| 1. |
- Lundquist, Patrik, et al.
(författare)
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Barriers to the Intestinal Absorption of Four Insulin-Loaded Arginine-Rich Nanoparticles in Human and Rat
- 2022
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Ingår i: ACS Nano. - : American Chemical Society (ACS). - 1936-0851 .- 1936-086X. ; 16:9, s. 14210-14229
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Tidskriftsartikel (refereegranskat)abstract
- Peptide drugs and biologics provide opportunities for treatments of many diseases. However, due to their poor stability and permeability in the gastrointestinal tract, the oral bioavailability of peptide drugs is negligible. Nanoparticle formulations have been proposed to circumvent these hurdles, but systemic exposure of orally administered peptide drugs has remained elusive. In this study, we investigated the absorption mechanisms of four insulin-loaded arginine-rich nanoparticles displaying differing composition and surface characteristics, developed within the pan-European consortium TRANS-INT. The transport mechanisms and major barriers to nanoparticle permeability were investigated in freshly isolated human jejunal tissue. Cytokine release profiles and standard toxicity markers indicated that the nanoparticles were nontoxic. Three out of four nanoparticles displayed pronounced binding to the mucus layer and did not reach the epithelium. One nanoparticle composed of a mucus inert shell and cell-penetrating octarginine (ENCP), showed significant uptake by the intestinal epithelium corresponding to 28 ± 9% of the administered nanoparticle dose, as determined by super-resolution microscopy. Only a small fraction of nanoparticles taken up by epithelia went on to be transcytosed via a dynamin-dependent process. In situ studies in intact rat jejunal loops confirmed the results from human tissue regarding mucus binding, epithelial uptake, and negligible insulin bioavailability. In conclusion, while none of the four arginine-rich nanoparticles supported systemic insulin delivery, ENCP displayed a consistently high uptake along the intestinal villi. It is proposed that ENCP should be further investigated for local delivery of therapeutics to the intestinal mucosa.
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| 2. |
- Mårtensson, Ulrika, et al.
(författare)
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Deletion of the G protein-coupled receptor 30 impairs glucose tolerance, reduces bone growth, increases blood pressure, and eliminates estradiol-stimulated insulin release in female mice.
- 2009
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Ingår i: Endocrinology. - : The Endocrine Society. - 1945-7170 .- 0013-7227. ; 150:2, s. 687-98
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Tidskriftsartikel (refereegranskat)abstract
- In vitro studies suggest that the G protein-coupled receptor (GPR) 30 is a functional estrogen receptor. However, the physiological role of GPR30 in vivo is unknown, and it remains to be determined whether GPR30 is an estrogen receptor also in vivo. To this end, we studied the effects of disrupting the GPR30 gene in female and male mice. Female GPR30((-/-)) mice had hyperglycemia and impaired glucose tolerance, reduced body growth, increased blood pressure, and reduced serum IGF-I levels. The reduced growth correlated with a proportional decrease in skeletal development. The elevated blood pressure was associated with an increased vascular resistance manifested as an increased media to lumen ratio of the resistance arteries. The hyperglycemia and impaired glucose tolerance in vivo were associated with decreased insulin expression and release in vivo and in vitro in isolated pancreatic islets. GPR30 is expressed in islets, and GPR30 deletion abolished estradiol-stimulated insulin release both in vivo in ovariectomized adult mice and in vitro in isolated islets. Our findings show that GPR30 is important for several metabolic functions in female mice, including estradiol-stimulated insulin release.
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| 3. |
- T. Tegler, Lotta, et al.
(författare)
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A highly selective polypeptide binder for human Acetylcholine esterase
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- A highly selective high-affinity polypeptide conjugate binder for human Acetylcholine Esterase (hAChE) has been obtained by coupling a derivative of acridine, a known medium-affinity inhibitor of hAChE, to each member of a 16-membered set of 42-residue polypeptide scaffolds. The best candidate, 4-C10L17-Ac, was identified to have a KD of 10 nM or less in an assay where each polypeptide conjugate was titrated with hAChE in 50 mM phosphate buffer at pH 7.0 and 298K. It was found in a sandwich ELISA to have high selectivity for hAChE in cerebrospinal fluid. Targeting the active site of hAChE by a polypeptide conjugate binder presents a special problem as it is buried deep inside the protein in a cavity that is approximately 20 Å deep. In order to permit simultaneous and cooperative binding of the acridine and the polypeptide to the active site and the AChE surface a fourteen atom spacer was needed. The 9-aminoacridine group was linked to the side chain of a lysine residue in each polypeptide via a spacer prepared from two aminohexanoic acid fragments. The results reinforce the impression that polypeptide conjugates are excellent alternatives to currently used protein binder technologies in diagnostic and therapeutic applications and that the conjugation of enzyme inhibitors to polypeptide scaffolds is a strategy of general applicability in the design of high-affinity binders for enzymes.
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| 4. |
- Adriani, O., et al.
(författare)
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The PAMELA space mission
- 2008
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Ingår i: Astroparticle, Part. Space Phys., Detect. Med. Phys. Appl. - Proc. Conf.. - : WORLD SCIENTIFIC. - 9812819088 - 9789812819086 ; , s. 858-864
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Konferensbidrag (refereegranskat)abstract
- The PAMELA (a Payload for Antimatter-Matter Exploration and Light-nuclei Astrophysics) experiment, is a satellite-borne particle spectrometer. It was launched on 15th June 2006 from the Baikonur cosmodrome in Kazakhstan, is installed into the Russian Resurs-DK1 satellite. PAMELA is composed of a time-of-flight system, a magnetic spectrometer, a silicon-tungsten electromagnetic calorimeter, an anticoincidence system, a shower tail catcher scintillator and a neutron detector. Among the PAMELA major objectives are the study of charged particles in the cosmic radiation, the investigation of the nature of dark matter, by mean of the measure of the cosmic-ray antiproton and positron spectra over the largest energy range ever achieved. PAMELA has been in a nearly continuous data taking mode since llth July 2006. The status of the apparatus and performances will be presented.
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| 5. |
- Agback, Peter, et al.
(författare)
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Root morphology and cluster root formation by seabuckthorn (Hippophae rhamnoides L.) in response to nitrogen, phosphorus and iron deficiency
- 2015
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Ingår i: Plant and Soil. - : Springer Science and Business Media LLC. - 0032-079X .- 1573-5036. ; 397, s. 75-91
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Tidskriftsartikel (refereegranskat)abstract
- The aims were to investigate effects of availability of nitrogen (N), phosphorus (P) and iron (Fe) on root properties of seabuckthorn (HippophaA << rhamnoides) and to test the hypothesis that seabuckthorn is able to form cluster roots (CRs).Two sources of seabuckthorn were used: the seabuckthorn cultivar BHi10726 originating from a breeding programme based on H.r. ssp mongolica and carried out in rich agricultural field soil in the black earth (chernozem) region of Russia and the seabuckthorn accession named Pk originating in a natural population of H.r. ssp turkestanica in the mountainous region of northern Pakistan. Three cultivation systems giving different water availabilities were used at two levels each of N, P and Fe. Root morphology of seedlings and clones was characterized and metabolite content in extracts of young and old CRs of Pk was analyzed by proton nuclear magnetic resonance spectroscopy.Availability of N affected growth and distribution of biomass between shoot and root, while P and Fe deficiency modified root system architecture towards more lateral roots. Densely positioned rootlets with a determinate type of growth consistent with the definition of CR were observed under low P and low Fe. Pk formed on average 12 CRs per plant, which was 3 to 4-fold higher compared to BHi10726 also when normalized per root length. Malate and glycine were most abundant of the organic acids and amino acids, respectively, and decreased in old CRs.Seabuckthorn has the ability to form cluster roots especially in Pk and under deficiency of P and Fe. The two sources of seabuckthorn with different histories showed distinctly different root system architectures. The high contents of malate and glycine and their decrease in old CRs may reflect roles in CR metabolism.
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| 6. |
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| 7. |
- Andersson, Per Ola, et al.
(författare)
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A Novel ATR-FTIR Approach for Characterisation and Identification of Ex Situ Immobilised Species
- 2007
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Ingår i: ChemPhysChem. - : Wiley. - 1439-4235 .- 1439-7641. ; 8:5, s. 712-722
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Tidskriftsartikel (refereegranskat)abstract
- We demonstrate a novel method to analyse ex situ prepared protein chips by attenuated total reflection Fourier IR spectroscopy (ATR-FTIR), which circumvents tedious functionalisation steps of internal reflection elements (IREs), and simultaneously allows for complementary measurements by other analytical techniques. This concept is proven by utilising immobilised metal affinity capture (IMACTM) chips containing about 10 m thick films of copolymers coated with nitrilotriacetic acid (NTA) groups, which originally was manufactured for surface enhanced laser desorption ionisation (SELDI) spectrometry. Three immobilisation steps were analysed by ATR-FTIR spectroscopy: 1) NTA complexation with nickel(II) ions 2) binding of two histidine (His)-tagged synthetic peptides of 25 (25-His6) and 48 (48-His6) amino acids to the NTA-groups and 3) attachment of a ligand, mesyl amide, to the surface-bound 48-His6. Despite interference from H2O, both amide I and II were well resolved. Utilising peptide adsorption in the thick copolymer matrix yields a high saturation peptide concentration of 100 mg mL-1 and a dissociation constant of 116±11 M, as determined by a detailed analysis of the Langmuir adsorption isotherm. The mesyl amide ligand was directly seen in the raw ATR-FTIR spectrum with specific peaks in the fingerprint region at 1172 and 1350 cm-1. Several aspects of the fine structure of the amide I band of the peptide were analysed: influences from secondary structure, amino side chains and competing contamination product. We believe that this approach has great potential as a stand-alone or complementary analytical tool for determination of the chemical composition of functionalised surfaces. We emphasise further that with this approach no chemical treatment of IREs is needed; the chips can be regenerated and reused, and applied in other experimental set-ups.
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| 8. |
- Artursson, Per, et al.
(författare)
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A new opening for orally taken peptide drugs
- 2020
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Ingår i: Nature Biomedical Engineering. - : Springer Science and Business Media LLC. - 2157-846X. ; 4:1, s. 12-13
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- Negatively charged nanoparticles of about 50 nm in size permit the oral delivery of insulin and other peptide drugs by temporally enhancing the permeability of the intestinal wall.
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| 9. |
- Belfrage, Per, et al.
(författare)
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Methods for the determination in the nanomole range of lipids in liver fine-needle aspiration biopsies
- 1970
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Ingår i: Scandinavian Journal of Clinical & Laboratory Investigation. - : Informa UK Limited. - 1502-7686 .- 0036-5513. ; 26:1, s. 53-60
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Tidskriftsartikel (refereegranskat)abstract
- Triglycerides and phospholipids 0.5-2mg (wet weight) of human liver material obtained by the fine-needle aspiration biopsy technique can be estimated after chloroform:methanol extraction by the determination of lipid phosphorus and glyceride glycerol. The methods are based on the colorimetric determination of inorganic phosphate after oxidation of the phospholipids and the enzymatic fluorometric determination of glycerol after complete hydrolysis of the triglycerides, catalyzed by purified pancreatic lipase. The accumulation of lipids is then expressed as the increase in the molar ratio of triglycerides over phospholipids.
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| 10. |
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