Osteoarthritis Affects Mammalian Oogenesis: Effects of Collagenase-Induced Osteoarthritis on Oocyte Cytoskeleton in a Mouse Model

Known as a degenerative joint disorder of advanced age affecting predominantly females, osteoarthritis can develop in younger and actively working people because of activities involving loading and injuries of joints. Collagenase-induced osteoarthritis (CIOA) in a mouse model allowed us to investigate for the first time its effects on key cytoskeletal structures (meiotic spindles and actin distribution) of ovulated mouse oocytes. Their meiotic spindles, actin caps, and chromatin were analyzed by immunofluorescence. A total of 193 oocytes from mice with CIOA and 209 from control animals were obtained, almost all in metaphase I (M I) or metaphase II (MII). The maturation rate was lower in CIOA (26.42% M II) than in controls (55.50% M II). CIOA oocytes had significantly larger spindles (average 37 μm versus 25 μm in controls, p < 0.001 ), with a proportion of large spindles more than 64% in CIOA versus up to 15% in controls ( p < 0.001 ). Meiotic spindles were wider in 68.35% M I and 54.90% M II of CIOA oocytes (mean 18.04 μm M I and 17.34 μm M II versus controls: 11.64 μm M I and 12.64 μm M II), and their poles were approximately two times broader (mean 6.9 μm) in CIOA than in controls (3.6 μm). CIOA oocytes often contained disoriented microtubules. Actin cap was visible in over 91% of controls and less than 20% of CIOA oocytes. Many CIOA oocytes without an actin cap had a nonpolarized thick peripheral actin ring (61.87% of M I and 52.94% of M II). Chromosome alignment was normal in more than 82% in both groups. In conclusion, CIOA affects the cytoskeleton of ovulated mouse oocytes—meiotic spindles are longer and wider, their poles are broader and with disorganized fibers, and the actin cap is replaced by a broad nonpolarized ring. Nevertheless, meiotic spindles were successfully formed in CIOA oocytes and, even when abnormal, allowed correct alignment of chromosomes.

An Application of the Eigenproblem for Biochemical Similarity

Protein alignment finds its application in refining results of sequence alignment and understanding protein function. A previous study aligned single molecules, making use of the minimization of sums of the squares of eigenvalues, obtained for the antisymmetric Cartesian coordinate distance matrices Dx and Dy. This is used in our program to search for similarities between amino acids by comparing the sums of the squares of eigenvalues associated with the Dx, Dy, and Dz distance matrices. These matrices are obtained by removing atoms that could lead to low similarity. Candidates are aligned, and trilateration is used to attach all previously striped atoms. A TM-score is the scoring function that chooses the best alignment from supplied candidates. Twenty essential amino acids that take many forms in nature are selected for comparison. The correct alignment is taken into account most of the time by the alignment algorithm. It was numerically detected by the TM-score 70% of the time, on average, and 15% more cases with close scores can be easily distinguished by human observation.

A dual mechanism of action of AT-527 against SARS-CoV-2 polymerase

A worldwide effort is ongoing to discover drugs against the Severe Acute Respiratory Syndrome coronavirus type 2 (SARS-CoV-2), which has so far caused >3.5 million fatalities (https://covid19.who.int/). The virus essential RNA-dependent RNA polymerase complex is targeted by several nucleoside/tide analogues whose mechanisms of action and clinical potential are currently evaluated. The guanosine analogue AT-527, a double prodrug of its 5'-triphosphate AT-9010, is currently in phase III clinical trials as a COVID19 treatment. Here we report the cryo-EM structure at 2.98 Å resolution of the SARS-CoV-2 nsp12-nsp7-(nsp8)2 complex with RNA showing AT-9010 bound at three sites of nsp12. At the RdRp active-site, one AT-9010 is incorporated into the RNA product. Its 2'-methyl group prevents correct alignment of a second AT-9010 occupying the incoming NTP pocket. The 2'-F, 2'-methyl 3'-OH ribose scaffold explains the non-obligate RNA chain-termination potency of this NA series for both HCV NS5 and SARS-CoV RTCs. A third AT-9010 molecule 5'-diphosphate binds to a coronavirus-specific pocket in the nsp12 N-terminus NiRAN domain, a SelO pseudo-kinase structural and functional homologue. This unique binding mode impedes NiRAN-mediated UMPylation of SARS-CoV-2 nsp8 and nsp9 proteins. Our results suggest a mechanism of action for AT-527 in line with a therapeutic use for COVID19.

An Improved Procedure for Strongly Coupled Prediction of Sailing Yacht Performance

In this paper, an improved procedure for strongly coupled prediction of sailing yacht performance is developed. The procedure uses 3D RANS CFD to compute the hydrodynamic forces. When coupled to a rigid body motion solver and a sail force model, along with a rudder control algorithm, this allows sailing yacht performance to be predicted within CFD software. The procedure provides improved convergence when compared to a previously published method. The grid motion scheme, partially using overset grid techniques, means that correct alignment between the free surface and the background grid is ensured even at large heel angles. The capabilities are demonstrated with performance predictions for the SYRF 14 m yacht, at one true wind speed, over a range of true wind angles, with up- and downwind sailsets. The results are compared to predictions from the ORC-VPP for a yacht with similar main particulars.

A polarized nucleus-cytoskeleton-ECM connection in migrating cardioblasts controls heart tube formation

The formation of the cardiac tube is a remarkable example of complex morphogenetic processes conserved from invertebrates to humans. It involves coordinated collective migration of contralateral rows of cardiac cells. The molecular processes underlying the specification of cardioblasts (CBs) prior to migration are well established and significant advances have been made in understanding the process of lumen formation. However, the mechanisms of collective cardiac cells migration remain elusive. Here we identified CAP and MSP300 as novel actors involved during CBs migration. They both exhibit highly similar temporal and spatial expression patterns in migrating cardiac cells and are necessary for the correct number and alignment of CBs, a prerequisite for the coordination of their collective migration. Our data suggest that CAP and MSP300 are part of a protein complex linking focal adhesion sites to nuclei via the actin cytoskeleton that maintains post-mitotic state and correct alignment of CBs.

CHARACTERIZING DRIVER TAKE-OVER ACCURACY: EFFECT OF AGE, SEX, STARTLE, AND SECONDARY TASK

The Acoustic Startling Pre-stimulus (ASPS, i.e. a loud sound preceding a physical perturbation was previously found to accelerate take-over actions in adults but not teens in autonomous vehicle scenarios. It is not clear if the ASPS also influences the accuracy of the take-over response across ages and sexes. Therefore the aims of this study are: to characterize take-over accuracy across age/experience and sex and to examine the effect of the ASPS and a secondary task on steering wheel alignment in autonomous vehicle take-over scenarios. Fourteen adult (7 males and 13 teenage (6 males drivers volunteered for this study. Participants were instructed to align a marker on the steering wheel with a marker on a lateral post as fast as they could, when a sled perturbation started. Two of the conditions included the ASPS. Two of the conditions involved mobile texting while the sled started moving. The angle between the steering wheel and the lateral post was used to quantify overshooting, undershooting, or correct alignment during steering. Results showed that adult female subjects reached correct alignment slightly more frequently than any other group, while male adult drivers decreased their alignment error after the first trial. Both female and male adult drivers had a reduced alignment angle when the first trial had an ASPS compared to when the first trial had no ASPS while teen drivers performed similarly with ASPS or without. This study showed that take-over accuracy and steering control are influenced by sex, age/experience, and a startle-based warning.

On the determination of residual stresses in additively manufactured lattice structures

The determination of residual stresses becomes more complicated with increasing complexity of the structures investigated. Additive manufacturing techniques generally allow the production of `lattice structures' without any additional manufacturing step. These lattice structures consist of thin struts and are thus susceptible to internal stress-induced distortion and even cracks. In most cases, internal stresses remain locked in the structures as residual stress. The determination of the residual stress in lattice structures through nondestructive neutron diffraction is described in this work. It is shown how two difficulties can be overcome: (a) the correct alignment of the lattice structures within the neutron beam and (b) the correct determination of the residual stress field in a representative part of the structure. The magnitude and the direction of residual stress are discussed. The residual stress in the strut was found to be uniaxial and to follow the orientation of the strut, while the residual stress in the knots was more hydrostatic. Additionally, it is shown that strain measurements in at least seven independent directions are necessary for the estimation of the principal stress directions. The measurement directions should be chosen according to the sample geometry and an informed choice on the possible strain field. If the most prominent direction is not measured, the error in the calculated stress magnitude increases considerably.

Femoral Torsion in CT: Why We Should Add Some Trigonometry

Purpose: In our department, we routinely perform Computed Tomography (CT) or Magnetic Resonance Imaging (MRI) of the lower extremity to assess femoral torsion for preoperative planning prior correcting osteotomies. It might be difficult to assess the FA, because on axial images the depicted part of the femoral neck is too short to allow correct alignment of the axis of the femoral neck. Measurements can also be performed on oblique axial images with improvement of depiction of the femoral neck but are smaller than those of measurements on axial images, due to the fact that images depend on rules of trigonometry. The aim is to provide a trigonometrical formula to calculate the value for FA, allowing precise assessment of femoral anteversion on oblique axial images similar to those on axial images. Materials and Methods: Trigonometrical transformation is performed in three steps. Initial measurement of femoral anteversion on oblique images is transformed via a corresponding rectangular triangle from coronal images to calculated femoral anteversion on axial images. Results: The first triangle is labelled with a1, b1, c1 for the sides and alpha1, beta1for the angles. Second and third triangle is labelled correspondingly. Length of both cathetus a1 and b1 are calculated as follows: cathetus a1=sin alpha1*c1 and cathetus b1=cos alpha 1*c1. Cathetus b2 is calculated as follows: b2=cos alpha2*c2. alpha3=tan-1 (a3/b3). Initial calculation of angle alpha3=tan-1 (a3/b3)=tan-1 (a1/b2), with a3=a1=sin alpha1*c1 and b3=b2=cos alpha2*b1*c, with b1=cos alpha1*c1 and b3=cos alpha2*cos alpha1*c1². The final formula is then: alpha3=tan-1 ((sin alpha1*c1)/(cos alpha²*cos alpha1*c12)) = tan-1 ((sin alpha1/(alpha2*cos alpha1*c1)) Conclusion: In this study we can provide a formula: alpha3=tan-1 ((sin alpha1/(cos alpha2*cos alpha1*c1)), which allows to calculate the femoral anteversion for true axial reconstructed images with the increased accuracy of measurements on oblique images and the ability to use the already known reference values from the literature.

THE EFFECTIVENESS OF MYOFASCIAL RELEASE IN THE CERVICAL REGION FOR TENSION HEADACHE RELIEF

Myofascial release is a manual thrust applied to the body face to treat the facial system. Myofascial release restores the flexibility and elasticity of the fascia, thus having a broad ability to alter the body's range of motion and dissolve restrictions in the fascial tissues. Sedentary lifestyle and positions that cause pain can gradually cause muscle and fascial restrictions and shortening. This study aims to identify the effectiveness of myofascial release in the cervical region for tension headache relief. This study is characterized as a bibliographical research of qualitative descriptive character, where the researcher is the one who describes the research object, assuming the role of observer and explorer, directly collecting the data in the place where the study phenomena occurred or appeared. It is concluded that through the use of this technique, it is possible to deactivate trigger points, promote a correct alignment of the body structure, minimize movement restriction and pain without compromising the myofascial structures.

Planning Brain Tumor Resection Using a Probabilistic Atlas of Cortical and Subcortical Structures Critical for Functional Processing: A Proof of Concept

BACKGROUND Functional preoperative planning for resection of intrinsic brain tumors in eloquent areas is still a challenge. Predicting subcortical functional framework is especially difficult. Direct electrical stimulation (DES) is the recommended technique for resection of these lesions. A reliable probabilistic atlas of the critical cortical epicenters and subcortical framework based on DES data was recently published. OBJECTIVE To propose a pipeline for the automated alignment of the corticosubcortical maps of this atlas with T1-weighted MRI. METHODS To test the alignment, we selected 10 patients who underwent resection of brain lesions by using DES. We aligned different cortical and subcortical functional maps to preoperative volumetric T1 MRIs (with/without gadolinium). For each patient we quantified the quality of the alignment, and we calculated the match between the location of the functional sites found at DES and the functional maps of the atlas. RESULTS We found an accurate brain extraction and alignment of the functional maps with both the T1 MRIs of each patient. The matching analysis between functional maps and functional responses collected during surgeries was 88% at cortical and, importantly, 100% at subcortical level, providing a further proof of the correct alignment. CONCLUSION We demonstrated quantitatively and qualitatively the reliability of this tool that may be used for presurgical planning, providing further functional information at the cortical level and a unique probabilistic prevision of distribution of the critical subcortical structures. Finally, this tool offers the chance for multimodal planning through integrating this functional information with other neuroradiological and neurophysiological techniques.