High Efficiency RNA Extraction From Sperm Cells Using Guanidinium Thiocyanate Supplemented With Tris(2-Carboxyethyl)Phosphine

The extraction of high-quality ribonucleic acid (RNA) from tissues and cells is a key step in many biological assays. Guanidinium thiocyanate-phenol-chloroform (AGPC) is a widely used and efficient method to obtain pure RNA from most tissues and cells. However, it is not efficient with some cells like sperm cells because they are resistant to chaotropic lysis solutions containing guanidinium thiocyanate such as Buffer RLT+ and Trizol. Here, we show that disulfide bonds are responsible for the chemical resistance of sperm cells to RNA extraction reagents. We show that while β-mercaptoethanol (βME) can increase sperm lysis in Buffer RLT+, it has no effect in Trizol and leaves sperm cells intact. We measured the reduction of disulfide bonds in 2,2′-dithiodipyridine (DTDP) and observed that βME has a pH-dependent activity in chaotropic solutions, suggesting that pH is a limiting factor. We identified tris(2-carboxyethyl)phosphine (TCEP) as an efficient lysis enhancer of AGPC solutions that can retain reducing activity even at acidic pH. Trizol supplemented with TCEP allows the complete and rapid lysis of sperm cells, increasing RNA yield by 100-fold and resulting in RNA with optimal quality for reverse transcription and polymerase chain reaction. Our findings highlight the importance of efficient cell lysis and extraction of various macromolecules for bulk and single-cell assays, and can be applied to other lysis-resistant cells and vesicles, thereby optimizing the amount of required starting material and animals

Upper and Lower Limits on Neutralino WIMP Mass and Spin--Independent Scattering Cross Section, and Impact of New (g-2)_{mu} Measurement

We derive the allowed ranges of the spin--independent interaction cross section \sigsip for the elastic scattering of neutralinos on proton for wide ranges of parameters of the general Minimal Supersymmetric Standard Model. We investigate the effects of the lower limits on Higgs and superpartner masses from colliders, as well as the impact of constraints from \bsgamma and the new measurement of \gmtwo on the upper and lower limits on \sigsip. We further explore the impact of the neutralino relic density, including coannihilation, and of theoretical assumptions about the largest allowed values of the supersymmetric parameters. For $\mu>0$, requiring the latter to lie below 1\tev leads to \sigsip\gsim 10^{-11}\pb at \mchi\sim100\gev and \sigsip\gsim 10^{-8}\pb at \mchi\sim1\tev. When the supersymmetric parameters are allowed above 1\tev, for 440\gev \lsim \mchi\lsim 1020 \gev we derive a {\em parameter--independent lower limit} of \sigsip \gsim 2\times 10^{-12}\pb. (No similar lower limits can be set for $\mu<0$ nor for 1020\gev\lsim\mchi\lsim2.6\tev.) Requiring \abundchi<0.3 implies a {\em parameter--independent upper limit} \mchi\lsim2.6\tev. The new \epem--based measurement of $(g-2)_{\mu}$ restricts \mchi\lsim 350\gev at $1 \sigma$ CL and \mchi\lsim515\gev at $2 \sigma$ CL, and implies $\mu>0$. The largest allowed values of \sigsip have already become accessible to recent experimental searches.Comment: LaTeX, 17 pages, 9 eps figures. Version to appear in JHE

Single-Cell Multiomics Techniques: From Conception to Applications

Recent advances in methods for single-cell analyses and barcoding strategies have led to considerable progress in research. The development of multiplexed assays offers the possibility to conduct parallel analyses of multiple factors and processes for comprehensive characterization of cellular and molecular states in health and disease. These technologies have expanded extremely rapidly in the past years and constantly evolve and provide better specificity, precision and resolution. This review summarizes recent progress in single-cell multiomics approaches, and focuses, in particular, on the most innovative techniques that integrate genome, epigenome and transcriptome profiling. It describes the methodologies, discusses their advantages and limitations, and explains how they have been applied to studies on cell heterogeneity and differentiation, and epigenetic reprogramming. Keywords: chromatin accessibility; epigenomics; genomics; multiomics; single-cell; transcriptomic

Dark matter in gravity-mediated supersymmetry breaking

In R parity conserving supersymmetric theories the lightest superpartner (LSP) is stable. The LSPs may comprise a large fraction of the energy density of the current universe, which would lead to dramatic astrophysical consequences. In this talk, I discuss some of the main lessons we have learned about supersymmetric models from relic abundance considerations of the LSP.Comment: 6 pages, LaTeX with espcrc2.sty, to be published in the "Proceedings of the 5th International Conference on Supersymmetries in Physics (SUSY 97)", Philadelphia, Pennsylvania, May 27-31, 199

Signatures of Axinos and Gravitinos at Colliders

The axino and the gravitino are well-motivated candidates for the lightest supersymmetric particle (LSP) and also for cold dark matter in the Universe. Assuming that a charged slepton is the next-to-lightest supersymmetric particle (NLSP), we show how the NLSP decays can be used to probe the axino LSP scenario in hadronic axion models as well as the gravitino LSP scenario at the Large Hadron Collider and the International Linear Collider. We show how one can identify experimentally the scenario realized in nature. In the case of the axino LSP, the NLSP decays will allow one to estimate the value of the axino mass and the Peccei-Quinn scale.Comment: 20 pages, 5 figures, revised version as published in Phys.Lett.B (comments on the experimental feasibility added

Bayesian approach and Naturalness in MSSM analyses for the LHC

The start of LHC has motivated an effort to determine the relative probability of the different regions of the MSSM parameter space, taking into account the present, theoretical and experimental, wisdom about the model. Since the present experimental data are not powerful enough to select a small region of the MSSM parameter space, the choice of a judicious prior probability for the parameters becomes most relevant. Previous studies have proposed theoretical priors that incorporate some (conventional) measure of the fine-tuning, to penalize unnatural possibilities. However, we show that such penalization arises from the Bayesian analysis itself (with no ad hoc assumptions), upon the marginalization of the mu-parameter. Furthermore the resulting effective prior contains precisely the Barbieri-Giudice measure, which is very satisfactory. On the other hand we carry on a rigorous treatment of the Yukawa couplings, showing in particular that the usual practice of taking the Yukawas "as required", approximately corresponds to taking logarithmically flat priors in the Yukawa couplings. Finally, we use an efficient set of variables to scan the MSSM parameter space, trading in particular B by tan beta, giving the effective prior in the new parameters. Beside the numerical results, we give accurate analytic expressions for the effective priors in all cases. Whatever experimental information one may use in the future, it is to be weighted by the Bayesian factors worked out here.Comment: LaTeX, 19 pages, 3 figure

The complete radiative corrections to the gaugino and Higgsino masses in the Minimal Supersymmetric Model

We determine the radiative corrections to the masses of the gauginos and Higgsinos in the MSSM, including all sectors of the theory in a one-loop calculation in the on-mass-shell renormalization scheme. We find that a gluino which is massless at tree level receives a mass of between 0 and 3 GeV, primarily due to the top/stop contribution. This radiatively generated mass depends directly on the off-diagonal element of the squark mass matrix. In the case of a massive gluino, its mass receives typically large corrections, as large as 40\% for a 125 GeV gluino. We find that the contributions to the neutralino and chargino mass corrections from the gauge/Higgs/gaugino/Higgsino sector are typically +-1%. The lightest neutralino, which can receive corrections larger than 25%, receives 5% corrections over most of the parameter space. We combine our results with the results of LEP and CDF searches to obtain the lower bounds on the neutralino and chargino masses at one-loop. We also demonstrate how the radiative corrections affect the presently excluded region of parameter space.Comment: 16 pages, JHU-TIPAC-940001, PURD-TH-94-04. 6 uuencoded postscript figures included. Uncomment the line "%\input epsf" to embed the figures in the tex

A description of the neutralino observables in terms of projectors

Applying Jarlskog's treatment of the CKM matrix, to the neutralino mass matrix in MSSM for real soft gaugino SUSY breaking and $\mu$-parameters, we construct explicit analytical expressions for the four projectors which acting on any neutralino state project out the mass eigenstates. Analytical expressions for the neutralino mass eigenvalues in terms of the various SUSY parameters, are also given. It is shown that these projectors and mass eigenvalues are sufficient to describe any physical observable involving neutralinos, to any order of perturbation theory. As an example, the $e^-e^+ \to \tilde \chi^0_i \tilde \chi^0_j$ cross section at tree level is given in terms of these projectors. The expected magnitude of their various matrix elements in plausible SUSY scenarios is also discussed.Comment: 14 pages, no figures. Version to appear in Phys. Rev. D. e-mail: [email protected]

Gravitino Dark Matter and Cosmological Constraints

The gravitino is a promising candidate for cold dark matter. We study cosmological constraints on scenarios in which the gravitino is the lightest supersymmetric particle and a charged slepton the next-to-lightest supersymmetric particle (NLSP). We obtain new results for the hadronic nucleosynthesis bounds by computing the 4-body decay of the NLSP slepton into the gravitino, the associated lepton, and a quark-antiquark pair. The bounds from the observed dark matter density are refined by taking into account gravitinos from both late NLSP decays and thermal scattering in the early Universe. We examine the present free-streaming velocity of gravitino dark matter and the limits from observations and simulations of cosmic structures. Assuming that the NLSP sleptons freeze out with a thermal abundance before their decay, we derive new bounds on the slepton and gravitino masses. The implications of the constraints for cosmology and collider phenomenology are discussed and the potential insights from future experiments are outlined. We propose a set of benchmark scenarios with gravitino dark matter and long-lived charged NLSP sleptons and describe prospects for the Large Hadron Collider and the International Linear Collider.Comment: 51 pages, 20 figures, revised version matches published version (results unchanged, JHEP style used, figures replaced with new high-quality figures, typos corrected, references added

Exploring the Local Grammar of Evaluation: The Case of Adjectival Patterns in American and Italian Judicial Discourse

Based on a 2-million word bilingual comparable corpus of American and Italian judgments, this paper tests the applicability of a local grammar to study evaluative phraseology in judicial discourse in English and Italian. In particular, the study compares the use of two patterns: v-link + ADJ + that pattern / copula + ADJ + che and v-link + ADJ + to-infinitive pattern / copula + ADJ + verbo all’infinito in the disciplinary genre of criminal judgments delivered by the US Supreme Court and the Italian Corte Suprema di Cassazione. It is argued that these two patterns represent a viable and efficient diagnostic tool for retrieving instances of evaluative language and they represent an ideal starting point and a relevant unit of analysis for a cross-language analysis of evaluation in domainrestricted specialised discourse. Further, the findings provided shed light on important interactions occurring among major interactants involved in the judicial discourse