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Radionuclide imaging of HER2 expression in tumours may enable stratification of patients with breast, ovarian, and gastroesophageal cancers for HER2-targeting therapies. A first-generation HER2-binding affibody molecule [Tc-99m]Tc-ZHER2:V2 demonstrated favorable imaging properties in preclinical studies. Thereafter, the affibody scaffold has been extensively modified, which increased its melting point, improved storage stability, and increased hydrophilicity of the surface. In this study, a second-generation affibody molecule (designated ZHER2:41071) with a new improved scaffold has been prepared and characterized. HER2-binding, biodistribution, and tumour-targeting properties of [Tc-99m]Tc-labelled ZHER2:41071 were investigated. These properties were compared with properties of the first-generation affibody molecules, [Tc-99m]Tc-ZHER2:V2 and [Tc-99m]Tc-ZHER2:2395. [Tc-99m]Tc-ZHER2:41071 bound specifically to HER2 expressing cells with an affinity of 58 +/- 2 pM. The renal uptake for [Tc-99m]Tc-ZHER2:41071 and [Tc-99m]Tc-ZHER2:V2 was 25-30 fold lower when compared with [Tc-99m]Tc-ZHER2:2395. The uptake in tumour and kidney for [Tc-99m]Tc-ZHER2:41071 and [Tc-99m]Tc-ZHER2:V2 in SKOV-3 xenografts was similar. In conclusion, an extensive re-engineering of the scaffold did not compromise imaging properties of the affibody molecule labelled with Tc-99m using a GGGC chelator. The new probe, [Tc-99m]Tc-ZHER2:41071 provided the best tumour-to-blood ratio compared to HER2-imaging probes for single photon emission computed tomography (SPECT) described in the literature so far. [Tc-99m]Tc-ZHER2:41071 is a promising candidate for further clinical translation studies.

OBJECTIVE: To study: (i) acute computed tomography (CT) characteristics and clinical outcome; (ii) clinical course and (iii) Corticosteroid Randomisation after Significant Head Injury acute calculator protocol (CRASH) model and clinical outcome in patients with severe traumatic brain injury (sTBI). METHODS: Initial CT (CTi) and CT 24 hours post-trauma (CT24) were evaluated according to Marshall and Rotterdam classifications. Rancho Los Amigos Cognitive Scale-Revised (RLAS-R) and Glasgow Outcome Scale Extended (GOSE) were assessed at three months and one year post-trauma. The prognostic value of the CRASH model was evaluated. RESULTS: Thirty-seven patients were included. Marshall CTi and CT24 were significantly correlated with RLAS-R at three months. Rotterdam CT24 was significantly correlated with GOSE at three months. RLAS-R and the GOSE improved significantly from three months to one year. CRASH predicted unfavourable outcome at six months for 81% of patients with bad outcome and for 85% of patients with favourable outcome according to GOSE at one year. CONCLUSION: Neither CT nor CRASH yielded clinically useful predictions of outcome at one year post-injury. The study showed encouragingly many instances of significant recovery in this population of sTBI. The combination of lack of reliable prognostic indicators and favourable outcomes supports the case for intensive acute management and rehabilitation as the default protocol in the cases of sTBI.

The overexpression and excessive signaling of platelet-derived growth factor receptor β (PDGFRβ) has been detected in cancers, atherosclerosis, and a variety of fibrotic diseases. Radionuclide in vivo visualization of PDGFRβ expression might help to select PDGFRβ targeting treatment for these diseases. The goal of this study was to evaluate the feasibility of in vivo radionuclide imaging of PDGFRβ expression using an Affibody molecule, a small nonimmunoglobulin affinity protein. Methods The PDGFRβ-binding Z09591 Affibody molecule was site-specifically conjugated with a maleimido derivative of DOTA and labeled with 111In. Targeting of the PDGFRβ-expressing U-87 MG glioblastoma cell line using 111In-DOTA-Z09591 was evaluated in vitro and in vivo. Results DOTA-Z09591 was stably labeled with 111In with preserved specific binding to PDGFRβ-expressing cells in vitro. The dissociation constant for 111In-DOTA-Z09591 binding to U-87 MG cells was determined to be 92 ± 10 pM. In mice bearing U-87 MG xenografts, the tumor uptake of 111In-DOTA-Z09591 was 7.2 ± 2.4 percentage injected dose per gram and the tumor-to-blood ratio was 28 ± 14 at 2 h after injection. In vivo receptor saturation experiments demonstrated that targeting of U-87 MG xenografts in mice was PDGFRβ-specific. U-87 MG xenografts were clearly visualized using small-animal SPECT/CT at 3 h after injection. Conclusion This study demonstrates the feasibility of in vivo visualization of PDGFRβ-expressing xenografts using an Affibody molecule. Further development of radiolabeled Affibody molecules might provide a useful clinical imaging tool for PDGFRβ expression during various pathologic conditions.

INTRODUCTION: This is the first report on the simultaneous successful treatment of a large ruptured saccular aneurysm and stenotic parent artery with a single flow-diverting stent. CASE PRESENTATION: We report the case of a 68-year-old Caucasian man with occlusion of the right vertebral artery and a ruptured aneurysm at the junction of the left posterior inferior cerebellar artery-left vertebral artery that was successfully treated by the deployment of a single flow-diverting stent in the stenotic left vertebral artery. Stent deployment was complicated by thrombotic occlusion of the basilar artery, which was successfully reopened. The patient recovered completely, and follow-up angiography at 4 months and 1 year showed patent vertebral artery with gradual shrinkage of the aneurysm. CONCLUSIONS: This report contributes to the literature on treatment of large ruptured aneurysms localized in stenotic arteries and in areas of the endocranium where a mass of embolic material in the aneurysm (coils) might compromise the circulation in the parent blood vessel or compress vital brain structures.

Basilaristrombos är ett akut och livshotande tillstånd. Intravenös trombolys är förstahandsbehandling, men ibland kan det vara en fördel att kombinera eller ersätta behandlingen med trombektomi. Lyckade behandlingsresultat med trombektomi finns beskrivna ända upp till 12–24 timmar efter insjuknandet. I fallbeskrivningarna diskuteras basilaristrombos hos två patienter som insjuknat med oklar medvetandesänkning. DT-angiografi gav diagnosen, och trombektomi kunde utföras 10 respektive 13 timmar efter insjuknandet. Långtidsresultatet var gott. Slutsatsen är att DT alltid ska kompletteras med DT-angiografi vid utredning av en akut medvetandepåverkad patient. Trombektomi bör övervägas om basilaristrombos påvisas., [Thrombectomy gave good results in basilar thrombosis. Prolonged time window for the intervention is proposed]. Basilar stroke is one of the most devastating acute cerebral vascular events. Intravascular thrombectomy may be performed beyond the 4.5 hours intravenous thrombolysis time window. However, the time window for thrombectomy in basilar stroke remains unclear. In our clinics, two cases of coma of unknown origin were diagnosed through CT angiography as distal basilar occlusion and treated with thromb­ectomy. Recanalization was achieved at 10 hours after stroke onset in case 1 and at 13 hours in case 2. The first patient regained consciousness shortly after operation and walked unlimited at 2 months after stroke. The second patient woke up slowly with cortical blindness, cerebellar ataxia, and unilateral weakness. At 10 months post stroke, he moved unlimited with mRS 2. Therefore, acute brain CT scan should always be accompanied with CT angiography in cases of coma of unknown origin for the differential diagnosis of basilar stroke where thrombectomy may be considered up to 13 hours after stroke onset.

BACKGROUND: Our aim was to evaluate the long-term treatment results in patients with intracranial aneurysms treated with endovascular techniques. METHODS: Forty-four patients treated due to intracranial aneurysms between 1996 and 2002 were investigated with a time-of-flight sequence magnetic resonance angiography (TOF MRA). RESULTS: Depending on the assessment, 47% to 51% of the treated aneurysms had a residual neck at the last digital subtraction angiography follow-up. There was filling of the aneurysm base (2%) in only 1 patient, whereas the remaining aneurysms were totally occluded. A TOF MRA performed 6 to 14 (mean 9.68) years after the last procedure showed a stable result in 93.9% of the treated aneurysms. There were no de novo aneurysms and previously untreated aneurysms were unchanged in size. CONCLUSION: Our long-term follow-up showed a stable result in previously coiled intracranial aneurysms.

Affibody molecules are non-immunoglobulin-derived affinity proteins based on a three-helical bundle protein domain. Here, we describe the design process of an optimized Affibody molecule scaffold with improved properties and a surface distinctly different from that of the parental scaffold. The improvement was achieved by applying an iterative process of amino acid substitutions in the context of the human epidermal growth factor receptor 2 (HER2)-specific Affibody molecule Z(HER2:342). Replacements in the N-terminal region, loop 1, helix 2 and helix 3 were guided by extensive structural modeling using the available structures of the parent Z domain and Affibody molecules. The effect of several single substitutions was analyzed followed by combination of up to 11 different substitutions. The two amino acid substitutions N23T and S33K accounted for the most dramatic improvements, including increased thermal stability with elevated melting temperatures of up to +12 degrees C. The optimized scaffold contains 11 amino acid substitutions in the nonbinding surface and is characterized by improved thermal and chemical stability, as well as increased hydrophilicity, and enables generation of identical Affibody molecules both by chemical peptide synthesis and by recombinant bacterial expression. A HER2-specific Affibody tracer, [MMA-DOTA-Cys61]-Z(HER2:2891)-Cys (ABY-025), was produced by conjugating MMA-DOTA (maleimide-monoamide-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) to the peptide produced either chemically or in Escherichia coli. ABY-025 showed high affinity and specificity for HER2 (equilibrium dissociation constant, K(D), of 76 pM) and detected HER2 in tissue sections of SKOV-3 xenograft and human breast tumors. The HER2-binding capacity was fully retained after three cycles of heating to 90 degrees C followed by cooling to room temperature. Furthermore, the binding surfaces of five Affibody molecules targeting other proteins (tumor necrosis factor alp

Syftet med denna studie var att försöka få en inblick i hur det är att leva i den komplexa kontexten i att ha en hbt (homo-bi-trans) läggning och att vara kristen. Vi ville på individnivå försöka tolka och förstå genom ett queerteoretiskt perspektiv. Vi har gjort en kvalitativ studie som baseras på sex halvstrukturerade intervjuer. Vidare ligger studiens fokus på personer som har ett aktivt liv inom den kristna kontexten. De begrepp som genomsyrar hela studien är heteronormativitet, hbt och stigmatisering. Analysen är baserad på en analysmodell som heter Interpretative phenomenological analysis och bygger på att lyfta fram teman. Totalt lyfter vi fram sju teman som går under namnen; Socialt rum, Personligt rum, Anpassning, Tyst konsensus, Diskriminering, Erkännande och Framtid. I varje tema lyfts intervjupersonernas egna upplevelser fram genom citat som sen knyts an till vald teori och begrepp. Slutsatser man kan dra av analysen är att det är en komplex miljö som dock börjar bli mer inkluderande för hbt personer. Det komplexa ligger i att olika kristna församlingar ser olika på dessa frågor och det finns ingen klar gemensam hållning. Detta ger olika grader på positiva och negativa sidor med en hbt läggning i den kristna kontexten., The aim with this study was to try getting an insight in how it is to live in the complex context in having it hbt (homo-bi-transsexual) disposition and to be Christian. We wanted on an individual level try to interpret and to understand through a queertheoretical perspective. We have done a qualitative study that we have based on 6st semi-structured interviews. Furthermore, the study's focus lies on persons who have an active life within the Christian context. The concepts that inform the entire study are heteronormativity, hbt and stigma. The analysis is based on an analysis model that is called Interpretative phenomenological analysis, and builds on lifting forward themes. A total of seven themes are lifted and they go by the names; Social rooms, Personal rooms, Adaptation, Quiet consensus, Discrimination, Admission and Future. In each theme, the interview persons' own experiences are lifted forward through quotations and are linked to our selected theory and concepts. Conclusions one can deduct of the analysis, is that it is a complex environment that however begins to become more including for hbt persons. The complex lies in that differently Christian parishes see differently on these questions and there is no clear common attitude. This gives various degrees on positive and negative sides with an hbt disposition in the Christian context.

Background context: Intradural disc herniation is relatively rare complication of the spinal degenerative process that occurs most frequently in the lumbar part of the spine. Both myelographic and magnetic resonance features of this entity have been described, and the mechanism of intradural herniation has already been proposed and generally accepted. In this article, we present a case of spontaneous resorption of an intradural, fragmented intervertebral disc. Spontaneous resorption of intradural disc fragments has not been previously reported. Purpose: To discuss a possible mechanism of spontaneous resorption of the subdural disc fragments. Study design: Case report and literature review. Methods: Radiological follow-up of a 46-year-old man with the intradural herniation of disc fragments. Conclusion: The reaction generated by the meninges might lead to the complete resorption of intrathecally localized disc fragments.

An Affibody® (Affibody) ligand with specific binding to human transferrin was selected by phage display technology from a combinatorial protein library based on the staphylococcal protein A (SpA)-derived Z domain. Strong and selective binding of the selected Affibody ligand to transferrin was demonstrated using biosensor technology and dot blot analysis. Impressive specificity was demonstrated as transferrin was the only protein recovered by affinity chromatography from human plasma. Efficient Affibody-mediated capture of transferrin, combined with IgG- and HSA-depletion, was demonstrated for human plasma and cerebrospinal fluid (CSF). For plasma, 85% of the total transferrin content in the samples was depleted after only two cycles of transferrin removal, and for CSF, 78% efficiency was obtained in single-step depletion. These results clearly suggest a potential for the development of Affibody-based resins for the removal of abundant proteins in proteomics analyses., QC 20100728

Interaktiv 3d-grafik är ett område som i dagsläget genomgår stora förändringar med ständigt ökad efterfrågan. Tack vare förbättrad mjuk- och hårdvara går det idag att åstadkomma saker som bara för ett par år sedan var fiktion. Syftet med rapporten är att dokumentera dessa möjligheter genom att undersöka grunderna för hantering och manipulering av 3d-modeller i programvaran Macromedia Director 8.5, samt implementera denna kunskap i en interaktiv 3d- applikation. 3d-grafik som används i Director är polygonbaserad och renderas i realtid på användarens dator. Moduleringen av 3d-grafiken kan utföras på två sätt, antingen i programmeringsspråket Lingo eller via fristående program. Director erbjuder goda möjligheter för manipulering av 3d-modeller vilket har utnyttjas i arbetets praktiska del. Den praktiska delen av vårt arbete har resulterat i en applikation där användaren kan inreda ett virtuellt rum i 3d-miljö. All interaktion med denna sker i realtid. Applikationen kan användas som plattform för projekt där företag vill visualisera sina produkter i en interaktiv 3d-miljö. För att skapa en sådan interaktiv 3d-miljö krävs det i dagsläget goda kunskaper om hur 3d-grafik är uppbyggd och hur den hanteras i programmeringsspråket Lingo. Denna rapport är ämnad att ge läsaren en inblick av detta.

Genetic strategies have become important in the design ofefficient processes for production of recombinant proteins.When constructing a production scheme, inherent properties,purity and quality requirements, as well as the final use ofthe protein, have to be considered. In this thesis, geneticdesign has been applied to differentEscherichia coliproduction processes for human peptidehormones, facilitating the down-stream processing andincreasing the expression yields. UponE. coliproduction of insulin-like growth factor I(IGF-I), misfolded and aggregated forms of IGF-I have reducedthe yield of correctly folded IGF-I. Extensive in vitrorefolding schemes have thus been considered necessary forobtaining acceptable yields. We investigated whethercoexpression of the IGF binding protein type 1 (IGFBP-1) wouldincrease the yield of correctly folded IGF-I. In this study,the two fusion proteins BB-IGFBP-1 and Z-IGF-I were used,containing the serum albumin binding affinity tag BB and theIgG-binding affinity tag Z, respectively. It was demonstratedthat correctly folded IGF-I could be recovered by twosubsequent affinity chromatography steps, verifying thatZ-IGF-I/BB-IGFBP-1 heterodimers were formed in vivo.Furthermore, the addition of a glutathione redox buffer duringcultivation significantly improved the relative yields ofcorrectly folded IGF-I, suggesting that affinity-assisted invivo folding could be considered as an attractive strategy forrecombinant proteins secreted to theE. coliperiplasm. Aiming for an efficient process for production of both humaninsulin and proinsulin C-peptide, the possibility to integratethe removal of an affinity handle with the processing ofproinsulin to insulin and C-peptide, was investigated.Expression vectors encoding three different ZZ-proinsulinfusion proteins were constructed. Between the two IgG-binding Zdomains and proinsulin, trypsin-sensitive cleavage sites,consisting of either arginine, lysine-arginine or lysine wereengineered. A study of cl, NR 20140805

The invention relates to a method for the preparation of insulin C-peptide which includes expressing in a host cell of a multimer polypeptide containing copies of the insulin C-peptide, and degradation of the expressed polypeptide for the release of individual copies of the insulin C-peptide. The invention also relates to molecules of the nucleinic acid, to vectors of expression and host cells for the application of this method and to multimer insulin C-polypeptide expressed and degraded in accordance with the method. 21 claims, US 6558924-B1 (2003-05-06)