Automated analysis of rabbit knee calcified cartilage morphology using micro-computed tomography and deep learning

Structural dynamics of calcified cartilage (CC) are poorly understood. Conventionally, CC structure is analyzed using histological sections. Micro-computed tomography (µCT) allows for three-dimensional (3D) imaging of mineralized tissues; however, the segmentation between bone and mineralized cartilage is challenging. Here, we present state-of-the-art deep learning segmentation for µCT images to assess 3D CC morphology. The sample includes 16 knees from 12 New Zealand White rabbits dissected into osteochondral samples from six anatomical regions: lateral and medial femoral condyles, lateral and medial tibial plateaus, femoral groove, and patella (n = 96). The samples were imaged with µCT and processed for conventional histology. Manually segmented CC from the images was used to train segmentation models with different encoder–decoder architectures. The models with the greatest out-of-fold evaluation Dice score were selected. CC thickness was compared across 24 regions, co-registered between the imaging modalities using Pearson correlation and Bland–Altman analyses. Finally, the anatomical CC thickness variation was assessed via a Linear Mixed Model analysis. The best segmentation models yielded average Dice of 0.891 and 0.807 for histology and µCT segmentation, respectively. The correlation between the co-registered regions was strong (r = 0.897, bias = 21.9 µm, standard deviation = 21.5 µm). Finally, both methods could separate the CC thickness between the patella, femoral, and tibial regions (p < 0.001). As a conclusion, the proposed µCT analysis allows for ex vivo 3D assessment of CC morphology. We demonstrated the biomedical relevance of the method by quantifying CC thickness in different anatomical regions with a varying mean thickness. CC was thickest in the patella and thinnest in the tibial plateau. Our method is relatively straightforward to implement into standard µCT analysis pipelines, allowing the analysis of CC morphology. In future research, µCT imaging might be preferable to histology, especially when analyzing dynamic changes in cartilage mineralization. It could also provide further understanding of 3D morphological changes that may occur in mineralized cartilage, such as thickening of the subchondral plate in osteoarthritis and other joint diseases.     

Melatonin and Metformin Failed to Modify the Effect of Dacarbazine in Melanoma

Lessons Learned

• Melatonin did not increase the efficacy of systemic chemotherapy in melanoma.
• Metformin did not increase the efficacy of systemic chemotherapy in melanoma.

     

Electrochemical functionalization of gold and silicon surfaces by a maleimide group as a biosensor for immunological application

In the present study we investigated the preparation of biofunctionalized surfaces using the direct electrochemical grafting of maleimidophenyl molecules with subsequent covalent immobilization of specific peptide to detect target antibody, thereby extending the application of the biosensing systems towards immunodiagnostics. Para-maleimidophenyl (p-MP) functional groups were electrochemically grafted on gold and silicon surfaces from solutions of the corresponding diazonium salt. A specially synthesized peptide modified with cysteine (Cys-peptide) was then immobilized on the p-MP grafted substrates by cross-linking between the maleimide groups and the sulfhydryl group of the cysteine residues. Accordingly, the Cys-peptide worked as an antigen that was able to bind specifically the target antibody (anti-GST antibody), while it was non-sensitive to a negative contrast antibody (i.e. anti-Flag β). The immobilization of both specific and non-specific antibodies on the Cys-peptide-modified surfaces was monitored by infrared spectroscopic ellipsometry, a quartz crystal microbalance integrated in flow injection analysis system and potentiometric response. The results obtained clearly demonstrated that the direct modification of a surface with maleimidophenyl provides a very simple and reliable way of preparing biofunctionalized surfaces suitable for the construction of immunological biosensors.     

Impaired presynaptic function and elimination of synapses at premature stages during postnatal development of the cerebellum in the absence of CALEB (CSPG5/neuroglycan C)

Chicken acidic leucine‐rich EGF‐like domain‐containing brain protein (CALEB), also known as chondroitin sulfate proteoglycan (CSPG)5 or neuroglycan C, is a neural chondroitin sulfate‐containing and epidermal growth factor (EGF)‐domain‐containing transmembrane protein that is implicated in synaptic maturation. Here, we studied the role of CALEB within the developing cerebellum. Adult CALEB‐deficient mice displayed impaired motor coordination in Rota‐Rod experiments. Analysis of the neuronal connectivity of Purkinje cells by patch‐clamp recordings demonstrated impairments of presynaptic maturation of inhibitory synapses. GABAergic synapses on Purkinje cells revealed decreased evoked amplitudes, altered paired‐pulse facilitation and reduced depression after repetitive stimulation at early postnatal but not at mature stages. Furthermore, the elimination of supernumerary climbing fiber synapses on Purkinje cells was found to occur at earlier developmental stages in the absence of CALEB. For example, at postnatal day 8 in wild‐type mice, 54% of Purkinje cells had three or more climbing fiber synapses in contrast to mutants where this number was decreased to less than 25%. The basic properties of the climbing fiber Purkinje cell synapse remained unaffected. Using Sholl analysis of dye‐injected Purkinje cells we revealed that the branching pattern of the dendritic tree of Purkinje cells was not impaired in CALEB‐deficient mice. The alterations observed by patch‐clamp recordings correlated with a specific pattern and timing of expression of CALEB in Purkinje cells, i.e. it is dynamically regulated during development from a high chondroitin sulfate‐containing form to a non‐chondroitin sulfate‐containing form. Thus, our results demonstrated an involvement of CALEB in the presynaptic differentiation of cerebellar GABAergic synapses and revealed a new role for CALEB in synapse elimination in Purkinje cells.     

Research on the Grinding Energy Density in a Jet Mill

Raw materials are used in many industrial technologies. The raw material frequently has to be prepared as an intermediate with an appropriate particle size distribution, which requires the use of grinding. In grinding processes, energy consumption is a very important profitability criterion for the applied particular size reduction technology. The paper describes the comminution process that takes place in the jet mill using a modified form of the thermodynamic theory of grinding. In this theory, new material characteristics have been added: the surface and volumetric density of grinding energy. The thermodynamic theory is a combination of the classical Kick’s theory and the modified form of Rittinger’s theory. The tested physical magnitudes are a measure of the energy consumption of the grinding process. They describe the energy that must be provided in the grinding process to overcome interactions between particles related to the volume and surface of the material. Knowledge of these magnitudes is necessary to model thermomechanical phenomena in the solid state. The paper presents the results of research on comminution in a jet mill, on the basis of which the values of the tested material magnitudes were determined. It is graphically shown how the values of the tested magnitudes depend on the grain size of the ground samples.     

The fluid handling performance of the curea P1 multipurpose dressing against superabsorbent and foam dressing technologies

Using a novel, automated robotic phantom system containing multiple wound simulants, we determined the fluid handling performance of the curea P1 multipurpose dressing vs market‐leading comparator superabsorbent and foam‐based dressings (FBDs). Specifically, we measured the retained, residual, evaporated, and (potentially occurring) spillover fluid shares for high‐ vs low‐viscosity exudate‐simulant test fluids, at 12, 24, and 30 hours postapplication of the dressings. These experiments were conducted for off‐loaded (‘prone’), non‐off‐loaded (‘supine’), and vertical (‘side‐lying’) simulated body positions. We found that the multipurpose dressing exhibited the best and most robust fluid handling performance across all the test configurations, for both the low‐ and high‐viscosity fluids. The FBD consistently showed the poorest performance compared to the other dressings, rendering it unlikely to be able to manage viscous exudates in ambulant patients (such as when applied to venous leg ulcers) as effectively as the other dressings. The superabsorbent dressing performed better than the foam dressing, but its fluid handling metrics were inferior to those of the multipurpose dressing. The current comparative quantification of the shares of retained, residual, evaporated, and spillover fluid, acquired through standardised laboratory tests, should help decision‐makers to select dressings that best meet their patient needs.     

Postal survey on asthma, chronic bronchitis and respiratory symptoms among adult Estonians and non-Estonians (FinEsS-study)

BACKGROUND: According to the data of the population census held in Estonia in 1989, the resident population consisted of 61.5% Estonians and 38.5% non-Estonians, the majority of them being Russians--30.3%. The aim of this study was to investigate the prevalence of asthma, chronic bronchitis and respiratory symptoms among the adults in these ethnic groups. METHODS: The study was a postal survey, similar in design to parallel studies currently in progress in Finland, Estonia and Sweden (FinEsS). RESULTS: The prevalence of physician-diagnosed asthma was 2.0%, being similar in men and women and in both Estonians and non-Estonians. The prevalence of physician-diagnosed chronic bronchitis was 10.5%. Compared with the non-Estonian population, the prevalence was much lower among Estonians (7.9% vs. 13.5%, p<0.001). Such a difference in prevalence was observed in all age groups. The high prevalence of chronic bronchitis in non-Estonians could be related to more extensive smoking habits, smoking indoors and probably ETS exposure. The defined OR between non-Estonians vs. Estonians after correcting for the influence of smoking habits, area of domicile, age, gender, and socioeconomic group was 1.30. A similar pattern of differences between Estonians and non-Estonians was observed in respiratory symptoms. CONCLUSION: This study revealed that the prevalence of physician-diagnosed asthma in Estonians and non-Estonians was the same. Physician-diagnosed chronic bronchitis was more frequent in non-Estonians than in Estonians. A significant difference was determined in the prevalence of respiratory symptoms, especially among those common to chronic bronchitis. The differences remained significant after correction for possible confounders.