Protocol for labeling and fixation of intact lysosomes with esterified amino acid analogs to assess lysosomal expansion in living eukaryotic cells

The lysosomal compartment is a key hub for cell metabolism, and morphological alterations have been described in several pathological conditions. Here, we describe the use of amino acid analogs modified by the presence of a methyl ester group that accumulates within lysosomes. This generates an intraluminal osmotic effect able to trigger a rapid and selective expansion of the lysosomal compartment within 2 h of treatment. We also present protocols to preserve lysosomal morphology, which yields a more accurate size measurement. For complete details on the use and execution of this protocol, please refer to Scerra et al. (2021)

Two efficient procedures to correct the positioning errors in the plane-polar scanning

Two techniques to effectively compensate known positioning errors in a plane-polar near-field–far-field (NF–FF) transformation, using a minimum number of NF data and adopting an oblate ellipsoid to shape the considered antenna, are proposed and validated through experimental proofs. The former makes use of the singular value decomposition method to recover the voltage samples which would be acquired by the probe at the points fixed by the non-redundant sampling representation from the collected positioning error affected ones, whereas the latter employs an iterative scheme. The NF data required by the classical NF–FF transformation with plane-rectangular scanning are then efficiently evaluated via a two-dimensional optimal sampling interpolation formula. The effectiveness of the proposed techniques is assessed by experimental tests performed at the Antenna Characterisation Lab of the University of Salerno

Near-Field/Far-Field Transformation with Helicoidal Scanning from Irregularly Spaced Data

A fast and accurate technique for the compensation of the probe positioning errors in the near-field/far-field transformation with helicoidal scanning is proposed in this paper. It relies on a nonredundant sampling representation using a spherical modelling of the antenna under test and employs an iterative scheme to evaluate the near-field data at the points fixed by the helicoidal nonredundant representation from the acquired irregularly distributed ones. Once these helicoidal data have been recovered, those required by a classical cylindrical near-field/far-field transformation are efficiently determined by using an optimal sampling interpolation algorithm. Some numerical tests assessing the effectiveness of the proposed approach and its stability with respect to random errors affecting the near-field data are shown

Sequencing in orthognathic bimaxillary surgery: which jaw should be operated first? A scoping review

Bimaxillary orthognathic surgery is widely used for the correction of dentoskeletal deformities. Surgery sequencing (maxilla or mandible first) remains debated, and guidelines and consensus are lacking. This scoping review summarizes the state of the art and compares the advantages and disadvantages of both approaches. The review was conducted following PRISMA-ScR guidelines. Three electronic databases (PubMed, Scopus, Web of Science) were searched using the PICO protocol and key words in orthognathic surgical sequencing. Four reviewers screened the records independently, and disagreement was resolved by consensus. A total of 23 records met the inclusion criteria. The advantages and disadvantages of the two approaches were compared and assessed for accuracy of reporting. Within the limitations of the present study, available evidence for the intrinsic advantages and the accuracy of the mandible-first sequence supports the choice of this approach in most cases. Nevertheless, each clinical case needs to be evaluated individually, as no dogmatic recommendations can be given for sequencing in bimaxillary orthognathic surger

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An Innovative Direct NF-FF Transformation Technique with Helicoidal Scanning

A direct near-field-far-field transformation with helicoidal scanning is developed. It is based on the nonredundant sampling representation of electromagnetic fields and uses a spherical antenna modelling to determine the number of helix turns. Moreover, the number of voltage samples on each of them is fixed by the maximum transverse dimension of the antenna, both to simplify the mechanical scanning and to reduce the computational effort. This technique allows the evaluation of the antenna far field directly from a minimum set of near-field data without interpolating them. Although the number of near-field data employed by the developed technique is slightly increased with respect to that required by rigorously applying the nonredundant sampling representation on the helix, it is still remarkably smaller than that needed by the standard near-field-far-field transformation with cylindrical scanning. The effectiveness of the technique is assessed by numerical and experimental results

Competitive binding of extracellular accumulated heparan sulfate reduces lysosomal storage defects and triggers neuronal differentiation in a model of Mucopolysaccharidosis IIIB.

Mucopolysaccharidoses (MPSs) are a group of inherited lysosomal storage disorders associated with the deficiency of lysosomal enzymes involved in glycosaminoglycan (GAG) degradation. The resulting cellular accumulation of GAGs is responsible for widespread tissue and organ dysfunctions. The MPS III, caused by mutations in the genes responsible for the degradation of heparan sulfate (HS), includes four subtypes (A, B, C, and D) that present significant neurological manifestations such as progressive cognitive decline and behavioral disorders. The established treatments for the MPS III do not cure the disease but only ameliorate non-neurological clinical symptoms. We previously demonstrated that the natural variant of the hepatocyte growth factor NK1 reduces the lysosomal pathology and reactivates impaired growth factor signaling in fibroblasts from MPS IIIB patients. Here, we show that the recombinant NK1 is effective in rescuing the morphological and functional dysfunctions of lysosomes in a neuronal cellular model of the MPS IIIB. More importantly, NK1 treatment is able to stimulate neuronal differentiation of neuroblastoma SK-NBE cells stable silenced for the NAGLU gene causative of the MPS IIIB. These results provide the basis for the development of a novel approach to possibly correct the neurological phenotypes of the MPS IIIB as well as of other MPSs characterized by the accumulation of HS and progressive neurodegeneration

Near-Field to Far-Field Transformation Techniques with Spiral Scannings: A Comprehensive Review

An overview of the near-field-far-field (NF-FF) transformation techniques with innovative spiral scannings, useful to derive the radiation patterns of the antennas commonly employed in the modern wireless communication systems, is provided in this paper. The theoretical background and the development of a unified theory of the spiral scannings for quasi-spherical and nonspherical antennas are described, and an optimal sampling interpolation expansion to evaluate the probe response on a quite arbitrary rotational surface from a nonredundant number of its samples, collected along a proper spiral wrapping it, is presented. This unified theory can be applied to spirals wrapping the conventional scanning surfaces and makes it possible to accurately reconstruct the NF data required by the NF-FF transformation employing the corresponding classical scanning. A remarkable reduction of the measurement time is so achieved, due to the use of continuous and synchronized movements of the positioning systems and to the reduced number of needed NF measurements. Some numerical and experimental results relevant to the spherical spiral scanning case when dealing with quasi-planar and electrically long antennas are shown