Lampbrush Chromosomes and Fragile X Mental Retardation Protein, FMRP

Fragile X Syndrome is the most common form of inherited mental retardation, occurring in 1/4000 males and 1/8000 females. This syndrome is caused by the lack of expression of the Fragile X mental retardation protein, or FMRP. FMRP is an RNA binding protein, involved in translational regulation of specific mRNA messages. However it is still relatively unknown where FMRP binds its mRNA cargo. Here we show endogenous FMRP associating with nascent transcripts along the actively transcribed loops of a Xenopus laevis oocyte chromosome. Nuclei were isolated from oocytes and prepared for fluorescence microscopy by fixing onto glass slides. Panels A and B show a phase contrast image of the chromosomes followed by fluorescent imaging where red is a DAPI stain for nucleic acid and green is FMRP. Panels C and D show a magnification of one especially large loop of single stranded DNA extending from the double stranded DNA axis. The scale bar represents 2 mm

Kinetics and Steric Hindrance Effects of Carbon Dioxide Absorption into Aqueous Potassium Alaninate Solutions

Kinetic studies of carbon dioxide absorption into aqueous potassium alaninate (PA) solutions were performed using a stirred-cell reactor at concentrations ranging from 1.0 to 3.0 M and temperatures from 293.15 to 313.15 K. In the present work, the potassium salt of alanine was suggested as an energy-efficient absorbent for CO₂ capture. Densities and viscosities of the aqueous PA solution and physical solubilities of CO₂ in the solution were also measured to evaluate the reaction rate constant. The reaction rate was determined as the following equation:−rCO2=4.518×108exp(−3845T)exp(0.5706Cs)CsCCO2­(−<i>r</i>_CO₂: rate of reaction between absorbent and CO₂ gas [mol dm^–3s^–1]; <i>T</i>: temperature [K]; <i>C</i>_s: concentration of PA solution [mol dm^–3]; <i>C</i>_CO₂: concentration of CO₂ in PA solution [mol dm^–3]). Furthermore, the effect of the sterically hindered structure of PA was studied by comparing the diffusivity, physical solubility, and reaction rate constant with literature values of potassium glycinate

Making new media : creative production and digital literacies

hNSCs express diverse trophic factors. (A) In vitro proliferating and differentiated hNSCs expressed BDNF, NTF3, NTF4, NGF, VEGF, FGF2, and GDNF. (B) Western blotting analysis showed that hNSCs secreted higher levels of BDNF, NTF3, NTF4, NGF, and VEGF into the culture medium than human foreskin fibroblasts secrete. (TIFF 127 kb

Additional file 9: Table S2. of Human neural stem cells alleviate Alzheimer-like pathology in a mouse model

Sequences of primers used for reverse transcription (RT) and quantitative (q) PCR. (DOCX 25 kb

Additional file 8: Table S1. of Human neural stem cells alleviate Alzheimer-like pathology in a mouse model

List of primary antibodies used in immunohistochemistry (IHC) and western blot (WB). (DOCX 28 kb

Additional file 4: Figure S4. of Human neural stem cells alleviate Alzheimer-like pathology in a mouse model

The expression of Aβ-degrading enzymes in both in vivo and in vitro. (A) Transplantation of hNSCs (NSC, n = 7) did not significantly alter the levels of Mme and Ide expression in NSE/APPsw transgenic mice compared with vehicle injection (Veh, n = 6). (B) In vitro expression of Aβ-degrading enzymes in hNSCs. hNSCs under proliferation and differentiation conditions expressed IDE, MME, ECE1 (endothelin converting enzyme 1), ECE2 (endothelin converting enzyme 2), MMP2 (matrix metalloproteinase 2), PLAT (plasminogen activator, tissue), PLAU (plasminogen activator, urokinase), ACE (angiotensin 1 converting enzyme), and CTSB (cathepsin B). On western blot, there were no differences in the levels of Aβ42 in the media containing 1 μM soluble Aβ42 peptides between wells with and without incubation with hNSCs for 2 days. The number of mice (n) in A is indicated. All data represent mean ± SEM. Error bars indicate ± SEM. (TIFF 3170 kb

Additional file 1: Figure S1. of Human neural stem cells alleviate Alzheimer-like pathology in a mouse model

GFP expression by lenti-GFP-transduced hNSCs. Proliferating lenti-GFP-transduced hNSCs form neurospheres in culture dishes (A) and express GFP (B). Flow cytometry analysis using FlowJo (version 9.3.3) software showed that 94.5 % of all cells are GFP-positive (blue line histogram in C). The red line histogram is the negative control. Scale bar, 100 μm. (TIFF 310 kb

Additional file 6: Figure S6. of Human neural stem cells alleviate Alzheimer-like pathology in a mouse model

NSE/APPsw transgenic mouse-derived brain slices treated with BDNF-depleted CM. (A) Western blot showed that hNSCs secreted high level of BDNF into the cultured medium (lane 1; CM), and anti-BDNF antibody-mediated immunoprecipitation effectively removed BDNF in CM (lane 2; BDNF-depleted CM). Another SDS-PAGE gel was in parallel performed with silver staining to verify even loading. (B) Brain slices treated with DMEM (Ctr), CM, BDNF-depleted CM. Western blot analysis of phosphorylated tau (AT180) and BACE1 (n = 3 per group, where n is the number of experiments). All data represent mean ± SEM. Error bars indicate ± SEM. Mann–Whitney U-test, *p < 0.05. (TIFF 2110 kb

Image_1_Macrophage Lamin A/C Regulates Inflammation and the Development of Obesity-Induced Insulin Resistance.TIF

<p>Obesity-induced chronic low-grade inflammation, in particular in adipose tissue, contributes to the development of insulin resistance and type 2 diabetes. However, the mechanism by which obesity induces adipose tissue inflammation has not been completely elucidated. Recent studies suggest that alteration of the nuclear lamina is associated with age-associated chronic inflammation in humans and fly. These findings led us to investigate whether the nuclear lamina regulates obesity-mediated chronic inflammation. In this study, we show that lamin A/C mediates inflammation in macrophages. The gene and protein expression levels of lamin A/C are significantly increased in epididymal adipose tissues from obese rodent models and omental fat from obese human subjects compared to their lean controls. Flow cytometry and gene expression analyses reveal that the protein and gene expression levels of lamin A/C are increased in adipose tissue macrophages (ATMs) by obesity. We further show that ectopic overexpression of lamin A/C in macrophages spontaneously activates NF-κB, and increases the gene expression levels of proinflammatory genes, such as Il6, Tnf, Ccl2, and Nos2. Conversely, deletion of lamin A/C in macrophages reduces LPS-induced expression of these proinflammatory genes. Importantly, we find that myeloid cell-specific lamin A/C deficiency ameliorates obesity-induced insulin resistance and adipose tissue inflammation. Thus, our data suggest that lamin A/C mediates the activation of ATM inflammation by regulating NF-κB, thereby contributing to the development of obesity-induced insulin resistance.</p

Table_2_Macrophage Lamin A/C Regulates Inflammation and the Development of Obesity-Induced Insulin Resistance.PDF

<p>Obesity-induced chronic low-grade inflammation, in particular in adipose tissue, contributes to the development of insulin resistance and type 2 diabetes. However, the mechanism by which obesity induces adipose tissue inflammation has not been completely elucidated. Recent studies suggest that alteration of the nuclear lamina is associated with age-associated chronic inflammation in humans and fly. These findings led us to investigate whether the nuclear lamina regulates obesity-mediated chronic inflammation. In this study, we show that lamin A/C mediates inflammation in macrophages. The gene and protein expression levels of lamin A/C are significantly increased in epididymal adipose tissues from obese rodent models and omental fat from obese human subjects compared to their lean controls. Flow cytometry and gene expression analyses reveal that the protein and gene expression levels of lamin A/C are increased in adipose tissue macrophages (ATMs) by obesity. We further show that ectopic overexpression of lamin A/C in macrophages spontaneously activates NF-κB, and increases the gene expression levels of proinflammatory genes, such as Il6, Tnf, Ccl2, and Nos2. Conversely, deletion of lamin A/C in macrophages reduces LPS-induced expression of these proinflammatory genes. Importantly, we find that myeloid cell-specific lamin A/C deficiency ameliorates obesity-induced insulin resistance and adipose tissue inflammation. Thus, our data suggest that lamin A/C mediates the activation of ATM inflammation by regulating NF-κB, thereby contributing to the development of obesity-induced insulin resistance.</p