98 results on '"Carina Sihlbom"'
Search Results
2. Age and sex effects across the blood proteome after ionizing radiation exposure can bias biomarker screening and risk assessment
- Author
-
Britta Langen, Egor Vorontsov, Johan Spetz, John Swanpalmer, Carina Sihlbom, Khalil Helou, and Eva Forssell-Aronsson
- Subjects
Medicine ,Science - Abstract
Abstract Molecular biomarkers of ionizing radiation (IR) exposure are a promising new tool in various disciplines: they can give necessary information for adaptive treatment planning in cancer radiotherapy, enable risk projection for radiation-induced survivorship diseases, or facilitate triage and intervention in radiation hazard events. However, radiation biomarker discovery has not yet resolved the most basic features of personalized medicine: age and sex. To overcome this critical bias in biomarker identification, we quantitated age and sex effects and assessed their relevance in the radiation response across the blood proteome. We used high-throughput mass spectrometry on blood plasma collected 24 h after 0.5 Gy total body irradiation (15 MV nominal photon energy) from male and female C57BL/6 N mice at juvenile (7-weeks-old) or adult (18-weeks-old) age. We also assessed sex and strain effects using juvenile male and female BALB/c nude mice. We showed that age and sex created significant effects in the proteomic response regarding both extent and functional quality of IR-induced responses. Furthermore, we found that age and sex effects appeared non-linear and were often end-point specific. Overall, age contributed more to differences in the proteomic response than sex, most notably in immune responses, oxidative stress, and apoptotic cell death. Interestingly, sex effects were pronounced for DNA damage and repair pathways and associated cellular outcome (pro-survival vs. pro-apoptotic). Only one protein (AHSP) was identified as a potential general biomarker candidate across age and sex, while GMNN, REG3B, and SNCA indicated some response similarity across age. This low yield advocated that unisex or uniage biomarker screening approaches are not feasible. In conclusion, age- and sex-specific screening approaches should be implemented as standard protocol to ensure robustness and diagnostic power of biomarker candidates. Bias-free molecular biomarkers are a necessary progression towards personalized medicine and integral for advanced adaptive cancer radiotherapy and risk assessment.
- Published
- 2022
- Full Text
- View/download PDF
3. Identification of Novel Glycans in the Mucus Layer of Shark and Skate Skin
- Author
-
Etty Bachar-Wikstrom, Kristina A. Thomsson, Carina Sihlbom, Lisa Abbo, Haitham Tartor, Sara K. Lindén, and Jakob D. Wikstrom
- Subjects
elasmobranchs ,sharks ,skin ,mucus layer ,mucin ,glycans ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
The mucus layer covering the skin of fish has several roles, including protection against pathogens and mechanical damage. While the mucus layers of various bony fish species have been investigated, the composition and glycan profiles of shark skin mucus remain relatively unexplored. In this pilot study, we aimed to explore the structure and composition of shark skin mucus through histological analysis and glycan profiling. Histological examination of skin samples from Atlantic spiny dogfish (Squalus acanthias) sharks and chain catsharks (Scyliorhinus retifer) revealed distinct mucin-producing cells and a mucus layer, indicating the presence of a functional mucus layer similar to bony fish mucus albeit thinner. Glycan profiling using liquid chromatography–electrospray ionization tandem mass spectrometry unveiled a diverse repertoire of mostly O-glycans in the mucus of the two sharks as well as little skate (Leucoraja erinacea). Elasmobranch glycans differ significantly from bony fish, especially in being more sulfated, and some bear resemblance to human glycans, such as gastric mucin O-glycans and H blood group-type glycans. This study contributes to the concept of shark skin having unique properties and provides a foundation for further research into the functional roles and potential biomedical implications of shark skin mucus glycans.
- Published
- 2023
- Full Text
- View/download PDF
4. Postprandial metabolism of apolipoproteins B48, B100, C-III, and E in humans with APOC3 loss-of-function mutations
- Author
-
Marja-Riitta Taskinen, Elias Björnson, Niina Matikainen, Sanni Söderlund, Joel Rämö, Mari-Mia Ainola, Antti Hakkarainen, Carina Sihlbom, Annika Thorsell, Linda Andersson, Per-Olof Bergh, Marcus Henricsson, Stefano Romeo, Martin Adiels, Samuli Ripatti, Markku Laakso, Chris J. Packard, and Jan Borén
- Subjects
Metabolism ,Medicine - Abstract
Background Apolipoprotein C-III (apoC-III) is a regulator of triglyceride (TG) metabolism, and due to its association with risk of cardiovascular disease, is an emergent target for pharmacological intervention. The impact of substantially lowering apoC-III on lipoprotein metabolism is not clear.Methods We investigated the kinetics of apolipoproteins B48 and B100 (apoB48 and apoB100) in chylomicrons, VLDL1, VLDL2, IDL, and LDL in patients heterozygous for a loss-of-function (LOF) mutation in the APOC3 gene. Studies were conducted in the postprandial state to provide a more comprehensive view of the influence of this protein on TG transport.Results Compared with non-LOF variant participants, a genetically determined decrease in apoC-III resulted in marked acceleration of lipolysis of TG-rich lipoproteins (TRLs), increased removal of VLDL remnants from the bloodstream, and substantial decrease in circulating levels of VLDL1, VLDL2, and IDL particles. Production rates for apoB48-containing chylomicrons and apoB100-containing VLDL1 and VLDL2 were not different between LOF carriers and noncarriers. Likewise, the rate of production of LDL was not affected by the lower apoC-III level, nor were the concentration and clearance rate of LDL-apoB100.Conclusion These findings indicate that apoC-III lowering will have a marked effect on TRL and remnant metabolism, with possibly significant consequences for cardiovascular disease prevention.Trial registration ClinicalTrials.gov NCT04209816 and NCT01445730.Funding Swedish Heart-Lung Foundation, Swedish Research Council, ALF grant from the Sahlgrenska University Hospital, Novo Nordisk Foundation, Sigrid Juselius Foundation, Helsinki University Hospital Government Research funds, Finnish Heart Foundation, and Finnish Diabetes Research Foundation.
- Published
- 2022
- Full Text
- View/download PDF
5. Increased expression and accumulation of GDF15 in IPF extracellular matrix contribute to fibrosis
- Author
-
Agata Radwanska, Christopher Travis Cottage, Antonio Piras, Catherine Overed-Sayer, Carina Sihlbom, Ramachandramouli Budida, Catherine Wrench, Jane Connor, Susan Monkley, Petra Hazon, Holger Schluter, Matthew J. Thomas, Cory M. Hogaboam, and Lynne A. Murray
- Subjects
Cell biology ,Pulmonology ,Medicine - Abstract
Idiopathic pulmonary fibrosis (IPF) is a chronic disease of unmet medical need. It is characterized by formation of scar tissue leading to a progressive and irreversible decline in lung function. IPF is associated with repeated injury, which may alter the composition of the extracellular matrix (ECM). Here, we demonstrate that IPF patient–derived pulmonary ECM drives profibrotic response in normal human lung fibroblasts (NHLF) in a 3D spheroid assay. Next, we reveal distinct alterations in composition of the diseased ECM, identifying potentially novel associations with IPF. Growth differentiation factor 15 (GDF15) was identified among the most significantly upregulated proteins in the IPF lung–derived ECM. In vivo, GDF15 neutralization in a bleomycin-induced lung fibrosis model led to significantly less fibrosis. In vitro, recombinant GDF15 (rGDF15) stimulated α smooth muscle actin (αSMA) expression in NHLF, and this was mediated by the activin receptor-like kinase 5 (ALK5) receptor. Furthermore, in the presence of rGDF15, the migration of NHLF in collagen gel was reduced. In addition, we observed a cell type–dependent effect of GDF15 on the expression of cell senescence markers. Our data suggest that GDF15 mediates lung fibrosis through fibroblast activation and differentiation, implicating a potential direct role of this matrix-associated cytokine in promoting aberrant cell responses in disease.
- Published
- 2022
- Full Text
- View/download PDF
6. Nitrogen limitation reveals large reserves in metabolic and translational capacities of yeast
- Author
-
Rosemary Yu, Kate Campbell, Rui Pereira, Johan Björkeroth, Qi Qi, Egor Vorontsov, Carina Sihlbom, and Jens Nielsen
- Subjects
Science - Abstract
Cells maintain reserves in their metabolic and translational capacities enabling fast response to changing environments. Here, the authors quantify reserves in yeast by stepwise reduction in nitrogen availability and a combination of multi-omic analysis and metabolic modelling.
- Published
- 2020
- Full Text
- View/download PDF
7. Lipid droplet-associated kinase STK25 regulates peroxisomal activity and metabolic stress response in steatotic liver[S]
- Author
-
Annika Nerstedt, Yeshwant Kurhe, Emmelie Cansby, Mara Caputo, Lei Gao, Egor Vorontsov, Marcus Ståhlman, Esther Nuñez-Durán, Jan Borén, Hanns-Ulrich Marschall, Douglas G. Mashek, Darren N. Saunders, Carina Sihlbom, Andrew J. Hoy, and Margit Mahlapuu
- Subjects
protein kinases ,nonalcoholic fatty liver disease ,steatohepatitis ,serine/threonine kinase 25 ,Biochemistry ,QD415-436 - Abstract
Nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) are emerging as leading causes of liver disease worldwide and have been recognized as one of the major unmet medical needs of the 21st century. Our recent translational studies in mouse models, human cell lines, and well-characterized patient cohorts have identified serine/threonine kinase (STK)25 as a protein that coats intrahepatocellular lipid droplets (LDs) and critically regulates liver lipid homeostasis and progression of NAFLD/NASH. Here, we studied the mechanism-of-action of STK25 in steatotic liver by relative quantification of the hepatic LD-associated phosphoproteome from high-fat diet-fed Stk25 knockout mice compared with their wild-type littermates. We observed a total of 131 proteins and 60 phosphoproteins that were differentially represented in STK25-deficient livers. Most notably, a number of proteins involved in peroxisomal function, ubiquitination-mediated proteolysis, and antioxidant defense were coordinately regulated in Stk25−/− versus wild-type livers. We confirmed attenuated peroxisomal biogenesis and protection against oxidative and ER stress in STK25-deficient human liver cells, demonstrating the hepatocyte-autonomous manner of STK25's action. In summary, our results suggest that regulation of peroxisomal function and metabolic stress response may be important molecular mechanisms by which STK25 controls the development and progression of NAFLD/NASH.
- Published
- 2020
- Full Text
- View/download PDF
8. Liver Graft Proteomics Reveals Potential Incipient Mechanisms behind Early Renal Dysfunction after Liver Transplantation
- Author
-
Åsa Norén, Mihai Oltean, Styrbjörn Friman, Antonio Molinaro, Johan Mölne, Carina Sihlbom, Gustaf Herlenius, and Annika Thorsell
- Subjects
liver transplantation ,ischemia–reperfusion injury ,renal failure ,proteomics ,outcome ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Acute kidney injury (AKI) is frequent after liver transplantation (LT) and correlates with later development of chronic kidney disease. Its etiology is multifactorial and combines pre-, intra-, and postoperative factors. Additionally, the liver graft itself seems an important element in the development of AKI, yet the detailed mechanisms remain unclear. We hypothesized that grafts of LT recipients developing significant early AKI may show distinct proteomic alterations, and we set out to identify proteome differences between LT recipients developing moderate or severe AKI (n = 7) and LT recipients without early renal injury (n = 7). Liver biopsies obtained one hour after reperfusion were assessed histologically and using quantitative proteomics. Several cytokines and serum amyloid A2 (SAA2) were analyzed in serum samples obtained preoperatively, 2–4 h, and 20–24 h after graft reperfusion, respectively. LT induced mild histological alterations without significant differences between groups but uniformly altered liver function tests peaking on postoperative day 1, with a trend towards more severe alterations in patients developing AKI. Global quantitative proteomic analysis revealed 136 proteins differing significantly in their expression levels (p < 0.05, FC 20%): 80 proteins had higher and 56 had lower levels in the AKI group. Most of these proteins were related to immune and inflammatory responses, host defense, and neutrophil degranulation. No differences between the studied pro- and anti-inflammatory cytokines or SAA2 between groups were found at any moment. Our results suggest that grafts of LT patients who develop early AKI reveal a distinct proteome dominated by an early yet prominent activation of the innate immunity. These findings support the hypothesis that AKI after LT may be favored by certain graft characteristics.
- Published
- 2022
- Full Text
- View/download PDF
9. Quantifying absolute gene expression profiles reveals distinct regulation of central carbon metabolism genes in yeast
- Author
-
Rosemary Yu, Egor Vorontsov, Carina Sihlbom, and Jens Nielsen
- Subjects
gene expression ,growth rate ,nitrogen metabolism ,central carbon metabolism ,amino acid metabolism ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
In addition to controlled expression of genes by specific regulatory circuits, the abundance of proteins and transcripts can also be influenced by physiological states of the cell such as growth rate and metabolism. Here we examine the control of gene expression by growth rate and metabolism, by analyzing a multi-omics dataset consisting of absolute-quantitative abundances of the transcriptome, proteome, and amino acids in 22 steady-state yeast cultures. We find that transcription and translation are coordinately controlled by the cell growth rate via RNA polymerase II and ribosome abundance, but they are independently controlled by nitrogen metabolism via amino acid and nucleotide availabilities. Genes in central carbon metabolism, however, are distinctly regulated and do not respond to the cell growth rate or nitrogen metabolism as all other genes. Understanding these effects allows the confounding factors of growth rate and metabolism to be accounted for in gene expression profiling studies.
- Published
- 2021
- Full Text
- View/download PDF
10. Platelet proteome and function in X−linked thrombocytopenia with thalassemia and in silico comparisons with gray platelet syndrome
- Author
-
Daniel Bergemalm, Sofia Ramström, Caroline Kardeby, Kjell Hultenby, Anna Göthlin Eremo, Carina Sihlbom, Jörgen Bergström, Jan Palmblad, and Maria Åström
- Subjects
Diseases of the blood and blood-forming organs ,RC633-647.5 - Abstract
In X-linked thrombocytopenia with thalassemia (XLTT; OMIM 314050), caused by the mutation p.R216Q in exon 4 of the GATA1 gene, male hemizygous patients display macrothrombocytopenia, bleeding diathesis and a b-thalassemia trait. Herein, we describe findings in two unrelated Swedish XLTT families with a bleeding tendency exceeding what is expected from the thrombocytopenia. Blood tests revealed low P-PAI-1 and P-factor 5, and elevated S-thrombopoietin levels. Transmission electron microscopy showed diminished numbers of platelet a- and dense granules. The proteomes of isolated blood platelets from five male XLTT patients, compared to five sex- and agematched controls, were explored. Quantitative mass spectrometry showed alterations of 83 proteins (fold change ≥±1.2, q
- Published
- 2020
- Full Text
- View/download PDF
11. The Proteomic Signature of Intestinal Acute Rejection in the Mouse
- Author
-
Mihai Oltean, Jasmine Bagge, George Dindelegan, Diarmuid Kenny, Antonio Molinaro, Mats Hellström, Ola Nilsson, Carina Sihlbom, Anna Casselbrant, Marcela Davila, and Michael Olausson
- Subjects
intestinal transplantation ,rejection ,biomarkers ,enzymes ,chromogranin A ,Microbiology ,QR1-502 - Abstract
Intestinal acute rejection (AR) lacks a reliable non-invasive biomarker and AR surveillance is conducted through frequent endoscopic biopsies. Although citrulline and calprotectin have been suggested as AR biomarkers, these have limited clinical value. Using a mouse model of intestinal transplantation (ITx), we performed a proteome-wide analysis and investigated rejection-related proteome changes that may eventually be used as biomarkers. ITx was performed in allogenic (Balb/C to C57Bl) and syngeneic (C57Bl) combinations. Graft samples were obtained three and six days after transplantation (n = 4/time point) and quantitative proteomic analysis with iTRAQ-labeling and mass spectrometry of whole tissue homogenates was performed. Histology showed moderate AR in all allografts post-transplantation at day six. Nine hundred and thirty-eight proteins with at least three unique peptides were identified in the intestinal grafts. Eighty-six proteins varying by >20% between time points and/or groups had an alteration pattern unique to the rejecting allografts: thirty-seven proteins and enzymes (including S100-A8 and IDO-1) were significantly upregulated whereas forty-nine (among other chromogranin, ornithine aminotransferase, and arginase) were downregulated. Numerous proteins showed altered expression during intestinal AR, several of which were previously identified to be involved in acute rejection, although our results also identified previously unreported proteome changes. The metabolites and downstream metabolic pathways of some of these proteins and enzymes may become potential biomarkers for intestinal AR.
- Published
- 2021
- Full Text
- View/download PDF
12. Low Concentrations of Vitamin C Reduce the Synthesis of Extracellular Polymers and Destabilize Bacterial Biofilms
- Author
-
Santosh Pandit, Vaishnavi Ravikumar, Alyaa M. Abdel-Haleem, Abderahmane Derouiche, V. R. S. S. Mokkapati, Carina Sihlbom, Katsuhiko Mineta, Takashi Gojobori, Xin Gao, Fredrik Westerlund, and Ivan Mijakovic
- Subjects
biofilms ,exopolymeric matrix ,quantitative proteomics ,Bacillus subtilis ,vitamin C ,Microbiology ,QR1-502 - Abstract
Extracellular polymeric substances (EPS) produced by bacteria form a matrix supporting the complex three-dimensional architecture of biofilms. This EPS matrix is primarily composed of polysaccharides, proteins and extracellular DNA. In addition to supporting the community structure, the EPS matrix protects bacterial biofilms from the environment. Specifically, it shields the bacterial cells inside the biofilm, by preventing antimicrobial agents from getting in contact with them, thereby reducing their killing effect. New strategies for disrupting the formation of the EPS matrix can therefore lead to a more efficient use of existing antimicrobials. Here we examined the mechanism of the known effect of vitamin C (sodium ascorbate) on enhancing the activity of various antibacterial agents. Our quantitative proteomics analysis shows that non-lethal concentrations of vitamin C inhibit bacterial quorum sensing and other regulatory mechanisms underpinning biofilm development. As a result, the EPS biosynthesis in reduced, and especially the polysaccharide component of the matrix is depleted. Once the EPS content is reduced beyond a critical point, bacterial cells get fully exposed to the medium. At this stage, the cells are more susceptible to killing, either by vitamin C-induced oxidative stress as reported here, or by other antimicrobials or treatments.
- Published
- 2017
- Full Text
- View/download PDF
13. Comparative Analysis of two Helicobacter pylori Strains using Genomics and Mass Spectrometry-Based Proteomics
- Author
-
Roger Karlsson, Kaisa Thorell, Shaghayegh Hosseini, Diarmuid Kenny, Carina Sihlbom, Åsa Sjöling, Anders Karlsson, and Intawat Nookaew
- Subjects
Genomics ,Helicobacter pylori ,Proteomics ,prognosis ,TMT ,MaxQuant ,Microbiology ,QR1-502 - Abstract
Helicobacter pylori, a gastroenteric pathogen believed to have co-evolved with humans over 100,000 years, shows significant genetic variability. This motivates the study of different H. pylori strains and the diseases they cause in order to identify determinants for disease evolution. In this study, we used proteomics tools to compare two H. pylori strains. Nic25_A was isolated in Nicaragua from a patient with intestinal metaplasia, and P12 was isolated in Europe from a patient with duodenal ulcers. Differences in the abundance of surface proteins between the two strains were determined with two mass spectrometry-based methods, label-free quantification (MaxQuant) or the use of tandem mass tags (TMT). Each approach used a lipid-based protein immobilization (LPI™) technique to enrich peptides of surface proteins. Using the MaxQuant software, we found 52 proteins that differed significantly in abundance between the two strains (up- or downregulated by a factor of 1.5); with TMT, we found 18 proteins that differed in abundance between the strains. Strain P12 had a higher abundance of proteins encoded by the cag pathogenicity island, while levels of the acid response regulator ArsR and its regulatory targets (KatA, AmiE, and proteins involved in urease production) were higher in strain Nic25_A. Our results show that differences in protein abundance between H. pylori strains can be detected with proteomic approaches; this could have important implications for the study of disease progression.
- Published
- 2016
- Full Text
- View/download PDF
14. Recombinant Glycoprotein E of Varicella Zoster Virus Contains Glycan-Peptide Motifs That Modulate B Cell Epitopes into Discrete Immunological Signatures
- Author
-
Rickard Nordén, Jonas Nilsson, Ebba Samuelsson, Christian Risinger, Carina Sihlbom, Ola Blixt, Göran Larson, Sigvard Olofsson, and Tomas Bergström
- Subjects
antibody binding ,B cell epitope ,glycoprotein E ,glycosylation ,vaccine ,varicella zoster virus ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
A recombinant subunit vaccine (Shingrix®) was recently licensed for use against herpes zoster. This vaccine is based on glycoprotein E (gE) of varicella zoster virus (VZV), the most abundantly expressed protein of VZV, harboring sites for N- and O-linked glycosylation. The subunit vaccine elicits stronger virus-specific CD4+ T cell response as well as antibody B cell response to gE, compared to the currently used live attenuated vaccine (Zostavax®). This situation is at variance with the current notion since a live vaccine, causing an active virus infection, should be far more efficient than a subunit vaccine based on only one single viral glycoprotein. We previously found gE to be heavily glycosylated, not least by numerous clustered O-linked glycans, when it was produced in human fibroblasts. However, in contrast to Zostavax®, which is produced in fibroblasts, the recombinant gE of Shingrix® is expressed in Chinese hamster ovary (CHO) cells. Hence, the glycan occupancy and glycan structures of gE may differ considerably between the two vaccine types. Here, we aimed at (i) defining the glycan structures and positions of recombinant gE and (ii) identifying possible features of the recombinant gE O-glycosylation pattern contributing to the vaccine efficacy of Shingrix®. Firstly, recombinant gE produced in CHO cells (“Shingrix situation„) is more scarcely decorated by O-linked glycans than gE from human fibroblasts (“Zostavax situation„), with respect to glycan site occupancy. Secondly, screening of immunodominant B cell epitopes of gE, using a synthetic peptide library against serum samples from VZV-seropositive individuals, revealed that the O-linked glycan signature promoted binding of IgG antibodies via a decreased number of interfering O-linked glycans, but also via specific O-linked glycans enhancing antibody binding. These findings may, in part, explain the higher protective efficacy of Shingrix®, and can also be of relevance for development of subunit vaccines to other enveloped viruses.
- Published
- 2019
- Full Text
- View/download PDF
15. Identification of Potential Plasma Biomarkers for Abdominal Aortic Aneurysm Using Tandem Mass Tag Quantitative Proteomics
- Author
-
Anders E. Henriksson, Markus Lindqvist, Carina Sihlbom, Jörgen Bergström, and Dan Bylund
- Subjects
aortic aneurysm ,biomarker ,bleomycin hydrolase ,clinical proteomics ,mass spectrometry ,proteomics ,Microbiology ,QR1-502 - Abstract
Plasma biomarkers that identify abdominal aortic aneurysm (AAA) rupture risk would greatly assist in stratifying patients with small aneurysms. Identification of such biomarkers has hitherto been unsuccessful over a range of studies using different methods. The present study used an alternative proteomic approach to find new, potential plasma AAA biomarker candidates. Pre-fractionated plasma samples from twelve patients with AAA and eight matched controls without aneurysm were analyzed by mass spectrometry applying a tandem mass tag (TMT) technique. Eight proteins were differentially regulated in patients compared to controls, including decreased levels of the enzyme bleomycin hydrolase. The down-regulation of this enzyme was confirmed in an extended validation study using an enzyme-linked immunosorbent assay (ELISA). The TMT-based proteomic approach thus identified novel potential plasma biomarkers for AAA.
- Published
- 2018
- Full Text
- View/download PDF
16. Pharmacokinetic Properties of the Nephrotoxin Orellanine in Rats
- Author
-
Deman Najar, Börje Haraldsson, Annika Thorsell, Carina Sihlbom, Jenny Nyström, and Kerstin Ebefors
- Subjects
orellanine ,clearance ,fungal toxin ,half-life ,Medicine - Abstract
Orellanine is a nephrotoxin found in mushrooms of the Cortinarius family. Accidental intake of this substance may cause renal failure. Orellanine is specific for proximal tubular cells and could, therefore, potentially be used as treatment for metastatic renal cancer, which originates from these cells. However, more information is needed about the distribution and elimination of orellanine from the body to understand its potential use for therapy. In this study, 5 mg/kg orellanine (unlabeled and 3H-labeled) was injected intravenously in rats (Wistar and Sprague Dawley). Distribution was measured (Wistar rats, n = 10, n = 12) using radioluminography and the highest amount of orellanine was found in the kidney cortex and bladder at all time-points investigated. The pharmacokinetic properties of orellanine was investigated using LC-MS/MS and β-scintillation to measure the amount of orellanine in plasma. Three groups of rats were investigated: control rats with intact kidneys (n = 10) and two groups with bilateral renal artery ligation (n = 7) where animals in one of these groups were treated with peritoneal dialysis (n = 8). Using LC-MS/MS, the half-life of orellanine was found to be 109 ± 6 min in the controls. In the groups with ligated renal arteries, orellanine had a half-life of 756 ± 98 min without and 238 ± 28 min with dialysis. Thus, orellanine was almost exclusively eliminated by glomerular filtration as well as by peritoneal dialysis.
- Published
- 2018
- Full Text
- View/download PDF
17. A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization
- Author
-
Jafar Mahdavi, Necmettin Pirinccioglu, Neil J. Oldfield, Elisabet Carlsohn, Jeroen Stoof, Akhmed Aslam, Tim Self, Shaun A. Cawthraw, Liljana Petrovska, Natalie Colborne, Carina Sihlbom, Thomas Borén, Karl G. Wooldridge, and Dlawer A. A. Ala'Aldeen
- Subjects
campylobacter jejuni ,histo-blood group antigens ,flaa ,major outer membrane protein ,o-glycosylation ,biofilm ,Biology (General) ,QH301-705.5 - Abstract
Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. Significantly, the MOMP was shown to be O-glycosylated at Thr268; previously only flagellin proteins were known to be O-glycosylated in C. jejuni. Substitution of MOMP Thr268 led to significantly reduced binding to BgAgs. The O-glycan moiety was characterized as Gal(β1–3)-GalNAc(β1–4)-GalNAc(β1–4)-GalNAcα1-Thr268; modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. Glycosylation of MOMP at Thr268 promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis.
- Published
- 2014
- Full Text
- View/download PDF
18. Pro-inflammatory cytokines can act as intracellular modulators of commensal bacterial virulence
- Author
-
Jafar Mahdavi, Pierre-Joseph Royer, Hong S. Sjölinder, Sheyda Azimi, Tim Self, Jeroen Stoof, Lee M. Wheldon, Kristoffer Brännström, Raymond Wilson, Joanna Moreton, James W. B. Moir, Carina Sihlbom, Thomas Borén, Ann-Beth Jonsson, Panos Soultanas, and Dlawer A. A. Ala'Aldeen
- Subjects
neisseria meningitidis ,type iv pili ,glycosylation ,cytokines ,patho-adaptation ,transcriptional regulators ,Biology (General) ,QH301-705.5 - Abstract
Interactions between commensal pathogens and hosts are critical for disease development but the underlying mechanisms for switching between the commensal and virulent states are unknown. We show that the human pathogen Neisseria meningitidis, the leading cause of pyogenic meningitis, can modulate gene expression via uptake of host pro-inflammatory cytokines leading to increased virulence. This uptake is mediated by type IV pili (Tfp) and reliant on the PilT ATPase activity. Two Tfp subunits, PilE and PilQ, are identified as the ligands for TNF-α and IL-8 in a glycan-dependent manner, and their deletion results in decreased virulence and increased survival in a mouse model. We propose a novel mechanism by which pathogens use the twitching motility mode of the Tfp machinery for sensing and importing host elicitors, aligning with the inflamed environment and switching to the virulent state.
- Published
- 2013
- Full Text
- View/download PDF
19. A macrophage-collagen fragment axis mediates subcutaneous adipose tissue remodeling in mice.
- Author
-
Vujičić, Milica, Broderick, Isabella, Salmantabar, Pegah, Perian, Charlène, Nilsson, Jonas, Wallem, Carina Sihlbom, and Asterholm, Ingrid Wernstedt
- Subjects
TISSUE remodeling ,ADIPOSE tissues ,WEIGHT gain ,MICE ,METABOLIC disorders - Abstract
Efficient removal of fibrillar collagen is essential for adaptive subcutaneous adipose tissue (SAT) expansion that protects against ectopic lipid deposition during weight gain. Here, we used mice to further define the mechanism for this collagenolytic process. We show that loss of collagen type-1 (CT1) and increased CT1-fragment levels in expanding SAT are associated with proliferation of resident M2-like macrophages that display increased CD206-mediated engagement in collagen endocytosis compared to chow-fed controls. Blockage of CD206 during acute high-fat diet-induced weight gain leads to SAT CT1-fragment accumulation associated with elevated inflammation and fibrosis markers. Moreover, these SAT macrophages’ engagement in collagen endocytosis is diminished in obesity associated with elevated levels collagen fragments that are too short to assemble into triple helices. We show that such short fragments provoke M2-macrophage proliferation and fibroinflammatory changes in fibroblasts. In conclusion, our data delineate the importance of a macrophage-collagen fragment axis in physiological SAT expansion. Therapeutic targeting of this process may be a means to prevent pathological adipose tissue remodeling, which in turn may reduce the risk for obesity-related metabolic disorders. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
20. Site‐specific N ‐glycan profiles of α 5 β 1 integrin from rat liver
- Author
-
Ekaterina Mirgorodskaya, Estelle Dransart, Massiullah Shafaq‐Zadah, Daniel Roderer, Carina Sihlbom, Hakon Leffler, and Ludger Johannes
- Subjects
Cell Biology ,General Medicine - Published
- 2022
21. m6A modification of TERRA RNA is required for telomere maintenance and is a therapeutic target for ALT positive Neuroblastoma
- Author
-
Roshan Vaid, Ketan Thombare, Akram Mendez, Rebeca Burgos-Panadero, Anna Djos, Daniel Jachimowicz, Christoph Bartenhagen, Navinder Kumar, Carina Sihlbom, Susanne Fransson, John Inge Johnsen, Per Kogner, Tommy Martinsson, Matthias Fischer, and Tanmoy Mondal
- Abstract
Telomerase-negative tumors can maintain telomere length by alternative lengthening of telomeres (ALT) but the mechanism behind ALT is poorly understood. Aggressive Neuroblastoma (NB), in particular, relapsed tumors are positive for ALT (ALT+) which suggests that better dissection of the ALT mechanism could provide novel therapeutic opportunities. TERRA long non-coding RNA (lncRNA) which is derived from the telomere ends is localized to telomeres in R-loop dependent manner and is essential for telomere maintenance. In the present study, we provide evidence that RNA modification at theN6position of internal adenosine (m6A) in TERRA RNA by methyltransferase METTL3 is essential for telomere maintenance in ALT+ cells and that loss of TERRA m6A/METTL3 leads to telomere damage. We observed that R-loop enriched TERRA is abundantly m6A modified and m6A mediated recruitment of hnRNPA2B1 to TERRA RNA is essential for R-loop formation. Our data suggest that m6A drives telomere targeting of TERRA via R-loop and this m6A mediated R-loop formation could be a widespread mechanism utilized by other chromatin-interacting lncRNAs. Furthermore, treating ALT+ NB cells with METTL3 inhibitor leads to compromised telomere targeting of TERRA and accumulation of DNA damage over telomere, suggesting METTL3 inhibition could be a therapeutic opportunity for ALT+ NB.
- Published
- 2022
22. Role of endogenous incretins in the regulation of postprandial lipoprotein metabolism
- Author
-
Marja-Riitta Taskinen, Niina Matikainen, Elias Björnson, Sanni Söderlund, Mari Ainola, Antti Hakkarainen, Nina Lundbom, Carina Sihlbom, Annika Thorsell, Linda Andersson, Martin Adiels, Bolette Hartmann, Carolyn F Deacon, Jens J Holst, Chris J Packard, and Jan Borén
- Subjects
Male ,Endocrinology ,Glucagon-Like Peptide 1 ,Endocrinology, Diabetes and Metabolism ,Lipoproteins ,Chylomicrons ,Humans ,General Medicine ,Gastric Inhibitory Polypeptide ,Apolipoprotein B-48 ,Postprandial Period ,Incretins ,Triglycerides - Abstract
Objective Incretins are known to influence lipid metabolism in the intestine when administered as pharmacologic agents. The aggregate influence of endogenous incretins on chylomicron production and clearance is less clear, particularly in light of opposing effects of co-secreted hormones. Here, we tested the hypothesis that physiological levels of incretins may impact on production or clearances rates of chylomicrons and VLDL. Design and methods A group of 22 overweight/obese men was studied to determine associations between plasma levels of glucagon-like peptides 1 and 2 (GLP-1 and GLP-2) and glucose-dependent insulinotropic polypeptide (GIP) after a fat-rich meal and the production and clearance rates of apoB48- and apoB100-containing triglyceride-rich lipoproteins. Subjects were stratified by above- and below-median incretin response (area under the curve). Results Stratification yielded subgroups that differed about two-fold in incretin response. There were neither differences in apoB48 production rates in chylomicrons or VLDL fractions nor in apoB100 or triglyceride kinetics in VLDL between men with above- vs below-median incretin responses. The men with above-median GLP-1 and GLP-2 responses exhibited higher postprandial plasma and chylomicron triglyceride levels, but this could not be related to altered kinetic parameters. No differences were found between incretin response subgroups and particle clearance rates. Conclusion We found no evidence for a regulatory effect of endogenous incretins on contemporaneous chylomicron or VLDL metabolism following a standardised fat-rich meal. The actions of incretins at pharmacological doses may not be reflected at physiological levels of these hormones.
- Published
- 2022
23. Lung macrophages utilize unique cathepsin K-dependent phagosomal machinery to degrade intracellular collagen
- Author
-
Ivo Fabrik, Orsolya Bilkei-Gorzo, Maria Öberg, Daniela Fabrikova, Johannes Fuchs, Carina Sihlbom, Melker Göransson, and Anetta Härtlova
- Subjects
Ecology ,Health, Toxicology and Mutagenesis ,Plant Science ,Biochemistry, Genetics and Molecular Biology (miscellaneous) - Abstract
Resident tissue macrophages (RTMs) are organ-specialized phagocytes responsible for the maintenance and protection of tissue homeostasis. It is well established that tissue diversity is reflected by the heterogeneity of RTMs origin and phenotype. However, much less is known about tissue-specific phagocytic and proteolytic macrophage functions. Here, using quantitative proteomics approach, we identify cathepsins as key determinants of phagosome maturation in primary peritoneal, lung and brain resident macrophages. The data further uncover cathepsin K (CtsK) as a molecular marker for lung phagosomes required for intracellular protein and collagen degradation. Pharmacological blockade of CtsK activity diminished phagosomal proteolysis and collagenolysis in lung resident macrophages. Furthermore, pro-fibrotic TGF-β negatively regulated CtsK-mediated phagosomal collagen degradation independently from classical endocytic proteolytic pathways. In humans, phagosomal CtsK activity was reduced in COPD lung macrophages and non-COPD lung macrophages exposed to cigarette smoke extract. Taken together, this study provides a comprehensive map of how peritoneal, lung and brain tissue environment shapes phagosomal composition, revealing CtsK as a key molecular determinant of lung phagosomes contributing to phagocytic collagen clearance in lungs.
- Published
- 2022
24. Platelet proteome and function in X−linked thrombocytopenia with thalassemia and in silico comparisons with gray platelet syndrome
- Author
-
Jörgen Bergström, Caroline Kardeby, Kjell Hultenby, Carina Sihlbom, Daniel Bergemalm, Maria Åström, Anna Göthlin Eremo, Jan Palmblad, and Sofia Ramström
- Subjects
medicine.medical_specialty ,Chemistry ,Fibrinogen binding ,Hematology ,medicine.disease ,Protein ubiquitination ,Collagen receptor ,Gray platelet syndrome ,Bleeding diathesis ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Internal medicine ,medicine ,Platelet ,GPVI ,Dense granule ,030215 immunology - Abstract
In X-linked thrombocytopenia with thalassemia (XLTT; OMIM 314050), caused by the mutation p.R216Q in exon 4 of the GATA1 gene, male hemizygous patients display macrothrombocytopenia, bleeding diathesis and a b-thalassemia trait. Herein, we describe findings in two unrelated Swedish XLTT families with a bleeding tendency exceeding what is expected from the thrombocytopenia. Blood tests revealed low P-PAI-1 and P-factor 5, and elevated S-thrombopoietin levels. Transmission electron microscopy showed diminished numbers of platelet a- and dense granules. The proteomes of isolated blood platelets from five male XLTT patients, compared to five sex- and agematched controls, were explored. Quantitative mass spectrometry showed alterations of 83 proteins (fold change ≥±1.2, q
- Published
- 2020
25. Proteomic analysis of follicular fluid during human ovulation
- Author
-
Carina Sihlbom, Farnosh Zakerkish, Mats Brännström, Elisabet Carlsohn, Asgeir Thoroddsen, and Sjoerd van der Post
- Subjects
Adult ,Ovulation ,Proteomics ,media_common.quotation_subject ,Ovary ,Mass Spectrometry ,Andrology ,03 medical and health sciences ,Follicle ,0302 clinical medicine ,Ovarian Follicle ,Follicular phase ,medicine ,Humans ,Prospective Studies ,030212 general & internal medicine ,Menstrual cycle ,media_common ,Sweden ,030219 obstetrics & reproductive medicine ,business.industry ,Proteins ,Obstetrics and Gynecology ,General Medicine ,Oocyte ,Follicular fluid ,Follicular Fluid ,medicine.anatomical_structure ,Follicular Phase ,Proteome ,Female ,business - Abstract
Introduction Human ovulation is a biologically complex process that involves several biochemical factors, promoting follicular rupture and release of a fertilizable oocyte. Proteins which are present in follicular fluid at high concentrations during ovulation are likely to be active participants in the biochemical pathways of ovulation. The aim of the study was to identify, by use of a modern proteomic technique, proteins of human follicular fluid which are differentially regulated during ovulation of the natural menstrual cycle. Material and methods This prospective experimental study over 3 years included women planned for laparoscopic sterilization. During surgery, retrieval of the dominant follicle was performed either at the preovulatory stage or during ovulation. Four women of preovulatory phase and four women of ovulatory phase met the predetermined criteria of hormone levels for respective phases, and samples of these were finally included out of the 15 women operated. Follicular fluid was aspirated from the excised follicle and subjected to mass spectrometry with the isobaric tags for relative and absolute quantification (iTRAQ) technology for isobaric tagging of peptides. This enables simultaneous identification and quantification of proteins. The protein profiles of the follicular fluid of the preovulatory phase and the ovulatory phase were analyzed, and proteins that were present were identified. Results A total of 502 proteins were identified, several of which previously have not been identified in human follicular fluid. Of the 115 proteins that were found in all samples, 20 proteins were at higher levels during ovulation. These were inflammatory-related proteins, coagulation factors, proteins in lipid metabolism, complement factors and antioxidants. Five proteins were present in lower levels during ovulation, with three being enzymes and the other two proteins of lipid metabolism and iron transport. Conclusions Twenty-five follicular fluid proteins, with differential regulation during ovulation, were identified in human follicular fluid of the natural menstrual cycle. These proteins may have essential roles in the ovulatory cascade.
- Published
- 2020
26. Nitrogen limitation reveals large reserves in metabolic and translational capacities of yeast
- Author
-
Jens Nielsen, Egor Vorontsov, Rosemary Yu, Johan Björkeroth, Kate Campbell, Qi Qi, Carina Sihlbom, and Rui Pedro Gomes Pereira
- Subjects
0301 basic medicine ,Proteomics ,Ribosomal Proteins ,Proteome ,Nitrogen ,Molecular biology ,Systems biology ,Science ,General Physics and Astronomy ,Chemostat ,Saccharomyces cerevisiae ,Biology ,Ribosome ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Bioreactors ,Ribosomal protein ,Gene Expression Regulation, Fungal ,Protein biosynthesis ,lcsh:Science ,Cell Engineering ,Multidisciplinary ,General Chemistry ,Yeast ,Cell biology ,030104 developmental biology ,Glucose ,Protein Biosynthesis ,Fermentation ,lcsh:Q ,Protein Processing, Post-Translational ,030217 neurology & neurosurgery ,Metabolic Networks and Pathways - Abstract
Cells maintain reserves in their metabolic and translational capacities as a strategy to quickly respond to changing environments. Here we quantify these reserves by stepwise reducing nitrogen availability in yeast steady-state chemostat cultures, imposing severe restrictions on total cellular protein and transcript content. Combining multi-omics analysis with metabolic modeling, we find that seven metabolic superpathways maintain >50% metabolic capacity in reserve, with glucose metabolism maintaining >80% reserve capacity. Cells maintain >50% reserve in translational capacity for 2490 out of 3361 expressed genes (74%), with a disproportionately large reserve dedicated to translating metabolic proteins. Finally, ribosome reserves contain up to 30% sub-stoichiometric ribosomal proteins, with activation of reserve translational capacity associated with selective upregulation of 17 ribosomal proteins. Together, our dataset provides a quantitative link between yeast physiology and cellular economics, which could be leveraged in future cell engineering through targeted proteome streamlining., Cells maintain reserves in their metabolic and translational capacities enabling fast response to changing environments. Here, the authors quantify reserves in yeast by stepwise reduction in nitrogen availability and a combination of multi-omic analysis and metabolic modelling.
- Published
- 2020
27. NIST Interlaboratory Study on Glycosylation Analysis of Monoclonal Antibodies
- Author
-
Miyako Nakano, Alena Wiegandt, Yunli Hu, Viv Lindo, Paulina A. Urbanowicz, Zsuzsanna Lakos, Cassie Caron, Song Klapoetke, Niels Christian Reichardt, Niclas Chiang Tan, Sandra Maier, Rene Hennig, Marton Szigeti, Ju Yeon Lee, Ying Qing Yu, Gregory O. Staples, Sachin Patil, Jolanta Jaworek, Waltraud Evers, Benjamin G. Kremkow, Youngsuk Seo, Kathirvel Alagesan, Yuetian Chen, Gordan Lauc, David L. Duewer, Yang Yang, Daniele Menard, Hyun Joo An, Tim Kelly, Stephen E. Stein, Joseph W. Leone, Anja Wiechmann, Ravi Amunugama, Peng George Wang, Clemens Grunwald-Grube, Maria Lorna A. De Leoz, Göran Larson, Rob Haselberg, Samanta Cajic, Stephanie A. Archer-Hartmann, Maja Pučić-Baković, Edward D. Bodnar, Pauline M. Rudd, Anja Resemann, Daniel Kolarich, Akira Harazono, Jeffrey S. Rohrer, Juan Echevarria Ruiz, Stuart Pengelley, Jong Shin Yoo, Arun V. Everest-Dass, Nicolle H. Packer, Steven W. Mast, William R. Alley, Erika Lattová, Anne Zeck, Corné J.M. Stroop, Radoslaw P. Kozak, Chun Shao, Alain Beck, Joseph Zaia, Erdmann Rapp, Lily Liu, Jennie Truong, Yaojun Wang, Christopher W. Cairo, Roisin O'Flaherty, Radka Saldova, Kudrat Goswami, Emy Komatsu, Jessica Örnros, Taiki Sugiyama, Prachi Bhoskar, Pralima Pradhan, Carlito B. Lebrilla, András Guttman, Christine Merle, Brian Kasper, Oscar G. Potter, Soo Kyung Suh, Li Phing Liew, Ranjan Chakrabarti, Terry D. Cyr, Sohei Funaoka, Masaaki Toyoda, Pui King Amy Leung, Toyin Kasali, Jerko Štambuk, Yanming An, Wolfgang Jabs, Bernd Meyer, Chunxia Zou, John F. Cipollo, Sa Rang Kim, Aaron Shafer, Randy M. Whittal, Jichao Kang, Albert J. R. Heck, Yehia Mechref, Hoi Kei Yau, Guinevere S. M. Lageveen-Kammeijer, Shiwei Sun, Kenichiro Furuki, Richard B. Jones, Béla Reiz, Niclas G. Karlsson, Mohammedazam Lahori, Xu Li, Barbara Adamczyk, Rui Cao, Lauren Wu, Koichi Kato, Detlev Suckau, Paweł Link-Lenczowski, Kelvin H. Lee, Xiaomin Song, Noortje de Haan, Ruth Frenkel, Adam Fung, Friedrich Altmann, Manfred Wuhrer, David Falck, Andreas Bock, Paula Magnelli, Brian Gau, Sachiko Kondo, Robert J. Emery, Chunsheng Jin, Louise Royle, David C. Muddiman, Hélène Perreault, John W. Froehlich, Disha Dadke, Peiqing Zhang, Lara K. Mahal, Takashi Nishikaze, Andrew Saati, Chuncui Huang, Hui Zhang, Carina Sihlbom, Parastoo Azadi, Jonas Nilsson, Yaming Liu, Yannis-Nicolas François, Nassur Said, Jin Young Kim, C. T. Yuen, Shuang Yang, Emmanuelle Leize-Wagner, David Harvey, Xiaofeng Shi, Yan Li, Hirokazu Yagi, Zoran Sosic, Elizabeth M. Hecht, Hua Yuan, Marybeth Creskey, Hyun Kyoung Lee, Sadanori Sekiya, Peter de Vreugd, Len Bell, Sam Tep, BioAnalytical Chemistry, AIMMS, Department of Plant and Microbial Biology, University of California, Laboratory of Infrared Material and Devices, Ningbo University (NBU), University of Natural Resources and Life Sciences (BOKU), Bruker Daltonik GmbH, Bruker Daltonik, Centre d'Immunologie Pierre Fabre, Xinjiang Agriculture University, Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Instituto de Salud Carlos III [Madrid] (ISC)-ministerio de ciencia e innovacion, Complex Carbohydrate Research Center, University of Georgia [USA], GENOS, Universität Duisburg-Essen [Essen], Max Planck Institute for Dynamics of Complex Technical Systems, Max-Planck-Gesellschaft, Section de mathématiques [Genève], Université de Genève (UNIGE), Department of Computer Science [York] (CS-YORK), University of York [York, UK], State Key Laboratory of Hybrid Rice, Department of Genetics, College of Life Sciences, Wuhan University, LeidenUniversity Medical Center, University College Dublin [Dublin] (UCD), Texas A&M University System, College of Engineering and Computer Science, Australian National University (ANU), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Institute of cardiometabolism and nutrition (ICAN), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), University of Edinburgh, University of Alberta, Department of Biological Sciences, Mass Spectrometry Facility, University of Alberta-Department of Chemistry, Volvo Car Corporation, Centre for Research in Intelligent Systems, Monash University [Clayton], Department of Chemistry [Winnipeg, MB, Canada], University of Manitoba [Winnipeg], Department of Chemistry [Winnipeg, Manitoba, Canada], Université de Strasbourg (UNISTRA), Laboratoire de synthèses métallo-induites, Dynamique et structure moléculaire par spectrométrie de masse (LDSM2), School of Mechanics and Engineering [Chengdu], Southwest Jiaotong University (SWJTU), School of Management and Economics [University of Electronic Science and Technology of China], and University of Electronic Science and Technology of China (UESTC)
- Subjects
Proteomics ,PROTEIN ,fluerescence ,Biochemistry ,reference antibody ,THERAPEUTIC ANTIBODIES ,Biopharmaceutics ,Analytical Chemistry ,chemistry.chemical_compound ,Biological sciences ,Glycomics ,NISTaAb ,Analysis method ,ComputingMilieux_MISCELLANEOUS ,glycoproteins ,mass spectrometry ,chemistry.chemical_classification ,0303 health sciences ,glycan ,interlaboratory study ,030302 biochemistry & molecular biology ,Glycopeptides ,Antibodies, Monoclonal ,3. Good health ,glycomics ,fluorescence ,glycosylation ,glycopeptide ,NISTmAb ,lipids (amino acids, peptides, and proteins) ,Protein glycosylation ,Glycan ,Glycosylation ,QUANTITATION ,medicine.drug_class ,Computational biology ,Biology ,Monoclonal antibody ,03 medical and health sciences ,GLYCOMIC ANALYSIS ,SDG 3 - Good Health and Well-being ,Polysaccharides ,[CHIM.ANAL]Chemical Sciences/Analytical chemistry ,Report ,medicine ,Humans ,LC-MS/MS ,Molecular Biology ,030304 developmental biology ,Biological Products ,IDENTIFICATION ,MASS-SPECTROMETRY ,PROFILES ,QUANTIFICATION ,carbohydrates (lipids) ,chemistry ,biology.protein ,Laboratories ,Glycoprotein ,Protein Processing, Post-Translational - Abstract
A broad-based interlaboratory study of glycosylation profiles of a reference and modified IgG antibody involving 103 reports from 76 laboratories., Graphical Abstract Highlights A broad-based interlaboratory study of the glycosylation of a reference antibody: NISTmAb. 103 reports were received from 76 diverse laboratories worldwide. Analysis involved two samples, the NISTmAb and an enzymatically modified sample, enabling within-lab separation of random and systematic errors using the “Youden two-sample” method. Consensus values were derived and similar performance across all experimental methods was noted., Glycosylation is a topic of intense current interest in the development of biopharmaceuticals because it is related to drug safety and efficacy. This work describes results of an interlaboratory study on the glycosylation of the Primary Sample (PS) of NISTmAb, a monoclonal antibody reference material. Seventy-six laboratories from industry, university, research, government, and hospital sectors in Europe, North America, Asia, and Australia submitted a total of 103 reports on glycan distributions. The principal objective of this study was to report and compare results for the full range of analytical methods presently used in the glycosylation analysis of mAbs. Therefore, participation was unrestricted, with laboratories choosing their own measurement techniques. Protein glycosylation was determined in various ways, including at the level of intact mAb, protein fragments, glycopeptides, or released glycans, using a wide variety of methods for derivatization, separation, identification, and quantification. Consequently, the diversity of results was enormous, with the number of glycan compositions identified by each laboratory ranging from 4 to 48. In total, one hundred sixteen glycan compositions were reported, of which 57 compositions could be assigned consensus abundance values. These consensus medians provide community-derived values for NISTmAb PS. Agreement with the consensus medians did not depend on the specific method or laboratory type. The study provides a view of the current state-of-the-art for biologic glycosylation measurement and suggests a clear need for harmonization of glycosylation analysis methods.
- Published
- 2020
28. Subtyping of cardiac amyloidosis by mass spectrometry-based proteomics of endomyocardial biopsies
- Author
-
Fredrik Noborn, Christer Thomsen, Egor Vorontsov, Emanuele Bobbio, Carina Sihlbom, Jonas Nilsson, Christian L. Polte, Entela Bollano, Kristina Vukusic, Joakim Sandstedt, Göran Dellgren, Kristjan Karason, Anders Oldfors, and Göran Larson
- Subjects
Internal Medicine - Abstract
Cardiac amyloidosis is a severe condition leading to restrictive cardiomyopathy and heart failure. Mass spectrometry-based methods for cardiac amyloid subtyping have become important diagnostic tools but are currently used only in a few reference laboratories. Such methods include laser-capture microdissection to ensure the specific analysis of amyloid deposits. Here we introduce a direct proteomics-based method for subtyping of cardiac amyloidosis. Endomyocardial biopsies were retrospectively analysed from fresh frozen material of 78 patients with cardiac amyloidosis and from 12 biopsies of unused donor heart explants. Cryostat sections were digested with trypsin and analysed with liquid chromatography - mass spectrometry, and data were evaluated by proteomic software. With a diagnostic threshold set to 70% for each of the four most common amyloid proteins affecting the heart (LC κ, LC λ, TTR and SAA), 65 of the cases (87%) could be diagnosed, and of these, 61 cases (94%) were in concordance with the original diagnoses. The specimens were also analysed for the summed intensities of the amyloid signature proteins (ApoE, ApoA-IV and SAP). The intensities were significantly higher (p < 0.001) for all assigned cases compared with controls. Cardiac amyloidosis can be successfully subtyped without the prior enrichment of amyloid deposits with laser microdissection.
- Published
- 2022
- Full Text
- View/download PDF
29. Non-random organization of flux control mechanisms in yeast central metabolic pathways
- Author
-
Rosemary Yu, Egor Vorontsov, Carina Sihlbom, and Jens Nielsen
- Abstract
Metabolic flux can be regulated by a variety of different mechanisms, but the organization of these mechanisms within the metabolic network has remained unknown. Here we test the hypothesis that flux control mechanisms are not distributed randomly in the metabolic network, but rather organized according to pathway. Combining proteomics, phosphoproteomics, and metabolic modeling, we report the largest collection of flux-enzyme-phosphoenzyme relationships to date in Saccharomyces cerevisiae. In support of the hypothesis, we show that (i) amino acid metabolic pathways are predominantly regulated by enzyme abundance stemming from transcriptional regulation; (ii) upper glycolysis and associated pathways, by inactivating enzyme phosphorylation; (iii) lower glycolysis and associated pathways, by activating enzyme phosphorylation; and (iv) glycolipid/glycophospholipid pathways, by a combination of enzyme phosphorylation and metabolic compartmentalization. We delineate the evolutionary history for the observed organization of flux control mechanisms in yeast central metabolic pathways, furthering our understanding of the regulation of metabolism and its evolution.
- Published
- 2021
30. Site-specific N-glycan profiles of α
- Author
-
Ekaterina, Mirgorodskaya, Estelle, Dransart, Massiullah, Shafaq-Zadah, Daniel, Roderer, Carina, Sihlbom, Hakon, Leffler, and Ludger, Johannes
- Subjects
Glycosylation ,Liver ,Polysaccharides ,Integrin beta1 ,Animals ,Integrin alpha5 ,Glycoproteins ,Rats - Abstract
Like most other cell surface proteins, αHere, we have established the first comprehensive site-specific glycan map of αWe expect that the glycoproteomics data of the current study will serve as a resource for the exploration of structural mechanisms by which glycans control αGlycosylation of α
- Published
- 2021
31. Age and Sex Effects across the Blood Proteome after Lonizing Radiation Exposure: Consequences for Biomarker Screening and Risk Assessment
- Author
-
Britta Langen, Egor Vorontsov, Johan Spetz, John Swanpalmer, Carina Sihlbom, Khalil Helou, and Eva Forssell-Aronsson
- Abstract
Molecular biomarkers of ionizing radiation (IR) exposure are a promising new tool in various disciplines: they can give necessary information for adaptive treatment planning in cancer radiotherapy, enable risk projection for radiation-induced survivorship diseases, or facilitate triage and intervention in radiation hazard events. However, radiation biomarker discovery has not yet resolved the most basic features of personalized medicine: age and sex. To overcome this critical bias in biomarker identification, we quantitated age and sex effects and assessed their relevance in the radiation response across the blood proteome. We used high-throughput mass spectrometry on blood plasma collected 24 hours after 0.5 Gy total body irradiation (15 MV nominal photon energy) from male and female C57BL/6N mice at juvenile (7-weeks-old) or adult (18-weeks-old) age. We also assessed sex and strain effects using juvenile male and female BALB/c nude mice. We showed that age and sex created significant effects in the proteomic response regarding both extent and functional quality of IR-induced responses. Furthermore, we found that age and sex effects appeared non-linear and were often end-point specific. Overall, age contributed more to differences in the proteomic response than sex, most notably in immune responses, oxidative stress, and apoptotic cell death. Interestingly, sex effects were pronounced for DNA damage & repair pathways and associated cellular outcome (pro-survival vs. pro-apoptotic). Only one protein (AHSP) was identified as a potential general biomarker candidate across age and sex, while GMNN, REG3B, and SNCA indicated some response similarity across age. This low yield advocated that unisex or uniage biomarker screening approaches are not feasible. In conclusion, age- and sex-specific screening approaches should be implemented as standard protocol to ensure robustness and diagnostic power of biomarker candidates. Bias-free molecular biomarkers are a necessary progression towards personalized medicine and integral for advanced adaptive cancer radiotherapy and risk assessment.
- Published
- 2021
32. Age and sex effects across the blood proteome after ionizing radiation exposure can bias biomarker screening and risk assessment
- Author
-
Britta Langen, Egor Vorontsov, Johan Spetz, John Swanpalmer, Carina Sihlbom, Khalil Helou, and Eva Forssell-Aronsson
- Subjects
Male ,Proteomics ,Multidisciplinary ,Proteome ,Mice, Nude ,Risk Assessment ,Mice, Inbred C57BL ,Mice ,Neoplasms ,Radiation, Ionizing ,Animals ,Female ,Radiation Injuries ,Biomarkers - Abstract
Molecular biomarkers of ionizing radiation (IR) exposure are a promising new tool in various disciplines: they can give necessary information for adaptive treatment planning in cancer radiotherapy, enable risk projection for radiation-induced survivorship diseases, or facilitate triage and intervention in radiation hazard events. However, radiation biomarker discovery has not yet resolved the most basic features of personalized medicine: age and sex. To overcome this critical bias in biomarker identification, we quantitated age and sex effects and assessed their relevance in the radiation response across the blood proteome. We used high-throughput mass spectrometry on blood plasma collected 24 h after 0.5 Gy total body irradiation (15 MV nominal photon energy) from male and female C57BL/6 N mice at juvenile (7-weeks-old) or adult (18-weeks-old) age. We also assessed sex and strain effects using juvenile male and female BALB/c nude mice. We showed that age and sex created significant effects in the proteomic response regarding both extent and functional quality of IR-induced responses. Furthermore, we found that age and sex effects appeared non-linear and were often end-point specific. Overall, age contributed more to differences in the proteomic response than sex, most notably in immune responses, oxidative stress, and apoptotic cell death. Interestingly, sex effects were pronounced for DNA damage and repair pathways and associated cellular outcome (pro-survival vs. pro-apoptotic). Only one protein (AHSP) was identified as a potential general biomarker candidate across age and sex, while GMNN, REG3B, and SNCA indicated some response similarity across age. This low yield advocated that unisex or uniage biomarker screening approaches are not feasible. In conclusion, age- and sex-specific screening approaches should be implemented as standard protocol to ensure robustness and diagnostic power of biomarker candidates. Bias-free molecular biomarkers are a necessary progression towards personalized medicine and integral for advanced adaptive cancer radiotherapy and risk assessment.
- Published
- 2021
33. A Glycoproteomic Approach to Identify Novel Proteoglycans
- Author
-
Carina Sihlbom, Jonas Nilsson, Göran Larson, Andrea Persson, Fredrik Noborn, and Mahnaz Nikpour
- Subjects
chemistry.chemical_compound ,Chromatography ,chemistry ,Liquid chromatography–mass spectrometry ,Disaccharide ,Tetrasaccharide ,Heparan sulfate ,Chondroitin sulfate ,Tandem mass spectrometry ,Glycopeptide ,Glycoproteomics - Abstract
In this chapter, we describe a glycoproteomic approach for the identification of novel chondroitin sulfate proteoglycans (CSPGs) using a combination of biochemical enrichments, enzymatic digestions, and nanoscale liquid chromatography tandem mass spectrometry (nLC-MS/MS) analysis. The identification is achieved by trypsin digestion of CSPG-containing samples, followed by enrichment of chondroitin sulfate (CS) glycopeptides by strong anion exchange chromatography (SAX). The enriched CS glycopeptides are then digested with chondroitinase ABC to depolymerize the CS polysaccharides, generating a residual hexasaccharide structure, composed of the linkage region tetrasaccharide extended with a terminal dehydrated disaccharide, still attached to the peptide. The obtained CS glycopeptides are analyzed by nLC-MS/MS, and the generated data sets are evaluated through proteomic software with adjustment in the settings to allow for glycopeptide identification. This approach has enabled the identification of several novel core proteins in human samples and in Caenorhabditis elegans. Here we specifically describe the procedure for the enrichment and characterization of CS glycopeptides from human cerebrospinal fluid (CSF).
- Published
- 2021
34. A Glycoproteomic Approach to Identify Novel Proteoglycans
- Author
-
Fredrik, Noborn, Mahnaz, Nikpour, Andrea, Persson, Carina, Sihlbom, Jonas, Nilsson, and Göran, Larson
- Subjects
Proteomics ,Tandem Mass Spectrometry ,Chondroitin Sulfates ,Glycopeptides ,Animals ,Humans ,Proteoglycans ,Caenorhabditis elegans ,Chromatography, Liquid - Abstract
In this chapter, we describe a glycoproteomic approach for the identification of novel chondroitin sulfate proteoglycans (CSPGs) using a combination of biochemical enrichments, enzymatic digestions, and nanoscale liquid chromatography tandem mass spectrometry (nLC-MS/MS) analysis. The identification is achieved by trypsin digestion of CSPG-containing samples, followed by enrichment of chondroitin sulfate (CS) glycopeptides by strong anion exchange chromatography (SAX). The enriched CS glycopeptides are then digested with chondroitinase ABC to depolymerize the CS polysaccharides, generating a residual hexasaccharide structure, composed of the linkage region tetrasaccharide extended with a terminal dehydrated disaccharide, still attached to the peptide. The obtained CS glycopeptides are analyzed by nLC-MS/MS, and the generated data sets are evaluated through proteomic software with adjustment in the settings to allow for glycopeptide identification. This approach has enabled the identification of several novel core proteins in human samples and in Caenorhabditis elegans. Here we specifically describe the procedure for the enrichment and characterization of CS glycopeptides from human cerebrospinal fluid (CSF).
- Published
- 2021
35. Silencing of STE20-type kinase STK25 in human aortic endothelial and smooth muscle cells is atheroprotective
- Author
-
Emmelie Cansby, Sima Kumari, Mara Caputo, Ying Xia, Rando Porosk, Jonathan Robinson, Hao Wang, Britt-Marie Olsson, Josefine Vallin, Julie Grantham, Ursel Soomets, L. Thomas Svensson, Carina Sihlbom, Hanns-Ulrich Marschall, Andreas Edsfeldt, Isabel Goncalves, and Margit Mahlapuu
- Subjects
Myocytes, Smooth Muscle ,Intracellular Signaling Peptides and Proteins ,Medicine (miscellaneous) ,Humans ,Protein Serine-Threonine Kinases ,General Agricultural and Biological Sciences ,Atherosclerosis ,Lipid Metabolism ,Lipids ,General Biochemistry, Genetics and Molecular Biology - Abstract
Recent studies highlight the importance of lipotoxic damage in aortic cells as the major pathogenetic contributor to atherosclerotic disease. Since the STE20-type kinase STK25 has been shown to exacerbate ectopic lipid storage and associated cell injury in several metabolic organs, we here investigate its role in the main cell types of vasculature. We depleted STK25 by small interfering RNA in human aortic endothelial and smooth muscle cells exposed to oleic acid and oxidized LDL. In both cell types, the silencing of STK25 reduces lipid accumulation and suppresses activation of inflammatory and fibrotic pathways as well as lowering oxidative and endoplasmic reticulum stress. Notably, in smooth muscle cells, STK25 inactivation hinders the shift from a contractile to a synthetic phenotype. Together, we provide several lines of evidence that antagonizing STK25 signaling in human aortic endothelial and smooth muscle cells is atheroprotective, highlighting this kinase as a new potential therapeutic target for atherosclerotic disease.
- Published
- 2021
36. Role of apolipoprotein C‐III overproduction in diabetic dyslipidaemia
- Author
-
Nina Lundbom, Linda Andersson, Sanni Söderlund, Juhani Kahri, Haihong Zhou, Martin Adiels, Marja-Riitta Taskinen, Annika Thorsell, Kirsi H. Pietiläinen, Antti Hakkarainen, Niina Matikainen, Elias Björnson, Carina Sihlbom, Chris J. Packard, Jan Borén, Research Programs Unit, CAMM - Research Program for Clinical and Molecular Metabolism, Faculty of Medicine, HUS Abdominal Center, Endokrinologian yksikkö, HUS Internal Medicine and Rehabilitation, Marja-Riitta Taskinen Research Group, Department of Medicine, HUS Medical Imaging Center, Department of Diagnostics and Therapeutics, University of Gothenburg, University of Helsinki, Department of Neuroscience and Biomedical Engineering, Merck, University of Glasgow, Aalto-yliopisto, and Aalto University
- Subjects
Blood Glucose ,Male ,Very low-density lipoprotein ,Apolipoprotein B ,Endocrinology, Diabetes and Metabolism ,apolipoprotein C-III ,Type 2 diabetes ,030204 cardiovascular system & hematology ,CORONARY EVENTS ,chemistry.chemical_compound ,0302 clinical medicine ,Endocrinology ,OF-FUNCTION MUTATIONS ,Glucose homeostasis ,RISK ,2. Zero hunger ,PLASMA ,biology ,Area under the curve ,Middle Aged ,Postprandial Period ,REMNANT CHOLESTEROL ,TRIGLYCERIDE-RICH LIPOPROTEIN ,3. Good health ,Postprandial ,Female ,lipids (amino acids, peptides, and proteins) ,type 2 diabetes ,VLDL ,medicine.drug ,medicine.medical_specialty ,LOW-DENSITY-LIPOPROTEIN ,stable isotopes ,030209 endocrinology & metabolism ,METABOLISM ,03 medical and health sciences ,Internal medicine ,Internal Medicine ,medicine ,Humans ,Hypoglycemic Agents ,Triglycerides ,ta217 ,Aged ,Dyslipidemias ,Apolipoprotein C-III ,Triglyceride ,Liraglutide ,business.industry ,medicine.disease ,lipoproteins ,Diabetes Mellitus, Type 2 ,chemistry ,kinetics ,3121 General medicine, internal medicine and other clinical medicine ,biology.protein ,business ,CIII - Abstract
openaire: EC/H2020/305707/EU//RESOLVE Aims: To investigate how apolipoprotein C-III (apoC-III) metabolism is altered in subjects with type 2 diabetes, whether the perturbed plasma triglyceride concentrations in this condition are determined primarily by the secretion rate or the removal rate of apoC-III, and whether improvement of glycaemic control using the glucagon-like peptide-1 analogue liraglutide for 16 weeks modifies apoC-III dynamics. Materials and Methods: Postprandial apoC-III kinetics were assessed after a bolus injection of [5,5,5- 2 H 3 ]leucine using ultrasensitive mass spectrometry techniques. We compared apoC-III kinetics in two situations: in subjects with type 2 diabetes before and after liraglutide therapy, and in type 2 diabetic subjects with matched body mass index (BMI) non-diabetic subjects. Liver fat content, subcutaneous abdominal and intra-abdominal fat were determined using proton magnetic resonance spectroscopy. Results: Improved glycaemic control by liraglutide therapy for 16 weeks significantly reduced apoC-III secretion rate (561 ± 198 vs. 652 ± 196 mg/d, P = 0.03) and apoC-III levels (10.0 ± 3.8 vs. 11.7 ± 4.3 mg/dL, P = 0.035) in subjects with type 2 diabetes. Change in apoC-III secretion rate was significantly associated with the improvement in indices of glucose control (r = 0.67; P = 0.009) and change in triglyceride area under the curve (r = 0.59; P = 0.025). In line with this, the apoC-III secretion rate was higher in subjects with type 2 diabetes compared with BMI-matched non-diabetic subjects (676 ± 208 vs. 505 ± 174 mg/d, P = 0.042). Conclusions: The results reveal that the secretion rate of apoC-III is associated with elevation of triglyceride-rich lipoproteins in subjects with type 2 diabetes, potentially through the influence of glucose homeostasis on the production of apoC-III.
- Published
- 2019
37. Protein kinase MST3 modulates lipid homeostasis in hepatocytes and correlates with nonalcoholic steatohepatitis in humans
- Author
-
Carina Sihlbom, Marcus Ståhlman, Margit Mahlapuu, Annika Nerstedt, Emmelie Cansby, Jan Borén, Hanns-Ulrich Marschall, Nagaraj M. Kulkarni, Elin Magnusson, Yeshwant Kurhe, Manoj Amrutkar, and Matthias Blüher
- Subjects
Male ,0301 basic medicine ,Mitochondria, Liver ,Biochemistry ,Mice ,0302 clinical medicine ,Non-alcoholic Fatty Liver Disease ,Lipid droplet ,Nonalcoholic fatty liver disease ,Homeostasis ,RNA, Small Interfering ,Cells, Cultured ,Gene knockdown ,Chemistry ,Cell biology ,Pyruvate carboxylase ,Liver ,Organ Specificity ,Gene Knockdown Techniques ,Female ,RNA Interference ,lipids (amino acids, peptides, and proteins) ,medicine.symptom ,Oxidation-Reduction ,Biotechnology ,Inflammation ,Protein Serine-Threonine Kinases ,Diet, High-Fat ,03 medical and health sciences ,Genetics ,medicine ,Animals ,Humans ,Protein kinase A ,Molecular Biology ,Triglycerides ,Catabolism ,Lipid Droplet Associated Proteins ,Lipid metabolism ,Lipid Droplets ,Lipid Metabolism ,medicine.disease ,Dietary Fats ,Cell Compartmentation ,Rats ,Mice, Inbred C57BL ,Oxidative Stress ,030104 developmental biology ,Hepatocytes ,030217 neurology & neurosurgery - Abstract
Ectopic lipid storage in the liver is considered the main risk factor for nonalcoholic steatohepatitis (NASH). Understanding the molecular networks controlling hepatocellular lipid deposition is therefore essential for developing new strategies to effectively prevent and treat this complex disease. Here, we describe a new regulator of lipid partitioning in human hepatocytes: mammalian sterile 20-like (MST) 3. We found that MST3 protein coats lipid droplets in mouse and human liver cells. Knockdown of MST3 attenuated lipid accumulation in human hepatocytes by stimulating β-oxidation and triacylglycerol secretion while inhibiting fatty acid influx and lipid synthesis. We also observed that lipogenic gene expression and acetyl-coenzyme A carboxylase protein abundance were reduced in MST3-deficient hepatocytes, providing insight into the molecular mechanisms underlying the decreased lipid storage. Furthermore, MST3 expression was positively correlated with key features of NASH (i.e., hepatic lipid content, lobular inflammation, and hepatocellular ballooning) in human liver biopsies. In summary, our results reveal a role of MST3 in controlling the dynamic metabolic balance of liver lipid catabolism vs. lipid anabolism. Our findings highlight MST3 as a potential drug target for the prevention and treatment of NASH and related complex metabolic diseases.-Cansby, E., Kulkarni, N. M., Magnusson, E., Kurhe, Y., Amrutkar, M., Nerstedt, A., Stahlman, M., Sihlbom, C., Marschall, H.-U., Boren, J., Bluher, M., Mahlapuu, M. Protein kinase MST3 modulates lipid homeostasis in hepatocytes and correlates with nonalcoholic steatohepatitis in humans.
- Published
- 2019
38. Mice exposed to maternal androgen excess and diet-induced obesity have altered phosphorylation of catechol-O-methyltransferase in the placenta and fetal liver
- Author
-
Mattias Carlström, Elisabet Stener-Victorin, Angelica Lindén Hirschberg, Maria Manti, Elisabet Jerlhag, Manuel Maliqueo, Romina Fornes, Xiaojuan Qi, Anna Benrick, Egor Vorontsov, Jenny Nyström, and Carina Sihlbom
- Subjects
Male ,medicine.medical_specialty ,Dietary Sugars ,Placenta ,Endocrinology, Diabetes and Metabolism ,Medicine (miscellaneous) ,030209 endocrinology & metabolism ,Catechol O-Methyltransferase ,Diet, High-Fat ,Androgen Excess ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Fetus ,0302 clinical medicine ,Pregnancy ,Internal medicine ,medicine ,Animals ,Obesity ,030212 general & internal medicine ,Phosphorylation ,Nutrition and Dietetics ,Catechol-O-methyl transferase ,Triglyceride ,business.industry ,Dihydrotestosterone ,medicine.disease ,Endocrinology ,medicine.anatomical_structure ,Liver ,chemistry ,Female ,Liver function ,business - Abstract
Maternal obesity together with androgen excess in mice negatively affects placental function and maternal and fetal liver function as demonstrated by increased triglyceride content with dysfunctional expression of enzymes and transcription factors involved in de novo lipogenesis and fat storage. To identify changes in molecular pathways that might promote diseases in adulthood, we performed a global proteomic analysis using a liquid-chromatography/mass-spectrometry system to investigate total and phosphorylated proteins in the placenta and fetal liver in a mouse model that combines maternal obesity with maternal androgen excess.After ten weeks on a control diet (CD) or high fat/high sugar-diet, dams were mated with males fed the CD. Between gestational day (GD) 16.5 and GD 18.5, mice were injected with vehicle or dihydrotestosterone (DHT) and sacrificed at GD 18.5 prior to dissection of the placentas and fetal livers. Four pools of female placentas and fetal livers were subjected to a global proteomic analysis. Total and phosphorylated proteins were filtered by ANOVA q 0.05, and this was followed by two-way ANOVA to determine the effect of maternal obesity and/or androgen exposure.In placenta, phosphorylated ATP-citrate synthase was decreased due to maternal obesity, and phosphorylated catechol-O-methyltransferase (COMT) was differentially expressed due to the interaction between maternal diet and DHT exposure. In fetal liver, five total proteins and 48 proteins phosphorylated in one or more sites, were differentially expressed due to maternal obesity or androgen excess. In fetal liver, phosphorylated COMT expression was higher in fetus exposed to maternal obesity.These results suggest a common regulatory mechanism of catecholamine metabolism in the placenta and the fetal liver as demonstrated by higher phosphorylated COMT expression in the placenta and fetal liver from animals exposed to diet-induced maternal obesity and lower expression of phosphorylated COMT in animals exposed to maternal androgen excess.
- Published
- 2019
39. Author response: Quantifying absolute gene expression profiles reveals distinct regulation of central carbon metabolism genes in yeast
- Author
-
Egor Vorontsov, Carina Sihlbom, Rosemary Yu, and Jens Nielsen
- Subjects
Biochemistry ,Gene expression ,Biology ,Central carbon metabolism ,Gene ,Yeast - Published
- 2021
40. Differential fate of acellular vascular scaffolds in vivo involves macrophages and T-cell subsets
- Author
-
Nikhil B. Nayakawde, Deepti Antony, Uzair Ul Haq, Michael Olausson, Carina Sihlbom, Sudip Ghosh, Meghshree Deshmukh, Debashish Banerjee, and Evelin Berger
- Subjects
Extracellular matrix ,Scaffold ,medicine.anatomical_structure ,Immune system ,Decellularization ,In vivo ,Chemistry ,T cell ,Cell ,medicine ,Macrophage ,Cell biology - Abstract
Biological scaffold or implant is a popular choice for the preparation of tissue-engineered organs and has the potential to address donor shortage in the clinics. However, biological scaffolds prepared by physical or chemical agents cause damage to the extracellular matrix by potentially inducing immune responses after implantation. The current study explores an alternative route for the preparation of acellular scaffolds and explores the fate of the prepared scaffolds in a milieu of immune cells following implantation without using immunosuppressant. Using the syngeneic (Lewis male-Lewis female) and allogeneic (Brown Norway male-Lewis female) models and different tissue routes (subcutaneous vs omentum) for transplantation, normal blood vascular scaffolds were implanted which was converted to acellular vascular scaffolds by in vivo natural decellularization at the end of 2 months of observation. We also prepared chemically decellularized acellular scaffolds from normal untreated blood vascular scaffolds using a cocktail of chemicals which was also similarly placed in subcutaneous and omentum sites. Here, we applied in-depth quantitative proteomics along with histology and image analysis to comprehensively describe and compare the proteome of the natural and chemically decellularized scaffold. Our data confirm that site-specific advantages exist in modulating the ECM and regulating the immune responses (macrophage and T cells) following implantation, which possibly led to the production of an acellular scaffold (natural decellularization) under in vivo conditions. The current approach opens up the possibility to create tailor-made acellular scaffolds to build functional blood vessels. In addition, the identification of different tissue sites facilitates differential immune response against the scaffolds. This study provides a rich resource aimed toward an enhanced mechanistic understanding to study immune responses under similar settings in the field of transplantation and regenerative medicine.Impact statementThe development of a scaffold helps in the preparation of a functional organ in the clinics. In the current study, we prepared an acellular vascular scaffold by utilizing site specific tissue changes and vis-à-vis compared with a conventionally chemically prepared biological scaffold at genomic and protein level, which helped us to identify immunological trigger following implantation. The current study which was carried out without any immunosuppressive agents could help to establish (a) alternative strategies for preparing biological scaffolds as well as (b) implantable sites as potential bioreactors to circumvent any adverse immune reactions for acceptance of the scaffold/implant post implantation.
- Published
- 2020
41. Revealing interspecies transmission barriers of avian influenza A viruses
- Author
-
Britt-Marie Olsson, Robert H. S. Kraus, Carina Sihlbom, Elinor Jax, Caroline Bröjer, Göran Larson, Mahmoud M. Naguib, Josef D. Järhult, Jan Nilsson, Cecilia Lindskog, Bjorn R. Olsen, Åke Lundkvist, Patrik Ellström, Michelle Wille, Per Eriksson, and J. Waldenstroem
- Subjects
Innate immune system ,Host (biology) ,Receptor expression ,Biology ,medicine.disease_cause ,Influenza A virus subtype H5N1 ,law.invention ,Interspecies transmission ,Transmission (mechanics) ,Evolutionary biology ,law ,Pandemic ,Influenza A virus ,medicine - Abstract
Influenza A virus (IAV) pandemics result from interspecies transmission events within the avian reservoir and further to mammals including humans. Investigating molecular virus–host interactions dictating this process and the adaptations to the new hosts that follow is vital to understand zoonotic IAV spread. Receptor incompatibility has been suggested to limit zoonotic IAV transmission from the wild bird reservoir. Other barriers to interspecies transmission, particularly within the avian system, largely remain elusive. Through assessment of infection dynamics of mallard origin IAV in two different avian hosts, coupled with studies of receptor expression and host response we aimed to reveal the host-pathogen interactions in a cross-species transmission event. We found that shedding patterns and innate immune responses were highly dependent on viral genotypes, host species and inoculation routes, but less dependent on receptor expression. Further, in contrary to the prevailing dogma we demonstrate that birds can produce a wide range of different sialylated structures also found in mammals, e.g. extended N- and O-linked Neu5Acα2,6 terminated glycans. Overall, receptor incompatibility is not the sole transmission barrier for IAV between birds and to humans, but other host-pathogen factors deserve dedicated studies to achieve proper pandemic preparedness.
- Published
- 2020
42. Apolipoprotein B48 metabolism in chylomicrons and very low‐density lipoproteins and its role in triglyceride transport in normo‐ and hypertriglyceridemic human subjects
- Author
-
Antti Hakkarainen, Linda Andersson, Carina Sihlbom, Sanni Söderlund, Martin Adiels, Juhani Kahri, Haihong Zhou, Jan Borén, Jesper Lundbom, Chris J. Packard, Elias Björnson, Niina Matikainen, Nina Lundbom, Marja-Riitta Taskinen, Annika Thorsell, HUS Abdominal Center, Staff Services, CAMM - Research Program for Clinical and Molecular Metabolism, Research Programs Unit, University of Helsinki, Endokrinologian yksikkö, Clinicum, Department of Medicine, HUS Internal Medicine and Rehabilitation, Marja-Riitta Taskinen Research Group, HUS Medical Imaging Center, and Department of Diagnostics and Therapeutics
- Subjects
Male ,0301 basic medicine ,Very low-density lipoprotein ,Apolipoprotein B ,Lipoproteins, VLDL ,030204 cardiovascular system & hematology ,apoB100 ,B-48 TRANSPORT ,chemistry.chemical_compound ,0302 clinical medicine ,apoB48 ,OF-FUNCTION MUTATIONS ,Chylomicrons ,RISK ,Hypertriglyceridemia ,education.field_of_study ,PLASMA ,biology ,digestive, oral, and skin physiology ,STABLE-ISOTOPE ,Middle Aged ,REMNANT CHOLESTEROL ,3. Good health ,Protein Transport ,CARDIOVASCULAR-DISEASE ,Apolipoprotein B-100 ,lipids (amino acids, peptides, and proteins) ,medicine.medical_specialty ,C-III ,Lipolysis ,Lipoproteins ,Dietary lipid ,Population ,postpranidal lipid metabolism ,stable isotopes ,03 medical and health sciences ,Internal medicine ,Internal Medicine ,medicine ,Humans ,education ,Triglycerides ,Triglyceride ,Triglyceride transport ,business.industry ,medicine.disease ,030104 developmental biology ,Endocrinology ,chemistry ,kinetics ,FAT ,Heart Disease Risk Factors ,3121 General medicine, internal medicine and other clinical medicine ,RICH LIPOPROTEINS ,biology.protein ,Apolipoprotein B-48 ,business ,Chylomicron - Abstract
Background: \ud Renewed interest in triglyceride-rich lipoproteins as causative agents in cardiovascular disease mandates further exploration of the integrated metabolism of chylomicrons and very low-density lipoproteins (VLDL).\ud \ud Methods: \ud Novel tracer techniques and an integrated multi-compartmental model were used to determine the kinetics of apoB48- and apoB100-containing particles in the chylomicron and VLDL density intervals in 15 subjects with a wide range of plasma triglyceride levels.\ud \ud Results: \ud Following a fat-rich meal, apoB48 appeared in the chylomicron, VLDL1 and VLDL2 fractions in all subjects. Chylomicrons cleared rapidly from the circulation but apoB48-containing VLDL accumulated, and over the day were 3-fold higher in those with high versus low plasma triglyceride. ApoB48-containing particles were secreted directly into both the chylomicron and VLDL fractions at rates that were similar across the plasma triglyceride range studied. During fat absorption, whilst most triglyceride entered the circulation in chylomicrons, the majority of apoB48 particles were secreted into the VLDL density range.\ud \ud Conclusion: \ud The intestine secretes apoB48-containing particles not only as chylomicrons but also directly into the VLDL1 and VLDL2 density ranges both in the basal state and during dietary lipid absorption. Over the day, apoB48-containing particles appear to comprise about 20–25% of circulating VLDL and, especially in those with elevated triglycerides, form part of a slowly cleared ‘remnant’ particle population, thereby potentially increasing CHD risk. These findings provide a metabolic understanding of the potential consequences for increased CHD risk when slowed lipolysis leads to the accumulation of remnants, especially in individuals with hypertriglyceridemia.
- Published
- 2020
43. Profiling of airway epithelial proteome reveals sex differences in early stage COPD linked to xenobiotic metabolism and ER stress
- Author
-
Mingxing Yang, Reza Karimi, Tina Heyder, Chuanxing Li, Maxie Kohler, Riitta Kaarteenaho, Carina Sihlbom, Åsa M. Wheelock, Johan Grunewald, C. Magnus Sköld, Heta Merikallio, and Helena Forsslund
- Subjects
COPD ,business.industry ,Proteome ,Unfolded protein response ,Medicine ,Airway ,Bioinformatics ,business ,medicine.disease ,Drug metabolism - Published
- 2020
44. Differential Activation of Immune Cells for Genetically Different Decellularized Cardiac Tissues
- Author
-
Christopher Agbajogu, Galyna Travnikova, Michael Olausson, Carina Sihlbom, Debashish Banerjee, Nikhil B. Nayakawde, and Ketaki Methe
- Subjects
Proteomics ,Swine ,T cell ,0206 medical engineering ,Biomedical Engineering ,Bioengineering ,Biocompatible Materials ,02 engineering and technology ,Biology ,CD8-Positive T-Lymphocytes ,Biochemistry ,Immune tolerance ,Biomaterials ,Extracellular matrix ,03 medical and health sciences ,Mice ,Immune system ,medicine ,Immune Tolerance ,Animals ,030304 developmental biology ,0303 health sciences ,Decellularization ,Immunogenicity ,Heart ,Acquired immune system ,020601 biomedical engineering ,Cell biology ,Extracellular Matrix ,medicine.anatomical_structure ,Heterografts ,CD8 - Abstract
The immunogenicity of the extracellular matrix (ECM) from genetically similar (syngeneic) and dissimilar (allogeneic and xenogeneic) species has puzzled the scientific community for many years. After implantation, the literature describes an absorption of ECM material since it is biodegradable. However, no clear insight really exists to substantiate how the underlying immune and biological responses result in absorption of ECM materials. In this context, it is important to characterize infiltrating cells and identify dominant cell populations in the infiltrate. We have studied the immune response in mice after implantation of decellularized (DC) cardiac scaffolds derived from pig and mouse. The polymorphism of the infiltrate into the implanted material signifies the importance of the adaptive immune response that is distinct for xenoimplants and alloimplants. Matrix resorption takes place mainly through phagocytic cells such as mast cells, dendritic cells, and macrophages. Histochemical observations show that innate CD8+ T cells develop immune tolerance, whereas proteomic analysis predicts the different T cell progenies for alloscaffolds and xenoscaffolds. The amalgamation of graft tolerance and involvement of both B and T cell populations in the vicinity of the graft could be decisive in wound remodeling and survival of the graft. This challenging area presents potential targets for the development of immune-privileged biomaterials, immune tolerant cells, and therapeutic agents in the future. Impact statement In this study, we have characterized the allogeneic and xenogeneic immune responses for decellularized (DC) cardiac scaffolds. We postulate that although the T cells are important players for immune tolerance of DC graft, the mechanism of their differentiation inside the host is donor specific. In this study, we have reported the distinct immune responses for syngeneic DC scaffolds than allogeneic and xenogeneic scaffolds. This distinct response provides the bases for the different immune responses reported for DC homografts in the literature. This study can provide the greater insight for modification of postimplant strategies to achieve host acceptance of donor extracellular matrix scaffolds.
- Published
- 2020
45. Vitellogenin offsets oxidative costs of reproduction in female painted dragon lizards
- Author
-
Mark R. Wilson, Christopher R. Friesen, Jörgen Bergström, Mats Olsson, Willow R. Lindsay, Evelin Berger, and Carina Sihlbom
- Subjects
0106 biological sciences ,food.ingredient ,Physiology ,media_common.quotation_subject ,Oxidative phosphorylation ,Aquatic Science ,Biology ,medicine.disease_cause ,010603 evolutionary biology ,01 natural sciences ,Andrology ,Vitellogenins ,03 medical and health sciences ,Vitellogenin ,food ,Yolk ,biology.animal ,medicine ,Animals ,Molecular Biology ,Ovulation ,Ecology, Evolution, Behavior and Systematics ,030304 developmental biology ,media_common ,chemistry.chemical_classification ,0303 health sciences ,Reactive oxygen species ,Lizard ,Reproduction ,Vitellogenesis ,Lizards ,Oxidative Stress ,chemistry ,Insect Science ,biology.protein ,Female ,Animal Science and Zoology ,Oxidative stress - Abstract
Vitellogenesis (“yolking” of follicles) is a bioenergetically costly stage of reproduction requiring enlargement of the liver to produce vitellogenin (VTG) yolk precursor proteins, which are transported and deposited at the ovary. VTG may, however, serve non-nutritive antioxidant functions, a hypothesis supported by empirical work on aging and other life-history transitions in several taxa. We test this hypothesis in female painted dragon lizards (Ctenophorus pictus) by examining covariation in VTG with the ovarian cycle, and relative to reactive oxygen species (ROS) including baseline superoxide (bSO). Plasma VTG decreased prior to ovulation, when VTG is deposited into follicles. VTG, however, remained elevated post-ovulation when no longer necessary for yolk provisioning and was unrelated to reproductive investment. Instead, VTG was strongly and positively predicted by prior bSO. ROS, in turn, was negatively predicted by prior VTG, while simultaneously sampled VTG was a positive predictor. These findings are consistent with the hypothesis that VTG functions as an antioxidant to counteract oxidative stress associated with vitellogenesis. The relationship between bSO and VTG was strongest in post-ovulatory females, indicating its function may be largely antioxidant at this time. In conclusion, VTG may be under selection to offset oxidative costs of reproduction in egg-producing species.
- Published
- 2020
46. Effective Assignment of α2,3/α2,6-Sialic Acid Isomers by LC-MS/MS-Based Glycoproteomics
- Author
-
Ulrika Westerlind, Manuel Schorlemer, Vanessa Caixeta, Christian Pett, Waqas Nasir, Carina Sihlbom, René P. Zahedi, Jonas Nilsson, Göran Larson, and Britt-Marie Olsson
- Subjects
0301 basic medicine ,Proteomics ,Glycosylation ,viruses ,Mass spectrometry ,010402 general chemistry ,01 natural sciences ,Catalysis ,03 medical and health sciences ,chemistry.chemical_compound ,Tandem Mass Spectrometry ,Lc ms ms ,Carbohydrate Conformation ,chemistry.chemical_classification ,010401 analytical chemistry ,Glycosidic bond ,Stereoisomerism ,General Chemistry ,General Medicine ,Glycopeptide ,Glycoproteomics ,Sialic acid ,0104 chemical sciences ,030104 developmental biology ,Biochemistry ,chemistry ,Sialic Acids ,sense organs ,Glycoprotein ,Chromatography, Liquid - Abstract
Distinct structural changes of the α2,3/α2,6-sialic acid glycosidic linkages on glycoproteins are of importance in cancer biology, inflammatory diseases, and virus tropism. Current glycoproteomic methodologies are, however, not amenable toward high-throughput characterization of sialic acid isomers. To enable such assignments, a mass spectrometry method utilizing synthetic model glycopeptides for the analysis of oxonium ion intensity ratios was developed. This method was successfully applied in large-scale glycoproteomics, thus allowing the site-specific structural characterization of sialic acid isomers.
- Published
- 2018
47. Cracking the Sugar Code by Mass Spectrometry
- Author
-
Carina Sihlbom, Ekaterina Mirgorodskaya, Carol L. Nilsson, Niclas G. Karlsson, and Göran Larson
- Subjects
0301 basic medicine ,03 medical and health sciences ,030104 developmental biology ,Structural Biology ,Chemistry ,Honor ,Context (language use) ,Mass spectrometric ,Spectroscopy ,Code (semiotics) ,Genealogy - Abstract
The structural study of glycans and glycoconjugates is essential to assign their roles in homeostasis, health, and disease. Once dominated by nuclear magnetic resonance spectroscopy, mass spectrometric methods have become the preferred toolbox for the determination of glycan structures at high sensitivity. The patterns of such structures in different cellular states now allow us to interpret the sugar codes in health and disease, based on structure-function relationships. Dr. Catherine E. Costello was the 2017 recipient of the American Society for Mass Spectrometry’s Distinguished Contribution Award. In this Perspective article, we describe her seminal work in a historical and geographical context and review the impact of her research accomplishments in the field. 8
- Published
- 2018
48. Orellanine specifically targets renal clear cell carcinoma
- Author
-
Heidi Hedman, Lovisa Bergwall, Emelie Roos, Kerstin Ebefors, Deman Najar, Anders Herrmann, Börje Haraldsson, Sven Lundstam, Alina Khramova, Jan Törnell, Martin Johansson, Ulf Nilsson, Jenny Nyström, Hanna Wallentin, Carina Sihlbom, and Lisa Buvall
- Subjects
0301 basic medicine ,Programmed cell death ,Pathology ,medicine.medical_specialty ,Necrosis ,clear cell renal cell carcinoma ,nephrotoxin ,necrosis ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Renal cell carcinoma ,medicine ,Kidney ,business.industry ,Orellanine ,apoptosis ,medicine.disease ,Clear cell renal cell carcinoma ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,chemistry ,Apoptosis ,030220 oncology & carcinogenesis ,medicine.symptom ,business ,Clear cell ,Research Paper ,anti-carcinogenic treatment - Abstract
Renal cell carcinoma (RCC), arising from the proximal tubule in the kidney, accounts for approximately 85% of kidney cancers and causes over 140,000 annual deaths worldwide. In the last decade, several new therapies have been identified for treatment of metastatic RCC. Although these therapies increase survival time compared to standard care, none of them has curative properties. The nephrotoxin orellanine specifically targets proximal tubular epithelial cells, leaving other organs unaffected. We therefore hypothesized that the selective toxicity of orellanine extends to clear cell RCC (ccRCC) cells since they emanate from proximal tubular cells. Orellanine would thus target both primary and metastatic ccRCC in vitro and in vivo. We found that orellanine induces dose-dependent cell death in proximal tubular cells and in all ccRCC cells tested, both primary and cell lines, with no toxicity detected in control cells. The toxic action of orellanine involve decreased protein synthesis, disrupted cell metabolism and induction of apoptosis. In nude rats carrying human ccRCC xenografts, brief orellanine treatment eliminated more than 90% of viable tumor mass compared to control rats. This identifies orellanine as a potential treatment concept for ccRCC patients on dialysis, due to its unique selective toxicity towards ccRCC.
- Published
- 2017
49. Cartilage oligomeric matrix protein neoepitope in the synovial fluid of horses with acute lameness: A new biomarker for the early stages of osteoarthritis
- Author
-
Emilia Svala, S. Ekman, L. Mattsson Hultén, Carina Sihlbom, Ulla Rüetschi, Anders Lindahl, Eva Skiöldebrand, Annika Lundqvist, Patrik Önnerfjord, and K. Björkman
- Subjects
0301 basic medicine ,musculoskeletal diseases ,Pathology ,medicine.medical_specialty ,lameness ,medicine.drug_class ,cartilage oligomeric matrix protein neoepitope ,Lameness, Animal ,Osteoarthritis ,Cartilage Oligomeric Matrix Protein ,Monoclonal antibody ,Article ,03 medical and health sciences ,synovial fluid ,medicine ,Synovial fluid ,Animals ,Matrilin Proteins ,Horses ,Glycoproteins ,Cartilage oligomeric matrix protein ,Antiserum ,biology ,business.industry ,General Medicine ,medicine.disease ,musculoskeletal system ,Experimental and Basic Research Studies ,horse ,osteoarthritis ,030104 developmental biology ,Polyclonal antibodies ,Lameness ,biology.protein ,Biomarker (medicine) ,biomarker ,Horse Diseases ,business ,Biomarkers - Abstract
Background: Clinical tools to diagnose the early changes of osteoarthritis (OA) that occur in the articular cartilage are lacking. Objectives: We sought to identify and quantify a novel cartilage oligomeric matrix protein (COMP) neoepitope in the synovial fluid from the joints of healthy horses and those with different stages of OA. Study design: In vitro quantitative proteomics and assay development with application in synovial fluids samples obtained from biobanks of well-characterised horses. Methods: Articular cartilage explants were incubated with or without interleukin-1β for 25 days. Media were analysed via quantitative proteomics. Synovial fluid was obtained from either normal joints (n = 15) or joints causing lameness (n = 17) or with structural OA lesions (n = 7) and analysed for concentrations of the COMP neoepitope using a custom-developed inhibition enzyme-linked immunosorbent assay (ELISA). Explants were immunostained with polyclonal antibodies against COMP and the COMP neoepitopes. Results: Semitryptic COMP peptides were identified and quantified in cell culture media from cartilage explants. A rabbit polyclonal antibody was raised against the neoepitope of the N-terminal portion of one COMP fragment (sequence SGPTHEGVC). An inhibition ELISA was developed to quantify the COMP neoepitope in synovial fluid. The mean concentration of the COMP neoepitope significantly increased in the synovial fluid from the joints responsible for acute lameness compared with normal joints and the joints of chronically lame horses and in joints with chronic structural OA. Immunolabelling for the COMP neoepitope revealed a pericellular staining in the interleukin-1β-stimulated explants. Main limitations: The ELISA is based on polyclonal antisera rather than a monoclonal antibody. Conclusions: The increase in the COMP neoepitope in the synovial fluid from horses with acute lameness suggests that this neoepitope has the potential to be a unique candidate biomarker for the early molecular changes in articular cartilage associated with OA. (Less)
- Published
- 2017
50. Anaesthetic-induced cardioprotection in an experimental model of the Takotsubo syndrome - isoflurane vs. propofol
- Author
-
Sven-Erik Ricksten, Elmir Omerovic, Carina Sihlbom, Jonatan Oras, Helen Seeman-Lodding, Annika Thorsell, Björn Redfors, Joel Lundgren, and Anwar Ali
- Subjects
Male ,Hemodynamics ,030204 cardiovascular system & hematology ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Takotsubo Cardiomyopathy ,Isoprenaline ,medicine ,Animals ,Ketamine ,Propofol ,Ejection fraction ,Isoflurane ,business.industry ,Isoproterenol ,Heart ,General Medicine ,Rats ,Disease Models, Animal ,Anesthesiology and Pain Medicine ,Blood pressure ,Echocardiography ,Anesthesia ,Anesthetics, Inhalation ,Midazolam ,business ,Anesthetics, Intravenous ,030217 neurology & neurosurgery ,medicine.drug - Abstract
Background Takotsubo syndrome (TS) is an acute cardiac condition with a substantial mortality for which no specific treatment is available. We have previously shown that isoflurane attenuates the development of left ventricular (LV) dysfunction in an experimental TS-model. We compared the effects of equi-anaesthetic doses of isoflurane, propofol and ketamine+midazolam on haemodynamics, global and regional LV systolic function and the activation of intracellular metabolic pathways in experimental TS. We hypothesized that cardioprotection in experimental TS is specific for isoflurane. Methods Forty-five rats were randomized to isoflurane (0.6 MAC, n = 15), propofol (bolus 200 mg/kg+360 mg/kg/h, n = 15) or ketamine (100 mg/kg)+midazolam (10 mg/kg, n = 15) anaesthesia. Arterial pressure, heart rate and body temperature were continuously measured and arterial blood gas analysis was performed intermittently. TS was induced by intraperitoneal injection of isoprenaline, 50 mg/kg. LV echocardiography was performed 90 min after isoprenaline injection. Apical cardiac tissue was analysed by global discovery proteomics and pathway analysis. Results Isoprenaline-induced changes in arterial blood pressure, heart rate or body temperature did not differ between groups. LV ejection fraction was higher and extent of LV akinesia was lower with isoflurane, when compared with the propofol and the ketamine+midazolam groups. In this TS-model, the proteomic analysis revealed an up-regulation of pathways involved in inflammation, coagulation, endocytosis and lipid metabolism. This up-regulation was clearly attenuated with isoflurane compared to propofol. Conclusion In an experimental model of TS, isoflurane, but not propofol, exerts a cardioprotective effect. The proteomic analysis suggests that inflammation might be involved in pathogenesis of TS.
- Published
- 2017
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.