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Diagnostics (Basel, Switzerland) Jun 2023Ensuring a robust and reliable evaluation of coma deepness and prognostication of neurological outcome is challenging. We propose to develop PET neuroimaging as a new...
Ensuring a robust and reliable evaluation of coma deepness and prognostication of neurological outcome is challenging. We propose to develop PET neuroimaging as a new diagnostic and prognosis tool for comatose patients using a recently published methodology to perform functional PET (fPET). This exam permits the quantification of task-specific changes in neuronal metabolism in a single session. The aim of this protocol is to determine whether task-specific changes in glucose metabolism during the acute phase of coma are able to predict recovery at 18 months. Participation will be proposed for all patients coming for a standard PET-CT in our center in order to evaluate global cerebral metabolism during the comatose state. Legally appointed representative consent will be obtained to slightly modify the exam protocol: (1) F-fluorodeoxyglucose (F-FDG) bolus plus continuous infusion instead of a simple bolus and (2) more time under camera to perform dynamic acquisition. Participants will undergo a 55-min fPET session with a 20% bolus + 80% infusion protocol. Two occurrences of three block (5-min rest, 10-min auditory stimulation and 10-min emotional auditory stimulation) will be performed after reaching equilibrium of FDG arterial concentration. We will compare the regional brain metabolism at rest and during the sessions of auditory and emotional auditory stimulation to search for a determinant of coma recovery (18 months of follow-up after the exam). Emotional auditory stimulation should induce an activation of: the auditory cortex, the consciousness areas and the neural circuitry for emotion (function to coma deepness). An activation analysis will be carried out to highlight regional brain activation using dedicated custom-made software based on Python statistical and image processing toolboxes. The association between activation levels and the Coma Recovery Scale-Revisited (CRS-R) will be assessed using multivariate analysis. If successful, the results from this study will help improve coma prognosis evaluation based on the pattern of neuronal metabolism at the onset of the pathology. The study protocol, rationale and methods are described in this paper.
PubMed: 37370921
DOI: 10.3390/diagnostics13122026 -
Frontiers in Chemistry 2019Trace elements play a crucial role in many biochemical processes, mainly as components of vitamins and enzymes. Although small amounts of metal ions have protective... (Review)
Review
Trace elements play a crucial role in many biochemical processes, mainly as components of vitamins and enzymes. Although small amounts of metal ions have protective properties, excess metal levels result in oxidative injury, which is why metal ion homeostasis is crucial for the proper functioning of the brain. The changes of their level in the brain have been proven to be a risk factor for Alzheimer's, Parkinson's, and Huntington's diseases, as well as amyotrophic lateral sclerosis. Therefore, it is currently an important application of various analytical methods. This review covers the most important of them: inductively coupled ground mass spectrometry (ICP-MS), flame-induced atomic absorption spectrometry (FAAS), electrothermal atomic absorption spectrometry (GFAAS), optical emission spectrometry with excitation in inductively coupled plasma (ICP-OES), X-ray fluorescence spectrometry (XRF), and neutron activation analysis (NAA). Additionally, we present a summary of concentration values found by different research groups.
PubMed: 30891444
DOI: 10.3389/fchem.2019.00115 -
PloS One 2012Cardiac transcription factors are master regulators during heart development. Some were shown to transdifferentiate tail tip and cardiac fibroblasts into cardiomyocytes....
Cardiac transcription factors are master regulators during heart development. Some were shown to transdifferentiate tail tip and cardiac fibroblasts into cardiomyocytes. However, recent studies have showed that controversies exist. Potential difference in tail tip and cardiac fibroblast isolation may possibly confound the observations. Moreover, due to the use of a cardiac reporter (Myh6) selection strategy for induced cardiomyocyte enrichment, and the lack of tracking signals for each transcription factors, individual roles of each transcription factors in activating cardiac gene expression in mammalian non-myoblastic cells have never been elucidated. Answers to these questions are an important step toward cardiomyocyte regeneration. Because mouse 10T1/2 fibroblasts are non-myoblastic in nature and can be induced to express genes of all three types of muscle cells, they are an ideal model for the analysis of cardiac and non-cardiac gene activation after induction. We constructed bi-cistronic lentiviral vectors, capable of expressing cardiac transcription factors along with different fluorescent tracking signals. By infecting 10T1/2 fibroblasts with Nkx2-5, Tbx5, Gata4 or Myocd cardiac transcription factor lentivirus alone or different combinations, we found that only Tbx5+Myocd and Tbx5+Gata4+Myocd combinations induced Myh6 and Tnnt2 cardiac marker protein expression. Microarray-based gene ontology analysis revealed that Tbx5 alone activated genes involved in the Wnt receptor signaling pathway and inhibited genes involved in a number of cardiac-related processes. Myocd alone activated genes involved in a number of cardiac-related processes and inhibited genes involved in the Wnt receptor signaling pathway and non-cardiac processes. Gata4 alone inhibited genes involved in non-cardiac processes. Tbx5+Gata4+Myocd was the most effective activator of genes associated with cardiac-related processes. Unlike Tbx5, Gata4, Myocd alone or Tbx5+Myocd, Tbx5+Gata4+Myocd activated the fewest genes associated with non-cardiac processes. Conclusively, Tbx5, Gata4 and Myocd play different roles in cardiac gene activation in mammalian non-myoblastic cells. Tbx5+Gata4+Myocd activates the most cardiac and the least non-cardiac gene expression.
Topics: Animals; Cell Line; Doxycycline; Fibroblasts; GATA4 Transcription Factor; Gene Expression Profiling; Gene Ontology; Genetic Vectors; HEK293 Cells; Homeobox Protein Nkx-2.5; Homeodomain Proteins; Humans; Lentivirus; Mice; Microscopy, Confocal; Myocardium; Myosin Heavy Chains; Nuclear Proteins; Oligonucleotide Array Sequence Analysis; Reverse Transcriptase Polymerase Chain Reaction; T-Box Domain Proteins; Trans-Activators; Transcription Factors; Transcriptional Activation; Troponin T
PubMed: 23144723
DOI: 10.1371/journal.pone.0048028 -
International Journal of Molecular... Mar 2021Plant growth and development are challenged by biotic and abiotic stresses including salinity and heat stresses. For as an important greening and economic tree species... (Comparative Study)
Comparative Study
Plant growth and development are challenged by biotic and abiotic stresses including salinity and heat stresses. For as an important greening and economic tree species in China, increasing soil salinization and global warming have become major environmental challenges. We aim to unravel the molecular mechanisms underlying tree tolerance to salt stress and high temprerature (HT) stress conditions. Transcriptomics revealed that a transcription factor (TF) was significantly induced by salt stress in . This study focuses on addressing the biological functions of . The gene was cloned, and its temporal and spatial expression was analyzed under different stresses. was significantly upregulated under 150 mM NaCl and 37 °C for 12 h. The result is consistent with the presence of stress responsive -elements in the promoter. Subcellular localization analysis showed that was targeted to the nucleus. Additionally, was highly expressed in the leaves and roots. To investigate the core activation region of protein and its potential regulatory factors and targets, we conducted trans-activation analysis and the result indicates that the C-terminal region of 191-343 amino acids of the was a potent activation domain. Furthermore, overexpression of stimulated plant growth and enhanced salinity and HT tolerance. Moreover, 14 stress-related genes upregulated in the transgenic plants under high salt and HT conditions may be potential targets of the . All the results demonstrate that plays an important role in salt and HT stress tolerance.
Topics: Amino Acid Sequence; Chlorophyll; Crosses, Genetic; Gene Expression Regulation, Plant; Genes, Plant; Heat-Shock Response; Hot Temperature; Nuclear Proteins; Plant Leaves; Plant Proteins; Plant Roots; Plants, Genetically Modified; Populus; Promoter Regions, Genetic; Salinity; Salt Stress; Salt-Tolerant Plants; Sequence Alignment; Sequence Homology, Amino Acid; Species Specificity; Subcellular Fractions; Nicotiana; Transcription Factors; Transcriptional Activation
PubMed: 33800795
DOI: 10.3390/ijms22052656 -
Journal of Radioanalytical and Nuclear... Oct 2017A new γγ coincidence system has been set up at NIST. It is operated with a digital data finder supported by new software developed at NIST. The system is used to...
A new γγ coincidence system has been set up at NIST. It is operated with a digital data finder supported by new software developed at NIST. The system is used to explore possible enhancements in instrumental neutron activation analysis (INAA) and study applicability to neutron capture prompt gamma activation analysis (PGAA). The performance of the system is tested with certified reference materials for efficiency calibration and quantitative performance. Comparisons of INAA results based on conventional gamma-ray spectrometry data with INAA results based on coincidence data obtained from the same samples show improvements in the counting uncertainties and demonstrates the quantitative accuracy of the new system.
PubMed: 29056809
DOI: 10.1007/s10967-017-5342-7 -
Acta Orthopaedica Et Traumatologica... Jul 2023The aim of this study was to present an analysis of platelet-rich plasma obtained from patients with knee osteoarthritis and reveal the factors affecting its features.
OBJECTIVE
The aim of this study was to present an analysis of platelet-rich plasma obtained from patients with knee osteoarthritis and reveal the factors affecting its features.
METHODS
A total of 62 patients (mean age: 56.68 ± 7.13 years) with symptomatic knee osteoarthritis were included in this study. Age (years), gender, height (m), weight (kg), body mass index (kg/m2), duration of symptoms, smoking status, smoking index, general health status, and physical activity scores were recorded. Whole blood and platelet-rich plasma cell counts were performed with a hematology analyzer. White blood cell, red blood cell, and platelet counts were recorded. According to the dose of injected platelets, efficiency of the procedure, purity of platelet-rich plasma, and activation classification, dose of platelets, efficiency of the procedure (platelet recovery rate, %), and purity of the obtained platelet-rich plasma product (relative composition in platelets, %) were calculated. Correlation analysis between the features of platelet-rich plasma and the patient-related variables, including age, gender, body mass index, smoking status, smoking index, presence of other health conditions, physical activity scores, duration of symptoms, and pain levels, was performed.
RESULTS
Dose of injected platelets, efficiency of the procedure, purity of platelet-rich plasma, and activation analysis showed that the dose of injected platelets was 3.25 billion, the efficiency of the process was 77%, and the purity rate of the platelet-rich plasma was 98.4%. Platelet-rich plasma platelet count was correlated with whole blood platelet count (r = 0.81, P < .001), whole blood white blood cell count (r = 0.39, P = .002), smoking status (r = 0.56, P = .03), smoking index (r = -0.63, P = .002), and the presence of hypertension (r = -0.31, P=.04). Platelet-rich plasma white blood cell and purity of platelet-rich plasma were correlated with the smoking status of the patients (r = 0.52, P = .01; r = 0.64, P = .003, respectively).
CONCLUSION
This study has demonstrated that high dose and very pure platelet-rich plasma with medium efficiency was yielded with this platelet-rich plasma preparation procedure; whole blood platelet count, the presence of hypertension, and the smoking status of patients affect the features of the obtained platelet-rich plasma.
LEVEL OF EVIDENCE
Level IV, Diagnostic Study.
Topics: Humans; Middle Aged; Osteoarthritis, Knee; Platelet Count; Body Mass Index; Hypertension
PubMed: 37670448
DOI: 10.5152/j.aott.2023.22077 -
Scientific Reports Mar 2021Potassium is known for its effect on modifiable chronic diseases like hypertension, cardiac disease, diabetes (type-2), and bone health. In this study, a new method,...
Potassium is known for its effect on modifiable chronic diseases like hypertension, cardiac disease, diabetes (type-2), and bone health. In this study, a new method, neutron generator based neutron activation analysis (NAA), was utilized to measure potassium (K) in mouse carcasses. A DD110 neutron generator based NAA assembly was used for irradiation.Thirty-two postmortem mice (n= 16 males and 16 females, average weight [Formula: see text] and [Formula: see text] g) were employed for this study. Soft-tissue equivalent mouse phantoms were prepared for the calibration. All mice were irradiated for 10 minutes, and the gamma spectrum with 42K was collected using a high efficiency, high purity germanium (HPGe) detector. A lead shielding assembly was designed and developed around the HPGe detector to obtain an improved detection limit. Each mouse sample was irradiated and measured twice to reduce uncertainty. The average potassium concentration was found to be significantly higher in males [Formula: see text] compared to females [Formula: see text]. We also observed a significant correlation between potassium concentration and the weight of the mice. The detection limit for potassium quantification with the NAA system was 46 ppm. The radiation dose to the mouse was approximately 56 [Formula: see text] mSv for 10-min irradiation. In conclusion, this method is suitable for estimating individual potassium concentration in small animals. The direct evaluation of total body potassium in small animals provides a new way to estimate potassium uptake in animal models. This method can be adapted later to quantify potassium in the human hand and small animals in vivo. When used in vivo, it is also expected to be a valuable tool for longitudinal assessment, kinetics, and health outcomes.
Topics: Animals; Bone and Bones; Disease Models, Animal; Gamma Rays; Germanium; Ion Transport; Male; Mice; Monte Carlo Method; Neutron Activation Analysis; Neutrons; Phantoms, Imaging; Potassium; Radiation Dosage; Whole-Body Irradiation
PubMed: 33737537
DOI: 10.1038/s41598-021-85233-2 -
International Journal of Molecular... Apr 2024The pathogenesis of carcinoma is believed to come from the combined effect of polygenic variation, and the initiation and progression of malignant tumors are closely...
The pathogenesis of carcinoma is believed to come from the combined effect of polygenic variation, and the initiation and progression of malignant tumors are closely related to the dysregulation of biological pathways. Quantifying the alteration in pathway activation and identifying coordinated patterns of pathway dysfunction are the imperative part of understanding the malignancy process and distinguishing different tumor stages or clinical outcomes of individual patients. In this study, we have conducted in silico pathway activation analysis using Riemannian manifold (RiePath) toward pan-cancer personalized characterization, which is the first attempt to apply the Riemannian manifold theory to measure the extent of pathway dysregulation in individual patient on the tangent space of the Riemannian manifold. RiePath effectively integrates pathway and gene expression information, not only generating a relatively low-dimensional and biologically relevant representation, but also identifying a robust panel of biologically meaningful pathway signatures as biomarkers. The pan-cancer analysis across 16 cancer types reveals the capability of RiePath to evaluate pathway activation accurately and identify clinical outcome-related pathways. We believe that RiePath has the potential to provide new prospects in understanding the molecular mechanisms of complex diseases and may find broader applications in predicting biomarkers for other intricate diseases.
Topics: Humans; Neoplasms; Precision Medicine; Biomarkers, Tumor; Gene Expression Regulation, Neoplastic; Signal Transduction; Gene Expression Profiling; Algorithms; Computational Biology; Gene Regulatory Networks; Computer Simulation
PubMed: 38673997
DOI: 10.3390/ijms25084411 -
Control of Platelet CLEC-2-Mediated Activation by Receptor Clustering and Tyrosine Kinase Signaling.Biophysical Journal Jun 2020Platelets are blood cells responsible for vascular integrity preservation. The activation of platelet receptor C-type lectin-like receptor II-type (CLEC-2) could...
Platelets are blood cells responsible for vascular integrity preservation. The activation of platelet receptor C-type lectin-like receptor II-type (CLEC-2) could partially mediate the latter function. Although this receptor is considered to be of importance for hemostasis, the rate-limiting steps of CLEC-2-induced platelet activation are not clear. Here, we aimed to investigate CLEC-2-induced platelet signal transduction using computational modeling in combination with experimental approaches. We developed a stochastic multicompartmental computational model of CLEC-2 signaling. The model described platelet activation beginning with CLEC-2 receptor clustering, followed by Syk and Src family kinase phosphorylation, determined by the cluster size. Active Syk mediated linker adaptor for T cell protein phosphorylation and membrane signalosome formation, which resulted in the activation of Bruton's tyrosine kinase, phospholipase and phosphoinositide-3-kinase, calcium, and phosphoinositide signaling. The model parameters were assessed from published experimental data. Flow cytometry, total internal reflection fluorescence and confocal microscopy, and western blotting quantification of the protein phosphorylation were used for the assessment of the experimental dynamics of CLEC-2-induced platelet activation. Analysis of the model revealed that the CLEC-2 receptor clustering leading to the membrane-based signalosome formation is a critical element required for the accurate description of the experimental data. Both receptor clustering and signalosome formation are among the rate-limiting steps of CLEC-2-mediated platelet activation. In agreement with these predictions, the CLEC-2-induced platelet activation, but not activation mediated by G-protein-coupled receptors, was strongly dependent on temperature conditions and cholesterol depletion. Besides, the model predicted that CLEC-2-induced platelet activation results in cytosolic calcium spiking, which was confirmed by single-platelet total internal reflection fluorescence microscopy imaging. Our results suggest a refined picture of the platelet signal transduction network associated with CLEC-2. We show that tyrosine kinase activation is not the only rate-limiting step in CLEC-2-induced activation of platelets. Translocation of receptor-agonist complexes to the signaling region and linker adaptor for T cell signalosome formation in this region are limiting CLEC-2-induced activation as well.
Topics: Blood Platelets; Cluster Analysis; Lectins, C-Type; Membrane Glycoproteins; Platelet Activation; Protein-Tyrosine Kinases; Signal Transduction
PubMed: 32396849
DOI: 10.1016/j.bpj.2020.04.023 -
Journal of Nuclear Materials. Journal... 2018Understanding the chemical durability of neutron shielding materials is necessary when assessing their long-term service potential. In this study, the chemical...
Understanding the chemical durability of neutron shielding materials is necessary when assessing their long-term service potential. In this study, the chemical durability of a Li enriched neutron shielding glass that has been exposed to natural, near-operational conditions is assessed by Prompt Gamma Activation Analysis (PGAA) and Neutron Depth Profiling (NDP). These non-destructive, nuclear analysis techniques are sensitive to Li, and PGAA is uniquely able to detect H in low quantities in solids. It was determined that the enriched alumino-silicate glass can alter within 2 months of exposure to the natural environment. This exposure resulted in an average surface alteration layer thickness of ≈22 μm. The alteration layer contained ≈47% less Li than the bulk glass. Alternatively, a 3 years exposed sample of the glass had a surface alteration depth of ≈30 μm and Li depletion levels in the alteration layer were between 47% and 75% less Li than the bulk glass. When the alteration layer on the 3 years sample was removed, the H content of the glass's surface was nearly eliminated. This sample also showed variable Li concentrations throughout the alteration volume, which contrasts with near static Li concentration in the alteration volume of the 2 months sample. From these findings it was determined that the depletion in Li at the surface of the glass will not affect the glass's neutron shielding properties, but it may change the mechanical stability of the glass's surface and, due to increased H content in the alteration layer, make it an inappropriate material for the lining of certain neutron analysis instruments.
PubMed: 31092955
DOI: 10.1016/j.jnucmat.2018.09.034