STUDY OF THE EFFECTS OF SHORT-CHAIN PFASs ON HUMAN CELLS WITH NMR-METABOLOMICS

Application of NMR-based Metabolomics to study the effects of shortchain PFASs on an in vitro mode

INVESTIGATING PHYSICAL PROPERTIES OF BIOPOLYMERS IN HAIR

Hair is of outmost importance for the human body. Hair is composed of 1–8% external hydrophobic lipid epidermis, 80–90% α-helix or β-sheet conformation of parallel polypeptide chains to form water-insoluble keratin, less than 3% melanin pigment, and 0.6–1.0% trace elements, 10–15% moisture (Figure 1) [1]. Keratin proteins are insoluble in water and play a protective role. They possess a heterogeneous morphology that classifies them to the fibrous structural proteins. We received samples of hair that had undergone treatment with a hair mask prepared by a cosmetic company (Figure 2). The samples were taken in different timelines (table 1) to assess not only the reconstruction of keratin induced by the cosmetic formulation but also the possibility of a long-term effect on the hai

Mycosporine-like Amino Acids and Other Phytochemicals Directly Detected by High-Resolution NMR on Klamath (Aphanizomenon flos-aquae) Blue-Green Algae

This study describes for the first time the use of high-resolution nuclear magnetic resonance (NMR) on Klamath (Aphanizomenon flos-aquae, AFA) blue-green algae directly on powder suspension. These algae are considered to be a "superfood", due to their complete nutritional profile that has proved to have important therapeutic effects. The main advantage of NMR spectroscopy is that it permits the detection of a number of metabolites all at once. The Klamath alga metabolome was revealed to be quite complex, and the most peculiar phytochemicals that can be detected directly on algae by NMR are mycosporine-like amino acids (porphyra-334, P334; shinorine, Shi) and low molecular weight glycosides (glyceryl \u3b2-d-galactopyranoside, GalpG; glyceryl 6-amino-6-deoxy-\u3b1-d-glucopyranoside, ADG), all compounds with a high nutraceutical value. The presence of cis-3,4-DhLys was revealed for the first time. This molecule could be involved in the anticancer properties ascribed to AFA

Molecular characterization and quantification using state of the art solid-state adiabatic TOBSY NMR in burn trauma

We describe a novel solid-state nuclear magnetic resonance (NMR) method that maximizes the advantages of high-resolution magic-angle-spinning (HRMAS), relative conventional liquid-state NMR approaches, when applied to intact biopsies of skeletal muscle specimens collected from burn trauma patients. This novel method, termed optimized adiabatic TOtal through Bond correlation SpectroscopY (TOBSY) solid-state NMR pulse sequence for two-dimensional (2D)1H-1H homonuclear scalar-coupling longitudinal isotropic mixing, was demonstrated to provide a 40–60% improvement in signal-to-noise ratio (SNR) relative to its liquid-state analogue TOCSY (TOtal Correlation SpectroscopY). Using 1-and 2-dimensional HRMAS NMR experiments, we identified several metabolites in burned tissues. Quantification of metabolites in burned tissues showed increased levels of lipid compounds, intracellular metabolites (e.g., taurine and phosphocreatine) and substantially decreased water-soluble metabolites (e.g., glutathione, carnosine, glucose, glutamine/glutamate and alanine). These findings demonstrate that HRMAS NMR Spectroscopy using TOBSY is a feasible technique that reveals new insights into the pathophysiology of burn trauma. Moreover, this method has applications that facilitate the development of novel therapeutic strategies

Magnetization transfer contrast MRI in GFP-tagged live bacteria

Green fluorescent protein (GFP) is a widely utilized molecular reporter of gene expression. However, its use in in vivo imaging has been restricted to transparent tissue mainly due to the tissue penetrance limitation of optical imaging. Magnetization transfer contrast (MTC) is a magnetic resonance imaging (MRI) methodology currently utilized to detect macromolecule changes such as decrease in myelin and increase in collagen content. MTC MRI imaging was performed to detect GFP in both in vitro cells and in an in vivo mouse model to determine if MTC imaging could be used to detect infection from Pseudomonas aeruginosa in murine tissues. It was demonstrated that the approach produces values that are protein specific and concentration dependent. This method provides a valuable, non-invasive imaging tool to study the impact of novel antibacterial therapeutics on bacterial proliferation and perhaps viability within the host system, and could potentially suggest the modulation of bacterial gene expression within the host when exposed to such compounds

High-resolution magic angle spinning magnetic resonance spectroscopy detects glycine as a biomarker in brain tumors

The non-essential amino acid neurotransmitter glycine (Gly) may serve as a biomarker for brain tumors. Using 36 biopsies from patients with brain tumors [12 glioblastoma multiforme (GBM); 10 low-grade (LG), including 7 schwannoma and 3 pylocytic astrocytoma; 7 meningioma (MN); 7 brain metastases (MT), including 3 adenocarcinoma and 4 breast cancer] and 9 control biopsies from patients undergoing surgery for epilepsy, we tested the hypothesis that the presence of glycine may distinguish among these brain tumor types. Using high-resolution magic angle spinning (HRMAS) 1H magnetic resonance spectroscopy (MRS), we determined a theoretically optimum echo time (TE) of 50 ms for distinguishing Gly signals from overlapping myo-inositol (Myo) signals and tested our methodology in phantom and biopsy specimens. Quantitative analysis revealed higher levels of Gly in tumor biopsies (all combined) relative to controls; Gly levels were significantly elevated in LG, MT and GBM biopsies (P≤0.05). Residual Myo levels were elevated in LG and MT and reduced in MN and GBM (P<0.05 vs. control levels). We observed higher levels of Gly in GBM as compared to LG tumors (P=0.05). Meanwhile, although Gly levels in GBM and MT did not differ significantly from each other, the Gly:Myo ratio did distinguish GBM from MT (P<0.003) and from all other groups, a distinction that has not been adequately made previously. We conclude from these findings that Gly can serve as a biomarker for brain tumors and that the Gly:Myo ratio may be a useful index for brain tumor classification

In vivo high-resolution magic angle spinning magnetic resonance spectroscopy of Drosophila melanogaster at 14.1 T shows trauma in aging and in innate immune-deficiency is linked to reduced insulin signaling

In vivo magnetic resonance spectroscopy (MRS), a non-destructive biochemical tool for investigating live organisms, has yet to be used in the fruit fly Drosophila melanogaster, a useful model organism for investigating genetics and physiology. We developed and implemented a high-resolution magic-angle-spinning (HRMAS) MRS method to investigate live Drosophila at 14.1 T. We demonstrated, for the first time, the feasibility of using HRMAS MRS for molecular characterization of Drosophila with a conventional MR spectrometer equipped with an HRMAS probe. We showed that the metabolic HRMAS MRS profiles of injured, aged wild-type (wt) flies and of immune deficient (imd) flies were more similar to chico flies mutated at the chico gene in the insulin signaling pathway, which is analogous to insulin receptor substrate 1–4 (IRS1–4) in mammals and less to those of adipokinetic hormone receptor (akhr) mutant flies, which have an obese phenotype. We thus provide evidence for the hypothesis that trauma in aging and in innate immune-deficiency is linked to insulin signaling. This link may explain the mitochondrial dysfunction that accompanies insulin resistance and muscle wasting that occurs in trauma, aging and immune system deficiencies, leading to higher susceptibility to infection. Our approach advances the development of novel in vivo non-destructive research approaches in Drosophila, suggests biomarkers for investigation of biomedical paradigms, and thus may contribute to novel therapeutic development

Distributed multi-touch virtual collaborative environments

Abstract – We present a Remote Multi-touch Collaborative Environment that integrates videoconferencing in an immersive shared space. The proposed solution supports synchronous remote and co-located interactions of two or more users sharing the same virtual workspace. Two applications, that incorporate 2D and 3D shared space, have been developed. I

The Promise of Participation and Decision-Making Power in Citizen Science

Citizen science is challenging professional researchers and their organizations to rethink the way they do science and connect with society. In any citizen science project, professional researchers are “making a promise” to the public about the level of participation and power in decision making that they are willing to provide to citizen scientists. Researchers should set expectations explicitly to ensure informed participation, trust, and motivation. Also, the design of tools for informed consent, information sharing, recognition, and privacy has to be adapted to the new power relations and distributed knowledge production. Based on fieldwork experiences and literature review about environmental and biomedical citizen science, this article examines the challenges and proposes solutions for: 1) setting expectations for informed participation; 2) addressing privacy concerns and adapting informed consent to evolving interests and networked environments; and 3) promoting citizen governance of research data. Citizen science has the potential to both increase scientific literacy and counteract mistrust and skepticism about scientific evidence of global problems (such as climate change) that need to be addressed. However, there are still many challenges to fulfilling the promise of citizen science – for example, empowering people and gaining trust. A few inspiring initiatives help us reflect on a facilitation model for engagement and informed participation; privacy by design; and new governance models for research data provided by citizen scientists