BOSS Constraints on Massive Particles during Inflation: The Cosmological Collider in Action

Massive particles leave imprints on primordial non-Gaussianity via couplings to the inflaton, even despite their exponential dilution during inflation: practically, the Universe acts as a Cosmological Collider. We present the first dedicated search for spin-zero particles using BOSS redshift-space galaxy power spectrum and bispectrum multipoles, as well as Planck CMB non-Gaussianity data. We demonstrate that some Cosmological Collider models are well approximated by the standard equilateral and orthogonal parametrization; assuming negligible inflaton self-interactions, this facilitates us translating Planck non-Gaussianity constraints into bounds on Collider models. Many models have signatures that are not degenerate with equilateral and orthogonal non-Gaussianity and thus require dedicated searches. Here, we constrain such models using BOSS three-dimensional redshift-space galaxy clustering data, focusing on spin-zero particles in the principal series and constraining their couplings to the inflaton at varying speed and mass, marginalizing over the unknown inflaton self-interactions. This is made possible through an improvement in Cosmological Bootstrap techniques and the combination of perturbation theory and halo occupation distribution models for galaxy clustering. Our work sets the standard for inflationary spectroscopy with cosmological observations, providing the ultimate link between physics on the largest and smallest scales.Comment: 35 pages, 15 figures, 6 table

Beneath the radar: immune-evasive cell sources for stroke therapy

Stem cell therapy is an emerging treatment paradigm for stroke patients with remaining neurological deficits. While allogeneic cell transplants overcome the manufacturing constraints of autologous grafts, they can be rejected by the recipient's immune system, which identifies foreign cells through the human leukocyte antigen (HLA) system. The heterogeneity of HLA molecules in the human population would require a very high number of cell lines, which may still be inadequate for patients with rare genetic HLAs. Here, we outline key progress in genetic HLA engineering in pluripotent stem and derived cells to evade the host's immune system, reducing the number of allogeneic cell lines required, and examine safety measures explored in both preclinical studies and upcoming clinical trials

Ultra-light axions and the S8S_8 tension: joint constraints from the cosmic microwave background and galaxy clustering

We search for ultra-light axions as dark matter (DM) and dark energy particle candidates, for axion masses $10^{-32}\,\mathrm{eV} \leq m_\mathrm{a} \leq 10^{-24}\,\mathrm{eV}$, by a joint analysis of cosmic microwave background (CMB) and galaxy clustering data -- and consider if axions can resolve the tension in inferred values of the matter clustering parameter $S_8$. We give legacy constraints from Planck 2018 CMB data, improving 2015 limits on the axion density $\Omega_\mathrm{a} h^2$ by up to a factor of three; CMB data from the Atacama Cosmology Telescope and the South Pole Telescope marginally weaken Planck bounds at $m_\mathrm{a} = 10^{-25}\,\mathrm{eV}$, owing to lower (and theoretically-consistent) gravitational lensing signals. We jointly infer, from Planck CMB and full-shape galaxy power spectrum and bispectrum data from the Baryon Oscillation Spectroscopic Survey (BOSS), that axions are, today, $< 10\%$ of the DM for $m_\mathrm{a} \leq 10^{-26}\,\mathrm{eV}$ and $< 1\%$ for $10^{-30}\,\mathrm{eV} \leq m_\mathrm{a} \leq 10^{-28}\,\mathrm{eV}$. BOSS data strengthen limits, in particular at higher $m_\mathrm{a}$ by probing high-wavenumber modes ($k < 0.4 h\,\mathrm{Mpc}^{-1}$). BOSS alone finds a preference for axions at $2.7 \sigma$, for $m_\mathrm{a} = 10^{-26}\,\mathrm{eV}$, but Planck disfavours this result. Nonetheless, axions in a window $10^{-28}\,\mathrm{eV} \leq m_\mathrm{a} \leq 10^{-25}\,\mathrm{eV}$ can improve consistency between CMB and galaxy clustering data, e.g., reducing the $S_8$ discrepancy from $2.7 \sigma$ to $1.6 \sigma$, since these axions suppress structure growth at the $8 h^{-1}\,\mathrm{Mpc}$ scales to which $S_8$ is sensitive. We expect improved constraints with upcoming high-resolution CMB and galaxy lensing and future galaxy clustering data, where we will further assess if axions can restore cosmic concordance.Comment: 52 pages, 22 figure

Efficient ssODN-Mediated Targeting by Avoiding Cellular Inhibitory RNAs through Precomplexed CRISPR-Cas9/sgRNA Ribonucleoprotein

CRISPR-Cas9がヒト細胞内のRNAで阻害されてしまう現象を発見し、iPS細胞での効率的な相同組み換えゲノム編集技術を実現. 京都大学プレスリリース. 2021-03-12.A simple step to enhance CRISPR-Cas9 genome editing. 京都大学プレスリリース. 2021-03-12.Combined with CRISPR-Cas9 technology and single-stranded oligodeoxynucleotides (ssODNs), specific single-nucleotide alterations can be introduced into a targeted genomic locus in induced pluripotent stem cells (iPSCs); however, ssODN knockin frequency is low compared with deletion induction. Although several Cas9 transduction methods have been reported, the biochemical behavior of CRISPR-Cas9 nuclease in mammalian cells is yet to be explored. Here, we investigated intrinsic cellular factors that affect Cas9 cleavage activity in vitro. We found that intracellular RNA, but not DNA or protein fractions, inhibits Cas9 from binding to single guide RNA (sgRNA) and reduces the enzymatic activity. To prevent this, precomplexing Cas9 and sgRNA before delivery into cells can lead to higher genome editing activity compared with Cas9 overexpression approaches. By optimizing electroporation parameters of precomplexed ribonucleoprotein and ssODN, we achieved efficiencies of single-nucleotide correction as high as 70% and loxP insertion up to 40%. Finally, we could replace the HLA-C1 allele with the C2 allele to generate histocompatibility leukocyte antigen custom-edited iPSCs

Review on metal complexes with 4N -methyl (thiosemicarbazone)

Diverse literature is available related to 4N-methyl (thiosemicarbazones) and their complexes. From previous decades to more recent times reports have discussed crystal structures of thiosemicarbazones. The properties and numerous applications of these compounds are also well documented. The chemistry of thiosemicarbazone is an important but vast area. This review is concerned specifically with the chemistry and structure of different derivatives of 4N-methyl (thiosemicarbazones) Schiff bases and their metal complexes. The chemical and physical properties, biological activities and catalytic applications of these compounds are discussed in detail

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

Prolonged expression of the CRISPR-Cas9 nuclease and gRNA from viral vectors may cause off-target mutagenesis and immunogenicity. Thus, a transient delivery system is needed for therapeutic genome editing applications. Here, we develop an extracellular nanovesicle-based ribonucleoprotein delivery system named NanoMEDIC by utilizing two distinct homing mechanisms. Chemical induced dimerization recruits Cas9 protein into extracellular nanovesicles, and then a viral RNA packaging signal and two self-cleaving riboswitches tether and release sgRNA into nanovesicles. We demonstrate efficient genome editing in various hard-to-transfect cell types, including human induced pluripotent stem (iPS) cells, neurons, and myoblasts. NanoMEDIC also achieves over 90% exon skipping efficiencies in skeletal muscle cells derived from Duchenne muscular dystrophy (DMD) patient iPS cells. Finally, single intramuscular injection of NanoMEDIC induces permanent genomic exon skipping in a luciferase reporter mouse and in mdx mice, indicating its utility for in vivo genome editing therapy of DMD and beyond

Determination of fungal activity in modified wood by means of micro-calorimetry and determination of total esterase activity

Beech and pine wood blocks were treated with 1,3-dimethylol-4,5-dihydroxyethylen urea (DMDHEU) to increasing weight percent gains (WPG). The resistance of the treated specimens against Trametes versicolor and Coniophora puteana, determined as mass loss, increased with increasing WPG of DMDHEU. Metabolic activity of the fungi in the wood blocks was assessed as total esterase activity (TEA) based on the hydrolysis of fluorescein diacetate and as heat or energy production determined by isothermal micro-calorimetry. Both methods revealed that the fungal activity was related with the WPG and the mass loss caused by the fungi. Still, fungal activity was detected even in wood blocks of the highest WPG and showed that the treatment was not toxic to the fungi. Energy production showed a higher consistency with the mass loss after decay than TEA; higher mass loss was more stringently reflected by higher heat production rate. Heat production did not proceed linearly, possibly due to the inhibition of fungal activity by an excess of carbon dioxide

A Parameter-Masked Mock Data Challenge for Beyond-Two-Point Galaxy Clustering Statistics

The last few years have seen the emergence of a wide array of novel techniques for analyzing high-precision data from upcoming galaxy surveys, which aim to extend the statistical analysis of galaxy clustering data beyond the linear regime and the canonical two-point (2pt) statistics. We test and benchmark some of these new techniques in a community data challenge "Beyond-2pt", initiated during the Aspen 2022 Summer Program "Large-Scale Structure Cosmology beyond 2-Point Statistics," whose first round of results we present here. The challenge dataset consists of high-precision mock galaxy catalogs for clustering in real space, redshift space, and on a light cone. Participants in the challenge have developed end-to-end pipelines to analyze mock catalogs and extract unknown ("masked") cosmological parameters of the underlying $\Lambda$CDM models with their methods. The methods represented are density-split clustering, nearest neighbor statistics, BACCO power spectrum emulator, void statistics, LEFTfield field-level inference using effective field theory (EFT), and joint power spectrum and bispectrum analyses using both EFT and simulation-based inference. In this work, we review the results of the challenge, focusing on problems solved, lessons learned, and future research needed to perfect the emerging beyond-2pt approaches. The unbiased parameter recovery demonstrated in this challenge by multiple statistics and the associated modeling and inference frameworks supports the credibility of cosmology constraints from these methods. The challenge data set is publicly available and we welcome future submissions from methods that are not yet represented.Comment: New submissions welcome! Challenge data available at https://github.com/ANSalcedo/Beyond2ptMoc

MECP2 Isoform-Specific Vectors with Regulated Expression for Rett Syndrome Gene Therapy

BACKGROUND:Rett Syndrome (RTT) is an Autism Spectrum Disorder and the leading cause of mental retardation in females. RTT is caused by mutations in the Methyl CpG-Binding Protein-2 (MECP2) gene and has no treatment. Our objective is to develop viral vectors for MECP2 gene transfer into Neural Stem Cells (NSC) and neurons suitable for gene therapy of Rett Syndrome. METHODOLOGY/PRINCIPAL FINDINGS:We generated self-inactivating (SIN) retroviral vectors with the ubiquitous EF1alpha promoter avoiding known silencer elements to escape stem-cell-specific viral silencing. High efficiency NSC infection resulted in long-term EGFP expression in transduced NSC and after differentiation into neurons. Infection with Myc-tagged MECP2-isoform-specific (E1 and E2) vectors directed MeCP2 to heterochromatin of transduced NSC and neurons. In contrast, vectors with an internal mouse Mecp2 promoter (MeP) directed restricted expression only in neurons and glia and not NSC, recapitulating the endogenous expression pattern required to avoid detrimental consequences of MECP2 ectopic expression. In differentiated NSC from adult heterozygous Mecp2(tm1.1Bird)+/- female mice, 48% of neurons expressed endogenous MeCP2 due to random inactivation of the X-linked Mecp2 gene. Retroviral MECP2 transduction with EF1alpha and MeP vectors rescued expression in 95-100% of neurons resulting in increased dendrite branching function in vitro. Insulated MECP2 isoform-specific lentiviral vectors show long-term expression in NSC and their differentiated neuronal progeny, and directly infect dissociated murine cortical neurons with high efficiency. CONCLUSIONS/SIGNIFICANCE:MeP vectors recapitulate the endogenous expression pattern of MeCP2 in neurons and glia. They have utility to study MeCP2 isoform-specific functions in vitro, and are effective gene therapy vectors for rescuing dendritic maturation of neurons in an ex vivo model of RTT