Urine Proteomics Analysis of Patients with Neuronal Ceroid Lipofuscinoses

The Neuronal Ceroid Lipofuscinoses (NCL) are a group of 13 rare neurodegenerative disorders characterised by accumulation of cellular storage bodies. There are few therapeutic options and existing tests do not monitor disease progression and treatment response. However, urine biomarkers could address this need. Proteomic analysis of CLN2 patient urine revealed activation of immune response pathways and pathways associated with the unfolded protein response. Analysis of CLN5 and CLN6 sheep model urine showed subtle changes. To confirm and investigate the relevance of candidate biomarkers a targeted LC-MS/MS proteomic assay was created. We applied this assay to additional CLN2 samples as well as other NCL patients, (CLN1, CLN3, CLN5, CLN6 and CLN7) and demonstrated that Hexosaminidase-A, Aspartate Aminotransferase-1 and LAMP1, are increased in NCL samples and betaine-homocysteine S-methyltransferase-1 was specifically increased in CLN2 patients. These proteins could be used to monitor effectiveness of future therapies aimed at treating systemic NCL disease

Fluid-phase endocytosis and lysosomal degradation of bovine lactoferrin in lung cells

The iron-binding protein lactoferrin and the cell-penetrating peptides derived from its sequence utilise endocytosis to enter different cell types. The full-length protein has been extensively investigated as a potential therapeutic against a range of pathogenic bacteria, fungi, and viruses, including SARS-CoV-2. As a respiratory antiviral agent, several activity mechanisms have been demonstrated for lactoferrin, at the extracellular and plasma membrane levels, but as a protein that enters cells it may also have intracellular antiviral activity. Characterisation of lactoferrin’s binding, endocytic traffic to lysosomes, or recycling endosomes for exocytosis is lacking, especially in lung cell models. Here, we use confocal microscopy, flow cytometry, and degradation assays to evaluate binding, internalisation, endocytic trafficking, and the intracellular fate of bovine lactoferrin in human lung A549 cells. In comparative studies with endocytic probes transferrin and dextran, we show that lactoferrin binds to negative charges on the cell surface and actively enters cells via fluid-phase endocytosis, in a receptor-independent manner. Once inside the cell, we show that it is trafficked to lysosomes where it undergoes degradation within two hours. These findings provide opportunities for investigating both lactoferrin and derived cell-penetrating peptides activities of targeting intracellular pathogens

Biological evaluation of calcium phosphate bone cements for spinal repair

The broad aim of this work was to investigate and optimise the properties of calcium phosphate bone cements (CPCs) for use in vertebroplasty to achieve effective primary fixation of spinal burst fractures. An in vitro sample production method was developed and the effect of CPCs on the pH and Ca concentration of culture medium assessed. The effect of collagen incorporation into CPCs was investigated in vitro, necessitating sterilisation of the samples. y irradiation was found to be the most effective technique. In vitro analysis of two CPCs found no significant difference in their biological properties, although their ability to promote differentiation was significantly higher than that seen in a PMMA control. Analysis of collagen augmented CPCs found that the incorporation of 1 wt% collagen resulted in slightly higher, though not clinically relevant, level of cytotoxicity. Despite this, cells were shown to proliferate equally well on each of the formulations. Incorporation of marine collagen did not significantly affect the level of differentiation observed; however, this level was significantly decreased when bovine collagen was considered. The CPCs investigated were better tolerated in vivo than PMMA. Significantly more bone apposition was observed on the surface of CPCs compared to PMMA and a small amount of remodelling was seen on the CPCs.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Biocompatibility of Calcium Phosphate Bone Cement with Optimised Mechanical Properties

The broad aim of this work was to investigate and optimize the properties of calcium phosphate bone cements (CPCs) for use in vertebroplasty to achieve effective primary fixation of spinal fractures. The incorporation of collagen, both bovine and from a marine sponge (Chondrosia reniformis), into a CPC was investigated. The biological properties of the CPC and collagen–CPC composites were assessed in vitro through the use of human bone marrow stromal cells. Cytotoxicity, proliferation, and osteoblastic differentiation were evaluated using lactate dehydrogenase, PicoGreen, and alkaline phosphatase activity assays, respectively. The addition of both types of collagen resulted in an increase in cytotoxicity, albeit not to a clinically relevant level. Cellular proliferation after 1, 7, and 14 days was unchanged. The osteogenic potential of the CPC was reduced through the addition of bovine collagen but remained unchanged in the case of the marine collagen. These findings, coupled with previous work showing that incorporation of marine collagen in this way can improve the physical properties of CPCs, suggest that such a composite may offer an alternative to CPCs in applications where low setting times and higher mechanical stability are important. © 2015 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials Published by Wiley Periodicals, Inc. 104B:308–315, 2015

Enhanced Release of Calcium Phosphate Additives from Bioresorbable Orthopaedic Devices using Irradiation Technology is Non-Beneficial in a Rabbit Model

Objectives Bioresorbable orthopaedic devices with calcium phosphate (CaP) fillers are commercially available on the assumption that increased calcium (Ca) locally drives new bone formation, but the clinical benefits are unknown. Electron beam (EB) irradiation of polymer devices has been shown to enhance the release of Ca. The aims of this study were to: 1) establish the biological safety of EB surface-modified bioresorbable devices; 2) test the release kinetics of CaP from a polymer device; and 3) establish any subsequent beneficial effects on bone repair in vivo. Methods ActivaScrew Interference (Bioretec Ltd, Tampere, Finland) and poly(L-lactide-co-glycolide) (PLGA) orthopaedic screws containing 10 wt% β-tricalcium phosphate (β-TCP) underwent EB treatment. In vitro degradation over 36 weeks was investigated by recording mass loss, pH change, and Ca release. Implant performance was investigated in vivo over 36 weeks using a lapine femoral condyle model. Bone growth and osteoclast activity were assessed by histology and enzyme histochemistry. Results Calcium release doubled in the EB-treated group before returning to a level seen in untreated samples at 28 weeks. Extensive bone growth was observed around the perimeter of all implant types, along with limited osteoclastic activity. No statistically significant differences between comparative groups was identified. Conclusion The higher than normal dose of EB used for surface modification did not adversely affect tissue response around implants in vivo. Surprisingly, incorporation of β-TCP and the subsequent accelerated release of Ca had no significant effect on in vivo implant performance, calling into question the clinical evidence base for these commercially available devices

Асинхронный электропривод ленточного конвейера

В работе проведен расчет электропривода ленточного конвейера. Описан технологический процесс работы установки, представлена кинематическая схема механизма привода конвейера с редуктором, даны его технические характеристики, сформулированы технические требования к электроприводу. Проведен расчет кинематической схемы ленточного конвейера и выбор мощности двигателя, проведен выбор электрооборудования на основе преобразователя частоты, расчет и построение статических характеристик. Выполнен синтез системы управления электроприводом. Динамические режимы исследованы на имитационной модели системы, которые подтвердили правильность выбранных технических решений.The paper calculated the belt drive. Described process plant operation, presented kinematics conveyor drive with gear mechanism, given its characteristics, formulated technical requirements for the electric drive. The calculation of the kinematic scheme of the belt conveyor and the choice of engine power, the election of the electrical equipment on the basis of the frequency converter, calculation and construction of static characteristics. Made synthesis of electric drive control system. Dynamic modes are investigated on a simulation model of the system, which confirmed the correctness of the chosen technical solutions