Structure Determination Feasibility of Three-Dimensional Electron Diffraction in Case of Limited Data

During the last two decades, three-dimensional electron diffraction (3D ED) has undergone a renaissance, starting with the introduction of precession (Precession Electron Diffraction Tomography, PEDT) that led to variations on the idea of collecting as much of the diffraction space as possible in order to solve crystal structures from sub-micron sized crystals. The most popular of these acquisition methods is based on the continuous tilting/rotation of the crystal (so-called Microcrystal Electron Diffraction, MicroED) akin to the oscillating crystal method in X-ray crystallography, which was enabled by the increase of sensitivity and acquisition speed in electron detectors. While 3D ED data is more complex than the equivalent X-ray data due to the higher proportion of dynamical scattering, the same basic principles of what is required in terms of data quality and quantity in order to solve a crystal structure apply; high completeness, high data resolution and good signal-to-noise statistics on measured reflection intensities. However, it may not always be possible to collect data in these optimum conditions, the most common limitations being the tilt range of the goniometer stage, often due to a small pole piece gap or the use of a non-tomography holder, or the position of the sample on the TEM grid, which may be too close to a grid bar and then the specimen of interest becomes occluded during tilting. Other factors that can limit the quality of the acquired data include the limited dynamic range of the detector, which can result on truncated intensities, or the sensitivity of the crystal to the electron beam, whereby the crystallinity of the particle is changing under the illumination of the beam. This limits the quality and quantity of the measured intensities and makes structure analysis of such data challenging. Under these circumstances, traditional approaches may fail to elucidate crystal structures, and global optimization methods may be used here as an alternative powerful tool. In this context, this work presents a systematic study on the application of a global optimization method to crystal structure determination from 3D ED data. The results are compared with known structure models and crystal phases obtained from traditional ab initio structure solution methods demonstrating how this strategy can be reliably applied to the analysis of partially complete 3D ED data

Isolation of osteogenic progenitors from human amniotic fluid using a single step culture protocol

<p>Abstract</p> <p>Background</p> <p>Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases. In the present study, we report a novel single step protocol for the osteoblastic differentiation of human amniotic fluid cells.</p> <p>Results</p> <p>The described protocol is able to provide osteoblastic cells producing nodules of calcium mineralization within 18 days from withdrawal of amniotic fluid samples. These cells display a complete expression of osteogenic markers (COL1, ONC, OPN, OCN, OPG, BSP, Runx2) within 30 days from withdrawal. In order to test the ability of these cells to proliferate on surfaces commonly used in oral osteointegrated implantology, we carried out cultures onto different test disks, namely smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium). Electron microscopy analysis evidenced the best cell growth on this latter surface.</p> <p>Conclusion</p> <p>The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology.</p

The non-inflammatory role of C1q during Her2/neu-driven mammary carcinogenesis

There is an ever increasing amount of evidence to support the hypothesis that complement C1q, the first component of the classical complement pathway, is involved in the regulation of cancer growth, in addition to its role in fighting infections. It has been demonstrated that C1q is expressed in the microenvironment of various types of human tumors, including breast adenocarcinomas. This study compares carcinogenesis progression in C1q deficient (neuT-C1KO) and C1q competent neuT mice in order to investigate the role of C1q in mammary carcinogenesis. Significantly accelerated autochthonous neu(+) carcinoma progression was paralleled by accelerated spontaneous lung metastases occurrence in C1q deficient mice. Surprisingly, this effect was not caused by differences in the tumor-infiltrating cells or in the activation of the complement classical pathway, since neuT-C1KO mice did not display a reduction in C3 fragment deposition at the tumor site. By contrast, a significant higher number of intratumor blood vessels and a decrease in the activation of the tumor suppressor WW domain containing oxidoreductase (WWOX) were observed in tumors from neuT-C1KO as compare with neuT mice. In parallel, an increase in Her2/neu expression was observed on the membrane of tumor cells. Taken together, our findings suggest that C1q plays a direct role both on halting tumor angiogenesis and on inducing apoptosis in mammary cancer cells by coordinating the signal transduction pathways linked to WWOX and, furthermore, highlight the role of C1q in mammary tumor immune surveillance regardless of complement system activation

Violent behaviors and Klinefelter syndrome: Two forensic cases from the past to the future

Klinefelter Syndrome (KS) (XXY) is the most common sex-chromosome aberration among men. The cognitive phenotype includes language learning problems, mental retardation, and psychiatric disorders. Patients can show criminal personality and psychosocial problems. The most common offences reported are sexual abuse, arson, burglary, homicide, drug-related crimes. KS very often goes under-diagnosed. The aim of our study is to verify the hypothesis of correlation between chromosomal abnormalities and criminal behaviors through the analysis of a forensic case of uxoricide/suicide. We report the case of an old man, found dead in his cottage with a gun in the right hand. Judicial inspection demonstrated suicidal single gunshot injuries. Data analysis highlighted a personal story of uxoricide. Autopsy investigation showed the typical KS phenotype. The post-mortem clinical diagnosis was confirmed through the genetic analysis of the karyotype. The retrospective analysis of literature with this case showed a possible correlation between KS and psychiatric traits, with criminal personality and sexual disorders

Gadoxetic acid uptake as a molecular imaging biomarker for sorafenib resistance in patients with hepatocellular carcinoma: a post hoc analysis of the SORAMIC trial

PURPOSE Gadoxetic acid uptake on hepatobiliary phase MRI has been shown to correlate with ß-catenin mutation in patients with HCC, which is associated with resistance to certain therapies. This study aimed to evaluate the prognostic value of gadoxetic acid uptake on hepatobiliary phase MRI in patients with advanced HCC receiving sorafenib. METHODS 312 patients with available baseline hepatobiliary phase MRI images received sorafenib alone or following selective internal radiation therapy (SIRT) within SORAMIC trial. The signal intensity of index tumor and normal liver parenchyma were measured on the native and hepatobiliary phase MRI images, and relative tumor enhancement higher than relative liver enhancement were accepted as high gadoxetic acid uptake, and its prognostic value was assessed using univariate and multivariate Cox proportional hazard models. RESULTS The median OS of the study population was 13.4 (11.8-14.5) months. High gadoxetic acid uptake was seen in 51 (16.3%) patients, and none of the baseline characteristics was associated with high uptake. In univariate analysis, high gadoxetic acid uptake was significantly associated with shorter overall survival (10.7 vs. 14.0~months, p = 0.005). Multivariate analysis confirmed independent prognostic value of high gadoxetic acid uptake (HR, 1.7 1.21-2.3, p = 0.002), as well as Child-Pugh class (p = 0.033), tumor diameter (p = 0.002), and ALBI grade (p = 0.015). CONCLUSION In advanced HCC patients receiving sorafenib (alone or combined with SIRT), high gadoxetic acid uptake of the tumor on pretreatment MRI, a surrogate of ß-catenin mutation, correlates with shorter survival. Gadoxetic acid uptake status might serve in treatment decision-making process

Prognostic value of baseline imaging and clinical features in patients with advanced hepatocellular carcinoma

AIMS To investigate the prognostic value of baseline imaging features for overall survival (OS) and liver decompensation (LD) in patients with hepatocellular carcinoma (HCC). DESIGN Patients with advanced HCC from the SORAMIC trial were evaluated in this post hoc analysis. Several radiological imaging features were collected from baseline computed tomography (CT) and magnetic resonance imaging (MRI) imaging, besides clinical values. The prognostic value of these features for OS and LD (grade 2 bilirubin increase) was quantified with univariate Cox proportional hazard models and multivariate Least Absolute Shrinkage and Selection Operator (LASSO) regression. RESULTS Three hundred and seventy-six patients were included in this study. The treatment arm was not correlated with OS. LASSO showed satellite lesions, atypical HCC, peritumoral arterial enhancement, larger tumour size, higher albumin-bilirubin (ALBI) score, liver-spleen ratio <1.5, ascites, pleural effusion and higher bilirubin values were predictors of worse OS, and higher relative liver enhancement, smooth margin and capsule were associated with better OS. LASSO analysis for LD showed satellite lesions, peritumoral hypointensity in hepatobiliary phase, high ALBI score, higher bilirubin values and ascites were predictors of LD, while randomisation to sorafenib arm was associated with lower LD. CONCLUSIONS Imaging features showing aggressive tumour biology and poor liver function, in addition to clinical parameters, can serve as imaging biomarkers for OS and LD in patients receiving sorafenib and selective internal radiation therapy for HCC

Correlation of liver enhancement in gadoxetic acid-enhanced MRI with liver functions: a multicenter-multivendor analysis of hepatocellular carcinoma patients from SORAMIC trial

OBJECTIVES To evaluate the correlation between liver enhancement on hepatobiliary phase and liver function parameters in a multicenter, multivendor study. METHODS A total of 359 patients who underwent gadoxetic acid-enhanced MRI using a standardized protocol with various scanners within a prospective multicenter phase II trial (SORAMIC) were evaluated. The correlation between liver enhancement on hepatobiliary phase normalized to the spleen (liver-to-spleen ratio, LSR) and biochemical laboratory parameters, clinical findings related to liver functions, liver function grading systems (Child-Pugh and Albumin-Bilirubin ALBI), and scanner characteristics were analyzed using uni- and multivariate analyses. RESULTS There was a significant positive correlation between LSR and albumin (rho = 0.193; p < 0.001), platelet counts (rho = 0.148; p = 0.004), and sodium (rho = 0.161; p = 0.002); and a negative correlation between LSR and total bilirubin (rho = -0.215; p < 0.001) and AST (rho = -0.191; p < 0.001). Multivariate analysis confirmed independent significance for each of albumin (p = 0.022), total bilirubin (p = 0.045), AST (p = 0.031), platelet counts (p = 0.012), and sodium (p = 0.006). The presence of ascites (1.47 vs. 1.69, p < 0.001) and varices (1.55 vs. 1.69, p = 0.006) was related to significantly lower LSR. Similarly, patients with ALBI grade 1 had significantly higher LSR than patients with grade 2 (1.74 ± 0.447 vs. 1.56 ± 0.408, p < 0.001); and Child-Pugh A patients had a significantly higher LSR than Child-Pugh B (1.67 ± 0.44 vs. 1.49 ± 0.33, p = 0.021). Also, LSR was negatively correlated with MELD-Na scores (rho = -0.137; p = 0.013). However, one scanner brand was significantly associated with lower LSR (p < 0.001). CONCLUSIONS The liver enhancement on the hepatobiliary phase of gadoxetic acid-enhanced MRI is correlated with biomarkers of liver functions in a multicenter cohort. However, this correlation shows variations between scanner brands. KEY POINTS • The correlation between liver enhancement on the hepatobiliary phase of gadoxetic acid-enhanced MRI and liver function is consistent in a multicenter-multivendor cohort. • Signal intensity-based indices (liver-to-spleen ratio) can be used as an imaging biomarker of liver function. • However, absolute values might change between vendors

Localizing Autonomous Underwater Vehicles: Experimental Evaluation of a Long Baseline Method

This work concerns underwater networking with mobile assets, like Autonomous Underwater Vehicles (AUVs), for advanced monitoring and exploration of submerged environments. Particularly, we are interested in enabling an AUV to localize itself while moving underwater by acoustically polling beacon nodes statically deployed at well-known location. Our method only relies on a model of the AUV dynamics, on an on-board depth sensor and on long baseline ranging information. The AUV applies an Extended Kalman Filter to estimate its position, without needing any further local measurements but those of depths. We have evaluated the accuracy of the proposed method via experiments at sea in the shallow waters around the Italian island of Ponza, computing the average distance between the estimated locations of the AUV and its positions as measured by GPS along its trajectory (localization error). In deployments with up to four beacons, our simple method enables AUVs to swiftly self localize with errors never exceeding 3.62m (using only two beacons), 2.65m (three beacons) and 2.45m (four beacons)