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Abstract Antibody engineering technology is at the forefront of therapeutic antibody development. The primary goal for engineering a therapeutic antibody is the generation of an antibody with a desired specificity, affinity, function, and developability profile. Mature antibodies are considered antigen specific, which may preclude their use as a starting point for antibody engineering. Here, we explore the plasticity of mature antibodies by engineering novel specificity and function to a pre-selected antibody template. Using a small, focused library, we engineered AAL160, an anti-IL-1β antibody, to bind the unrelated antigen IL-17A, with the introduction of seven mutations. The final redesigned antibody, 11.003, retains favorable biophysical properties, binds IL-17A with sub-nanomolar affinity, inhibits IL-17A binding to its cognate receptor and is functional in a cell-based assay. The epitope of the engineered antibody can be computationally predicted based on the sequence of the template antibody, as is confirmed by the crystal structure of the 11.003/IL-17A complex. The structures of the 11.003/IL-17A and the AAL160/IL-1β complexes highlight the contribution of germline residues to the paratopes of both the template and re-designed antibody. This case study suggests that the inherent plasticity of antibodies allows for re-engineering of mature antibodies to new targets, while maintaining desirable developability profiles.

The results shown in this paper extend our research group’s previous work, which presents the theoretically achievable hydrogen engine-out NO​xeo (H​2-NO​xeo) Pareto front of a hydrogen hybrid electric vehicle (H​2-HEV). While the Pareto front is calculated offline, which requires significant computing power and time, this work presents an online-capable algorithm to tackle the energy management of a H​2-HEV with explicit consideration of the H​2-NO​xeo trade-off. Through the inclusion of realistic predictive data on the upcoming driving mission, a model predictive control algorithm (MPC) is utilized to effectively tackle the conflicting goal of achieving low hydrogen consumption while simultaneously minimizing NO​xeo. In a case study, it is shown that MPC is able to satisfy user-defined NO​xeo limits over the course of various driving missions. Moreover, a comparison with the optimal Pareto front highlights MPC’s ability to achieve close-to-optimal fuel performance for any desired cumulated NO​xeo target on four realistic routes for passenger cars.

The modification of peptides and proteins for application in drug discovery and chemical biology is currently a rapidly growing field of research. Here, the authors report a photocatalytic diselenide contraction method for the dimerization and site-specific functionalisation of peptides and protein.

ObjectivesTo develop a biological diary (CoronaCal) that allows anyone in the community to collect and store serial saliva samples and chart symptoms on ordinary printer paper.MethodsDiaries were analyzed for the presence of SARS-CoV-2 RNA using established polymerase chain reaction (PCR) procedures. CoronaCal diaries were distributed to volunteer subjects in the community during the peak of the COVID-19 outbreak in New York. Volunteers collected their own daily saliva samples and self-reported symptoms.ResultsSARS-CoV-2 RNA extracted from CoronaCals was measured using qPCR and RNA levels were correlated with reported symptoms. SARS-CoV-2 RNA was detected in CoronaCals from nine of nine people with COVID-19 symptoms or exposure to someone with COVID-19, and not in one asymptomatic person. CoronaCals were stored for up to 70 days at room temperature during collection and then frozen for up to four months before analysis, suggesting that SARS-CoV-2 RNA is stable once dried onto paper.ConclusionsSampling saliva on simple paper provides a useful method to study the natural history and epidemiology of COVID-19. The CoronaCal collection and testing method is easy to implement, inexpensive, non-invasive and scalable. The approach can inform the historical and epidemiological understanding of infections in individuals and populations.

Abstract Background For recurrent glioblastoma (GB) patients, several therapy options have been established over the last years such as more aggressive surgery, re-irradiation or chemotherapy. Age and the Karnofsky Performance Status Scale (KPSS) are used to make decisions for these patients as these are established as prognostic factors in the initial diagnosis of GB. This study’s aim was to evaluate preoperative patient comorbidities by using the age-adjusted Charlson Comorbidity Index (ACCI) as a prognostic factor for recurrent GB patients. Methods In this retrospective analysis we could include 123 patients with surgery for primary recurrence of GB from January 2007 until December 2016 (43 females, 80 males, mean age 57 years (range 21–80 years)). Preoperative age, sex, ACCI, KPSS and adjuvant treatment regimes were recorded for each patient. Extent of resection (EOR) was recorded as a complete/incomplete resection of the contrast-enhancing tumor part. Results Median overall survival (OS) was 9.0 months (95% CI 7.1–10.9 months) after first re-resection. Preoperative KPSS > 80% (P

Abstract Backround Median overall survival (OS) after diagnosis of glioblastoma (GBM) remains 15 months amongst patients receiving aggressive surgical resection, chemotherapy and irradiation. Treatment of patients with a poor preoperative Karnofsky Performance Status Scale (KPSS) is still controversial. Therefore, we retrospectively assessed the outcome after surgical treatment in patients with a KPSS of ≤60%. Methods We retrospectively included patients with a de-novo glioblastoma WHO °IV and preoperative KPSS ≤60%, who underwent surgery at two neurosurgical centres between September 2006 and March 2016. We recorded pre- and postoperative tumour volume, pre- and postoperative KPSS, OS, age and MGMT promoter status. Results One hundred twenty-three patients (58 females/65 males, mean age 67.4 ± 13.4 years) met the inclusion criteria. Seventy-five of the 123 patients (61%) underwent surgical resection. 48/123 patients (39%) received a biopsy. The median preoperative and postoperative tumour volume of all patients was 33.0 ± 31.3 cm3 (IR 15.0–56.5cm3) and 3.1 ± 23.8 cm3 (IR 0.2–15.0 cm3), respectively. The median KPSS was 60% (range 20–60%) preoperatively and 50% (range 0–80%) postoperatively. Patients who received a biopsy showed a median OS for patients who received a biopsy only was 3.0 months (95% CI 2.0–4.0 months), compared to patients who had a resection and had a median OS of 8 months (95% CI 3.1–12.9 months). Age (p

Summary: The photoreceptor rhodopsin (Rho) becomes active when a tethered inverse agonist ligand (11CR) is photoconverted to an agonist (ATR). The ligand-binding pocket of inactive rhodopsin is completely enclosed, whereas active rhodopsin displays pores accessible from the lipid bilayer. Stabilization of active rhodopsin impedes 11CR binding and photoreceptor dark adaptation. Here, we used genetic code expansion and bioorthogonal labeling to engineer Rho mutants that serve as FRET sensors for measuring 11CR binding kinetics and energetics. We found that mutations that alter a channel between transmembrane helices 5 and 6 (TM5/6) dramatically affect 11CR binding kinetics but not agonist release kinetics. Our data provide direct experimental evidence for 11CR entry between TM5/6 in Rho that involves dynamic allosteric control of the ligand entry channel. Our findings provide a conceptual framework for understanding the function of G protein-coupled receptors with hydrophobic ligands that are hypothesized to enter their binding pockets through transmembrane pores.

Abstract Background Liquid biopsy based on cell-free DNA circulating in plasma has shown solid results as a non-invasive biomarker. In the present study we evaluated the utility of circulating free DNA (cfDNA) and the sub-type tumor DNA (ctDNA) in hepatocellular cancer (HCC) patients to assess therapy response and clinical outcome. Methods A cohort of 13 patients recruited in the context of the SORAMIC trial with unresectable, advanced HCC and different etiological and clinicopathological characteristics was included in this exploratory study. Plasma samples were collected between liver micro-intervention and beginning of sorafenib-based systemic therapy and then in correspondence of three additional follow-ups. DNA was isolated from plasma and next generation sequencing (NGS) was performed on a panel of 597 selected cancer-relevant genes. Results cfDNA levels showed a significant correlation with the presence of metastases and survival. In addition cfDNA kinetic over time revealed a trend with the clinical history of the patients, supporting its use as a biomarker to monitor therapy. NGS-based analysis on ctDNA identified 28 variants, detectable in different combinations at the different time points. Among the variants, HNF1A, BAX and CYP2B6 genes showed the highest mutation frequency and a significant association with the patients’ clinicopathological characteristics, suggesting a possible role as driver genes in this specific clinical setting. Conclusions Taken together, the results support the prognostic value of cfDNA/ctDNA in advanced HCC patients with the potential to predict therapy response. These findings support the clinical utility of liquid biopsy in advanced HCC improving individualized therapy and possible earlier identification of treatment responders.

Mushrooms are a potent source of dietary antioxidants and are often marketed for their capability of decreasing oxidative stress and preventing diseases. To date, however, little research has been done on the antioxidant activity of commercially available mushroom extractions, or their stability over time. Herein, the antioxidant capacity of six commercially available mushroom tinctures (hydro-alcoholic extracts of chaga, maitake, shiitake, reishi, lion's mane, and turkey tail) were evaluated using a panel of five antioxidant assays: Oxygen Radical Absorbance Capacity (ORAC), Nanoceria Reducing Antioxidant Capacity (NanoCerac), DPPH radical scavenging, Total Phenolic Content (TPC), and Ferric Reducing Antioxidant Power (FRAP). Commercial samples were compared to our own laboratory preparations of the same species, in a preliminary assessment of extraction technique and antioxidant stability. Mushroom extracts exhibited different antioxidant capacities depending on species and extraction method (e.g., 0.023 to 0.63 μmol TE/mg for commercial ORAC vs. 0.05 to 1.8 μmol TE/mg for laboratory ORAC for all species). In all cases, laboratory extracts exhibited considerably higher antioxidant activities than commercial extracts. In both extract types, chaga and maitake consistently demonstrated higher antioxidant activity than the other mushrooms. Preliminary antioxidant stability testing of our samples revealed stable ORAC values for one month at room temperature, followed by a 48% decrease after 4 months. Our results provide important insights regarding the effects of extraction techniques and storage conditions, on the antioxidant activity in mushroom extracts, particularly those made and sold commercially.

Summary: A recent phylogenetic analysis showed global co-evolution of G protein-coupled receptors (GPCRs) and receptor-activity-modifying proteins (RAMPs) suggesting global interactions between these two protein families. Experimental validation of these findings is challenging because in humans whereas there are only three genes encoding RAMPs, there are about 800 genes encoding GPCRs. Here, we report an experimental approach to evaluate GPCR-RAMP interactions. As a proof-of-concept experiment, we over-expressed RAMP2 in HEK293T cells and evaluated the effect on the transcriptional levels of 14 representative GPCRs that were selected based on the earlier phylogenetic analysis. We utilized a multiplexed error-correcting fluorescence in situ hybridization (MERFISH) method to detect message levels for individual GPCRs in single cells. The MERFISH results showed changes in GPCR message levels with RAMP2 over-expression in a concordant pattern that was predicted by the earlier phylogenetic analysis. These results provide additional evidence that GPCR-RAMP interactions are more widespread than previously appreciated and that these interactions have functional consequences. : Biological Sciences; Cell Biology; Molecular Biology Subject Areas: Biological Sciences, Cell Biology, Molecular Biology

The spleen is often involved in malignant lymphoma, which manifests on CT as either splenomegaly or focal, hypodense lymphoma lesions. This study aimed to investigate the diagnostic value of radiomics features of the spleen in classifying malignant lymphoma against non-lymphoma as well as the determination of malignant lymphoma subtypes in the case of disease presence—in particular Hodgkin lymphoma (HL), diffuse large B-cell lymphoma (DLBCL), mantle-cell lymphoma (MCL), and follicular lymphoma (FL). Spleen segmentations of 326 patients (139 female, median age 54.1 +/− 18.7 years) were generated and 1317 radiomics features per patient were extracted. For subtype classification, we created four different binary differentiation tasks and addressed them with a Random Forest classifier using 10-fold cross-validation. To detect the most relevant features, permutation importance was analyzed. Classifier results using all features were: malignant lymphoma vs. non-lymphoma AUC = 0.86 (p < 0.01); HL vs. NHL AUC = 0.75 (p < 0.01); DLBCL vs. other NHL AUC = 0.65 (p < 0.01); MCL vs. FL AUC = 0.67 (p < 0.01). Classifying malignant lymphoma vs. non-lymphoma was also possible using only shape features AUC = 0.77 (p < 0.01), with the most important feature being sphericity. Based on only shape features, a significant AUC could be achieved for all tasks, however, best results were achieved combining shape and textural features. This study demonstrates the value of splenic imaging and radiomic analysis in the diagnostic process in malignant lymphoma detection and subtype classification.

Secukinumab, a human monoclonal antibody that selectively neutralizes IL-17A, has consistently shown low anti-drug antibody responses in patients with psoriasis, psoriatic arthritis, and ankylosing spondylitis. Secukinumab has also shown lower in vitro immunogenicity potential compared with other monoclonal antibodies used to treat psoriasis and psoriatic arthritis, and a significantly lower in vitro T cell precursor frequency compared with ixekizumab, which targets the same antigen. Here, secukinumab and ixekizumab were further examined regarding their specific T cell epitopes. Secukinumab- or ixekizumab-specific CD4 T cell lines were generated from 31 healthy, treatment-naïve donors via 28-day co-culture with mature monocyte-derived dendritic cells exposed to either antibody. Consistent with previous data, the frequency of preexisting T cells to secukinumab was significantly lower as compared with ixekizumab. Only two T cell lines from two different donors could be derived for secukinumab, but no specific T cell epitope was identified. In contrast, 32 T cell lines from eight donors were obtained for ixekizumab. For 11 of these T cell lines, the specific T cell epitopes could be identified and confirmed by major histocompatibility complex–associated peptide proteomics as being naturally presented peptides. All identified T cell epitopes cluster in four main regions that are overlapping with the complementarity-determining regions HCDR3, LCDR1, LCDR2 and LCDR3. Interestingly, ixekizumab CDRs contain amino acids that are not found in any of the germline family members. These amino acids may be associated with the higher number of T cell epitopes identified for ixekizumab light chain and may contribute to the increased in vitro immunogenicity potential observed for ixekizumab vs. secukinumab.

BACKGROUND:Wavelet transformed reconstructions of dynamic susceptibility contrast (DSC) MR perfusion (wavelet-MRP) are a new and elegant way of visualizing vascularization. Wavelet-MRP maps yield a clear depiction of hypervascular tumor regions, as recently shown. OBJECTIVE:The aim of this study was to elucidate a possible connection of the wavelet-MRP power spectrum in glioblastoma (GBM) with local vascularity and cell proliferation. METHODS:For this IRB-approved study 12 patients (63.0+/-14.9y; 7m) with histologically confirmed IDH-wildtype GBM were included. Target regions for biopsies were prospectively marked on tumor regions as seen on preoperative 3T MRI. During subsequent neurosurgical tumor resection 43 targeted biopsies were taken from these target regions, of which all 27 matching samples were analyzed. All specimens were immunohistochemically analyzed for endothelial cell marker CD31 and proliferation marker Ki67 and correlated to the wavelet-MRP power spectrum as derived from DSC perfusion weighted imaging. RESULTS:There was a strong correlation between wavelet-MRP power spectrum (median = 4.41) and conventional relative cerebral blood volume (median = 5.97 ml/100g) in Spearman's rank-order correlation (κ = .83, p < .05). In a logistic regression model, the wavelet-MRP power spectrum showed a significant correlation to CD31 dichotomized to no or present staining (p = .04), while rCBV did not show a significant correlation to CD31 (p = .30). No significant association between Ki67 and rCBV or wavelet-MRP was found (p = .62 and p = .70, respectively). CONCLUSION:The wavelet-MRP power spectrum derived from existing DSC-MRI data might be a promising new surrogate for tumor vascularity in GBM.

Computed tomography (CT) scans represent the gold standard in the planning of functional endoscopic sinus surgeries (FESS). Yet, radiologists and otolaryngologists have different perspectives on these scans. In general, residents often struggle with aspects involved in both reporting and operation planning. The aim of this study was to compare the completeness of structured reports (SR) of preoperative CT images and structured operation planning (SOP) to conventional reports (CR) and conventional operation planning (COP) to potentially improve future treatment decisions on an individual level. In total, 30 preoperative CT scans obtained for surgical planning of patients scheduled for FESS were evaluated using SR and CR by radiology residents. Subsequently, otolaryngology residents performed a COP using free texts and a SOP using a specific template. All radiology reports and operation plannings were evaluated by two experienced FESS surgeons regarding their completeness for surgical planning. User satisfaction of otolaryngology residents was assessed by using visual analogue scales. Overall radiology report completeness was significantly higher using SRs regarding surgically important structures compared to CRs (84.4 vs. 22.0%, p

Finding prognostic biomarkers with high accuracy in patients with pancreatic cancer (PC) remains a challenging problem. To improve the prediction of survival and to investigate the relevance of quantitative imaging biomarkers (QIB) we combined QIB with established clinical parameters. In this retrospective study a total of 75 patients with metastatic PC and liver metastases were analyzed. Segmentations of whole liver tumor burden (WLTB) from baseline contrast-enhanced CT images were used to derive QIBs. The benefits of QIBs in multivariable Cox models were analyzed in comparison with two clinical prognostic models from the literature. To discriminate survival, the two clinical models had concordance indices of 0.61 and 0.62 in a statistical setting. Combined clinical and imaging-based models achieved concordance indices of 0.74 and 0.70 with WLTB volume, tumor burden score (TBS), and bilobar disease being the three WLTB parameters that were kept by backward elimination. These combined clinical and imaging-based models have significantly higher predictive performance in discriminating survival than the underlying clinical models alone (p < 0.003). Radiomics and geometric WLTB analysis of patients with metastatic PC with liver metastases enhances the modeling of survival compared with models based on clinical parameters alone.

Abstract Aim of this study was to determine if perioperative hemodynamics have an impact on perioperative infarct volume and patients’ prognosis. 201 cases with surgery for a newly diagnosed or recurrent glioblastoma were retrospectively analyzed. Clinical data and perioperative hemodynamic parameters, blood tests and time of surgery were recorded. Postoperative infarct volume was quantitatively assessed by semiautomatic segmentation. Mean diastolic blood pressure (dBP) during surgery (rho −0.239, 95% CI −0.11 – −0.367, p = 0.017), liquid balance (rho 0.236, 95% CI 0.1–0.373, p = 0.017) and mean arterial pressure (MAP) during surgery (rho −0.206, 95% CI −0.07 – −0.34, p = 0.041) showed significant correlation to infarct volume. A rank regression model including also age and recurrent surgery as possible confounders revealed mean intraoperative dBP, liquid balance and length of surgery as independent factors for infarct volume. Univariate survival analysis showed mean intraoperative dBP and MAP as significant prognostic factors, length of surgery also remained as significant prognostic factor in a multivariate model. Perioperative close anesthesiologic monitoring of blood pressure and liquid balance is of high significance during brain tumor surgery and should be performed to prevent or minimize perioperative infarctions and to prolong survival.

Background: Our retrospective single-center study aims to evaluate the impact of structured reporting (SR) using a CEUS LI-RADS template on report quality compared to conventional free-text reporting (FTR) in contrast-enhanced ultrasound (CEUS) for the diagnosis of hepatocellular carcinoma (HCC). Methods: We included 50 patients who underwent CEUS for HCC staging. FTR created after these examinations were compared to SR retrospectively generated by using template-based online software with clickable decision trees. The reports were evaluated regarding report completeness, information extraction, linguistic quality and overall report quality by two readers specialized in internal medicine and visceral surgery. Results: SR significantly increased report completeness with at least one key feature missing in 31% of FTR vs. 2% of SR (p < 0.001). Information extraction was considered easy in 98% of SR vs. 86% of FTR (p = 0.004). The trust of referring physicians in the report was significantly increased by SR with a mean of 5.68 for SR vs. 4.96 for FTR (p < 0.001). SR received significantly higher ratings regarding linguistic quality (5.79 for SR vs. 4.83 for FTR (p < 0.001)) and overall report quality (5.75 for SR vs. 5.01 for FTR (p < 0.001)). Conclusions: Using SR instead of conventional FTR increases the overall quality of reports in CEUS examinations of HCC patients and may represent a valuable tool to facilitate clinical decision-making and improve interdisciplinary communication in the future.

Molecular details of how antidepressant drugs bind to the human serotonin transporter are not currently clear. Here, the authors introduce photo-cross-linkers into the protein and map the binding site of several antidepressants.

Purpose: After a percutaneous ablation of colorectal liver metastases (CRLM), follow-up investigations to evaluate potential tumor recurrence are necessary. The aim of this study was to analyze whether a combined 18F-Fluordesoxyglucose positron emission tomography-computed tomography (18F-FDG PET/CT) scan is cost-effective compared to a contrast-enhanced computed tomography (CE-CT) scan for detecting local tumor progression. Materials and Methods: A decision model based on Markov simulations that estimated lifetime costs and quality-adjusted life years (QALYs) was developed. Model input parameters were obtained from the recent literature. Deterministic sensitivity analysis of diagnostic parameters based on a Monte-Carlo simulation with 30,000 iterations was performed. The willingness-to-pay (WTP) was set to $100,000/QALY. Results: In the base-case scenario, CE-CT resulted in total costs of $28,625.08 and an efficacy of 0.755 QALYs, whereas 18F-FDG PET/CT resulted in total costs of $29,239.97 with an efficacy of 0.767. Therefore, the corresponding incremental cost-effectiveness ratio (ICER) of 18F-FDG PET/CT was $50,338.96 per QALY indicating cost-effectiveness based on the WTP threshold set above. The results were stable in deterministic and probabilistic sensitivity analyses. Conclusion: Based on our model, 18F-FDG PET/CT can be considered as a cost-effective imaging alternative for follow-up investigations after percutaneous ablation of colorectal liver metastases.

Radiomics is an emerging field of image analysis with potential applications in patient risk stratification. This study developed and evaluated machine learning models using quantitative radiomic features extracted from multiparametric magnetic resonance imaging (mpMRI) to detect and classify prostate cancer (PCa). In total, 191 patients that underwent prostatic mpMRI and combined targeted and systematic fusion biopsy were retrospectively included. Segmentations of the whole prostate glands and index lesions were performed manually in apparent diffusion coefficient (ADC) maps and T2-weighted MRI. Radiomic features were extracted from regions corresponding to the whole prostate gland and index lesion. The best performing combination of feature setup and classifier was selected to compare its predictive ability of the radiologist’s evaluation (PI-RADS), mean ADC, prostate specific antigen density (PSAD) and digital rectal examination (DRE) using receiver operating characteristic (ROC) analysis. Models were evaluated using repeated 5-fold cross-validation and a separate independent test cohort. In the test cohort, an ensemble model combining a radiomics model, with models for PI-RADS, PSAD and DRE achieved high predictive AUCs for the differentiation of (i) malignant from benign prostatic lesions (AUC = 0.889) and of (ii) clinically significant (csPCa) from clinically insignificant PCa (cisPCa) (AUC = 0.844). Our combined model was numerically superior to PI-RADS for cancer detection (AUC = 0.779; p = 0.054) as well as for clinical significance prediction (AUC = 0.688; p = 0.209) and showed a significantly better performance compared to mADC for csPCa prediction (AUC = 0.571; p = 0.022). In our study, radiomics accurately characterizes prostatic index lesions and shows performance comparable to radiologists for PCa characterization. Quantitative image data represent a potential biomarker, which, when combined with PI-RADS, PSAD and DRE, predicts csPCa more accurately than mADC. Prognostic machine learning models could assist in csPCa detection and patient selection for MRI-guided biopsy.

Purpose: Seizures related to tumor growth are common in glioma patients, especially in low-grade glioma patients this is often the first tumor manifestation. We hypothesize that there are associations between preoperative seizures and morphologic features (e.g., tumor size, location) and histogram features in patients with glioblastoma (GB). Methods: Retrospectively, 160 consecutive patients with initial diagnosis and surgery of GB (WHO IV) and preoperative MRI were analyzed. Preoperative MRI sequences were co-registered (T2-FLAIR, T1-contrast, DTI) and tumors were segmented by a neuroradiologist using the software ITK-snap blinded to the clinical data. Tumor volume (FLAIR, T1-contrast) and histogram analyses of ADC- and FA-maps were recorded in the contrast enhancing tumor part (CET) and the non-enhancing peritumoral edema (FLAIR). Location was determined after co-registration of the data with an atlas. Permutation-based multiple-testing adjusted t statistics were calculated to compare imaging variables between patients with and without seizures. Results: Patients with seizures showed significantly smaller tumors (CET, adj. p = 0.029) than patients without preoperative seizures. Less seizures were observed in patients with tumor location in the right cingulate gyrus (adj. p = 0.048) and in the right caudate nucleus (adj. p = 0.009). Significant differences of histogram analyses of FA in the contrast enhancing tumor part were observed between patients with and without seizures considering also tumor location and size. Conclusion: Preoperative seizures in GB patients are associated with lower preoperative tumor volume. The different histogram analyses suggest that there might be microstructural differences in the contrast enhancing tumor part of patients with seizures measured by fractional anisotropy. Higher variance of GB presenting without seizures might indicate a more aggressive growth of these tumors.

BackgroundStriatocapsular infarcts (SCIs) are defined as large subcortical infarcts involving the territory of more than one lenticulostriate artery. SCI without concomitant ischemia in the more distal middle cerebral artery (MCA) territory [isolated SCI (iSCI)] has been described as a rare infarct pattern. The purpose of this study was to assess the prevalence of iSCI in patients treated with endovascular thrombectomy (ET), to evaluate baseline and procedural parameters associated with this condition, and to describe the clinical course of iSCI patients.MethodsA retrospective analysis of 206 consecutive patients with an isolated MCA occlusion involving the lenticulostriate arteries and treated with ET was performed. Baseline patient and procedural characteristics and ischemic involvement of the striatocapsular and distal MCA territory [iSCI, as opposed to non-isolated SCI (niSCI)] were analyzed using multivariate logistic regression models. Prevalence of iSCI was assessed, and clinical course was determined with the rates of substantial neurological improvement and good functional short- and mid-term outcome (discharge/day 90 Modified Rankin Scale ≤2).ResultsiSCI was detected in 53 patients (25.7%), and niSCI was detected in 153 patients (74.3%). Successful reperfusion [thrombolysis in cerebral infarction (TICI) 2b/3] [adjusted odds ration (aOR) 8.730, 95% confidence interval (95% CI) 1.069–71.308] and good collaterals (aOR 2.100, 95% CI 1.119–3.944) were associated with iSCI. In successfully reperfused patients, TICI 3 was found to be an additional factor associated with iSCI (aOR 5.282, 1.759–15.859). Patients with iSCI had higher rates of substantial neurological improvement (71.7 vs. 37.9%, p

PURPOSE:Unambiguous evaluation of glioblastoma (GB) progression is crucial, both for clinical trials as well as day by day routine management of GB patients. 3D-volumetry in the follow-up of GB provides quantitative data on tumor extent and growth, and therefore has the potential to facilitate objective disease assessment. The present study investigated the utility of absolute changes in volume (delta) or regional, segmentation-based subtractions for detecting disease progression in longitudinal MRI follow-ups. METHODS:165 high resolution 3-Tesla MRIs of 30 GB patients (23m, mean age 60.2y) were retrospectively included in this single center study. Contrast enhancement (CV) and tumor-related signal alterations in FLAIR images (FV) were semi-automatically segmented. Delta volume (dCV, dFV) and regional subtractions (sCV, sFV) were calculated. Disease progression was classified for every follow-up according to histopathologic results, decisions of the local multidisciplinary CNS tumor board and a consensus rating of the neuro-radiologic report. RESULTS:A generalized logistic mixed model for disease progression (yes / no) with dCV, dFV, sCV and sFV as input variables revealed that only dCV was significantly associated with prediction of disease progression (P = .005). Delta volume had a better accuracy than regional, segmentation-based subtractions (79% versus 72%) and a higher area under the curve by trend in ROC curves (.83 versus .75). CONCLUSION:Absolute volume changes of the contrast enhancing tumor part were the most accurate volumetric determinant to detect progressive disease in assessment of GB and outweighed FLAIR changes as well as regional, segmentation-based image subtractions. This parameter might be useful in upcoming objective response criteria for glioblastoma.

Capturing conformational changes in proteins or protein-protein complexes is a challenge for both experimentalists and computational biologists. Solution nuclear magnetic resonance (NMR) is unique in that it permits structural studies of proteins under greatly varying conditions, and thus allows us to monitor induced structural changes. Paramagnetic effects are increasingly used to study protein structures as they give ready access to rich structural information of orientation and long-range distance restraints from the NMR signals of backbone amides, and reliable methods have become available to tag proteins with paramagnetic metal ions site-specifically and at multiple sites. In this study, we show how sparse pseudocontact shift (PCS) data can be used to computationally model conformational states in a protein system, by first identifying core structural elements that are not affected by the environmental change, and then computationally completing the remaining structure based on experimental restraints from PCS. The approach is demonstrated on a 27 kDa two-domain NS2B-NS3 protease system of the dengue virus serotype 2, for which distinct closed and open conformational states have been observed in crystal structures. By changing the input PCS data, the observed conformational states in the dengue virus protease are reproduced without modifying the computational procedure. This data driven Rosetta protocol enables identification of conformational states of a protein system, which are otherwise difficult to obtain either experimentally or computationally.

Abstract To assess the present status and distribution of the lynx in the Austrian Alps we analysed documented reports on lynx presence for the period 1995 to 1999. Records on lynx presence in Austria were collected by the hunter's associations of Carinthia, Styria, Salzburg and Upper Austria, regional NGO's and wildlife biologists. For this five-year period, 106 records of lynx presence were documented for the Austrian Alps. Of these we classified 100 as Quality 3 data (unconfirmed prey remains, tracks and observations) and 5 as Quality 2 data (confirmed prey remains). The only hard fact (Quality 1 data) in the Austrian Alps was a male lynx killed on a highway in southern Salzburg in 1995. From these reported signs of lynx presence we concluded that 20 years after the first reintroductions there still is no established lynx population in the Austrian Alps. The reports rather indicate a few individuals scattered over a wide area. As the monitoring of the Austrian lynx population is more a passive and arbitrary collection of reports than a systematic monitoring, we believe that the distribution reported here is likely underestimating the actual distribution of the lynx in the Austrian Alps. Therefore we emphasise the need to establish a new systematic monitoring system covering the entire potential lynx habitat in Austria.

Abstract Nine wild lynx were translocated from the Carpathian Mountains to Styria between 1977 and 1979 in order to reintroduce this species into the Austrian Alps. Field projects continued until 1982, when monitoring of the released animals ceased. After five years the lynx had spread as far as 120 km from the site of re-introduction, but observations were few and seldom confirmed. Only in the district of Carinthia, where lynx activity was strongest, did the hunters' association gather data by means of observation report forms. Lynx reports became scarce during the 1980s, and re-introduction seemed to have failed, when in 1989 a series of sheep kills by lynx in Carinthia re-activated the interest of the public in the lynx, and more observations were reported. The Carinthian Hunters' Association formed a lynx group to verify these reports. More intensive training of the menbers of the group since 1992 has revealed that much of the information collected was not valid proof of lynx presence. There is no established lynx population in the Austrian Alps at present. The most regular and reliable observations come from the original region of the releases and from southern Carinthia, where lynx immigrating from Slovenia may sustain the Austrian population.