this study was to demonstrate the existence of a relationship between Nasal Respiratory Insufficiency and a specific type of dental malocclusion using Angle's classification. Another purpose was to integrate the Dental and ORL fields in order to establish a reference system between both specialties. The study included 39 children of either sex, 6 to 12 years-old. They were treated during June 2005 at the ORL Department of the San Juan de Dios Hospital. They presented ORL conditions which also caused oral respiration. They were studied to determine the existence of dental malocclusion. The results did not show any relationship between Nasal Respiratory Insufficiency and malocclusion. The predominant malocclusion type was Class I and the main ORL pathology causing Nasal Respiratory Insufficiency was Adenotonsilar Hypertrophy.

Acetylcholine increases the free intracellular calcium concentration in podocytes in intact rat glomeruli via muscarinic M(5) receptors.

The effects of acetylcholine (ACh) on the free intracellular calcium concentration ([Ca2+](i)) of microdissected glomeruli were investigated using fura-2 fluorescence digital imaging and two-photon confocal microscopy. ACh caused a concentration-dependent [Ca2+](i) increases with an initial peak followed by a sustained plateau, which was suppressed by reduced extracellular Ca2+ concentrations. The [Ca2+](i) plateau was not affected by the L-type Ca2+ channel blocker nicardipine, whereas gadolinium and lanthanum (both at 1 microM) blocked the plateau. Diphenylacetoxy-N:-methylpiperidine methiodide (100 nM), an M(3)/M(5) receptor antagonist, and pirenzepine (1 microM), an M(1) receptor antagonist, completely inhibited the effect of ACh. [Ca2+](i) measurements using two-photon excitation of fluo-3 and staining of the cells with calcein/acetoxymethyl ester, for observation of the capillary network together with the glomerular cells, showed that [Ca2+](i) was increased in single podocytes. Immunohistochemical studies did not demonstrate M(3) receptor expression in glomerular cells. M(1) receptors could be detected only in the parietal sheet of Bowman's capsule, whereas M(5) receptors were found only in podocytes. The data show that ACh increases [Ca2+](i) in podocytes of intact glomeruli, most likely via muscarinic M(5) receptors.