Proof-of-concept of a robotic-driven photogrammetric scanner for intra-operative knee cartilage repair.

This work presents a proof-of-concept of a robotic-driven intra-operative scanner designed for knee cartilage lesion repair, part of a system for direct in vivo bioprinting. The proposed system is based on a photogrammetric pipeline, which reconstructs the cartilage and lesion surfaces from sets of photographs acquired by a robotic-handled endoscope, and produces 3D grafts for further printing path planning. A validation on a synthetic phantom is presented, showing that, despite the cartilage smooth and featureless surface, the current prototype can accurately reconstruct osteochondral lesions and their surroundings with mean error values of 0.199 ± 0.096 mm but with noticeable concentration on areas with poor lighting or low photographic coverage. The system can also accurately generate grafts for bioprinting, although with a slight tendency to underestimate the actual lesion sizes, producing grafts with coverage errors of -12.2 ± 3.7, -7.9 ± 4.9, and -15.2 ± 3.4% for the medio-lateral, antero-posterior, and craneo-caudal directions, respectively. Improvements in lighting and acquisition for enhancing reconstruction accuracy are planned as future work, as well as integration into a complete bioprinting pipeline and validation with ex vivo phantoms.

Fast and versatile platform for pedicle screw insertion planning.

PURPOSE: Computer-assisted surgical planning methods help to reduce the risks and costs in transpedicular fixation surgeries. However, most methods do not consider the speed and versatility of the planning as factors that improve its overall performance. In this work, we propose a method able to generate surgical plans in minimal time, within the required safety margins and accounting for the surgeon's personal preferences. METHODS: The proposed planning module takes as input a CT image of the patient, initial-guess insertion trajectories provided by the surgeon and a reduced set of parameters, delivering optimal screw sizes and trajectories in a very reduced time frame. RESULTS: The planning results were validated with quantitative metrics and feedback from surgeons. The whole planning pipeline can be executed at an estimated time of less than 1 min per vertebra. The surgeons remarked that the proposed trajectories remained in the safe area of the vertebra, and a Gertzbein-Robbins ranking of A or B was obtained for 95 % of them. CONCLUSIONS: The planning algorithm is safe and fast enough to perform in both pre-operative and intra-operative scenarios. Future steps will include the improvement of the preprocessing efficiency, as well as consideration of the spine's biomechanics and intervertebral rod constraints to improve the performance of the optimisation algorithm.

Exergaming delivery of a balance and fall prevention program for older adults: A feasibility study.

Objective: Older adults are at high risk of falls and this problem calls for efficient and scalable interventions. This study investigated whether a motion capture system paired with balance training exergaming software is a feasible strategy to deliver therapeutic exercise to older adults in an aged care facility. Methods: This study analyzed data from a quality improvement rehabilitation initiative. Two convenience samples of older adults were included: a usual care group (n = 12), admitted to a rehabilitation hospital and receiving standard in-patient therapy 5×/week and the Evolv group (n = 12), admitted to an aged care facility, prescribed exergaming 3×/week. All participants performed 30-minute exercise sessions based on a fall prevention program over 3 months. The Short Physical Performance Battery (SPPB) and Tinetti Performance Oriented Mobility Assessment test were administered pre- and post-treatment. Results: No adverse events were recorded during the interventions. Mean SPPB increase for Evolv participants was 2.25 ± 1.35 (p < .001, CI for mean = 1.39 to 3.11, d = 1.66), compared with a non-significant change in the usual care group (mean increase = 2.25 ± 3.82, p = .066, CI for mean = -0.18 to 4.68, d = 0.59). Tinetti improvement was significant for the individuals receiving usual care (3.83 ± 2.82, p = .012, CI for mean = 1.01 to 6.66, d = 0.86) but there were no significant between-group differences in outcomes. Conclusions: Exergaming with the Evolv system for balance and strength training may be a feasible strategy to improve physical function for older adults recovering in an aged care facility.

Expression of the DYRK1A gene correlates with its 3D positioning in the interphase nucleus of Down syndrome cells.

Down syndrome is a common birth defect caused by trisomy of chromosome 21. Chromosomes occupy distinct territories in interphase nuclei, and their distribution within the nuclear space is nonrandom. In humans with Down syndrome, two chromosomes 21 frequently localize proximal to one another and distant from the third chromosome. Here, we investigated the nuclear organization of DYRK1A and SOD1, two genes mapping to chromosome 21 that greatly contribute to the pathology. We found that DYRK1A conserves its central positioning between normal and trisomic cells, whereas SOD1 adopts more peripheral distribution in trisomic cells. We also found that the relative position of these genes with respect to each other varies among the different copies of chromosome territories 21 within a cell, and that this distinct distribution is associated with differences in their expression levels. All together, our results may explain, at least in part, the difference in the expression level of these two genes implicated in the pathogenesis of Down syndrome.

Tetramerization-defects of p53 result in aberrant ubiquitylation and transcriptional activity.

The tumor suppressor p53 regulates the expression of genes involved in cell cycle progression, senescence and apoptosis. Here, we investigated the effect of single point mutations in the oligomerization domain (OD) on tetramerization, transcription, ubiquitylation and stability of p53. As predicted by docking and molecular dynamics simulations, p53 OD mutants show functional defects on transcription, Mdm2-dependent ubiquitylation and 26S proteasome-mediated degradation. However, mutants unable to form tetramers are well degraded by the 20S proteasome. Unexpectedly, despite the lower structural stability compared to WT p53, p53 OD mutants form heterotetramers with WT p53 when expressed transiently or stably in cells wild type or null for p53. In consequence, p53 OD mutants interfere with the capacity of WT p53 tetramers to be properly ubiquitylated and result in changes of p53-dependent protein expression patterns, including the pro-apoptotic proteins Bax and PUMA under basal and adriamycin-induced conditions. Importantly, the patient derived p53 OD mutant L330R (OD1) showed the more severe changes in p53-dependent gene expression. Thus, in addition to the well-known effects on p53 stability, ubiquitylation defects promote changes in p53-dependent gene expression with implications on some of its functions.

Combined fluorescent-chromogenic in situ hybridization for identification and laser microdissection of interphase chromosomes.

Chromosome territories constitute the most conspicuous feature of nuclear architecture, and they exhibit non-random distribution patterns in the interphase nucleus. We observed that in cell nuclei from humans with Down Syndrome two chromosomes 21 frequently localize proximal to one another and distant from the third chromosome. To systematically investigate whether the proximally positioned chromosomes were always the same in all cells, we developed an approach consisting of sequential FISH and CISH combined with laser-microdissection of chromosomes from the interphase nucleus and followed by subsequent chromosome identification by microsatellite allele genotyping. This approach identified proximally positioned chromosomes from cultured cells, and the analysis showed that the identity of the chromosomes proximally positioned varies. However, the data suggest that there may be a tendency of the same chromosomes to be positioned close to each other in the interphase nucleus of trisomic cells. The protocol described here represents a powerful new method for genome analysis.

trans-10,cis-12 Conjugated linoleic acid inhibits lipoprotein lipase but increases the activity of lipogenic enzymes in adipose tissue from hamsters fed an atherogenic diet.

The aim of the present work was to investigate the effects of trans-10,cis-12 conjugated linoleic acid (CLA) on the activity and expression of lipogenic enzymes and lipoprotein lipase (LPL), as well as on the expression of transcriptional factors controlling these enzymes, in adipose tissue from hamsters, and to evaluate the involvement of these changes in the body fat-reducing effect of this CLA isomer. Thirty male hamsters were divided into three groups and fed atherogenic diets supplemented with 0 (linoleic group), 5 or 10 g trans-10,cis-12 CLA/kg diet, for 6 weeks. Body and adipose tissue weights, food intake and serum insulin were measured. Total and heparin-releasable LPL and lipogenic enzyme activities (acetyl-CoA carboxylase (ACC); fatty acid synthase (FAS); glucose-6-phosphate dehydrogenase (G6PDH); and malic enzyme (ME)) were assessed. ACC, FAS, LPL, sterol regulatory element-binding proteins (SREBP-1a), SREBP-1c and PPARgamma mRNA levels were also determined by real-time PCR. CLA did not modify food intake, body weight and serum insulin level. CLA feeding reduced adipose tissue weight, LPL activity and expression, and increased lipogenic enzyme activities, despite a significant reduction in ACC and FAS mRNA levels. The expression of the three transcriptional factors analysed (SREBP-1a, SREBP-1c and PPARgamma) was also reduced. These results appear to provide a framework for partially understanding the reduction in body fat induced by CLA. Inhibition of LPL activity seems to be an important mechanism underlying body fat reduction in hamsters. Further research is needed to better characterize the effects of CLA on lipogenesis and the role of these effects in CLA action.

Effects of conjugated linoleic acid on skeletal muscle triacylglycerol metabolism in hamsters.

OBJECTIVE: The present work evaluated the effects of conjugated linoleic acid (CLA) on various aspects of triacylglycerol metabolism in skeletal muscle to determine the potential involvement of this tissue in the effect of CLA to decrease body fat. METHODS: Animals were randomized to three groups that were fed atherogenic diets that provided different amounts of trans-10,cis-12 CLA (0%, 0.5%, or 1%) for 6 wk. Muscle triacylglycerol, protein, water, glycogen, and DNA contents and fatty acid profile in triacylglycerols were analyzed. Lipoprotein lipase and carnitine palmitoyltransferase-I (CPT-I) activities were assessed. Triacylglycerol, glucose, and insulin concentrations were evaluated in serum. RESULTS: The high dose of CLA increased food efficiency and gastrocnemius muscle weight. CLA feeding resulted in decreased muscle triacylglycerol content without changes in protein, water, glycogen, and DNA contents or in cell size (protein/DNA ratio) and produced decreased lipoprotein lipase activity and increased CPT-I activity. No differences were found between CLA doses. CLA feeding led to the saturation of stored triacylglycerols. CONCLUSIONS: Decreased fatty acid uptake and increased fatty acid oxidation can contribute to the decreased muscle triacylglycerol content observed in hamsters fed the CLA diets. The increase in muscle fatty acid beta-oxidation might ultimately prevent storage of triacylglycerols in adipose tissue. Nevertheless, the lack of matching of lipoprotein lipase and CPT-I modifications makes it difficult to ensure that skeletal muscle is responsible, at least in part, for the effect of CLA on decreasing body fat; thus, further research is needed.

Effects of conjugated linoleic acid on liver composition and fatty acid oxidation are isomer-dependent in hamster.

OBJECTIVE: The present work was designed to study the effects of the two main isomers of conjugated linoleic acid (CLA), cis-9,trans-11 and trans-10,cis-12, on liver composition and hepatic fatty acid oxidation in hamsters. METHODS: Animals were divided into three groups that were fed atherogenic diets supplemented with 0.5% linoleic acid, cis-9,trans-11 CLA, or trans-10,cis-12 CLA for 6 wk. Liver lipids, protein, water and DNA contents, and histologic structure were analyzed. Hepatic carnitine palmitoyltransferase-I and acyl coenzyme A oxidase activities were assessed. Triacylglycerol concentration, and aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, and alkaline phosphatase activities were evaluated in serum. CLA isomer contents were analyzed by gas chromatography in hepatic triacylglycerols. Peroxisome proliferator-activated receptor-alpha mRNA was determined by reverse transcriptase polymerase chain reaction. RESULTS: Trans-10,cis-12 CLA led to significantly greater weight, lower levels of triacylglycerol, cholesterol, and phospholipid, and larger total cell number in liver. Carnitine palmitoyltransferase-I and acyl coenzyme A oxidase activities were significantly increased by this isomer. No changes were induced by cis-9,trans-11 CLA. Trans-10,cis-12 CLA was recovered in significantly lower proportions than cis-9,trans-11 in liver triacylglycerols. Histopathologic analysis showed no abnormalities. No significant differences in serum aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, and alkaline phosphatase activities or in hepatic mRNA peroxisome proliferator-activated receptor-alpha expression were found among the three experimental groups. CONCLUSIONS: These results suggest that the addition of 0.5% of these CLA isomers to the diet do not induce toxic effects in liver after 6 wk of feeding. Intake of trans-10,cis-12 isomer but not of cis-9,trans-11 CLA increases liver fatty acid oxidation. This effect leads to decreased hepatic and serum triacylglycerols.

The trans-10,cis-12 isomer of conjugated linoleic acid reduces hepatic triacylglycerol content without affecting lipogenic enzymes in hamsters.

Conjugated linoleic acid (CLA) refers to the positional and geometric dienoic isomers of linoleic acid. The dietary intake of CLA has been associated with changes in lipid metabolism. The aim of the present work was to assess the effects of the two main isomers of CLA on sterol regulatory element binding protein (SREBP)-1a and SREBP-1c mRNA levels, as well as on mRNA levels and the activities of several lipogenic enzymes in liver. For this purpose hamsters were fed an atherogenic diet supplemented with 5 g linoleic acid, cis-9,trans-11 or trans-10,cis-12 CLA/kg diet for 6 weeks. The trans-10,cis-12 isomer intake produced significantly greater liver weight, but also significantly decreased liver fat accumulation. No changes in mRNA levels of SREBP-1a, SREBP-1c and lipogenic enzymes, or in the activities of these enzymes, were observed. There was no effect of feeding cis-9,trans-11 CLA. These results suggest that increased fat accumulation in liver does not occur on the basis of liver enlargement produced by feeding the trans-10,cis-12 isomer of CLA in hamsters. The reduction in hepatic triacylglycerol content induced by this isomer was not attributable to changes in lipogenesis.