Congruence between NOTCH3 mutations and GOM in 131 CADASIL patients

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common hereditary subcortical vascular dementia. It is caused by mutations in NOTCH3 gene, which encodes a large transmembrane receptor Notch3. The key pathological finding is the accumulation of granular osmiophilic material (GOM), which contains extracellular domains of Notch3, on degenerating vascular smooth muscle cells (VSMCs). GOM has been considered specifically diagnostic for CADASIL, but the reports on the sensitivity of detecting GOM in patients’ skin biopsy have been contradictory. To solve this contradiction, we performed a retrospective investigation of 131 Finnish, Swedish and French CADASIL patients, who had been adequately examined for both NOTCH3 mutation and presence of GOM. The patients were examined according to the diagnostic practice in each country. NOTCH3 mutations were assessed by restriction enzyme analysis of specific mutations or by sequence analysis. Presence of GOM was examined by electron microscopy (EM) in skin biopsies. Biopsies of 26 mutation-negative relatives from CADASIL families served as the controls. GOM was detected in all 131 mutation positive patients. Altogether our patients had 34 different pathogenic mutations which included three novel point mutations (p.Cys67Ser, p.Cys251Tyr and p.Tyr1069Cys) and a novel duplication (p.Glu434_Leu436dup). The detection of GOM by EM in skin biopsies was a highly reliable diagnostic method: in this cohort the congruence between NOTCH3 mutations and presence of GOM was 100%. However, due to the retrospective nature of this study, exact figure for sensitivity cannot be determined, but it would require a prospective study to exclude possible selection bias. The identification of a pathogenic NOTCH3 mutation is an indisputable evidence for CADASIL, but demonstration of GOM provides a cost-effective guide for estimating how far one should proceed with the extensive search for a new or an uncommon mutations among the presently known over 170 different NOTCH3 gene defects. The diagnostic skin biopsy should include the border zone between deep dermis and upper subcutis, where small arterial vessels of correct size are located. Detection of GOM requires technically adequate biopsies and distinction of true GOM from fallacious deposits. If GOM is not found in the first vessel or biopsy, other vessels or additional biopsies should be examined

Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy: A Genetic Cause of Cerebral Small Vessel Disease

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a single-gene disorder of the cerebral small blood vessels caused by mutations in the Notch3 gene. The exact prevalence of this disorder was unknown currently, and the number of reported CADASIL families is steadily increasing as the clinical picture and diagnostic examinations are becoming more widely known. The main clinical manifestations are recurrent stroke, migraine, psychiatric symptoms, and progressive cognitive impairment. The clinical course of CADASIL is highly variable, even within families. The involvement of the anterior temporal lobe and the external capsule on brain magnetic resonance imaging was found to have high sensitivity and specificity in differentiating CADASIL from the much more common sporadic cerebral small-vessel disease (SVD). The pathologic hallmark of the disease is the presence of granular osmiophilic material in the walls of affected vessels. CADASIL is a prototype single-gene disorder that has evolved as a unique model for studying the mechanisms underlying cerebral SVD. At present, the incidence and prevalence of CADASIL seem to be underestimated due to limitations in clinical, neuroradiological, and genetic diagnoses of this disorder

A long term numerical solution for the insolation quantities of the Earth

22 may 2004We present here a new solution for the astronomical computation of the insolation quantities on Earth spanning from -250 Myr to 250 Myr. This solution has been improved with respect to La93 (Laskar et al, 1993) by using a direct integration of the gravitational equations for the orbital motion, and by improving the dissipative contributions, in particular in the evolution of the Earth-Moon System. The orbital solution has been used for the calibration of the Neogene period (Lourens \etal, 2004), and is expected to be used for age calibrations of paleoclimatic data over 40 to 50 Myr, eventually over the full Palaeogene period (65 Myr) with caution. Beyond this time span, the chaotic evolution of the orbits prevents a precise determination of the Earth motion. However, the most regular components of the orbital solution could still be used over a much longer time span, which is why we provide here the solution over 250 Myr. Over this time interval, the most striking feature of the obliquity solution, apart from a secular global increase due to tidal dissipation, is a strong decrease of about $0.38$ degree in the next few millions of years, due to the crossing of the $s_6+g_5-g_6$ resonance (Laskar et al, 1993). For the calibration of the Mesozoic time scale (about 65 to 250 Myr), we propose to use the term of largest amplitude in the eccentricity, related to $g_2-g_5$, with a fixed frequency of $3.200$ ''/yr, corresponding to a period of 405000 yr. The uncertainty of this time scale over 100 Myr should be about $0.1\%$, and $0.2\%$ over the full Mesozoic era

Long term evolution and chaotic diffusion of the insolation quantities of Mars.

As the obliquity of Mars is strongly chaotic, it is not possible to give a solution for its evolution over more than a few million of years. Using the most recent data for the rotational state of Mars, and a new numerical integration of the Solar System, we provide here a precise solution for the evolution of Mars spin over 10 to 20 Myr.Over 250 Myr, we present a statistical study of its possible evolution, when considering the uncertainties in the present rotational state. Over much longer time span, reaching 5 Gyr, the chaotic diffusion prevails, and we have performed an extensive statistical analysis of the orbital and rotational evolution of Mars, relying on Laskar's secular solution of the Solar System, based on more than 600 orbital and 200 000 obliquity solutions over 5 Gyr.The density function of the eccentricity and obliquity are explicited with simple analytical formulas.We found an averaged eccentricity of Mars over 5 Gyr of e=0.0690 with standard deviation s_e=0.0299, while the averaged value of the obliquity is 37.62 deg with a standard deviation of 13.82 deg and a maximal value of 82.035 deg.We find that the probability for Mars obliquity to have reached more than 60 degin the past 1 Gyr is 63.0%, and 89.3% in 3 Gyr. Over 4 Gyr, the position of Mars axis is given by a uniform distribution on a spherical cap limited by the obliquity 58.62 deg, with the addition of a random noise allowing a slow diffusion beyond this limit. We can also define a standard model of Mars insolation parameters over 4 Gyr with the most probable values for the eccentricity 0.068 and 41.80 deg for the obliquity

Deep-learning based segmentation of challenging myelin sheaths

The segmentation of axons and myelin in electron microscopy images allows neurologists to highlight the density of axons and the thickness of the myelin surrounding them. These properties are of great interest for preventing and anticipating white matter diseases. This task is generally performed manually, which is a long and tedious process. We present an update of the methods used to compute that segmentation via machine learning. Our model is based on the architecture of the U-Net network. Our main contribution consists in using transfer learning in the encoder part of the UNet network, as well as test time augmentation when segmenting. We use the SE-Resnet50 backbone weights which was pre-trained on the ImageNet 2012 dataset. We used a data set of 23 images with the corresponding segmented masks, which also was challenging due to its extremely small size. The results show very encouraging performances compared to the state-of-the-art with an average precision of 92% on the test images. It is also important to note that the available samples were taken from elderly mices in the corpus callosum. This represented an additional difficulty, compared to related works that had samples taken from the spinal cord or the optic nerve of healthy individuals, with better contours and less debri

Characterization of early ultrastructural changes in the cerebral white matter of CADASIL small vessel disease using high pressure freezing/freeze-substitution

AIMS: The objective of this study was to elucidate the early white matter changes in CADASIL small vessel disease. METHODS: We used high pressure freezing and freeze substitution (HPF/FS) in combination with high resolution electron microscopy (EM), immunohistochemistry and confocal microscopy of brain specimens from control and CADASIL (TgNotch3R169C ) mice aged 4 to 15 months to study white matter lesions in the corpus callosum. RESULTS: We first optimized the HPF/FS protocol in which samples were chemically prefixed, frozen in a sample carrier filled with 20% polyvinylpyrrolidone and freeze-substituted in a cocktail of tannic acid, osmium tetroxide and uranyl acetate dissolved in acetone. EM analysis showed that CADASIL mice exhibit significant splitting of myelin layers and enlargement of the inner tongue of small calibre axons from the age of 6 months, then vesiculation of the inner tongue and myelin sheath thinning at 15 months of age. Immunohistochemistry revealed an increased number of oligodendrocyte precursor cells, although only in older mice, but no reduction in the number of mature oligodendrocytes at any age. The number of Iba1 positive microglial cells was increased in older but not in younger CADASIL mice, but the number of activated microglial cells (Iba1 and CD68 positive) was unchanged at any age. CONCLUSION: We conclude that early WM lesions in CADASIL affect first and foremost the myelin sheath and the inner tongue, suggestive of a primary myelin injury. We propose that those defects are consistent with a hypoxic/ischaemic mechanism