Huntington disease (HD) is a dominantly inherited disorder caused by a CAG expansion mutation in the huntingtin (HTT) gene, which results in the HTT protein that contains an expanded polyglutamine tract. The adult form of HD exhibits a late onset of the fully symptomatic phase. However, there is also a long presymptomatic phase, which has been increasingly investigated and recognized as important for the disease development. Moreover, the juvenile form of HD, evoked by a higher number of CAG repeats, resembles a neurodevelopmental disorder and has recently been the focus of additional interest. Multiple lines of data, such as the developmental necessity of HTT, its role in the cell cycle and neurogenesis, and findings from pluripotent stem cells, suggest the existence of a neurodevelopmental component in HD pathogenesis. Therefore, we discuss the early molecular pathogenesis of HD in pluripotent and neural stem cells, with respect to the neurodevelopmental aspects of HD.Electronic supplementary materialThe online version of this article (doi:10.1007/s12035-017-0477-7) contains supplementary material, which is available to authorized users.
Patients with Sjögren's syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced saliva production. Similarly, saliva production is severely compromised in patients receiving radiation treatment for head and neck cancers. Rodent models, developed to mimic these clinical conditions, facilitate an understanding of the disease pathogenesis and allow for the development of new therapeutic strategies. Therefore, the ability to accurately, reproducibly, and repeatedly measure salivary gland function in animal models is critical. Building on procedures previously described in the literature, a method was developed that meets these criteria and was used to evaluate salivary gland function in mice. An additional advantage of this new method is that it is easily mastered, and has little inter-operator variation. Salivary gland function is evaluated as the amount (weight or volume) or rate (mL/min) of saliva produced in response to pilocarpine stimulation. The collected saliva is a good source for the analyses of protein content, immunoglobulin concentrations, and other biomolecules.
Huntington disease (HD) is an incurable neurodegenerative disorder caused by expansion of CAG repeats in huntingtin (HTT) gene, resulting in expanded polyglutamine tract in HTT protein. Although, HD has its common onset in adulthood, subtle symptoms in patients may occur decades before diagnosis, and molecular and cellular changes begin much earlier, even in cells that are not yet lineage committed such as stem cells. Studies in induced pluripotent stem cell (iPSC) HD models have demonstrated that multiple molecular processes are altered by the mutant HTT protein and suggested its silencing as a promising therapeutic strategy. Therefore, we aimed to generate HD iPS cells with stable silencing of HTT and further to investigate the effects of HTT knock-down on deregulations of signaling pathways e.g., p53 downregulation, present in cells already in pluripotent state. We designed a gene silencing strategy based on RNAi cassette in piggyBAC vector for constant shRNA expression. Using such system we delivered and tested several shRNA targeting huntingtin in mouse HD YAC128 iPSC and human HD109, HD71, and Control iPSC. The most effective shRNA (shHTT2) reagent stably silenced HTT in all HD iPS cells and remained active upon differentiation to neural stem cells (NSC). When investigating the effects of HTT silencing on signaling pathways, we found that in mouse HD iPSC lines expressing shRNA the level of mutant HTT inversely correlated with p53 levels, resulting in p53 level normalization upon silencing of mutant HTT. We also found that p53 deregulation continues into the NSC developmental stage and it was reversed upon HTT silencing. In addition, we observed subtle effects of silencing on proteins of Wnt/β-catenin and ERK1/2 signaling pathways. In summary, we successfully created the first mouse and human shRNA-expressing HD iPS cells with stable and continuous HTT silencing. Moreover, we demonstrated reversal of HD p53 phenotype in mouse HD iPSC, therefore, the stable knockdown of HTT is well-suited for investigation on HD cellular pathways, and is potentially useful as a stand-alone therapy or component of cell therapy. In addition, the total HTT knock-down in our human cells has further implications for mutant allele selective approach in iPSC.
Autoantibodies reactive against Ro52 are present in 70% of Sjögren’s syndrome patients and are associated with higher disease severity. However, their role in causing aqueous deficient dry eye, a major cause for morbidity in Sjögren’s syndrome, is unclear. To investigate whether immune responses targeting Ro52 contribute towards the dry eye, male and female NZM2758 mice were immunized with recombinant Ro52. Tear production was measured by the phenol red thread test. Sera were analyzed for anti-Ro52 levels by immunoprecipitation. Lacrimal glands were evaluated for inflammatory foci and IgG deposits. Our results showed that, although all mice generated anti-Ro52 antibodies, only females developed a significant drop in tear production. None of the mice developed severe lacrimal gland inflammation, and female mice with anti-Ro52 showed higher levels of IgG deposits within their glands. Passive transfer of anti-Ro52 sera caused reduced tear production in female mice, but not in males. This study demonstrates for the first time that immune responses initiated by Ro52 induce aqueous dry eye, and this may be driven by anti-Ro52 antibodies. Furthermore, the sexual dimorphism in glandular dysfunction suggests that the lacrimal glands in females are more susceptible to autoantibody-mediated injury.
StreszczenieCel pracy. Celem pracy była ocena porównawcza postawy ciała dzieci w zależności od wieku rozpoczęcia edukacji szkolnej. Materiał i metody. Przebadano 64 uczniów z klas I-II, w tym 22 sześciolatków oraz 42 siedmiolatków. Materiał badawczy został podzielony na trzy grupy: grupa I -pierwszoklasiści w wieku 6 lat, grupa II -pierwszoklasiści w wieku 7 lat, grupa III -drugoklasiści w wieku 7 lat. Do analizy postawy ciała w płaszczyźnie strzałkowej zastosowano metodę fotogrametryczną. Wyniki. Przeprowadzone badania wykazały, iż dzieci w wieku 7 lat charakteryzowały się większymi odchyleniami parametrów w płaszczyźnie strzałkowej niż dzieci 6-letnie. Dzieci 7-letnie w drugiej klasie miały postawę bardziej zbliżoną do 6-latków niż do ich rówieśników z klasy pierwszej. Wnioski. Wykazano statystycznie istotne różnice w masie ciała między 6-i 7-latkami uczęszczającymi do pierwszej klasy. W płaszczyźnie strzałkowej wykazano istotne różnice w kącie nachylenia odcinka lędźwiowo-krzyżowego, długości całego kręgosłupa, długości kifozy piersiowej i kącie lordozy lędźwiowej.Słowa kluczowe: dzieci, krzywizny kręgosłupa, metoda fotogrametryczna, edukacja wczesnoszkolna Abstract Introduction. Th e aim of the study was to assess posture in children depending on the age of starting school education. Ma terial and methods. A total of 64 students from classes I-II, including 22 six year olds and 42 seven year olds. The research material was divided into three groups: group I -fi rst graders at the age of 6 years, group II -fi rst graders at the age of 7 years, group III -second graders at the age of 7 years. Photogrammetric method was used for the analysis of posture in the sagittal plane. R es ults. The study showed that the 7-year-old children have greater deviations of the parameters in the sagittal plane than 6-year-old children. 7-year-old children in the second grade had a posture closer to 6-year-olds who have just started attending school than their equals in the fi rst grade. Conclusions. Somatic analysis showed statistically signifi cant differences in body weight between 6 and 7 year olds attending the fi rst grade. Signifi cant differences in the sagittal plane were shown in the angle of the lumbar-sacral segment, entire spine length, thoracic kyphosis length and lumbar lordosis angle.Key words: children, spinal curvatures, photogrammetry method, primary education Wprowadzenie Z dniem 1 września 2014 roku wejdzie w życie ustawa, która wprowadza obowiązek szkolny dla dzieci sześcioletnich. Do tego czasu szkoły muszą przygotować się na ich przyjście poprzez wprowadzenie nowego programu nauczania, odpowiedniego usytuowania klas, dopasowania mebli do wysokości ciała dzieci, zakupienia nowych urządzeń, takich jak tabli-
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