Synthesis of optically-labelled vanadyl complexes and their evaluation as protein tyrosine phosphatase inhibitors and intracellular probes

Cysteine based phosphatases (CBPs) are a diverse family of enzymes that act to remove the phosphate group from proteins, phosphatidylinositols and other biomolecules, thereby counteracting the effects of kinases. Due to the wide range of roles they play in the cell, the CBPs are implicated in many diseases such as cancer, diabetes and neurodegenerative disorders, and therefore it is of interest to develop compounds to study these enzymes. Within our group, vanadium compounds have been shown to inhibit the CBPs with a potency and selectivity that can be tuned by varying the ligand and the oxidation state of the vanadium centre. One subset of interest is the vanadium complexes of 3-hydroxypicolinic acid which have an inhibition range against phosphatases of low nanomolar to high micromolar depending on the oxidation state of vanadium and the enzyme studied. A small selection of novel fluorescent vanadyl complexes based on the hydroxypicolinic acid motif have been synthesised as novel phosphatase inhibitors. Incorporation of an amine-functionalised spacer between the picolinate ligands provided a handle for incorporation of a fluorescent group away from the picolinate-vanadyl core. These compounds were then tested against a range of phosphatases to determine their potency and selectivity as inhibitors of CBPs. The complexes showed good inhibition properties against the CBPs investigated in a range similar to that of free vanadyl. The fluorophore was used to study the interaction of the vanadyl complexes with the enzymes, and to investigate cell uptake. Fluorescence quenching and anisotropy assays were carried out to look at the stability of complexes in assay conditions. From the data obtained it appears that the vanadyl complexes are stable in the presence of the enzymes and under assay conditions. Preliminary data also indicate that the complexes are taken up into cells and therefore can be investigated as intracellular probes.Open Acces

BHPR research: qualitative1. Complex reasoning determines patients' perception of outcome following foot surgery in rheumatoid arhtritis

Background: Foot surgery is common in patients with RA but research into surgical outcomes is limited and conceptually flawed as current outcome measures lack face validity: to date no one has asked patients what is important to them. This study aimed to determine which factors are important to patients when evaluating the success of foot surgery in RA Methods: Semi structured interviews of RA patients who had undergone foot surgery were conducted and transcribed verbatim. Thematic analysis of interviews was conducted to explore issues that were important to patients. Results: 11 RA patients (9 ♂, mean age 59, dis dur = 22yrs, mean of 3 yrs post op) with mixed experiences of foot surgery were interviewed. Patients interpreted outcome in respect to a multitude of factors, frequently positive change in one aspect contrasted with negative opinions about another. Overall, four major themes emerged. Function: Functional ability & participation in valued activities were very important to patients. Walking ability was a key concern but patients interpreted levels of activity in light of other aspects of their disease, reflecting on change in functional ability more than overall level. Positive feelings of improved mobility were often moderated by negative self perception ("I mean, I still walk like a waddling duck”). Appearance: Appearance was important to almost all patients but perhaps the most complex theme of all. Physical appearance, foot shape, and footwear were closely interlinked, yet patients saw these as distinct separate concepts. Patients need to legitimize these feelings was clear and they frequently entered into a defensive repertoire ("it's not cosmetic surgery; it's something that's more important than that, you know?”). Clinician opinion: Surgeons' post operative evaluation of the procedure was very influential. The impact of this appraisal continued to affect patients' lasting impression irrespective of how the outcome compared to their initial goals ("when he'd done it ... he said that hasn't worked as good as he'd wanted to ... but the pain has gone”). Pain: Whilst pain was important to almost all patients, it appeared to be less important than the other themes. Pain was predominately raised when it influenced other themes, such as function; many still felt the need to legitimize their foot pain in order for health professionals to take it seriously ("in the end I went to my GP because it had happened a few times and I went to an orthopaedic surgeon who was quite dismissive of it, it was like what are you complaining about”). Conclusions: Patients interpret the outcome of foot surgery using a multitude of interrelated factors, particularly functional ability, appearance and surgeons' appraisal of the procedure. While pain was often noted, this appeared less important than other factors in the overall outcome of the surgery. Future research into foot surgery should incorporate the complexity of how patients determine their outcome Disclosure statement: All authors have declared no conflicts of interes

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

Mutations in phospholipase C epsilon 1 are not sufficient to cause diffuse mesangial sclerosis

Diffuse mesangial sclerosis occurs as an isolated abnormality or as a part of a syndrome. Recently, mutations in phospholipase C epsilon 1 (PLCE1) were found to cause a nonsyndromic, autosomal recessive form of this disease. Here we describe three children from one consanguineous kindred of Pakistani origin with diffuse mesangial sclerosis who presented with congenital or infantile nephrotic syndrome. Homozygous mutations in PLCE1 (also known as KIAA1516, PLCE, or NPHS3) were identified following genome-wide mapping of single-nucleotide polymorphisms. All affected children were homozygous for a four-basepair deletion in exon 3, which created a premature translational stop codon. Analysis of the asymptomatic father of two of the children revealed that he was also homozygous for the same mutation. We conclude this nonpenetrance may be due to compensatory mutations at a second locus and that mutation within PLCE1 is not always sufficient to cause diffuse mesangial sclerosi

"Gênero" para um dicionário marxista: a política sexual de uma palavra "Gender" for a marxist dictionary: the sexual politics of a word

A autora conta suas desventuras teóricas após aceitar escrever um verbete sobre "gênero" para um dicionário marxista reputado. Em suas próprias palavras: "Além disso, mesmo se Marx e Engels - ou até Gayle Rubin - não se aventuraram pela sexologia, medicina ou biologia em suas discussões sobre sexo/gênero, ou sobre a questão da mulher, eu sabia que teria de fazê-lo. Ao mesmo tempo, estava claro que outras GRANDES correntes dos escritos feministas modernos sobre sexo, sexualidade e gênero se entrelaçavam constantemente mesmo com as mais modestas interpretações de minha encomenda. A maioria delas, talvez especialmente as correntes psicanalítica e literária do feminismo francês e inglês, não aparece em meu verbete sobre Geschlecht. De modo geral, o verbete abaixo focaliza os escritos das feministas norte-americanas. Este não é um escândalo trivial."<br>The misadventures of the Author in writing about a "key-word" for a famous marxist dictionary are recounted here. In her own words: "Also, even if Marx and Engels - or Gayle Rubin, for that matter - had not ventured into sexology, medicine, or biology for their discussions of sex/gender or the woman question, I knew I would have to do so. At the same time, it was clear that other BIG currents of modern feminist writing on sex, sexuality, and gender interlaced constantly with even the most modest interpretation of my assignment. Most of those, perhaps especially the French and British feminist psychoanalytic and literary currents, do not appear in my entry on Geschlecht. In general, the entry below focuses on writing by US feminists. This is not a trivial scandal.

The CMS Statistical Analysis and Combination Tool: COMBINE

International audienceThis paper describes the COMBINE software package used for statistical analyses by the CMS Collaboration. The package, originally designed to perform searches for a Higgs boson and the combined analysis of those searches, has evolved to become the statistical analysis tool presently used in the majority of measurements and searches performed by the CMS Collaboration. It is not specific to the CMS experiment, and this paper is intended to serve as a reference for users outside of the CMS Collaboration, providing an outline of the most salient features and capabilities. Readers are provided with the possibility to run COMBINE and reproduce examples provided in this paper using a publicly available container image. Since the package is constantly evolving to meet the demands of ever-increasing data sets and analysis sophistication, this paper cannot cover all details of COMBINE. However, the online documentation referenced within this paper provides an up-to-date and complete user guide

Search for long-lived heavy neutrinos in the decays of B mesons produced in proton-proton collisions at s\sqrt{s} = 13 TeV

International audienceA search for long-lived heavy neutrinos (N) in the decays of \PB mesons produced in proton-proton collisions at $\sqrt{s}$ = 13 TeV is presented. The data sample corresponds to an integrated luminosity of 41.6 fb$^{-1}$ collected in 2018 by the CMS experiment at the CERN LHC, using a dedicated data stream that enhances the number of recorded events containing B mesons. The search probes heavy neutrinos with masses in the range 1 $\lt$$m_\mathrm{N}$$\lt$ 3 GeV and decay lengths in the range 10$^{-2}$$\lt$$c\tau$$\lt$ 10$^{4}$ mm, where $\tau_\mathrm{N}$ is the N proper mean lifetime. Signal events are defined by the signature B $\to$$\ell_\mathrm{B}$NX; N $\to$$\ell^{\pm} \pi^{\mp}$, where the leptons $\ell_\mathrm{B}$ and $\ell$ can be either a muon or an electron, provided that at least one of them is a muon. The hadronic recoil system, X, is treated inclusively and is not reconstructed. No significant excess of events over the standard model background is observed in any of the $\ell^{\pm}\pi^{\mp}$ invariant mass distributions. Limits at 95% confidence level on the sum of the squares of the mixing amplitudes between heavy and light neutrinos, $\vert V_\mathrm{N}\vert^2$, and on $c\tau$ are obtained in different mixing scenarios for both Majorana and Dirac-like N particles. The most stringent upper limit $\vert V_\mathrm{N}\vert^2$ $\lt$ 2.0$\times$10$^{-5}$ is obtained at $m_\mathrm{N}$ = 1.95 GeV for the Majorana case where N mixes exclusively with muon neutrinos. The limits on $\vert V_\mathrm{N}\vert^2$ for masses 1 $\lt$ $m_\mathrm{N}$ $\lt$ 1.7 GeV are the most stringent from a collider experiment to date

Dark sector searches with the CMS experiment

Astrophysical observations provide compelling evidence for gravitationally interacting dark matter in the universe that cannot be explained by the standard model of particle physics. The extraordinary amount of data from the CERN LHC presents a unique opportunity to shed light on the nature of dark matter at unprecedented collision energies. This Report comprehensively reviews the most recent searches with the CMS experiment for particles and interactions belonging to a dark sector and for dark-sector mediators. Models with invisible massive particles are probed by searches for signatures of missing transverse momentum recoiling against visible standard model particles. Searches for mediators are also conducted via fully visible final states. The results of these searches are compared with those obtained from direct-detection experiments. Searches for alternative scenarios predicting more complex dark sectors with multiple new particles and new forces are also presented. Many of these models include long-lived particles, which could manifest themselves with striking unconventional signatures with relatively small amounts of background. Searches for such particles are discussed and their impact on dark-sector scenarios is evaluated. Many results and interpretations have been newly obtained for this Report.Astrophysical observations provide compelling evidence for gravitationally interacting dark matter in the universe that cannot be explained by the standard model of particle physics. The extraordinary amount of data from the CERN LHC presents a unique opportunity to shed light on the nature of dark matter at unprecedented collision energies. This Report comprehensively reviews the most recent searches with the CMS experiment for particles and interactions belonging to a dark sector and for dark-sector mediators. Models with invisible massive particles are probed by searches for signatures of missing transverse momentum recoiling against visible standard model particles. Searches for mediators are also conducted via fully visible final states. The results of these searches are compared with those obtained from direct-detection experiments. Searches for alternative scenarios predicting more complex dark sectors with multiple new particles and new forces are also presented. Many of these models include long-lived particles, which could manifest themselves with striking unconventional signatures with relatively small amounts of background. Searches for such particles are discussed and their impact on dark-sector scenarios is evaluated. Many results and interpretations have been newly obtained for this Report