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Stability of a Split Streptomycin Binding Aptamer
2016
Here we investigated the stability of an aptamer, which is formed by two RNA strands and binds the antibiotic streptomycin. Molecular dynamics simulations in aqueous solution confirmed the geometry and the pattern of hydrogen bond interactions that was derived from the crystal structure (1NTB). The result of umbrella sampling simulations indicated a favored streptomycin binding with a free energy of ΔGbind° = −101.7 kJ mol–1. Experimentally, the increase in oligonucleotide stability upon binding of streptomycin was probed by single-molecule force spectroscopy. Rate dependent force spectroscopy measurements revealed a decrease in the natural off-rate (koff-COMPLEX = 0.22 ± 0.16 s–1) for the …
Mutant p53 induces Golgi tubulo-vesiculation driving a prometastatic secretome
2020
TP53 missense mutations leading to the expression of mutant p53 oncoproteins are frequent driver events during tumorigenesis. p53 mutants promote tumor growth, metastasis and chemoresistance by affecting fundamental cellular pathways and functions. Here, we demonstrate that p53 mutants modify structure and function of the Golgi apparatus, culminating in the increased release of a pro-malignant secretome by tumor cells and primary fibroblasts from patients with Li-Fraumeni cancer predisposition syndrome. Mechanistically, interacting with the hypoxia responsive factor HIF1α, mutant p53 induces the expression of miR-30d, which in turn causes tubulo-vesiculation of the Golgi apparatus, leading …
Incomplete Timothy syndrome secondary to a mosaic mutation of the CACNA1C gene diagnosed using next-generation sequencing.
2016
Autosomal dominant genetic diseases can occur de novo and in the form of somatic mosaicism, which can give rise to a less severe phenotype, and make diagnosis more difficult given the sensitivity limits of the methods used. We report the case of female child with a history of surgery for syndactyly of the hands and feet, who was admitted at 6 years of age to a pediatric intensive care unit following cardiac arrest. The electrocardiogram (ECG) showed a long QT interval that on occasions reached 500 ms. Despite the absence of facial dysmorphism and the presence of normal psychomotor development, a diagnosis of Timothy syndrome was made given the association of syndactyly and the ECG features.…
Collective Infection of Cells by Viral Aggregates Promotes Early Viral Proliferation and Reveals a Cellular-Level Allee Effect
2018
In addition to the conventional release of free, individual virions, virus dispersal can involve multi-virion assemblies that collectively infect cells. However, the implications of collective infection for viral fitness remain largely unexplored. Using vesicular stomatitis virus, here, we compare the fitness of free versus saliva-aggregated viral particles. We find that aggregation has a positive effect on early progeny production, conferring a fitness advantage relative to equal numbers of free particles in most cell types. The advantage of aggregation resides, at least partially, in increasing the cellular multiplicity of infection. In mouse embryonic fibroblasts, the per capita, short-t…
Comparison between adenosine triphosphate bioluminescence and aerobic colony count to assess surface sanitation in the hospital environment
2022
Background: Adenosine triphosphate bioluminescence produced by the firefly luciferase has been successfullyintroduced to verify cleaning procedures in the food industry according to the Hazard Analysis Critical Control Pointprogram.Our aim was to evaluate the reliability of bioluminescence as a tool to monitor the effectiveness of sanitation in healthcaresettings, in comparison with the microbiological gold standard.Methods: 614 surfaces of various material were randomly sampled in Policlinico University Hospital units in Palermo,Italy, to detect adenosine triphosphate bioluminescence and aerobic colony count. Linear regression model andPearson correlation coefficient were used to estimate …
Differential binding cell-SELEX method to identify cell-specific aptamers using high-throughput sequencing
2018
AbstractAptamers have in recent years emerged as a viable alternative to antibodies. High-throughput sequencing (HTS) has revolutionized aptamer research by increasing the number of reads from a few (using Sanger sequencing) to millions (using an HTS approach). Despite the availability and advantages of HTS compared to Sanger sequencing, there are only 50 aptamer HTS sequencing samples available on public databases. HTS data in aptamer research are primarily used to compare sequence enrichment between subsequent selection cycles. This approach does not take full advantage of HTS because the enrichment of sequences during selection can be due to inefficient negative selection when using live…
Automatic detection of hemangiomas using unsupervised segmentation of regions of interest
2016
In this paper we compare the performances of three automatic methods of identifying hemangioma regions in images: 1) unsupervised segmentation using the Otsu method, 2) Fuzzy C-means clustering (FCM) and 3) an improved region growing algorithm based on FCM (RG-FCM). For each image, the starting point of the algorithms is a rectangular region of interest (ROI) containing the hemangioma. For computing the performances of each method, the ROIs had been manually labeled in 2 classes: pixels of hemangioma and pixels of non-hemangioma. The computed scores are given separately for each image, as well as global performances across all ROIs for both classes. The best classification of non-hemangioma…
Correlation between micro-hardness and mineral content in healthy human enamel
2016
Background Enamel is the hardest and the stiffest tissue in the human body. The enamel undergoes multidirectional stresses, withstands multimillion chewing cycles, all while protecting the internal dentin and pulp from damage due to mechanical overload and exposure to the harsh chemical environment of the mouth. Raman spectroscopy allows to study enamel mineral content in a non-destructive and site-specific way. While Raman spectroscopy has been applied in other studies to assess tooth mineralization, there are no studies that examine the relationship between micro-hardness and mineral content of the untreated enamel. An understanding of this relationship is extremely important in a clinica…
On the minimal number of singular fibers with non-compact Jacobians for families of curves over P1
2016
Abstract Let f : X → P 1 be a non-isotrivial family of semi-stable curves of genus g ≥ 1 defined over an algebraically closed field k. Denote by s nc the number of the singular fibers whose Jacobians are non-compact. We prove that s nc ≥ 5 if k = C and g ≥ 5 ; we also prove that s nc ≥ 4 if char ( k ) > 0 and the relative Jacobian of f is non-smooth.
Dynamic coarse-graining fills the gap between atomistic simulations and experimental investigations of mechanical unfolding
2017
We present a dynamic coarse-graining technique that allows to simulate the mechanical unfolding of biomolecules or molecular complexes on experimentally relevant time scales. It is based on Markov state models (MSM), which we construct from molecular dynamics simulations using the pulling coordinate as an order parameter. We obtain a sequence of MSMs as a function of the discretized pulling coordinate, and the pulling process is modeled by switching among the MSMs according to the protocol applied to unfold the complex. This way we cover seven orders of magnitude in pulling speed. In the region of rapid pulling we additionally perform steered molecular dynamics simulations and find excellen…