A polymorphism at codon 31 of gene p21 is not associated with primary open angle glaucoma in Caucasians

BACKGROUND: Primary open angle glaucoma (POAG) is considered to be a neurodegenerative optic neuropathy, in which cell death occurs by apoptosis. p21, is an important protective component of the apoptotic pathway, regulating cellular arrest in the presence of DNA damage. An unstable or altered p21 protein could modify the cellular response to genomic injury and abolish the effect of p21. A previous study on a Chinese cohort suggested that the p21 codon 31 polymorphism may alter the state of apoptosis in glaucomatous optic neuropathy, failing to protect the ganglion cells. The aim of this study was to test the hypothesis that a p21 codon 31 polymorphism is associated with POAG on a Caucasian cohort. METHODS: 140 POAG patients and a control group of 73 healthy individuals were included in the study. All the subjects were of Caucasian origin. Genomic DNA was amplified by polymerase chain reaction, followed by enzymatic restriction fragment length polymorphism technique (PCR-RFLP). Patients and controls were genotyped for a single nucleotide polymorphism (C/A transversion) in the third base of codon 31 of p21, which leads to a serine (Ser)/arginine (Arg) substitution. RESULTS: The distribution of the genotypes in the POAG patients showed 128 (91.4%) Ser homozygotes, 10 (7.1%) Ser/Arg heterozygotes and 2 (1.5%) Arg homozygotes. In the control cohort, there were 61 (83.6%) Ser homozygotes and 12 (16.4%) Ser/Arg heterozygotes. No Arg homozygotes were present amongst the control group. Both the allelic and genotypic frequencies of the Ser or Arg residues at codon 31 were not significantly different between POAG patients and controls (Fisher's exact test, P = 0.20 for alleles and P = 0.0561 for genotypes). CONCLUSION: This study suggests that the p21 codon 31 polymorphism does not contribute to the risk of POAG in the Caucasian population

Impaired neural development in a zebrafish model for Lowe syndrome

Lowe syndrome, which is characterized by defects in the central nervous system, eyes and kidneys, is caused by mutation of the phosphoinositide 5-phosphatase OCRL1. The mechanisms by which loss of OCRL1 leads to the phenotypic manifestations of Lowe syndrome are currently unclear, in part, owing to the lack of an animal model that recapitulates the disease phenotype. Here, we describe a zebrafish model for Lowe syndrome using stable and transient suppression of OCRL1 expression. Deficiency of OCRL1, which is enriched in the brain, leads to neurological defects similar to those reported in Lowe syndrome patients, namely increased susceptibility to heat-induced seizures and cystic brain lesions. In OCRL1-deficient embryos, Akt signalling is reduced and there is both increased apoptosis and reduced proliferation, most strikingly in the neural tissue. Rescue experiments indicate that catalytic activity and binding to the vesicle coat protein clathrin are essential for OCRL1 function in these processes. Our results indicate a novel role for OCRL1 in neural development, and support a model whereby dysregulation of phosphoinositide metabolism and clathrin-mediated membrane traffic leads to the neurological symptoms of Lowe syndrome

Dynamic Changes in Arabinogalactan-Protein, Pectin, Xyloglucan and Xylan Composition o the Cell Wall During Microspore Embryogenesis in Brassica napus

[EN] Microspore embryogenesis is a manifestation of plant cell totipotency whereby new cell walls are formed as a consequence of the embryogenic switch. In particular, the callose-rich subintinal layer created immediately upon induction of embryogenesis was recently related to protection against stress. However, little is currently known about the functional significance of other compositional changes undergone by the walls of embryogenic microspores. We characterized these changes in Brassica napus at different stages during induction of embryogenic microspores and development of microspore-derived embryos (MDEs) by using a series of monoclonal antibodies specific for cell wall components, including arabinogalactan-proteins (AGPs), pectins, xyloglucan and xylan. We used JIM13, JIM8, JIM14 and JIM16 for AGPs, CCRC-M13, LM5, LM6, JIM7, JIM5 and LM7 for pectins, CCRC-M1 and LM15 for xyloglucan, and LM11 for xylan. By transmission electron microscopy and quantification of immunogold labeling on high-pressure frozen, freeze-substituted samples, we profiled the changes in cell wall ultrastructure and composition at the different stages of microspore embryogenesis. As a reference to compare with, we also studied in vivo microspores and maturing pollen grains. We showed that the cell wall of embryogenic microspores is a highly dynamic structure whose architecture, arrangement and composition changes dramatically as microspores undergo embryogenesis and then transform into MDEs. Upon induction, the composition of the preexisting microspore intine walls is remodeled, and unusual walls with a unique structure and composition are formed. Changes in AGP composition were related to developmental fate. In particular, AGPs containing the JIM13 epitope were massively excreted into the cell apoplast, and appeared associated to cell totipotency. According to the ultrastructure and the pectin and xyloglucan composition of these walls, we deduced that commitment to embryogenesis induces the formation of fragile, plastic and deformable cell walls, which allow for cell expansion and microspore growth. We also showed that these special walls are transient, since cell wall composition in microspore-derived embryos was completely different. Thus, once adopted the embryogenic developmental pathway and far from the effects of heat shock exposure, cell wall biosynthesis would approach the structure, composition and properties of conventional cell walls.This work was supported by grant AGL2017-88135-R to JS-S from Spanish MINECO jointly funded by FEDER. AD would like to thank the University of Rouen and Normandie Regional Council (France) for financial support.Corral Martínez, P.; Driouich, A.; Seguí-Simarro, JM. (2019). Dynamic Changes in Arabinogalactan-Protein, Pectin, Xyloglucan and Xylan Composition o the Cell Wall During Microspore Embryogenesis in Brassica napus. Frontiers in Plant Science. 10:1-17. https://doi.org/10.3389/fpls.2019.00332S11710Barany, I., Fadon, B., Risueno, M. C., & Testillano, P. S. (2010). Cell wall components and pectin esterification levels as markers of proliferation and differentiation events during pollen development and pollen embryogenesis in Capsicum annuum L. Journal of Experimental Botany, 61(4), 1159-1175. doi:10.1093/jxb/erp392Daher, F. B., & Braybrook, S. A. (2015). How to let go: pectin and plant cell adhesion. Frontiers in Plant Science, 6. doi:10.3389/fpls.2015.00523Cavalier, D. M., Lerouxel, O., Neumetzler, L., Yamauchi, K., Reinecke, A., Freshour, G., … Keegstra, K. (2008). Disrupting Two Arabidopsis thaliana Xylosyltransferase Genes Results in Plants Deficient in Xyloglucan, a Major Primary Cell Wall Component. 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Differences in pectic polysaccharides between carrot embryogenic and non-embryogenic calli. Plant Cell Reports, 14(5). doi:10.1007/bf00232028Knox, J. P., Linstead, P., King, J., Cooper, C., & Roberts, K. (1990). Pectin esterification is spatially regulated both within cell walls and between developing tissues of root apices. Planta, 181(4). doi:10.1007/bf00193004Knox, J. ., Linstead, P. ., Cooper, J. P. C., & Roberts, K. (1991). Developmentally regulated epitopes of cell surface arabinogalactan proteins and their relation to root tissue pattern formation. The Plant Journal, 1(3), 317-326. doi:10.1046/j.1365-313x.1991.t01-9-00999.xLamport, D. T. A., & Várnai, P. (2012). Periplasmic arabinogalactan glycoproteins act as a calcium capacitor that regulates plant growth and development. New Phytologist, 197(1), 58-64. doi:10.1111/nph.12005Letarte, J., Simion, E., Miner, M., & Kasha, K. J. (2005). Arabinogalactans and arabinogalactan-proteins induce embryogenesis in wheat (Triticum aestivum L.) microspore culture. Plant Cell Reports, 24(12), 691-698. doi:10.1007/s00299-005-0013-5Majewska-Sawka, A., Münster, A., & Wisniewska, E. (2004). Temporal and Spatial Distribution of Pectin Epitopes in Differentiating Anthers and Microspores of Fertile and Sterile Sugar Beet. Plant and Cell Physiology, 45(5), 560-572. doi:10.1093/pcp/pch066Makowska, K., Kałużniak, M., Oleszczuk, S., Zimny, J., Czaplicki, A., & Konieczny, R. (2017). Arabinogalactan proteins improve plant regeneration in barley (Hordeum vulgare L.) anther culture. Plant Cell, Tissue and Organ Culture (PCTOC), 131(2), 247-257. doi:10.1007/s11240-017-1280-xMarcus, S. E., Verhertbruggen, Y., Hervé, C., Ordaz-Ortiz, J. J., Farkas, V., Pedersen, H. L., … Knox, J. P. (2008). Pectic homogalacturonan masks abundant sets of xyloglucan epitopes in plant cell walls. BMC Plant Biology, 8(1), 60. doi:10.1186/1471-2229-8-60McCartney, L., Marcus, S. E., & Knox, J. P. (2005). Monoclonal Antibodies to Plant Cell Wall Xylans and Arabinoxylans. Journal of Histochemistry & Cytochemistry, 53(4), 543-546. doi:10.1369/jhc.4b6578.2005McCartney, L., Ormerod, andrew P., Gidley, M. J., & Knox, J. P. (2000). Temporal and spatial regulation of pectic (14)-beta-D-galactan in cell walls of developing pea cotyledons: implications for mechanical properties. The Plant Journal, 22(2), 105-113. doi:10.1046/j.1365-313x.2000.00719.xMcCartney, L., Steele-King, C. G., Jordan, E., & Knox, J. P. (2003). Cell wall pectic (1→4)-β-d-galactan marks the acceleration of cell elongation in theArabidopsisseedling root meristem. The Plant Journal, 33(3), 447-454. doi:10.1046/j.1365-313x.2003.01640.xMicheli, F. (2001). Pectin methylesterases: cell wall enzymes with important roles in plant physiology. Trends in Plant Science, 6(9), 414-419. doi:10.1016/s1360-1385(01)02045-3MOHNEN, D. 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(2004). The Galactose Residues of Xyloglucan Are Essential to Maintain Mechanical Strength of the Primary Cell Walls in Arabidopsis during Growth. Plant Physiology, 134(1), 443-451. doi:10.1104/pp.103.027508Pennell, R. I., Janniche, L., Kjellbom, P., Scofield, G. N., Peart, J. M., & Roberts, K. (1991). Developmental Regulation of a Plasma Membrane Arabinogalactan Protein Epitope in Oilseed Rape Flowers. The Plant Cell, 1317-1326. doi:10.1105/tpc.3.12.1317Pereira, A. M., Pereira, L. G., & Coimbra, S. (2015). Arabinogalactan proteins: rising attention from plant biologists. Plant Reproduction, 28(1), 1-15. doi:10.1007/s00497-015-0254-6Pereira-Netto, A. B., Pettolino, F., Cruz-Silva, C. T. A., Simas, F. F., Bacic, A., Carneiro-Leão, A. M. dos A., … Maurer, J. B. B. (2007). Cashew-nut tree exudate gum: Identification of an arabinogalactan-protein as a constituent of the gum and use on the stimulation of somatic embryogenesis. 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Misbehaviour of XIST RNA in Breast Cancer Cells

A role of X chromosome inactivation process in the development of breast cancer have been suggested. In particular, the relationship between the breast cancer predisposing gene BRCA1 and XIST, the main mediator of X chromosome inactivation, has been intensely investigated, but still remains controversial. We investigated this topic by assessing XIST behaviour in different groups of breast carcinomas and in a panel of breast cancer cell lines both BRCA1 mutant and wild type. In addition, we evaluated the occurrence of broader defects of heterochromatin in relation to BRCA1 status in breast cancer cells. We provide evidence that in breast cancer cells BRCA1 is involved in XIST regulation on the active X chromosome, but not in its localization as previously suggested, and that XIST can be unusually expressed by an active X and can decorate it. This indicates that the detection of XIST cloud in cancer cell is not synonymous of the presence of an inactive X chromosome. Moreover, we show that global heterochromatin defects observed in breast tumor cells are independent of BRCA1 status. Our observations sheds light on a possible previously uncharacterized mechanism of breast carcinogenesis mediated by XIST misbehaviour, particularly in BRCA1-related cancers. Moreover, the significant higher levels of XIST-RNA detected in BRCA1-associated respect to sporadic basal-like cancers, opens the possibility to use XIST expression as a marker to discriminate between the two groups of tumors

Lowe Syndrome Protein OCRL1 Supports Maturation of Polarized Epithelial Cells

Mutations in the inositol polyphosphate 5-phosphatase OCRL1 cause Lowe Syndrome, leading to cataracts, mental retardation and renal failure. We noted that cell types affected in Lowe Syndrome are highly polarized, and therefore we studied OCRL1 in epithelial cells as they mature from isolated individual cells into polarized sheets and cysts with extensive communication between neighbouring cells. We show that a proportion of OCRL1 targets intercellular junctions at the early stages of their formation, co-localizing both with adherens junctional components and with tight junctional components. Correlating with this distribution, OCRL1 forms complexes with junctional components α-catenin and zonula occludens (ZO)-1/2/3. Depletion of OCRL1 in epithelial cells growing as a sheet inhibits maturation; cells remain flat, fail to polarize apical markers and also show reduced proliferation. The effect on shape is reverted by re-expressed OCRL1 and requires the 5′-phosphatase domain, indicating that down-regulation of 5-phosphorylated inositides is necessary for epithelial development. The effect of OCRL1 in epithelial maturation is seen more strongly in 3-dimensional cultures, where epithelial cells lacking OCRL1 not only fail to form a central lumen, but also do not have the correct intracellular distribution of ZO-1, suggesting that OCRL1 functions early in the maturation of intercellular junctions when cells grow as cysts. A role of OCRL1 in junctions of polarized cells may explain the pattern of organs affected in Lowe Syndrome

Author Correction: The landscape of viral associations in human cancers

Correction to: Nature Genetics https://doi.org/10.1038/s41588-019-0558-9, published online 05 February 2020

Dyadic plasticity in cardiomyocytes

Contraction of cardiomyocytes is dependent on sub-cellular structures called dyads, where invaginations of the surface membrane (t-tubules) form functional junctions with the sarcoplasmic reticulum (SR). Within each dyad, Ca2+ entry through t-tubular L-type Ca2+ channels (LTCCs) elicits Ca2+ release from closely apposed Ryanodine Receptors (RyRs) in the SR membrane. The efficiency of this process is dependent on the density and macroscale arrangement of dyads, but also on the nanoscale organization of LTCCs and RyRs within them. We presently review accumulating data demonstrating the remarkable plasticity of these structures. Dyads are known to form gradually during development, with progressive assembly of both t-tubules and junctional SR terminals, and precise trafficking of LTCCs and RyRs. While dyads can exhibit compensatory remodeling when required, dyadic degradation is believed to promote impaired contractility and arrythmogenesis in cardiac disease. Recent data indicate that this plasticity of dyadic structure/function is dependent on the regulatory proteins junctophilin-2, amphiphysin-2 (BIN1), and caveolin-3, which critically arrange dyadic membranes while stabilizing the position and activity of LTCCs and RyRs. Indeed, emerging evidence indicates that clustering of both channels enables “coupled gating”, implying that nanoscale localization and function are intimately linked, and may allow fine-tuning of LTCC-RyR crosstalk. We anticipate that improved understanding of dyadic plasticity will provide greater insight into the processes of cardiac compensation and decompensation, and new opportunities to target the basic mechanisms underlying heart disease

Genomic imbalances in 5918 malignant epithelial tumors: an explorative meta-analysis of chromosomal CGH data

BACKGROUND: Chromosomal abnormalities have been associated with most human malignancies, with gains and losses on some genomic regions associated with particular entities. METHODS: Of the 15429 cases collected for the Progenetix molecular-cytogenetic database, 5918 malignant epithelial neoplasias analyzed by chromosomal Comparative Genomic Hybridization (CGH) were selected for further evaluation. For the 22 clinico-pathological entities with more than 50 cases, summary profiles for genomic imbalances were generated from case specific data and analyzed. RESULTS: With large variation in overall genomic instability, recurring genomic gains and losses were prominent. Most entities showed frequent gains involving 8q2, while gains on 20q, 1q, 3q, 5p, 7q and 17q were frequent in different entities. Loss "hot spots" included 3p, 4q, 13q, 17p and 18q among others. Related average imbalance patterns were found for clinically distinct entities, e.g. hepatocellular carcinomas (ca.) and ductal breast ca., as well as for histologically related entities (squamous cell ca. of different sites). CONCLUSION: Although considerable case-by-case variation of genomic profiles can be found by CGH in epithelial malignancies, a limited set of variously combined chromosomal imbalances may be typical for carcinogenesis. Focus on the respective regions should aid in target gene detection and pathway deduction

Molecular Momentum Transport at Fluid-Solid Interfaces in MEMS/NEMS: A Review

This review is focused on molecular momentum transport at fluid-solid interfaces mainly related to microfluidics and nanofluidics in micro-/nano-electro-mechanical systems (MEMS/NEMS). This broad subject covers molecular dynamics behaviors, boundary conditions, molecular momentum accommodations, theoretical and phenomenological models in terms of gas-solid and liquid-solid interfaces affected by various physical factors, such as fluid and solid species, surface roughness, surface patterns, wettability, temperature, pressure, fluid viscosity and polarity. This review offers an overview of the major achievements, including experiments, theories and molecular dynamics simulations, in the field with particular emphasis on the effects on microfluidics and nanofluidics in nanoscience and nanotechnology. In Section 1 we present a brief introduction on the backgrounds, history and concepts. Sections 2 and 3 are focused on molecular momentum transport at gas-solid and liquid-solid interfaces, respectively. Summary and conclusions are finally presented in Section 4