41 results on '"U Boas"'
Search Results
2. Dendrimers in Medicine and Biotechnology: New Molecular Tools
- Author
-
U Boas, J B Christensen, P M H Heegaard and U Boas, J B Christensen, P M H Heegaard
- Published
- 2007
3. A dual center study to compare breath volatile organic compounds from smokers and non-smokers with and without COPD
- Author
-
AR Koczulla, Claus Vogelmeier, Stefan Zimmermann, Maria Allers, Jens M. Hohlfeld, Bianca Lavae-Mokhtari, Olaf Holz, Jens Langejuergen, Sven Schuchardt, A Gaida, L Kruse, U Boas, and Christoph Nell
- Subjects
Adult ,Male ,Pulmonary and Respiratory Medicine ,medicine.medical_specialty ,Pathology ,Copd patients ,01 natural sciences ,Gas Chromatography-Mass Spectrometry ,Pulmonary Disease, Chronic Obstructive ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,Disease severity ,Internal medicine ,medicine ,Humans ,ddc:610 ,Aged ,Volatile Organic Compounds ,COPD ,Non-invasive monitoring ,business.industry ,Smoking ,010401 analytical chemistry ,Breath sampling ,Reproducibility of Results ,Exhalation ,Breath analysis ,Exhaled breath ,Middle Aged ,medicine.disease ,Response to treatment ,Body Fluids ,0104 chemical sciences ,respiratory tract diseases ,Breath Tests ,030228 respiratory system ,Breath gas analysis ,Combustion products ,Dewey Decimal Classification::600 | Technik::610 | Medizin, Gesundheit ,business ,Airway inflammation - Abstract
There is increasing evidence that breath volatile organic compounds (VOC) have the potential to support the diagnosis and management of inflammatory diseases such as COPD. In this study we used a novel breath sampling device to search for COPD related VOCs. We included a large number of healthy controls and patients with mild to moderate COPD, recruited subjects at two different sites and carefully controlled for smoking. 222 subjects were recruited in Hannover and Marburg, and inhaled cleaned room air before exhaling into a stainless steel reservoir under exhalation flow control. Breath samples (2.5 l) were continuously drawn onto two Tenax(®) TA adsorption tubes and analyzed in Hannover using thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). Data of 134 identified VOCs from 190 subjects (52 healthy non-smokers, 52 COPD ex-smokers, 49 healthy smokers, 37 smokers with COPD) were included into the analysis. Active smokers could be clearly discriminated by higher values for combustion products and smoking related VOCs correlated with exhaled carbon monoxide (CO), indicating the validity of our data. Subjects from the study sites could be discriminated even after exclusion of cleaning related VOCs. Linear discriminant analysis correctly classified 89.4% of COPD patients in the non/ex-smoking group (cross validation (CV): 85.6%), and 82.6% of COPD patients in the actively smoking group (CV: 77.9%). We extensively characterized 134 breath VOCs and provide evidence for 14 COPD related VOCs of which 10 have not been reported before. Our results show that, for the utilization of breath VOCs for diagnosis and disease management of COPD, not only the known effects of smoking but also site specific differences need to be considered. We detected novel COPD related breath VOCs that now need to be tested in longitudinal studies for reproducibility, response to treatment and changes in disease severity.
- Published
- 2016
4. Erkennung von pulmonaler Graft-versus-Host-Krankheit (GvHD) in allogen stammzelltransplantierten Patienten durch elektronische Nase und Ionenmobilitätsspektrometer
- Author
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E Wollmer, AR Koczulla, A Hattesohl, A Burchert, C Wilhelm, Christoph Nell, Claus Vogelmeier, JC Groß, Jörg Ingo Baumbach, U Boas, and A Neubauer
- Subjects
Pulmonary and Respiratory Medicine - Published
- 2015
5. Dendrimers in Medicine and Biotechnology : New Molecular Tools
- Author
-
U Boas, J B Christensen, P M H Heegaard, U Boas, J B Christensen, and P M H Heegaard
- Subjects
- Chemotherapy, Polymers, Drugs--Dosage forms, Therapeutics, Drugs, Biotechnology, Dendrimers, Molecular structure, Nanoparticles, Drug carriers (Pharmacy), Manufactures
- Abstract
Dendrimers are a new class of macromolecule increasingly used in the fields of synthetic organic chemistry, biology, medicine and biotechnology. Dendrimers in Medicine and Biotechnology: New Molecular Tools looks at this exciting and rapidly growing area of science. Using an interdisciplinary approach with particular emphasis on biological applications, the book discusses the relationship between the dendrimer molecular motif and its biological properties. A general introduction to the subject of dendrimers, including definitions of terms and symbols, is provided. Subsequent sections discuss topics including dendrimers in biological systems, dendrimers as drug delivery devices, dendrimers in diagnostics and dendrimer drugs. Throughout the book examples from current research are also provided. This book will appeal to a wide range of scientists, including non specialists who require an introduction to dendrimers, as well as those wishing to know more about the application of dendrimers in the field of biology and medicine.
- Published
- 2006
6. Dendrimers: design, synthesis and chemical properties.
- Author
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U. Boas, J. B. Christensen, and P. M. H. Heegaard
- Published
- 2006
7. Serological detection of antibodies against Salmonella polysaccharides in ELISA employing a new method for coupling of polysaccharides
- Author
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Camilla Wiuff, K. Wredstrøm, Lars Andresen, B. Pedersen, U. Boas, Eva Irene Stenbaek Jauho, Peter M. H. Heegaard, and M. H. Jakobsen
- Subjects
chemistry.chemical_classification ,Coupling (electronics) ,Salmonella ,Chromatography ,biology ,Chemistry ,biology.protein ,medicine ,Antibody ,Polysaccharide ,medicine.disease_cause ,Serology
8. A dual center study to compare breath volatile organic compounds from smokers and non-smokers with and without COPD.
- Author
-
A Gaida, O Holz, C Nell, S Schuchardt, B Lavae-Mokhtari, L Kruse, U Boas, J Langejuergen, M Allers, S Zimmermann, C Vogelmeier, A R Koczulla, and J M Hohlfeld
- Published
- 2016
- Full Text
- View/download PDF
9. Specific molecular peak analysis by ion mobility spectrometry of volatile organic compounds in urine of COVID-19 patients: A novel diagnostic approach.
- Author
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Boeselt T, Terhorst P, Kroenig J, Nell C, Spielmanns M, Boas U, Veith M, Vogelmeier C, Greulich T, Koczulla AR, Beutel B, Huber J, and Heers H
- Subjects
- Humans, SARS-CoV-2, Ion Mobility Spectrometry, Sensitivity and Specificity, COVID-19 Testing, COVID-19 diagnosis, Volatile Organic Compounds analysis
- Abstract
Introduction: SARS-CoV-2 is usually diagnosed from naso-/oropharyngeal swabs which are uncomfortable and prone to false results. This study investigated a novel diagnostic approach to Covid-19 measuring volatile organic compounds (VOC) from patients' urine., Methods: Between June 2020 and February 2021, 84 patients with positive RT-PCR for SARS-CoV-2 were recruited as well as 54 symptomatic individuals with negative RT-PCR. Midstream urine samples were obtained for VOC analysis using ion mobility spectrometry (IMS) which detects individual molecular components of a gas sample based on their size, configuration, and charge after ionization., Results: Peak analysis of the 84 Covid and 54 control samples showed good group separation. In total, 37 individual specific peaks were identified, 5 of which (P134, 198, 135, 75, 136) accounted for significant differences between groups, resulting in sensitivities of 89-94% and specificities of 82-94%. A decision tree was generated from the relevant peaks, leading to a combined sensitivity and specificity of 98% each., Discussion: VOC-based diagnosis can establish a reliable separation between urine samples of Covid-19 patients and negative controls. Molecular peaks which apparently are disease-specific were identified. IMS is an additional non-invasive and cheap device for the diagnosis of this ongoing endemic infection. Further studies are needed to validate sensitivity and specificity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
10. Volatile Phases Derived from Serum, DC, or MLC Culture Supernatants to Deduce a VOC-Based Diagnostic Profiling Strategy for Leukemic Diseases.
- Author
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Baudrexler T, Boeselt T, Li L, Bohlscheid S, Boas U, Schmid C, Rank A, Schmohl J, Koczulla R, and Schmetzer HM
- Subjects
- Humans, Dendritic Cells, Prospective Studies, Lymphocyte Activation, Volatile Organic Compounds metabolism, Leukemia, Myeloid, Acute metabolism
- Abstract
Volatile organic compounds (VOCs) reflect the metabolism in healthy and pathological conditions, and can be collected easily in a noninvasive manner. They are directly measured using electronical nose (eNose), and may qualify as a systemic tool to monitor biomarkers related to disease. Myeloid leukemic blasts can be transformed into leukemia-derived dendritic cells (DC
leu ) able to improve (anti-leukemic) immune responses. To profile immunological changes in healthy and acute myeloid leukemic (AML) patients' ex vivo cell cultures, we correlated the cell biological data with the profiles of cell culture supernatant-derived VOCs. DC/DCleu from leukemic or healthy whole blood (WB) were generated without (Control) or with immunomodulatory Kit M (Granulocyte macrophage-colony-stimulating-factor (GM-CSF) + prostaglandin E1 (PGE1 )) in dendritic cell cultures (DC culture). Kit-pretreated/not pretreated WB was used to stimulate T cell-enriched immunoreactive cells in mixed lymphocyte cultures (MLC culture). Leukemia-specific adaptive and innate immune cells were detected with a degranulation assay (Deg) and an intracellular cytokine assay (InCyt). Anti-leukemic cytotoxicity was explored with a cytotoxicity fluorolysis assay (CTX). VOCs collected from serum or DC- and MLC culture supernatants (with vs. without Kit M pretreatment and before vs. after culture) were measured using eNose. Compared to the Control (without treatment), Kit M-pretreated leukemic and healthy WB gave rise to higher frequencies of mature (leukemia-derived) DC subtypes of activated and (memory) T cells after MLC. Moreover, antigen (leukemia)-specific cells of several lines (innate and adaptive immunity cells) were induced, giving rise to blast-lysing cells. The eNose could significantly distinguish between healthy and leukemic patients' serum, DC and MLC culture supernatant-derived volatile phases and could significantly separate several supernatant (with vs. without Kit M treatment, cultured vs. uncultured)-derived VOCs within subgroups (healthy DC or leukemic DC, or healthy MLC or leukemic MLC supernatants). Interestingly, the eNose could indicate a Kit M- and culture-associated effect. The eNose may be a prospective option for the deduction of a VOC-based profiling strategy using serum or cell culture supernatants and could be a useful diagnostic tool to recognize or qualify AML disease.- Published
- 2023
- Full Text
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11. A generic protocol to immobilize lipopolysaccharides on microbeads for multiplex analysis.
- Author
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Boas U, Sørensen MB, Andresen LO, and Berger SS
- Subjects
- Animals, Antibodies, Bacterial, Lipopolysaccharides, Microspheres, Swine, Actinobacillus pleuropneumoniae, Swine Diseases
- Abstract
Bead-based multiplex serodiagnostics enables simultaneous analysis of antibodies against several antigens. Binding of the antigens onto the surface of the bead, preserving the antigenicity of the antigen is a pivotal step to ensure high sensitivity and selectivity of the assay. Here, a generic method for immobilization of lipopolysaccharide (LPS) antigens from different Gram-negative bacteria to microbeads using non-covalent conjugation has been developed and tested. The method involves coupling of N,N-diethylethylenediamine (DEDA) and derivatives to microbeads. This enhances non-covalent interactions so that LPS is easily immobilized. LPS antigens from the Gram-negative bacteria Actinobacillus pleuropneumoniae (APP) and Salmonella enterica serogroup B (Sal. B) were immobilized on the DEDA-coupled microbeads. In parallel, the same LPS antigens were coupled to beads using two previously reported methods. The performance of microbeads coupled with antigen using the different methods was compared by measuring antibodies in positive and negative serum samples from pigs. DEDA-beads coupled with LPS detected pathogen specific serum antibodies with equal or higher sensitivity and specificity compared to the other coupling methods used in this study. Furthermore, derivatives of DEDA, where the tertiary amine was alkylated with a methyl (m-DEDA) and ethyl group (e-DEDA) to give a positively charged tetraalkylammonium group, were compared with DEDA for the binding of LPS antigens. Here, it was concluded that the DEDA-modified bead was most efficient in the binding of LPS antigens from two Actinobacillus pleuropneumoniae serovars and Salmonella enterica serogroup B.
- Published
- 2021
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12. Phosphocholine-Decorated PPI-Dendrimers Mimic Cell Membrane Phosphocholine Clusters and Tune the Innate Immune Activity of C-Reactive Protein.
- Author
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Boas U, Daoud MM, Meier S, Olsen TH, Gehring K, Mogensen DJ, and Heegaard PMH
- Subjects
- C-Reactive Protein, Cell Membrane, Immunity, Innate, Phosphorylcholine, Polypropylenes, Dendrimers
- Abstract
C-reactive protein (CRP) is widely used as biomarkers of infection and inflammation. It has a well-described ability to bind phosphocholine (PC), as well as PC-clusters from compromised and inflamed cell membranes and tissues. The binding of PC-clusters to CRP is of interest as this binding determines subsequent innate immune activity. We investigated PC-decorated dendrimers as mimics for PC-clusters. Five generations of poly(propylene imine) (PPI) dendrimers were modified with PC surface groups via a three-step synthetic sequence obtaining the PC-decorated dendrimers in high purity. The dendrimers were analyzed by NMR and infrared spectroscopy as well as HPLC. We developed immunoassays to show that dendrimer-PC binding to CRP was Ca
2+ -dependent with an apparent overall Kd of 11.9 nM for first generation (G1) PPI-PC, while G2-PPI-PC and G3-PPI-PC had slightly higher affinities, and G4-PPI-PC and G5-PPI-PC had slightly lower affinities. For all PC-dendrimers, the affinity was orders of magnitude higher than the affinity of free phosphocholine (PC), indicating a PC-cluster effect. Next, we investigated the binding of CRP:PPI-PC complexes to complement component C1q. C1q binding to CRP was dependent on the generation of PPI-PC bound to CRP, with second and third generation PPI-PCs leading to the highest affinity. The dendrimer-based approach to PC-cluster mimics and the simple binding assays presented here hold promise as tools to screen PC-compounds for their abilities to tune the innate immune activity of CRP.- Published
- 2021
- Full Text
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13. Antiparasitic activity of chicory (Cichorium intybus) and its natural bioactive compounds in livestock: a review.
- Author
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Peña-Espinoza M, Valente AH, Thamsborg SM, Simonsen HT, Boas U, Enemark HL, López-Muñoz R, and Williams AR
- Subjects
- Animals, Anthelmintics administration & dosage, Anthelmintics chemistry, Antiparasitic Agents chemistry, Cattle, Feces parasitology, Gastrointestinal Tract drug effects, Gastrointestinal Tract parasitology, Helminthiasis drug therapy, Humans, Intestinal Diseases, Parasitic drug therapy, Livestock anatomy & histology, Livestock parasitology, Parasite Egg Count, Sheep, Animal Feed analysis, Antiparasitic Agents administration & dosage, Cichorium intybus chemistry, Dietary Supplements, Nematoda drug effects
- Abstract
Increasing drug resistance in gastrointestinal (GI) parasites of livestock and concerns about chemical residues in animal products and the environment are driving the development of alternative control strategies that are less reliant on the use of synthetic drugs. An increasingly investigated approach is the use of bioactive forages with antiparasitic properties as part of the animal's diet (nutraceuticals) or as potential sources of novel, natural parasiticides. Chicory (Cichorium intybus) is a multi-purpose crop and one of the most promising bioactive forages in temperate regions, and numerous in vivo trials have explored its potential against parasitic nematodes in livestock. However, it is unclear whether chicory can induce a direct and broad activity against various GI parasites in different livestock species, and the levels of chicory in the diet that are required to exert an efficient antiparasitic effect. Moreover, the mechanisms leading to the reported parasiticidal activity of chicory are still largely unknown, and its bioactive phytochemicals have only recently been investigated. In this review, we summarise the progress in the study of the antiparasitic activity of chicory and its natural bioactive compounds against GI parasites in livestock, through examination of the published literature. The available evidence indicates that feeding chicory can reduce faecal egg counts and/or worm burdens of abomasal nematodes, but not infections with intestinal worms, in ruminants. Highly chicory-rich diets (≥ 70% of chicory dry matter in the diet) may be necessary to directly affect abomasal parasitism. Chicory is known to synthesise several bioactive compounds with potential antiparasitic activity, but most research has been devoted to the role of sesquiterpene lactones (SL). Recent in vitro studies have confirmed direct and potent activity of SL-rich extracts from chicory against different GI helminths of livestock. Chicory SL have also been reported to exhibit antimalarial properties and its potential antiprotozoal activity in livestock remains to be evaluated. Furthermore, the detailed identification of the main antiparasitic metabolites of chicory and their pharmacokinetics need further confirmation. Research gaps and perspectives on the potential use of chicory as a nutraceutical forage and a source of bioactive compounds for parasite control in livestock are discussed.
- Published
- 2018
- Full Text
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14. Simultaneous detection of antibodies to five Actinobacillus pleuropneumoniae serovars using bead-based multiplex analysis.
- Author
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Berger SS, Lauritsen KT, Boas U, Lind P, and Andresen LO
- Subjects
- Actinobacillus Infections blood, Actinobacillus Infections diagnosis, Actinobacillus Infections microbiology, Actinobacillus Infections veterinary, Actinobacillus pleuropneumoniae immunology, Animals, Complement Fixation Tests veterinary, Enzyme-Linked Immunosorbent Assay veterinary, Immunoassay, Sensitivity and Specificity, Swine, Swine Diseases blood, Swine Diseases diagnosis, Swine Diseases microbiology, Actinobacillus pleuropneumoniae classification, Antibodies, Bacterial blood, Serogroup
- Abstract
We developed and made a preliminary validation of a bead-based multiplexed immunoassay for simultaneous detection of porcine serum antibodies to Actinobacillus pleuropneumoniae serovars 1, 2, 6, 7, and 12. Magnetic fluorescent beads were coupled with A. pleuropneumoniae antigens and tested with a panel of serum samples from experimentally infected pigs and with serum samples from uninfected and naturally infected pigs. The multiplex assay was compared to in-house ELISAs and complement fixation (CF) tests, which have been used for decades as tools for herd classification in the Danish Specific Pathogen Free system. Assay specificities and sensitivities as well as the corresponding cutoff values were determined using receiver operating characteristic (ROC) curve analysis, and the A. pleuropneumoniae multiplex assay showed good correlation with the in-house ELISAs and CF tests with areas under ROC curves ≥ 0.988. Benefits of multiplexed assays compared to ELISAs and CF tests include reduced serum sample volumes needed for analysis, less labor, and shorter assay time.
- Published
- 2017
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15. Anthelmintic activity of chicory (Cichorium intybus): in vitro effects on swine nematodes and relationship to sesquiterpene lactone composition.
- Author
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Williams AR, Peña-Espinoza MA, Boas U, Simonsen HT, Enemark HL, and Thamsborg SM
- Subjects
- Animals, Anthelmintics chemistry, Anthelmintics isolation & purification, Anthelmintics pharmacology, Ascaris suum ultrastructure, Larva drug effects, Larva ultrastructure, Microscopy, Electron, Transmission, Oesophagostomum ultrastructure, Plant Extracts chemistry, Plant Extracts isolation & purification, Swine parasitology, Ascaris suum drug effects, Cichorium intybus chemistry, Oesophagostomum drug effects, Plant Extracts pharmacology
- Abstract
Chicory is a perennial crop that has been investigated as a forage source for outdoor-reared ruminants and pigs, and has been reported to have anthelmintic properties. Here, we investigated in vitro anthelmintic effects of forage chicory-extracts against the highly prevalent swine parasites Ascaris suum and Oesophagostomum dentatum. Methanol extracts were prepared and purified from two different cultivars of chicory (Spadona and Puna II). Marked differences were observed between the anthelmintic activity of extracts from the two cultivars. Spadona extracts had potent activity against A. suum third (L3) and fourth (L4) - stage larvae, as well as O. dentatum L4 and adults, whereas Puna II extracts had less activity against A. suum and no activity towards O. dentatum L4. Transmission-electron microscopy of A. suum L4 exposed to Spadona extracts revealed only subtle changes, perhaps indicative of a specific anthelmintic effect rather than generalized toxicity. Ultra-high liquid chromatography-mass spectrometry analysis revealed that the purified extracts were rich in sesquiterpene lactones (SL), and that the SL profile differed significantly between cultivars. This is the first report of anthelmintic activity of forage chicory towards swine nematodes. Our results indicate a significant anthelmintic effect, which may possibly be related to SL composition.
- Published
- 2016
- Full Text
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16. A dual center study to compare breath volatile organic compounds from smokers and non-smokers with and without COPD.
- Author
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Gaida A, Holz O, Nell C, Schuchardt S, Lavae-Mokhtari B, Kruse L, Boas U, Langejuergen J, Allers M, Zimmermann S, Vogelmeier C, Koczulla AR, and Hohlfeld JM
- Subjects
- Adult, Aged, Body Fluids chemistry, Breath Tests methods, Exhalation, Gas Chromatography-Mass Spectrometry methods, Humans, Male, Middle Aged, Pulmonary Disease, Chronic Obstructive diagnosis, Reproducibility of Results, Young Adult, Pulmonary Disease, Chronic Obstructive physiopathology, Smoking physiopathology, Volatile Organic Compounds analysis
- Abstract
There is increasing evidence that breath volatile organic compounds (VOC) have the potential to support the diagnosis and management of inflammatory diseases such as COPD. In this study we used a novel breath sampling device to search for COPD related VOCs. We included a large number of healthy controls and patients with mild to moderate COPD, recruited subjects at two different sites and carefully controlled for smoking. 222 subjects were recruited in Hannover and Marburg, and inhaled cleaned room air before exhaling into a stainless steel reservoir under exhalation flow control. Breath samples (2.5 l) were continuously drawn onto two Tenax(®) TA adsorption tubes and analyzed in Hannover using thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). Data of 134 identified VOCs from 190 subjects (52 healthy non-smokers, 52 COPD ex-smokers, 49 healthy smokers, 37 smokers with COPD) were included into the analysis. Active smokers could be clearly discriminated by higher values for combustion products and smoking related VOCs correlated with exhaled carbon monoxide (CO), indicating the validity of our data. Subjects from the study sites could be discriminated even after exclusion of cleaning related VOCs. Linear discriminant analysis correctly classified 89.4% of COPD patients in the non/ex-smoking group (cross validation (CV): 85.6%), and 82.6% of COPD patients in the actively smoking group (CV: 77.9%). We extensively characterized 134 breath VOCs and provide evidence for 14 COPD related VOCs of which 10 have not been reported before. Our results show that, for the utilization of breath VOCs for diagnosis and disease management of COPD, not only the known effects of smoking but also site specific differences need to be considered. We detected novel COPD related breath VOCs that now need to be tested in longitudinal studies for reproducibility, response to treatment and changes in disease severity.
- Published
- 2016
- Full Text
- View/download PDF
17. Sesquiterpene lactone containing extracts from two cultivars of forage chicory (Cichorium intybus) show distinctive chemical profiles and in vitro activity against Ostertagia ostertagi.
- Author
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Peña-Espinoza M, Boas U, Williams AR, Thamsborg SM, Simonsen HT, and Enemark HL
- Subjects
- Animals, Anthelmintics chemistry, Lactones chemistry, Plant Extracts chemistry, Sesquiterpenes chemistry, Anthelmintics pharmacology, Cichorium intybus chemistry, Lactones pharmacology, Ostertagia drug effects, Plant Extracts pharmacology, Sesquiterpenes pharmacology
- Abstract
The study investigated direct anthelmintic effects of sesquiterpene lactones (SL)-containing extracts from forage chicory against free-living and parasitic stages of Ostertagia ostertagi. Freeze-dried leaves from chicory cultivars 'Spadona' and 'Puna II' were extracted using methanol/water. Total SL were further fractionated by solid-phase extraction and resulting extracts were characterised by high-performance liquid chromatography (HPLC). O. ostertagi eggs from faeces of mono-infected calves were hatched and L1 were used in a larval feeding inhibition assay (LFIA), while cultured L3 were used in a larval exsheathment inhibition assay (LEIA). Adult worms were immediately recovered after slaughter and used for motility inhibition assays (AMIA). Electron microscopy (EM) was performed on adult O. ostertagi exposed to 1000 μg extract mL(-1) of both chicory cultivars. In all assays, decreasing concentrations of SL-containing extracts in PBS (1% DMSO) were tested in replicates with 1% DMSO in PBS as negative controls. HPLC demonstrated similar concentrations of most SL in both extracts. However, Spadona-extract contained significantly higher concentrations of 11, 13-dihydro-8-deoxylactucin (P = 0.01), while Puna II-extract had increased levels of 11, 13-dihydrolactucin (P < 0.0001). In the LFIA, both extracts reduced larval feeding at increasing concentrations, but Spadona-extract showed higher potency confirmed by significantly lower EC50 (P < 0.0001). In the LEIA, neither of the two extracts interfered with the exsheathment of L3 (P > 0.05). In the AMIA, both SL-containing extracts induced a dose-dependent effect but Spadona-extract showed greater activity and exerted faster worm paralysis than Puna II-extract with significantly lower EC50 (P < 0.0001). No cuticular damage was observed by EM in worms exposed to any of the extracts. We have demonstrated that SL-containing extracts from forage chicory can inhibit feeding of free-living larvae and exert direct effects against parasitic stages of O. ostertagi. Our results may contribute to the identification of natural anti-parasitic compounds and to interpret the in vivo anthelmintic effects of forage chicory., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2015
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18. Urea and thiourea modified polypropyleneimine dendrimers clear intracellular α-synuclein aggregates in a human cell line.
- Author
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Laumann K, Boas U, Larsen HM, Heegaard PM, and Bergström AL
- Subjects
- Cell Line, Cell Line, Tumor, Dendrimers chemistry, Humans, Polypropylenes chemistry, Thiourea chemistry, Urea chemistry, Dendrimers metabolism, Intracellular Membranes metabolism, Polypropylenes metabolism, Thiourea metabolism, Urea metabolism, alpha-Synuclein metabolism
- Abstract
Synucleinopathies are neurodegenerative pathologies in which disease progression is closely correlated to brain accumulation of insoluble α-synuclein, a small protein abundantly expressed in neural tissue. Here, two types of modified polypropyleneimine (PPI) dendrimers having either urea or methylthiourea (MTU) surface functional groups were investigated in a cellular model of synucleinopathy. Dendrimers are synthetic macromolecules that may be produced in a range of well-defined molecular sizes. Using cellomics array scan high-content screening, we show that both types of dendrimers are able to significantly reduce intracellular levels of α-synuclein aggregates dependent on the concentration, the type and molecular size of the dendrimer with the bigger size MTU-dendrimers having the highest potency. The intracellular clearance of α-synuclein aggregates by dendrimers was achieved at noncytotoxic concentrations.
- Published
- 2015
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19. Color test for selective detection of secondary amines on resin and in solution.
- Author
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Boas U and Mirsharghi S
- Subjects
- Acetaldehyde chemistry, Amines chemistry, Amino Acids chemistry, Fluorenes chemistry, Molecular Structure, Solid-Phase Synthesis Techniques, Solutions, Amines analysis, Colorimetry methods, Resins, Synthetic chemistry
- Abstract
Resins for solid-phase synthesis give orange to red-brown resin beads selectively when secondary amines are present on the resin when treated with a solution of acetaldehyde and an Fmoc-amino acid in NMP. The method shows good specificity and gives colorless beads when exposed to a variety of other functional groups. Furthermore, the acetaldehyde/Fmoc amino acid method can be used as a selective colorimetric test for secondary amines in solution.
- Published
- 2014
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20. Method to conjugate polysaccharide antigens to surfaces for the detection of antibodies.
- Author
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Boas U, Lind P, and Riber U
- Subjects
- Animals, Cattle, Mice, Polystyrenes chemistry, Antibodies, Bacterial chemistry, Antibodies, Monoclonal, Murine-Derived chemistry, Lipopolysaccharides chemistry, Salmonella typhimurium chemistry
- Abstract
A new generic method for the conjugation of lipopolysaccharide (LPS)-derived polysaccharide antigens from gram-negative bacteria has been developed using Salmonella as a model. After removal of lipid A from the LPS by mild acidolysis, the polysaccharide antigen was conjugated to polystyrene microbeads modified with N-alkyl hydroxylamine and N-alkyl-O-methyl hydroxylamine surface groups by incubation of antigen and beads for 16 h at 40 °C without the need for coupling agents. The efficiency of the new method was evaluated by flow cytometry in model samples and serum samples containing antibodies against Salmonella typhimurium and Salmonella dublin. The presented method was compared with a similar method for conjugation of Salmonella polysaccharide antigens to surfaces. Here, the new method showed higher antigen coupling efficiency by detecting low concentrations of antibodies. Furthermore, the polysaccharide-conjugated beads showed preserved bioactivity after 1 year of use., (Copyright © 2014 Elsevier Inc. All rights reserved.)
- Published
- 2014
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21. Enhancement of muramyldipeptide (MDP) immunostimulatory activity by controlled multimerization on dendrimers.
- Author
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Sorensen NS, Boas U, and Heegaard PM
- Subjects
- Acetylmuramyl-Alanyl-Isoglutamine pharmacology, Adjuvants, Immunologic pharmacology, Animals, B7 Antigens analysis, B7 Antigens biosynthesis, Dendrimers pharmacology, Flow Cytometry, Genes, MHC Class II immunology, Interleukin-12 biosynthesis, Interleukin-1beta biosynthesis, Interleukin-6 biosynthesis, Leukocytes, Mononuclear cytology, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear metabolism, Lymphocyte Activation immunology, Polymerization, Receptors, Pattern Recognition immunology, Receptors, Pattern Recognition metabolism, Swine, Acetylmuramyl-Alanyl-Isoglutamine chemical synthesis, Adjuvants, Immunologic chemical synthesis, Dendrimers chemical synthesis, Immunity, Innate, Leukocytes, Mononuclear drug effects, Lymphocyte Activation drug effects
- Abstract
Peptidoglycan is a widespread bacterial PAMP molecule and a powerful initiator of innate immune responses. It consists of repeating units of MDP, which as a monomer is only weakly immunostimulatory. Here, MDP-coupled dendrimers were prepared and investigated for stimulation of pig blood mononuclear cells. Compared to monomeric MDP, MDP-dendrimers induced a markedly enhanced production of IL-12 p40, IL-1β and IL-6 and completely down-regulated surface expression of B7 and MHC class II. These results suggest a possible novel strategy based on controlled multimerization of minimal PAMP motifs on dendrimers for preparing molecularly defined immunostimulators with predictable bioactivities., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2011
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22. Dendrimers for vaccine and immunostimulatory uses. A review.
- Author
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Heegaard PM, Boas U, and Sorensen NS
- Subjects
- Animals, Antigen-Antibody Reactions, Antigens chemistry, Antigens immunology, Biocompatible Materials chemistry, Cell Membrane chemistry, Cell Membrane immunology, Humans, Particle Size, Surface Properties, Vaccines immunology, Adjuvants, Immunologic chemistry, Dendrimers chemistry, Vaccines chemistry
- Abstract
Dendrimers are well-defined (monodisperse) synthetic globular polymers with a range of interesting chemical and biological properties. Chemical properties include the presence of multiple accessible surface functional groups that can be used for coupling biologically relevant molecules and methods that allow for precise heterofunctionalization of surface groups. Biologically, dendrimers are highly biocompatible and have predictable biodistribution and cell membrane interacting characteristics determined by their size and surface charge. Dendrimers have optimal characteristics to fill the need for efficient immunostimulating compounds (adjuvants) that can increase the efficiency of vaccines, as dendrimers can provide molecularly defined multivalent scaffolds to produce highly defined conjugates with small molecule immunostimulators and/or antigens. The review gives an overview on the use of dendrimers as molecularly defined carriers/presenters of small antigens, including constructs that have built-in immunostimulatory (adjuvant) properties, and as stand-alone adjuvants that can be mixed with antigens to provide efficient vaccine formulations. These approaches allow the preparation of molecularly defined vaccines with highly predictable and specific properties and enable knowledge-based vaccine design substituting the traditional empirically based approaches for vaccine development and production.
- Published
- 2010
- Full Text
- View/download PDF
23. Backbone amide linker in solid-phase synthesis.
- Author
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Boas U, Brask J, and Jensen KJ
- Subjects
- Amides chemistry, Cross-Linking Reagents chemistry, Glycopeptides chemical synthesis, Glycopeptides chemistry, Heterocyclic Compounds chemical synthesis, Heterocyclic Compounds chemistry, Oligosaccharides chemical synthesis, Oligosaccharides chemistry, Peptides chemical synthesis, Peptides chemistry, Peptides, Cyclic chemical synthesis, Peptides, Cyclic chemistry, Amides chemical synthesis, Cross-Linking Reagents chemical synthesis
- Published
- 2009
- Full Text
- View/download PDF
24. A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5'-diphosphocholine coupled dendrimers.
- Author
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Heegaard PM, Pedersen HG, Jensen AL, and Boas U
- Subjects
- Animals, Dendrimers chemistry, Humans, Sensitivity and Specificity, Swine, C-Reactive Protein analysis, Cytidine Diphosphate Choline chemistry, Enzyme-Linked Immunosorbent Assay methods
- Abstract
C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP. The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity.
- Published
- 2009
- Full Text
- View/download PDF
25. THAL, a sterically unhindered linker for the solid-phase synthesis of acid-sensitive protected peptide acids.
- Author
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Isidro-Llobet A, Boas U, Jensen KJ, Alvarez M, and Albericio F
- Subjects
- Carboxylic Acids chemistry, Crystallography, X-Ray, Models, Molecular, Molecular Structure, Peptides chemistry, Resins, Synthetic chemical synthesis, Resins, Synthetic chemistry, Sensitivity and Specificity, Stereoisomerism, Acids chemistry, Carboxylic Acids chemical synthesis, Heterocyclic Compounds, 2-Ring chemistry, Peptides chemical synthesis
- Abstract
The 5-(4-hydroxyphenyl)-3,4-ethylenedioxythienyl alcohol (THAL, Thiophene Acid Labile) is described as a new linker for the solid-phase synthesis of peptide carboxylic acids. It is based on the electron-rich 3,4-ethylenedioxythenyl (EDOTn) moiety and allows the obtention of free and tert-butyl-protected peptides by cleavage with 90% and 0.5% TFA, respectively. This very high acid lability makes it useful for the synthesis of sensitive peptides. Free and tert-butyl-protected Leu-enkephalins have been synthesized as models to demonstrate the utility of the linker.
- Published
- 2008
- Full Text
- View/download PDF
26. Dendrimers destabilize proteins in a generation-dependent manner involving electrostatic interactions.
- Author
-
Giehm L, Christensen C, Boas U, Heegaard PM, and Otzen DE
- Subjects
- Humans, Hydrogen-Ion Concentration, Static Electricity, Dendrimers chemistry, Polypropylenes chemistry, Proteins chemistry
- Abstract
Dendrimers are well-defined chemical polymers with a characteristic branching pattern that gives rise to attractive features such as antibacterial and antitumor activities as well as drug delivery properties. In addition, dendrimers can solubilize prion protein aggregates at very low concentrations, but their mode of action is unclear. We show that poly(propylene imine) dendrimers based on di-aminobutane (DAB) and modified with guanidinium surface groups reduce insulin thermostability and solubility considerably at microgram per microliter concentrations, while urea-modified groups have hardly any effect. Destabilization is markedly generation-dependent and is most pronounced for generation 3, which is also the most efficient at precipitating insulin. This suggests that proteins can interact with both dendrimer surface and interior. The pH-dependence reveals that interactions are mainly mediated by electrostatics, confirmed by studies on four other proteins. Ability to precipitate and destabilize are positively correlated, in contrast to conventional small-molecule denaturants and stabilizers, indicating that surface immobilization of denaturing groups profoundly affects its interactions with proteins.
- Published
- 2008
- Full Text
- View/download PDF
27. Guanidino- and urea-modified dendrimers as potent solubilizers of misfolded prion protein aggregates under non-cytotoxic conditions. dependence on dendrimer generation and surface charge.
- Author
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Cordes H, Boas U, Olsen P, and Heegaard PM
- Subjects
- Cell Line, Cell Survival drug effects, Dendrimers chemical synthesis, Dendrimers toxicity, Kinetics, Molecular Structure, Solubility, Surface Properties, Titrimetry, Dendrimers chemistry, Guanidines chemistry, Prions chemistry, Prions metabolism, Protein Folding, Urea chemistry
- Abstract
Amino-terminated dendrimers are well-defined synthetic hyperbranched polymers and have previously been shown to destabilize aggregates of the misfolded, pathogenic, and partially protease-resistant form of the prion protein (PrPSc), transforming it into a partially dissociated, protease-sensitive form with strongly reduced infectivity. The mechanism behind this is not known, but a low pH, creating multiple positively charged primary amines on the dendrimer surface, increases the efficiency of the reaction. In the present study, surface amines of the dendrimers were modified to yield either guanidino surface groups (being positively charged at neutral pH) or urea groups (uncharged). The ability of several generations of modified dendrimers and unmodified amino-terminated dendrimers to deplete PrPSc from persistently PrPSc-infected cells in culture (SMB cells) was studied. It was found that destabilization correlated with both the generation number of the dendrimer, with higher generations being more efficient, and the charge density of the surface groups. Urea-decorated dendrimers having an uncharged surface were less efficient than positively charged unmodified- (amino) and guanidino-modified dendrimers. The most efficient dendrimers (generation 4 (G4) and G5-unmodified and guanidino dendrimers) cleared PrPSc completely by incubation for 4 days at less than 50 nM. In contrast to both unmodified and guanidine-modified dendrimers, the uncharged urea dendrimers showed much lower cytotoxicity toward noninfected SMB cells. Therapeutic uses of modified dendrimers are indicated by the low concentrations of dendrimers needed.
- Published
- 2007
- Full Text
- View/download PDF
28. Dendrimer effects on peptide and protein fibrillation.
- Author
-
Heegaard PM, Boas U, and Otzen DE
- Subjects
- Hydrogen-Ion Concentration, Prions chemistry, Amyloid chemistry, Dendrimers chemistry, Peptides chemistry
- Abstract
Dendrimers are synthetic, symmetrically branched polymers that can be manufactured to a high degree of definition and therefore present themselves as monodisperse entities. Flexible and globular in shape and compartementalized into a partly inaccessible interior and a highly exposed surface, they offer numerous possibilities for interactions with and responses to biological macromolecules and biostructures including cell membranes and proteins. By way of their multiple functional surface groups, they allow the design of surfaces carrying a multitude of biological motifs and/or charges giving rise to quite significant biological and physico-chemical effects. Here we describe the surprising ability of dendrimers to interact with and perturb polypeptide conformations, particularly efficiently towards amyloid structures; that is, the structures of highly insoluble polypeptide aggregates involved in a range of serious and irreversibly progressive pathological conditions (protein-misfolding diseases). Interesting as this may be, the interaction of dendrimers with such generic peptidic aggregates also offers a new perspective on the molecular mechanisms governing assembly and disassembly of amyloid structures and thereby on determinants of protein and peptide folding. Despite the potent disaggregative nature of various dendrimers, they have variable effects on the stability of different proteins, suggesting that they do not act as generic denaturants, but rather exert their effects via specific interactions with individual parts of each protein.
- Published
- 2007
- Full Text
- View/download PDF
29. Carbocations in action. Design, synthesis, and evaluation of a highly acid-sensitive naphthalene-based backbone amide linker for solid-phase synthesis.
- Author
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Pittelkow M, Boas U, and Christensen JB
- Abstract
The design, synthesis, and properties of an extremely acid-labile backbone amide linker based on a regiospecifically substituted tetraalkoxy naphthaldehyde core are presented. This handle enables cleavage of peptide backbone amides (secondary amides) off a solid support using as little as 0.5% TFA in CH2Cl2. This proceeds without cleavage of tert-butyl ethers and tert-butyl esters. The design is based on a DFT study that predicted the most stabile alkoxy-substituted methyl naphthyl carbocation. [structure: see text]
- Published
- 2006
- Full Text
- View/download PDF
30. Dendrimer based anti-infective and anti-inflammatory drugs.
- Author
-
Heegaard PM and Boas U
- Subjects
- Animals, Disinfectants pharmacology, Humans, Vaccines, Anti-Infective Agents pharmacology, Anti-Inflammatory Agents pharmacology, Dendrimers pharmacology
- Abstract
Alternatives to traditional antibiotics and to antiviral and anti-inflammatory drugs are much in need and the molecular design and development of anti-infective compounds constitute a pivotal area in modern medicinal research. Dendrimers are a relatively new class of structurally well-defined, i.e. monodisperse, synthetic polymers with hyperbranched structures which enable a given molecular motif to be presented in a highly multivalent fashion. Several types of dendrimers with various structural elements and molecular dimensions are commercially available at an affordable price. The surface of dendrimers can be modified relatively easily and, depending on the surface motif, the pharmacological properties of the dendrimer such as cytotoxicity, bacteriocidal and virucidal effect, biodistribution and biopermeability may be modulated to fit a specific medicinal purpose. Dendrimers are thus highly suitable tools in drug discovery and they allow the synthesis of molecules with high and specific binding affinities to a wide variety of receptors, viruses and bacteria. Hence the use of dendrimers for the development of antiviral or antibacterial drugs, destroying the infective agent or disrupting multivalent binding interactions between the infective agent and cells of the host organism has become a highly active research field. The wide range of applications reported for the use of dendrimers as anti-infective and anti-inflammatory drugs in the patent literature demonstrates the general applicability of these molecules as drug candidates. The present review will briefly treat the intrinsic properties of dendrimers in biological systems, as well as general concerns regarding the treatment of infective diseases. The use of dendrimers as anti-infective and anti-inflammatory drugs will be based on a thorough review of the recent patent literature.
- Published
- 2006
- Full Text
- View/download PDF
31. Thiophene backbone amide linkers, a new class of easily prepared and highly acid-labile linkers for solid-phase synthesis.
- Author
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Jessing M, Brandt M, Jensen KJ, Christensen JB, and Boas U
- Subjects
- Dipeptides chemistry, Enkephalin, Leucine chemistry, Amides chemistry, Chemistry, Organic methods, Peptides chemical synthesis, Thiophenes chemistry
- Abstract
Solid-phase synthesis is of tremendous importance for small-molecule and biopolymer synthesis. Linkers (handles) that release amide-containing products after completion of solid-phase synthesis are widely used. Here we present a new class of highly acid-labile backbone amide linkers (BAL handles) based on 3,4-ethylenedioxythiophene (EDOT), which we have termed T-BAL. These thiophene linkers are synthesized in three convenient steps from commercially available EDOT. In the linker design, the spacer was introduced to the EDOT core either via a carbon-carbon bond or via a thioether linkage. Introduction of the spacer via a C-C bond was performed by a chemoselective Negishi coupling without transient protection of the aldehyde group to provide the T-BAL1 handle. Introduction via a thioether linkage was performed by a facile nucleophilic aromatic substitution between the brominated EDOT aldehyde and unprotected mercapto acids to provide T-BAL2 and T-BAL3 handles. The minimal use of protecting groups gave the corresponding linker molecules in few synthetic steps and in good yields. After anchoring of the linker to a polymeric support, introduction of the first amino acid was achieved by reductive amination, giving a secondary amine. A following acylation of the secondary amine with a symmetrical amino acid anhydride resulted in a backbone amide linkage between the handle and the growing substrate (e.g., peptide chain). After solid-phase synthesis, the substrates could be released from the resin by either low acid conditions using 1% TFA in CH2Cl2 or high acid conditions such as 50% TFA in CH2Cl2. Peptide thioesters could be released from the T-BAL1 handle under very mild conditions using aqueous acetic acid. Tert-butyl based protecting groups, tert-butyl esters, tert-butyl ethers, and Boc groups, as well as dimethyl acetals were relatively stable to these mild conditions for release of the peptides.
- Published
- 2006
- Full Text
- View/download PDF
32. Role of the peri-effect in synthesis and reactivity of highly substituted naphthaldehydes: a novel backbone amide linker for solid-phase synthesis.
- Author
-
Pittelkow M, Boas U, Jessing M, Jensen KJ, and Christensen JB
- Subjects
- Aldehydes chemical synthesis, Crystallography, X-Ray, Models, Molecular, Molecular Structure, Naphthalenes chemistry, Aldehydes chemistry, Amides chemistry, Naphthalenes chemical synthesis
- Abstract
Handles (linkers) with an aldehyde functionality that permits the anchoring of substrates by reductive amination have, since their first report in the mid-1990s, become widely-used tools in solid-phase synthesis. In the synthesis of peptides, they allow anchoring of the growing peptide chain through a backbone amide, thus giving easy access to C-terminal modified or cyclic peptides. Recently, we described two new handles (NAL-1 and NAL-2) with dialkoxynaphthaldehyde core structures. Here, we describe the design, synthesis and properties of a novel trialkoxynaphthalene-based backbone amide linker (NAL-3). The NAL-3 handle is based on a trialkoxynaphthaldehyde (NALdehyde-3) that was synthesized in nine high-yielding steps from 3-methoxyphenylacetic acid in 51% overall yield. The naphthalene ring system was constructed using a regioselective methanesulfonic acid-catalyzed ring-closing reaction. The tetra-substituted naphthalene derivative 1,3,6-trimethoxynaphthalene-2-carbaldehyde (7) was selectively demethylated in the 1 position using BBr(3). The selectivity of this reaction is discussed, based on the crystal structures of reactant and product, 1-hydroxy-3,6-dimethoxy-naphthalene-2-carbaldehyde (8), and in the context of the peri-effect. The new handle was anchored to an aminomethylated poly(styrene) solid support, followed by assembly of a model dipeptide, then a study of the cleavage properties under acidic conditions was carried out. Surprisingly, the trialkoxynaphthaldehyde-based handle proved less acid-labile than the dialkoxynaphthaldehyde handles, and this fact is discussed with respect to handle design.
- Published
- 2005
- Full Text
- View/download PDF
33. Amyloid aggregates of the prion peptide PrP106-126 are destabilised by oxidation and by the action of dendrimers.
- Author
-
Heegaard PM, Pedersen HG, Flink J, and Boas U
- Subjects
- Amino Acid Sequence, Molecular Sequence Data, Oxidation-Reduction, Prions chemistry, Recombinant Proteins, Amyloid metabolism, Prions metabolism
- Abstract
The prion protein (PrP) peptide 106-126 forms amyloid aggregates in vitro and this sequence is speculated to be involved in the formation of amyloid fibrils by the abnormally folded PrP protein (PrPSc) found in spongiform encephalopathies. It is shown here by incubation experiments in water using Thioflavin T (ThT) as a fluorescent probe for amyloid formation that changes in C-terminal charge, oxidation state and conformational stabilisation lead to large changes in amyloid forming behaviour (amyloidogenicity) of this peptide. Amyloid formation is favoured by a charged C-terminus and is strongly inhibited by oxidation. Furthermore, cationic dendrimers are shown to perturb peptide fibrillation in a process dependent on the nature of the charged groups on the dendrimer surface.
- Published
- 2004
- Full Text
- View/download PDF
34. Two dialkoxynaphthalene aldehydes as backbone amide linkers for solid-phase synthesis.
- Author
-
Boas U, Christensen JB, and Jensen KJ
- Subjects
- Acylation, Alanine chemistry, Amination, Chromatography, High Pressure Liquid, Glycine chemistry, Hydrogen-Ion Concentration, Molecular Structure, Peptides chemical synthesis, Peptides chemistry, Aldehydes chemical synthesis, Aldehydes chemistry, Amides chemistry, Naphthalenes chemistry
- Abstract
Two new solid-phase handles for backbone amide anchoring based on regioisomeric dialkoxynaphthalene aldehydes (NALdehydes) were synthesized in five convenient steps from the corresponding commercially available dihydroxynaphthalenes. The two NALdehydes were coupled to an aminomethyl polystyrene support, the first monomer attached by efficient reductive amination, and the secondary amine acylated to form naphthalene amide linker (NAL-1 and NAL-2) anchoring. After on-resin synthesis, release of peptides was effected with TFA/H(2)O (95:5), TFA/DCM (50:50), or low TFA concentrations. The properties of the NAL handles were evaluated in the solid-phase synthesis of a series of peptides, in which NAL-2 showed the best cleavage properties.
- Published
- 2004
- Full Text
- View/download PDF
35. Dendrimers in drug research.
- Author
-
Boas U and Heegaard PM
- Subjects
- Amines chemical synthesis, Animals, Anti-Bacterial Agents pharmacology, Antineoplastic Agents pharmacology, Antiviral Agents pharmacology, Esters chemical synthesis, Humans, Models, Chemical, Molecular Structure, Polymers chemical synthesis, Amines chemistry, Amines pharmacology, Esters chemistry, Esters pharmacology, Polymers chemistry, Polymers pharmacology
- Abstract
Dendrimers are versatile, derivatisable, well-defined, compartmentalised chemical polymers with sizes and physicochemical properties resembling those of biomolecules e.g. proteins. The present critical review (citing 158 references) briefly describes dendrimer design, nomenclature and divergent/convergent dendrimer synthesis. The characteristic physicochemical features of dendrimers are highlighted, showing the effect of solvent pH and polarity on their spatial structure. The use of dendrimers in biological systems are reviewed, with emphasis on the biocompatibility of dendrimers, such as in vitro and in vivo cytotoxicity, as well as biopermeability, biostability and immunogenicity. The review deals with numerous applications of dendrimers as tools for efficient multivalent presentation of biological ligands in biospecific recognition, inhibition and targeting. Dendrimers may be used as drugs for antibacterial and antiviral treatment and have found use as antitumor agents. The review highlights the use of dendrimers as drug or gene delivery devices in e.g. anticancer therapy, and the design of different host-guest binding motifs directed towards medical applications is described. Other specific examples are the use of dendrimers as 'glycocarriers' for the controlled multimeric presentation of biologically relevant carbohydrate moieties which are useful for targeting modified tissue in malignant diseases for diagnostic and therapeutic purposes. Finally, the use of specific types of dendrimers as scaffolds for presenting vaccine antigens, especially peptides, for use in vaccines is presented.
- Published
- 2004
- Full Text
- View/download PDF
36. New dendrimer-peptide host-guest complexes: towards dendrimers as peptide carriers.
- Author
-
Boas U, Söntjens SH, Jensen KJ, Christensen JB, and Meijer EW
- Subjects
- Amines chemistry, Amino Acids chemistry, Carboxylic Acids chemistry, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Conformation, Molecular Structure, Spectroscopy, Fourier Transform Infrared, Computer Simulation, Drug Carriers chemistry, Peptides chemical synthesis, Peptides chemistry
- Abstract
Adamantyl urea and adamantyl thiourea modified poly(propylene imine) dendrimers act as hosts for N-terminal tert-butoxycarbonyl (Boc)-protected peptides and form chloroform-soluble complexes. Investigations with NMR spectroscopy show that the peptide is bound to the dendrimer by ionic interactions between the dendrimer outer shell tertiary amines and the C-terminal carboxylic acid of the peptide, and also through host-urea to peptide-amide hydrogen bonding. The hydrogen-bonding nature of the peptide-dendrimer interactions was further confirmed by using Fourier transform IR spectroscopy, for which the NH- and CO-stretch signals of the peptide amide moieties shift towards lower wavenumbers upon complexation with the dendrimer. Spatial analysis of the complexes with NOESY spectroscopy generally shows close proximity of the N-terminal Boc group of the peptide to the peripheral adamantyl groups on the dendrimer host. The influence of side-chain motif on interactions with the host is analyzed by using seven different N-Boc-protected tripeptides as guests for the dendrimer. Downfield shifts of up to 1.3 ppm were observed for the guest amide NH-proton signals. These shifts decreased with increasing 'bulkiness' of the amino acid side chains. Despite this, the dendrimer was capable of making multiple peptide-dendrimer complexes when presented with a library of seven peptides. The different peptides were all present in the host, which did not show specific preferences, and could be released under mild acidic conditions. These results show the general nature of the peptide-dendrimer interactions in the formation of either single- or multiple-peptide-dendrimer complexes.
- Published
- 2002
- Full Text
- View/download PDF
37. The ortho backbone amide linker (o-BAL) is an easily prepared and highly acid-labile handle for solid-phase synthesis.
- Author
-
Boas U, Brask J, Christensen JB, and Jensen KJ
- Subjects
- Acids pharmacology, Amides chemistry, Amination, Benzaldehydes chemistry, Cross-Linking Reagents chemistry, Enkephalin, Leucine chemistry, Hydrolysis, Benzaldehydes chemical synthesis, Combinatorial Chemistry Techniques methods, Cross-Linking Reagents chemical synthesis
- Abstract
The tris(alkoxy)benzyl backbone amide linker (BAL) has found widespread application in solid-phase synthesis. The key intermediate for preparation of para BAL (p-BAL) is 2,6-dimethoxy-4-hydroxybenzaldehyde; several reports on its synthesis have appeared. However, the ortho analogue of the handle (o-BAL) has successfully been used by us for the synthesis of C-terminal-modified peptides, oligosaccharides, and substituted anilines. Here, we present a new and convenient synthesis of the key intermediate for o-BAL, 4,6-dimethoxy-2-hydroxybenzaldehyde, by a highly regioselective demethylation with BBr3, followed by purification through steam distillation. Cleavage studies of Leu-enkephalin anchored to either o-BAL or p-BAL handles revealed that both handles were surprisingly acid-labile and released the peptide with dilute TFA (5% and even 1% TFA in CH2Cl2). This useful property allowed the synthesis of fully protected Leu-enkephalin. The very convenient synthesis of 4,6-dimethoxy-2-hydroxybenzaldehyde combined with the benign properties of the o-BAL handle may make it the preferred regioisomer.
- Published
- 2002
- Full Text
- View/download PDF
38. Synthesis and properties of new thiourea-functionalized poly(propylene imine) dendrimers and their role as hosts for urea functionalized guests.
- Author
-
Boas U, Karlsson AJ, de Waal BF, and Meijer EW
- Abstract
Five generations of poly(propylene imine) dendrimers have been modified by palmityl and adamantyl endgroups via a thiourea linkage. The synthesis of the thiourea dendrimers DAB-dendr-(NHCSNHAd)(n) and DAB-dendr-(NHCSNHC(16)H(33))(n) (n = 4, 8, 16, 32, 64) proceeds smoothly via the amino-terminated DAB dendrimer and the adamantyl and palmityl isothiocyanates, respectively. The properties of the thiourea dendrimers have been studied by IR and (1)H NMR, including relaxation (T1, T2) measurements. The thiourea dendrimers are used as multivalent hosts for a number of guest molecules containing a terminal urea-glycine unit in organic solvents. The host-guest interactions have been investigated using 1D- and NOESY-NMR. These investigations show that the guest molecules bind to the dendritic host via thiourea (host)-urea (guest) hydrogen bonding, and ionic bonding between the terminal guest carboxylate moiety and the outer shell tertiary amines of the dendrimer. The ability to bind guest molecules of the adamantyl- and palmitylthiourea dendrimers has been compared with their respective urea containing dendrimer analogues, by NMR-titration, and competition experiments. Upon complexation, the thiourea dendrimer hosts show a larger downfield NH shift than the corresponding urea dendrimer hosts, indicative of stronger hydrogen bonding in the complexed state. Furthermore, microcalorimetry has been used to determine binding constants for formation of the host-guest complexes; the binding constants are typically in the order of 10(4) M(-1). Both NMR and microcalorimetric studies show that the thiourea dendrimers bind the urea containing guests with somewhat higher affinity than the corresponding urea dendrimers.
- Published
- 2001
- Full Text
- View/download PDF
39. New technology for regiospecific covalent coupling of polysaccharide antigens in ELISA for serological detection.
- Author
-
Jauho ES, Boas U, Wiuff C, Wredstrøm K, Pedersen B, Andresen LO, Heegaard PM, and Jakobsen MH
- Subjects
- Animals, Anthraquinones immunology, Antibodies, Bacterial immunology, Lipopolysaccharides analysis, Reproducibility of Results, Salmonella Infections, Animal blood, Salmonella typhimurium immunology, Swine, Ultraviolet Rays, Antibodies, Bacterial blood, Enzyme-Linked Immunosorbent Assay methods, Lipopolysaccharides immunology, Salmonella Infections, Animal immunology, Salmonella typhimurium isolation & purification
- Abstract
In this study we demonstrate a new UV irradiation technique for covalent coupling of bacterial polysaccharides derived from lipopolysaccharides to microtiter plates and the use of such plates in an enzyme linked immunosorbent assay (ELISA). Lipopolysaccharides were cleaved by mild acid hydrolysis into the lipid A part and the polysaccharide part. The polysaccharide was conjugated regiospecifically to a photochemically active compound, anthraquinone, resulting in a polysaccharide-anthraquinone conjugate. Anthraquinones forms active radicals when exposed to soft UV irradiation (350 nm) permitting the formation of stable covalent bonds to polymers e.g. microtiter plates. By this technique the polysaccharides are bound through the anthraquinone part of the polysaccharide-anthraquinone conjugates to the microtiter plates. This minimizes denaturation of O-antigen epitopes during binding to the microtiter plates and avoids cross-reactivity due to conserved domains in the lipid A. Furthermore, the covalent binding of the polysaccharide antigens are compatible with harsh assay conditions, such as extensive washing procedures and buffers with high salt concentrations with no risk of antigen leakage. Here we describe the use of this technique for the immobilization of lipopolysaccharide derived polysaccharides from Salmonella Typhimurium and Salmonella Choleraesuis lipopolysaccharides, representing the O-antigens 1, 4, 5, 6, 7, and 12. The functional polysaccharide surface gave similar ELISA results to plates coated passively with the corresponding unmodified lipopolysaccharide antigens. The plates were highly reproducible, showed very low inter- and intra-plate variation and were stable at room temperature for more than 8 months.
- Published
- 2000
- Full Text
- View/download PDF
40. Photochemical immobilization of anthraquinone conjugated oligonucleotides and PCR amplicons on solid surfaces.
- Author
-
Koch T, Jacobsen N, Fensholdt J, Boas U, Fenger M, and Jakobsen MH
- Subjects
- Base Sequence, DNA Primers, Magnetic Resonance Spectroscopy, Photochemistry, Polymerase Chain Reaction, Anthraquinones chemistry, DNA chemistry
- Abstract
Ligand immobilization on solid surfaces is an essential step in fields such as diagnostics, bio sensor manufacturing, and new material sciences in general. In this paper a photochemical approach based on anthraquinone as the chromophore is presented. Photochemical procedures offer special advantages as they are able to generate highly reactive species in an orientation specific manner. As presented here, anthraquinone (AQ) mediated covalent DNA immobilization appears to be superior to currently known procedures. A synthetic procedure providing AQ-phosphoramidites is presented. These reagents facilitate AQ conjugation during routine DNA synthesis, thus enabling the AQ-oligonucleotides to be immobilized in a very convenient and efficient manner. AQ-conjugated PCR primers can be used directly in PCR. When the PCR is performed in solution, the amplicons can be immobilized after the PCR. Moreover, when the primers are immobilized prior to the PCR, a solid-phase PCR can be performed and the amplicons are thus produced directly on the solid support.
- Published
- 2000
- Full Text
- View/download PDF
41. Evaluation of a novel enzyme-linked immunosorbent assay for detection of antibodies against Salmonella, employing a stable coating of lipopolysaccharide-derived antigens covalently attached to polystyrene microwells.
- Author
-
Wiuff C, Jauho ES, Stryhn H, Andresen LO, Thaulov K, Boas U, Jakobsen MH, and Heegaard PM
- Subjects
- Animals, Salmonella Infections, Animal immunology, Sensitivity and Specificity, Swine, Swine Diseases immunology, Swine Diseases microbiology, Antigens, Bacterial analysis, Enzyme-Linked Immunosorbent Assay veterinary, O Antigens analysis, Salmonella Infections, Animal diagnosis, Salmonella typhimurium immunology, Swine Diseases diagnosis
- Abstract
Polysaccharides derived from Salmonella typhimurium lipopolysaccharide (LPS) representing the O-antigen factors 1, 4, 5, and 12 and the O-antigen factors 6 and 7 from Salmonella choleraesuis LPS were derivatized with the photoreactive compound anthraquinone and subsequently covalently coupled to microtiter polystyrene plates by ultraviolet irradiation. Both polysaccharide antigens could be coupled simultaneously to the same microtiter plate. The coated surface was used in indirect ELISA for the determination of serum antibodies from pigs infected with bacteria of the two Salmonella groups and from uninfected pigs. This ELISA proved itself by having a good long-term durability and a high degree of reproducibility, including low day-to-day variations and low interplate variations. Furthermore, the ELISA showed good specificity and sensitivity when data were compared with the optical density levels of a panel of pig sera as determined by a conventional ELISA on the basis of passive coating of the two Salmonella LPS antigens (the mix-ELISA). The covalent anthraquinone mix-ELISA shows promise as a stable and durable alternative to the existing conventional ELISA for serological surveillance of Salmonella infections in pigs.
- Published
- 2000
- Full Text
- View/download PDF
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