scholarly journals Sampling interstitial fluid from human skin using a microneedle patch

2020 ◽  
Vol 12 (571) ◽  
pp. eaaw0285
Author(s):  
Pradnya P. Samant ◽  
Megan M. Niedzwiecki ◽  
Nicholas Raviele ◽  
Vilinh Tran ◽  
Juan Mena-Lapaix ◽  
...  
Keyword(s):  
Minimally Invasive ◽  
Human Skin ◽  
Interstitial Fluid ◽  
Mass Spectrometry Analysis ◽  
Limited Attention ◽  
Spectrometry Analysis ◽  
Tissue Interstitial Fluid ◽  
Exogenous Compounds ◽  
Continuous Glucose Monitors ◽  
Microneedle Patch

Tissue interstitial fluid (ISF) surrounds cells and is an underutilized source of biomarkers that complements conventional sources such as blood and urine. However, ISF has received limited attention due largely to lack of simple collection methods. Here, we developed a minimally invasive, microneedle-based method to sample ISF from human skin that was well tolerated by participants. Using a microneedle patch to create an array of micropores in skin coupled with mild suction, we sampled ISF from 21 human participants and identified clinically relevant and sometimes distinct biomarkers in ISF when compared to companion plasma samples based on mass spectrometry analysis. Many biomarkers used in research and current clinical practice were common to ISF and plasma. Because ISF does not clot, these biomarkers could be continuously monitored in ISF similar to current continuous glucose monitors but without requiring an indwelling subcutaneous sensor. Biomarkers distinct to ISF included molecules associated with systemic and dermatological physiology, as well as exogenous compounds from environmental exposures. We also determined that pharmacokinetics of caffeine in healthy adults and pharmacodynamics of glucose in children and young adults with diabetes were similar in ISF and plasma. Overall, these studies provide a minimally invasive method to sample dermal ISF using microneedles and demonstrate human ISF as a source of biomarkers that may enable research and translation for future clinical applications.

scholarly journals SAMHD1 Enhances Chikungunya and Zika Virus Replication in Human Skin Fibroblasts

Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 81
Author(s):  
Sineewanlaya Wichit
Keyword(s):  
Human Skin ◽  
Zika Virus ◽  
Antiviral Treatment ◽  
Skin Fibroblasts ◽  
Mass Spectrometry Analysis ◽  
Strong Increase ◽  
Human Skin Fibroblasts ◽  
Entry Site ◽  
Stable Isotope Labelling ◽  
Spectrometry Analysis

Chikungunya virus (CHIKV) and Zika virus (ZIKV) are emerging arboviruses that pose a worldwide threat to human health. Currently, neither vaccine nor antiviral treatment to control their infections is available. As the skin is a major viral entry site for arboviruses in the human host, we determined the global proteomic profile of CHIKV and ZIKV infections in human skin fibroblasts using stable isotope labelling by amino acids in cell culture (SILAC)-based mass spectrometry analysis. We show that the expressions of the interferon-stimulated proteins MX1, IFIT1, IFIT3 and ISG15, as well as expressions of defense response proteins DDX58, STAT1, OAS3, EIF2AK2, and SAMHD1 were significantly upregulated in these cells upon infection with either virus. Exogenous expression of IFITs proteins markedly inhibited CHIKV and ZIKV replication which, accordingly, was restored following the abrogation of IFIT1 or IFIT3. Overexpression of SAMHD1 in cutaneous cells or pretreatment of cells with the virus-like particles containing SAMHD1 restriction factor Vpx resulted in a strong increase or inhibition, respectively, in both CHIKV and ZIKV replication. Moreover, silencing of SAMHD1 by specific SAMHD1-siRNA resulted in a marked decrease in viral RNA levels. Together, these results suggest that IFITs are involved in the restriction of replication of CHIKV and ZIKV and provide, as yet unreported, evidence for a proviral role of SAMHD1 in arbovirus infection of human skin cells.

scholarly journals Skeletal muscle interstitial fluid metabolomics at rest and associated with an exercise bout: application in rats and humans

2019 ◽  
Vol 316 (1) ◽  
pp. E43-E53 ◽  
Author(s):  
Jie Zhang ◽  
Sudeepa Bhattacharyya ◽  
Robert C. Hickner ◽  
Alan R. Light ◽  
Christopher J. Lambert ◽  
...  
Keyword(s):  
Amino Acids ◽  
Interstitial Fluid ◽  
Vastus Lateralis ◽  
Exercise Bout ◽  
Male Rats ◽  
Mass Spectrometry Analysis ◽  
Maximal Heart Rate ◽  
Spectrometry Analysis ◽  
Flight Mass Spectrometry ◽  
Muscle Work

Blood or biopsies are often used to characterize metabolites that are modulated by exercising muscle. However, blood has inputs derived from multiple tissues, biopsies cannot discriminate between secreted and intracellular metabolites, and their invasive nature is challenging for frequent collections in sensitive populations (e.g., children and pregnant women). Thus, minimally invasive approaches to interstitial fluid (IF) metabolomics would be valuable. A catheter was designed to collect IF from the gastrocnemius muscle of acutely anesthetized adult male rats at rest or immediately following 20 min of exercise (~60% of maximal O2 uptake). Nontargeted, gas chromatography-time-of-flight mass spectrometry analysis was used to detect 299 metabolites, including nonannotated metabolites, sugars, fatty acids, amino acids, and purine metabolites and derivatives. Just 43% of all detected metabolites were common to IF and blood plasma, and only 20% of exercise-modified metabolites were shared in both pools, highlighting that the blood does not fully reflect the metabolic outcomes in muscle. Notable exercise patterns included increased IF amino acids (except leucine and isoleucine), increased α-ketoglutarate and citrate (which may reflect tricarboxylic acid cataplerosis or shifts in nonmitochondrial pathways), and higher concentration of the signaling lipid oleamide. A preliminary study of human muscle IF was conducted using a 20-kDa microdialysis catheter placed in the vastus lateralis of five healthy adults at rest and during exercise (65% of estimated maximal heart rate). Approximately 70% of commonly detected metabolites discriminating rest vs. exercise in rats were also changed in exercising humans. Interstitium metabolomics may aid in the identification of molecules that signal muscle work (e.g., exertion and fatigue) and muscle health.

scholarly journals SAMHD1 Enhances Chikungunya and Zika Virus Replication in Human Skin Fibroblasts

2019 ◽  
Vol 20 (7) ◽  
pp. 1695 ◽  
Author(s):  
Sineewanlaya Wichit ◽  
Rodolphe Hamel ◽  
Andreas Zanzoni ◽  
Fodé Diop ◽  
Alexandra Cribier ◽  
...  
Keyword(s):  
Human Skin ◽  
Zika Virus ◽  
Antiviral Treatment ◽  
Skin Fibroblasts ◽  
Mass Spectrometry Analysis ◽  
Strong Increase ◽  
Human Skin Fibroblasts ◽  
Entry Site ◽  
Stable Isotope Labelling ◽  
Spectrometry Analysis

Chikungunya virus (CHIKV) and Zika virus (ZIKV) are emerging arboviruses that pose a worldwide threat to human health. Currently, neither vaccine nor antiviral treatment to control their infections is available. As the skin is a major viral entry site for arboviruses in the human host, we determined the global proteomic profile of CHIKV and ZIKV infections in human skin fibroblasts using Stable Isotope Labelling by Amino acids in Cell culture (SILAC)-based mass-spectrometry analysis. We show that the expression of the interferon-stimulated proteins MX1, IFIT1, IFIT3 and ISG15, as well as expression of defense response proteins DDX58, STAT1, OAS3, EIF2AK2 and SAMHD1 was significantly up-regulated in these cells upon infection with either virus. Exogenous expression of IFITs proteins markedly inhibited CHIKV and ZIKV replication which, accordingly, was restored following the abrogation of IFIT1 or IFIT3. Overexpression of SAMHD1 in cutaneous cells, or pretreatment of cells with the virus-like particles containing SAMHD1 restriction factor Vpx, resulted in a strong increase or inhibition, respectively, of both CHIKV and ZIKV replication. Moreover, silencing of SAMHD1 by specific SAMHD1-siRNA resulted in a marked decrease of viral RNA levels. Together, these results suggest that IFITs are involved in the restriction of replication of CHIKV and ZIKV and provide, as yet unreported, evidence for a proviral role of SAMHD1 in arbovirus infection of human skin cells.

scholarly journals The molecular mechanism of N-acetylglucosamine side-chain attachment to the Lancefield group A Carbohydrate inStreptococcus pyogenes

2017 ◽  
Author(s):  
Jeffrey S. Rush ◽  
Rebecca J. Edgar ◽  
Pan Deng ◽  
Jing Chen ◽  
Haining Zhu ◽  
...  
Keyword(s):  
Cell Envelope ◽  
Mass Spectrometry Analysis ◽  
Side Chain ◽  
Antigenic Determinants ◽  
Limited Attention ◽  
Spectrometry Analysis ◽  
Gram Positive Bacteria ◽  
Essential Components ◽  
Group A ◽  
Lancefield Group

AbstractIn many Lactobacillales species (i.e. lactic acid bacteria), peptidoglycan is decorated by polyrhamnose polysaccharides that are critical for cell envelope integrity and cell shape and also represent key antigenic determinants. Despite the biological importance of these polysaccharides, their biosynthetic pathways have received limited attention. The important human pathogen,Streptococcus pyogenes, synthesizes a key antigenic surface polymer—the Lancefield group A carbohydrate (GAC). GAC is covalently attached to peptidoglycan and consists of a polyrhamnose polymer, with N-acetylglucosamine (GlcNAc) side chains, which is an essential virulence determinant. The molecular details of the mechanism of polyrhamnose modification with GlcNAc are currently unknown. In this report, using molecular genetics, analytical chemistry and mass spectrometry analysis, we demonstrated that GAC biosynthesis requires two distinct undecaprenol-linked GlcNAc-lipid intermediates: GlcNAc-pyrophosphorylundecaprenol (GlcNAc-P-P-Und) produced by the GlcNAc-phosphate transferase GacO and GlcNAc-phosphate-undecaprenol (GlcNAc-P-Und) produced by the glycosyltransferase GacI. Further investigations revealed that the GAC polyrhamnose backbone is assembled on GlcNAc-P-P-Und. Our results also suggested that a GT-C glycosyltranferase, GacL, transfers GlcNAc from GlcNAc-P-Und to polyrhamnose. Moreover, GacJ, a small membrane-associated protein, formed a complex with GacI and significantly stimulated its catalytic activity. Of note, we observed that GacI homologs perform a similar function inStreptococcus agalactiaeandEnterococcus faecalis. In conclusion, the elucidation of GAC biosynthesis inS. pyogenesreported here enhances our understanding of how other Gram-positive bacteria produce essential components of their cell wall.

Spectrophotometry Measurement of Polynuclear Aromatic Hydrocarbons in Hamilton Harbour Sediments

1991 ◽  
Vol 26 (1) ◽  
pp. 1-16 ◽  
Author(s):  
T.P. Murphy ◽  
H. Brouwer ◽  
M.E. Fox ◽  
E. Nagy
Keyword(s):  
Aromatic Hydrocarbons ◽  
Total Concentration ◽  
Coal Tar ◽  
Sediment Cores ◽  
Polynuclear Aromatic Hydrocarbons ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Near Surface ◽  
Spectrometry Analysis ◽  
Hamilton Harbour

Abstract Eighty-one sediment cores were collected to determine the extent of coal tar contamination in a toxic area of Hamilton Harbour. Over 800 samples were analyzed by a UV spectrophotometric technique that was standardized with gas chromatography/mass spectrometry analysis. The coal tar distribution was variable. The highest concentrations were near the Stelco outfalls and the Hamilton-Wentworth combined sewer outfalls. The total concentration of the 16 polynuclear aromatic hydrocarbons (PAHs) in 48,300 m3 of near-surface sediments exceeded 200 µg/g.

Sign in / Sign up

Export Citation Format

Share Document