Neuroserpin polymers cause oxidative stress in a neuronal model of the dementia FENIB

The serpinopathies are human pathologies caused by mutations that promote polymerisation and intracellular deposition of proteins of the serpin superfamily, leading to a poorly understood cell toxicity. The dementia FENIB is caused by polymerisation of the neuronal serpin neuroserpin (NS) within the endoplasmic reticulum (ER) of neurons. With the aim of understanding the toxicity due to intracellular accumulation of neuroserpin polymers, we have generated transgenic neural progenitor cell (NPC) cultures from mouse foetal cerebral cortex, stably expressing the control protein GFP (green fluorescent protein), or human wild type, G392E or delta NS. We have characterised these cell lines in the proliferative state and after differentiation to neurons. Our results show that G392E NS formed polymers that were mostly retained within the ER, while wild type NS was correctly secreted as a monomeric protein into the culture medium. Delta NS was absent at steady state due to its rapid degradation, but it was easily detected upon proteasomal block. Looking at their intracellular distribution, wild type NS was found in partial co-localisation with ER and Golgi markers, while G392E NS was localised within the ER only. Furthermore, polymers of NS were detected by ELISA and immunofluorescence in neurons expressing the mutant but not the wild type protein. We used control GFP and G392E NPCs differentiated to neurons to investigate which cellular pathways were modulated by intracellular polymers by performing RNA sequencing. We identified 747 genes with a significant upregulation (623) or downregulation (124) in G392E NS-expressing cells, and we focused our attention on several genes involved in the defence against oxidative stress that were up-regulated in cells expressing G392E NS (Aldh1b1, Apoe, Gpx1, Gstm1, Prdx6, Scara3, Sod2). Inhibition of intracellular anti-oxidants by specific pharmacological reagents uncovered the damaging effects of NS polymers. Our results support a role for oxidative stress in the cellular toxicity underlying the neurodegenerative dementia FENIB

Pengaruh Pemberian Bunga Rosella Terhadap Perubahan Tekanan Darah Penderita Hipertensi Dengan Terapi Captopril Di Desa Kamiwangi Kecamatan Toili Barat Kabupaten Luwuk Banggai

Hypertension is a cardiovasculer disturbance which is mostly founded in society. in healing hypertension, it needs continually Treatments. Rosella flower is a plant that has many good Components to heal hypertension. This research uses Quasy experiment design with control group. the amout of population in this research are 40 people. The method in taking sample uses purposive sampling and it got 30 respondents as the sample. The information of blood pressure was taken from the clients by using spygmomanometer, then it was displayed in table and processed by using SPSS program (statistic program for society science), used coupling T test, uncoupling T test with interpretation level 95%(a<0.05). The result of the research shows that the hypertension patients with captopril therapy which were given boiled dry rosella, got faster reduction of blood pressure after 2 hours than the hypertension patients with only captopril therapy. By using coupling T test, p-value was 0,00 . With mean value of the difference of reduction of sistolik blood pressure on the first day in intervention group was 28 mmHg while the control group was 11mmHg. on the second day, the reduction of sistolik blood pressure in intervention group was 13,33 l, and for control group, it was 5 mmHg. The diastolic blood pressure on the first day, the mean of the reduction of blood pressure in intervention group was 14mmHg while in control group was 6mmHg. On the second day, the mean of sistolic blood pressure in intervention group was 6 mmHg, and 2mmHg for control group. Key words: hypertension, rosella flower, captopril therapy Hipertensi merupakan penyakit gangguan kardiovaskuler. Bunga rosella adalah tumbuhan herbal untuk mengobati hipertensi. Tujuan penelitian melihat pengaruh bunga rosella terhadap tekanan darah penderita hipertensi dengan terapi captopril. Metode : quasi eksperiment design with control group, purposive sampling, 30 subyek (15 kelompok intervensi, 15 kelompok kontrol). Intervensi, subyek dengan captopril ditambahkan seduhan kering bunga rosella. Kontrol subyek mendapatkan dengan captopril saja. Sistolik pre kelompok intervensi, pada kategori hipertensi grade I, II, III. Sistolik post masuk pada kategori normal, pre hipertensi, hipertensi grade I, II, III. Diastolik pre kelompok intervensi terbesar kategori pre hipertensi, post diastolik terbesar berada pada kategori normal. Kelompok kontrol sistolik pre terbesar pada hipertensi grade II, post berada pada hipertensi grade I. Diastolik kelompok kontrol pre terbesar pada hipertensi grade I, post terbesar hipertensi grade I. Dianalisis dengan uji T berpasangan, dan uji T tidak berpasangan, α ≤ 0,05. Hasil Uji T berpasangan tekanan darah pre-post sistolik p-value 0,000, diastolik p-value 0,004, didapatkan pengaruh bunga rosella terhadap penderita hipertensi dengan captopril, penurunan tekanan darah kelompok intervensi sistolik 19,333, diastolik 10,000 mmHg, kelompok kontrol sistolik 9,000 mmHg, diastolik 4,333 mmHg. Hasil Uji T tidak berpasangan di dapatkan perbedaan penurunan tekanan darah kelompok intervensi lebih besar dibadingkan kelompok kontrol, dengan p-value, sistolik 0,000, diastolik 0,025. Perbedaan penurunan sistolik sebesar 12,333 mmHg, diastolik 6,333 mmHg. Kesimpulan didapatkan pengaruh pemberian bunga rosella terhadap Perubahan tekanan darah penderita hipertensi dengan terapi captopri

X-ray irradiated cultures of mouse cortical neural stem/progenitor cells recover cell viability and proliferation with dose-dependent kinetics

Exposure of the developing or adult brain to ionizing radiation (IR) can cause cognitive impairment and/ or brain cancer, by targeting neural stem/progenitor cells (NSPCs). IR effects on NSPCs include transient cell cycle arrest, permanent cell cycle exit/differentiation, or cell death, depending on the experimental conditions. In vivo studies suggest that brain age influences NSPC response to IR, but whether this is due to intrinsic NSPC changes or to niche environment modifications remains unclear. Here, we describe the dose-dependent, time-dependent effects of X-ray IR in NSPC cultures derived from the mouse foetal cerebral cortex. We show that, although cortical NSPCs are resistant to low/moderate IR doses, high level IR exposure causes cell death, accumulation of DNA double-strand breaks, activation of p53- related molecular pathways and cell cycle alterations. Irradiated NSPC cultures transiently upregulate differentiation markers, but recover control levels of proliferation, viability and gene expression in the second week post-irradiation. These results are consistent with previously described in vivo effects of IR in the developing mouse cortex, and distinct from those observed in adult NSPC niches or in vitro adult NSPC cultures, suggesting that intrinsic differences in NSPCs of different origins might determine, at least in part, their response to IR

The Action Mechanism of the Myc Inhibitor Termed Omomyc May Give Clues on How to Target Myc for Cancer Therapy

Recent evidence points to Myc – a multifaceted bHLHZip transcription factor deregulated in the majority of human cancers – as a priority target for therapy. How to target Myc is less clear, given its involvement in a variety of key functions in healthy cells. Here we report on the action mechanism of the Myc interfering molecule termed Omomyc, which demonstrated astounding therapeutic efficacy in transgenic mouse cancer models in vivo. Omomyc action is different from the one that can be obtained by gene knockout or RNA interference, approaches designed to block all functions of a gene product. This molecule – instead – appears to cause an edge-specific perturbation that destroys some protein interactions of the Myc node and keeps others intact, with the result of reshaping the Myc transcriptome. Omomyc selectively targets Myc protein interactions: it binds c- and N-Myc, Max and Miz-1, but does not bind Mad or select HLH proteins. Specifically, it prevents Myc binding to promoter E-boxes and transactivation of target genes while retaining Miz-1 dependent binding to promoters and transrepression. This is accompanied by broad epigenetic changes such as decreased acetylation and increased methylation at H3 lysine 9. In the presence of Omomyc, the Myc interactome is channeled to repression and its activity appears to switch from a pro-oncogenic to a tumor suppressive one. Given the extraordinary therapeutic impact of Omomyc in animal models, these data suggest that successfully targeting Myc for cancer therapy might require a similar twofold action, in order to prevent Myc/Max binding to E-boxes and, at the same time, keep repressing genes that would be repressed by Myc

From pluripotency to forebrain patterning: an in vitro journey astride embryonic stem cells

Embryonic stem cells (ESCs) have been used extensively as in vitro models of neural development and disease, with special efforts towards their conversion into forebrain progenitors and neurons. The forebrain is the most complex brain region, giving rise to several fundamental structures, such as the cerebral cortex, the hypothalamus, and the retina. Due to the multiplicity of signaling pathways playing different roles at distinct times of embryonic development, the specification and patterning of forebrain has been difficult to study in vivo. Research performed on ESCs in vitro has provided a large body of evidence to complement work in model organisms, but these studies have often been focused more on cell type production than on cell fate regulation. In this review, we systematically reassess the current literature in the field of forebrain development in mouse and human ESCs with a focus on the molecular mechanisms of early cell fate decisions, taking into consideration the specific culture conditions, exogenous and endogenous molecular cues as described in the original studies. The resulting model of early forebrain induction and patterning provides a useful framework for further studies aimed at reconstructing forebrain development in vitro for basic research or therapy

Restriction of Neural Precursor Ability to Respond to Nurr1 by Early Regional Specification

During neural development, spatially regulated expression of specific transcription factors is crucial for central nervous system (CNS) regionalization, generation of neural precursors (NPs) and subsequent differentiation of specific cell types within defined regions. A critical role in dopaminergic differentiation in the midbrain (MB) has been assigned to the transcription factor Nurr1. Nurr1 controls the expression of key genes involved in dopamine (DA) neurotransmission, e. g. tyrosine hydroxylase (TH) and the DA transporter (DAT), and promotes the dopaminergic phenotype in embryonic stem cells. We investigated whether cells derived from different areas of the mouse CNS could be directed to differentiate into dopaminergic neurons in vitro by forced expression of the transcription factor Nurr1. We show that Nurr1 overexpression can promote dopaminergic cell fate specification only in NPs obtained from E13.5 ganglionic eminence (GE) and MB, but not in NPs isolated from E13.5 cortex (CTX) and spinal cord (SC) or from the adult subventricular zone (SVZ). Confirming previous studies, we also show that Nurr1 overexpression can increase the generation of TH-positive neurons in mouse embryonic stem cells. These data show that Nurr1 ability to induce a dopaminergic phenotype becomes restricted during CNS development and is critically dependent on the region of NPs derivation. Our results suggest that the plasticity of NPs and their ability to activate a dopaminergic differentiation program in response to Nurr1 is regulated during early stages of neurogenesis, possibly through mechanisms controlling CNS regionalization. Citation: Soldati C, Cacci E, Biagioni S, Carucci N, Lupo G, et al. (2012) Restriction of Neural Precursor Ability to Respond to Nurr1 by Early Regional Specification. PLoS ONE 7(12): e51798. doi: 10.1371/journal.pone.005179

miR-21 is a negative modulator of T-cell activation

microRNAs (miRNAs) are a class of small non-coding RNAs acting as post-transcriptional regulators of gene expression and play fundamental roles in regulating immune response and autoimmunity. We show that memory T-lymphocytes express higher levels of miR-21 compared to naïve T-lymphocytes and that miR-21 expression is induced upon TCR engagement of naïve T-cells. We identify bona fide miR-21 targets by direct immuno-purification and profiling of AGO2-associated mRNAs in Jurkat cells over-expressing miR-21. Our analysis shows that, in T-lymphocytes, miR-21 targets genes are involved in signal transduction. Coherently, TCR signalling is dampened upon miR-21 over-expression in Jurkat cells, resulting in lower ERK phosphorylation, AP-1 activation and CD69 expression. Primary human lymphocytes in which we impaired miR-21 activity, display IFN-γ production enhancement and stronger activation in response to TCR engagement as assessed by CD69, OX40, CD25 and CD127 analysis. By intracellular staining of the endogenous protein in primary T-lymphocytes we validate three key regulators of lymphocyte activation as novel miR-21 targets. Our results highlight an unexpected function of miR-21 as a negative modulator of signal transduction downstream of TCR in T-lymphocytes