In experimental models of fetal alcohol spectrum disorder (FASD), cerebellar hypoplasia and hypofoliation are associated with insulin and insulin-like growth factor (IGF) resistance with impaired ...signaling through pathways that mediate growth, survival, plasticity, metabolism, and neurotransmitter function. To more directly assess the roles of impaired insulin and IGF signaling during brain development, we administered intracerebroventricular (ICV) injections of si-RNA targeting the insulin receptor, (InR), IGF-1 receptor (IGF-1R), or IGF-2R into postnatal day 2 (P2) Long Evans rat pups and examined the sustained effects on cerebellar function, structure, and neurotransmitter-related gene expression (P20).
Rotarod tests on P20 demonstrated significant impairments in motor function, and histological studies revealed pronounced cerebellar hypotrophy, hypoplasia, and hypofoliation in si-InR, si-IGF-1R, and si-IGF-2R treated rats. Quantitative RT-PCR analysis showed that si-InR, and to a lesser extent si-IGF-2R, broadly inhibited expression of insulin and IGF-2 polypeptides, and insulin, IGF-1, and IGF-2 receptors in the brain. ELISA studies showed that si-InR increased cerebellar levels of tau, phospho-tau and β-actin, and inhibited GAPDH. In addition, si-InR, si-IGF-1R, and si-IGF-2R inhibited expression of choline acetyltransferase, which mediates motor function. Although the ICV si-RNA treatments generally spared the neurotrophin and neurotrophin receptor expression, si-InR and si-IGF-1R inhibited NT3, while si-IGF-1R suppressed BDNF.
early postnatal inhibition of brain InR expression, and to lesser extents, IGF-R, causes structural and functional abnormalities that resemble effects of FASD. The findings suggest that major abnormalities in brains with FASD are mediated by impairments in insulin/IGF signaling. Potential therapeutic strategies to reduce the long-term impact of prenatal alcohol exposure may include treatment with agents that restore brain insulin and IGF responsiveness.
A neutral pentadentate ligand, di(pyrazolecarbimido)amine (Hdcadpz), and its adduct with HClO4, H2dcadpz+ClO4-, were for the first time isolated from our previously reported ...Cu3(dcadpz)2(Hpz)2(ClO4)2(ClO4)2.H2O by the use of (NH4)2S to remove the CuII ions and characterized by IR, EA, UV, NMR, MS, and X-ray crystallography. Reactions of copper(II) or nickel(II) nitrate with Hdcadpz in a 1:2 molar ratio generated two mononuclear precursors of Cu(dcadpz)2 (1) and Ni(dcadpz)2.2/3DMF (2). Furthermore, three new linear homo- and heterotrinuclear complexes of the same motif M{M'(dcadpz)2}M (M=CoII, NiII, M'=CuII, NiII), {Co(pdm)}2{Cu(dcadpz)2}(NO3)4 (3), {Ni(pdm)}2{Cu(dcadpz)2}(NO3)4 (4), and {Ni(MeOH)(H2O)2}2{Ni(dcadpz)2}(NO3)4 (5), were synthesized from these two precursors (pdm=2,6-pyridinedimethanol) and characterized by X-ray crystallography. Magnetic studies show that the central Cu(dcadpz)2 motif is antiferromagnetically coupled with both the terminal Co(II) atoms via the dcadpz- ligand in 3 with a J value of -5.27 cm(-1) and ferromagnetically coupled with both the terminal Ni(II) atoms in 4 with a J value of 2.50 cm(-1), while 5 behaves only as a Curie paramagnet between 2 and 300 K due to the diamagnetic character of the central square-planar Ni(II) atom.
BackgroundThe genetic diversity of human immunodeficiency virus type 1 (HIV-1) raises the question of whether vaccines that include a component to elicit antiviral T cell immunity based on a single ...viral genetic clade could provide cellular immune protection against divergent HIV-1 clades. Therefore, we quantified the cross-clade reactivity, among unvaccinated individuals, of anti–HIV-1 T cell responses to the infecting HIV-1 clade relative to other major circulating clades MethodsCellular immune responses to HIV-1 clades A, B, and C were compared by standardized interferon-γ enzyme-linked immunospot assays among 250 unvaccinated individuals, infected with diverse HIV-1 clades, from Brazil, Malawi, South Africa, Thailand, and the United States. Cross-clade reactivity was evaluated by use of the ratio of responses to heterologous versus homologous (infecting) clades of HIV-1 ResultsCellular immune responses were predominantly focused on viral Gag and Nef proteins. Cross-clade reactivity of cellular immune responses to HIV-1 clade A, B, and C proteins was substantial for Nef proteins (ratio, 0.97 95% confidence interval, 0.89–1.05) and lower for Gag proteins (ratio, 0.67 95% confidence interval, 0.62–0.73). The difference in cross-clade reactivity to Nef and Gag proteins was significant (P<.0001) ConclusionsCross-clade reactivity of cellular immune responses can be substantial but varies by viral protein
A high-oil maize (Zea mays L.) (HOM) population, KYHO, was developed across 10 generations by selective breeding. The objectives of this investigation were to evaluate the effects of selection for ...kernel oil on correlated traits and measure the genetic variance of kernel oil content (KOC). Kernel oil content, kernel protein content (KPC), and kernel starch content (KSC) were estimated by near-infrared reflectance spectroscopy (NIRS); palmitic, stearic, oleic, linoleic, linolenic, and arachidic fatty acids were analyzed by gas chromatography. Also measured at the appropriate time during the growth cycle were plant anthesis (AT), plant height (PH), ear height (EH), 100 kernels weight (KW), and red ear cob ratio (RCR). Sampling and measurement of the various parameters occurred under the same conditions for each generation for three years. Kernel oil content greatly increased from 3.53% in cycle 0 to 12.66% in cycle 10; the predicted selection response per cycle was 1.01 ± 0.05%. The realized heritability (h2R) and broad-sense heritability (h2B) of KOC in KYHO were 0.38 and 0.99 respectively, suggesting significant genetic variability, especially additive variance, across 11 selection cycles. The correlated traits of stearic, oleic, unsaturated fatty acid, and KPC and RCR increased steadily from the original generation to the final cycle (C10), whereas the remaining parameters decreased. There were significant (P 0.01) genotypic and phenotypic correlations between KOC and all measured traits, with the exception of arachidic acid. Red ear cob ratio was significantly positively correlated with KOC, and increased 7.41% with each 1% KOC increase.
Protein structures are decisive for their activities and interactions with other molecules. Global analysis of protein structures and conformational changes cannot be achieved by commonly used ...abundance-based proteomics. Here, we integrated cysteine covalent labeling, selective enrichment, and quantitative proteomics to study protein structures and structural changes on a large scale. This method was applied to globally investigate protein structures in HEK293T cells and protein structural changes in the cells with the tunicamycin (Tm)-induced endoplasmic reticulum (ER) stress. We quantified several thousand cysteine residues, which contain unprecedented and valuable information of protein structures. Combining this method with pulsed stable isotope labeling by amino acids in cell culture, we further analyzed the folding state differences between pre-existing and newly synthesized proteins in cells under the Tm treatment. Besides newly synthesized proteins, unexpectedly, many pre-existing proteins were found to become unfolded upon ER stress, especially those related to gene transcription and protein translation. Furthermore, the current results reveal that N-glycosylation plays a more important role in the folding process of the tertiary and quaternary structures than the secondary structures for newly synthesized proteins. Considering the importance of cysteine in protein structures, this method can be extensively applied in the biological and biomedical research fields.
Photochemistry: A series of emissive gold(III) complexes with fluorene‐containing cyclometalating ligands exhibits strong phosphorescence and long‐lived excited states with emission quantum yields ...and lifetimes up to 58 % and 305 μs, respectively. These complexes can sensitize energy up‐conversion of 9,10‐diphenylanthracene (DPA; see picture) and display rich two‐photon absorption properties (TPA; TTA=triplet–triplet annihilation).
Pre-clinical studies suggest that pan-inhibitors for histone deacetylases (HDACs) may benefit the treatment of pulmonary fibrosis (PF). However, HDAC3-specific roles or mechanisms during PF ...development are poorly understood.
The expression of HDAC3 and the impacts of RGFP966, a selective HDAC3 inhibitor, were examined in a bleomycin-induced PF mouse model and in TGF-β-challenged MRC-5 cells. H&E and Masson staining were employed to reveal the pathological changes in lung; Western blot to assess expressions of biomarkers related to fibrosis and epithelial-mesenchymal transition (EMT), activations of Notch1 and signal transducer and activator of transcription 1 (STAT1) signaling, and levels of inflammasome components, absent in melanoma 2 (AIM2) and apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC); ELISA to measure productions of proinflammatory interleukin (IL)-1β and IL-18; cycloheximide chase assay and coimmunoprecipitation to monitor the protein stability and acetylation of Notch intracellular domain 1 (NICD1) and STAT1, respectively.
HDAC3 was induced in in vitro and in vivo PF models, which was associated with elevated expressions of fibrosis-related biomarkers, EMT markers, and pro-inflammatory cytokines, activations of Notch1 and STAT1 signaling, and up-regulated inflammasome components, AIM2 and ASC. All the fibrotic phenotypes were potently inhibited by RGFP966, which boosted the acetylation of NICD1 and STAT1, accelerated the degradation of NICD1, and inhibited STAT1 phosphorylation. Overexpressing NICD1 or STAT1 restored the fibrotic phenotypes suppressed by RGFP966.
HDAC3 promoted EMT, inflammation, and PF development by activating Notch1 and STAT1 signaling. Therefore, targeting HDAC3, Notch1 or STAT1 signaling may ameliorate PF development.
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