Gene expression analyses determine two different subpopulations in KIT-negative GIST-like (KNGL) patients

Introduction: There are limited findings available on KIT-negative GIST-like (KNGL) population. Also, KIT expression may be post-transcriptionally regulated by miRNA221 and miRNA222. Hence, the aim of this study is to characterize KNGL population, by differential gene expression, and to analyze miRNA221/222 expression and their prognostic value in KNGL patients. Methods: KIT, PDGFRA, DOG1, IGF1R, MIR221 and MIR222 expression levels were determined by qRT-PCR. We also analyzed KIT and PDGFRA mutations, DOG1 expression, by immunohistochemistry, along with clinical and pathological data. Disease-free survival (DFS) and overall survival (OS) differences were calculated using Log-rank test. Results: Hierarchical cluster analyses from gene expression data identified two groups: group I had KIT, DOG1 and PDGFRA overexpression and IGF1R underexpression and group II had overexpression of IGF1R and low expression of KIT, DOG1 and PDGFRA. Group II had a significant worse OS (p = 0.013) in all the series, and showed a tendency for worse OS (p = 0.11), when analyzed only the localized cases. MiRNA222 expression was significantly lower in a control subset of KIT-positive GIST (p < 0.001). OS was significantly worse in KNGL cases with higher expression of MIR221 (p = 0.028) or MIR222 (p = 0.014). Conclusions: We identified two distinct KNGL subsets, with a different prognostic value. Increased levels of miRNA221/222, which are associated with worse OS, could explain the absence of KIT protein expression of most KNGL tumors

An ultra-sensitive electrochemical biosensor using the Spike protein for capturing antibodies against SARS-CoV-2 in point-of-care

Funding Information: The authors acknowledge funding through project TecniCov ( POCI-01-02B7-FEDER-069745 ), co-funded by FEDER through COMPETE2020 and Lisboa2020 and CY-Sensors ( POCI-01-0145-FEDER-032359 ) through Fundação para a Ciência e a Tecnologia (FCT) , Portugal. ARC acknowledge funding to National Foundation for Science and Technology , I.P., Portugal ( FCT ) through the PhD. Grant, reference SFRH/BD/130107/2017 . Publisher Copyright: © 2022 The AuthorsThis work presents an innovative ultra-sensitive biosensor having the Spike protein on carbon-based screen-printed electrodes (SPEs), for monitoring in point-of-care antibodies against SARS-CoV-2, a very important tool for epidemiological monitoring of COVID-19 infection and establishing vaccination schemes. In an innovative and simple approach, a highly conductive support is combined with the direct adsorption of Spike protein to enable an extensive antibody capture. The high conductivity was ensured by using carboxylated carbon nanotubes on the carbon electrode, by means of a simple and quick approach, which also increased the surface area. These were then modified with EDC/NHS chemistry to produce an amine layer and undergo Spike protein adsorption, to generate a stable layer capable of capturing the antibodies against SARS-CoV-2 in serum with great sensitivity. Electrochemical impedance spectroscopy was used to evaluate the analytical performance of this biosensor in serum. It displayed a linear response between 1.0 ​pg/mL and 10 ​ng/mL, with a detection limit of ∼0.7 ​pg/mL. The analysis of human positive sera containing antibody in a wide range of concentrations yielded accurate data, correlating well with the reference method. It also offered the unique ability of discriminating antibody concentrations in sera below 2.3 ​μg/mL, the lowest value detected by the commercial method. In addition, a proof-of-concept study was performed by labelling anti-IgG antibodies with quantum dots to explore a new electrochemical readout based on the signal generated upon binding to the anti-S protein antibodies recognised on the surface of the biosensor. Overall, the alternative serologic assay presented is a promising tool for assessing protective immunity to SARS-CoV-2 and a potential guide for revaccination.publishersversionpublishe

Substitute therapy of the sclera. Identification of the patologies and methodologies in the University Hospital of Granada (2014-2019)

Introducción: La esclera es un tejido avascular compuesto por tejido conectivo denso cuya función principal es proteger las estructuras intraoculares. Existen diversas patologías que ponen en riesgo la integridad de dicho tejido y, en consecuencia, pueden provocar terribles secuelas con un pronóstico nefasto. El tratamiento sustitutivo forma parte de la terapia de dicha patología, siendo el injerto de esclerótica cadáver una opción biocompatible y biomimética a valorar. El injerto de esclerótica de cadáver es un tejido de fácil acceso y con buenos resultados post-quirúrgicos por lo que no solo es usado en el ámbito de la patología escleral, sino que también es utilizado para cirugía de oculoplástica, glaucoma o de retina. No obstante, el peligro de transmisión de priones o el déficit de suministro en algunos países hacen que este tejido sea suplantado por otros tejidos o técnicas quirúrgicas. Métodos: Los objetivos de este estudio son identificar la patología esclerótica susceptible de terapia sustitutiva en los últimos 5 años en el complejo hospitalario universitario de Granada, identificar los métodos empleados como terapéutica sustitutiva en dicha patología y evaluar la práctica de dicha terapia en el contexto de la bibliografía existente al respecto. Para ello, se aplicó un diseño retrospectivo en el que se realizó una búsqueda en los archivos de la unidad de documentación clínica del Hospital Clínico San Cecilio y Hospital Virgen de las Nieves, de los cuales se obtuvieron 9 casos clínicos. Resultados: Entre los 9 casos clínicos, solo 4 fueron sometidos a tratamiento quirúrgico, y de estos, 3 de ellos a terapia sustitutiva por membrana amniótica o injerto escleral. En los 5 casos restantes no se llevó a cabo ninguna técnica quirúrgica, no obstante, podría ser de gran valor el uso del refuerzo con injerto de esclerótica en los casos con escleromalacia para evitar complicaciones futuras. La esclerótica procedente de cadáver se postula como una opción con unos resultados prometedores; sin embargo, las líneas de investigación actuales apuestan por el desarrollo de un sustituto basado en las ventajas de la fibrina agarosa que supla los puntos negativos del tejido donante.Introduction: The sclera is an avascular tissue composed of dense connective tissue whose main function is to protect intraocular structures. There are several pathologies that jeopardize the integrity of this tissue and, consequently, can cause terrible consequences with a dire prognosis. Substitute treatment is part of the therapy of this pathology, with the donor sclera graft being a biocompatible and biomimetic option to be evaluated. The donor sclera graft is an easily accessible tissue with good post-surgical results, so it is not only used in the field of scleral pathology, but also used for oculoplastic, glaucoma or retinal surgery. However, the danger of prion transmission or the supply deficit in some countries causes that, other tissues or surgical techniques supplant this tissue. Methods: The objectives of this study are to identify the sclerotic pathology susceptible of replacement therapy in the last 5 years in the university hospital of Granada, identify the methods used as a replacement therapy in this pathology and evaluate the practice of such therapy. A retrospective design was applied and a search was made in the archives of the clinical documentation unit of the San Cecilio Clinical Hospital and Virgen de las Nieves Hospital. There were 9 clinical cases found. Results: Among the nine clinical cases, only 4 were subjected to surgical treatment, and of these, three of them for replacement therapy of donor sclera or amniotic membrane. In the remaining 5 cases did not carry out any surgical technique, however, it could be of great value using reinforcement sclera graft in cases with scleromalacia to avoid further complications. Sclera from cadaver is postulated as an option with promising results; however, current research committed to the development of a substitute based on the advantages of fibrin agarose that mitigates the negative points of the donor tissue

Counteracting gradients of light and soil nutrients in the understorey of Mediterranean oak forests.

The forest canopy modifies the availability of resources (light, water, and soil nutrients) in the understorey. In this paper we analyze the relationships between woody canopy density, litter accumulation, and topsoil N and P availability in the understorey of two oak forests: one in southern Portugal and the other in southern Spain. Both forests persist on low-nutrient soils, particularly poor in P. We hypothesize that direct and indirect effects of the canopy overstorey cause opposite gradients in the availability of essential resources (light and key soil nutrients) in the understorey. In both studied forests we found significant relationships between the overall canopy density, light availability, topsoil litter accumulation, and the availability of N and P, which frequently limit plant growth. Path analysis (by Shipley’s d-sep method) showed that the available data were consistent with the proposed causal model. The average values of soil variables at the end quartiles of the light-availability gradient were compared. Results showed large differences in litter accumulation (~30×) and available-N and -P topsoil concentrations (~3×) in the Spanish forest (with the wider environmental gradient). Furthermore, P increased from the ‘very low’ range to the ‘low’ or even the ‘optimum’ range of availability (according to standard plant growth criteria), which suggests potential effects on the growth of the understorey plant species. We conclude that the counteracting gradients of the essential resources -light and nutrients- in the forest understorey resulted from direct and indirect effects of the canopy overstorey, respectively. We suggest that these counteracting effects of the woody canopy on essential resources of different nature must be considered when interpreting the patterns of understorey plant populations and communities.The spanish MEC (CGL2005-05830-C03-01-BOS, DINAMED project) and the Portuguese FCT(SFRH/BD/8322/2002 grant to SMM)supported the research.Peer reviewe

Observation of two new Ξb−\Xi_b^- baryon resonances

Two structures are observed close to the kinematic threshold in the $\Xi_b^0 \pi^-$ mass spectrum in a sample of proton-proton collision data, corresponding to an integrated luminosity of 3.0 fb$^{-1}$ recorded by the LHCb experiment. In the quark model, two baryonic resonances with quark content $bds$ are expected in this mass region: the spin-parity $J^P = \frac{1}{2}^+$ and $J^P=\frac{3}{2}^+$ states, denoted $\Xi_b^{\prime -}$ and $\Xi_b^{*-}$. Interpreting the structures as these resonances, we measure the mass differences and the width of the heavier state to be $m(\Xi_b^{\prime -}) - m(\Xi_b^0) - m(\pi^{-}) = 3.653 \pm 0.018 \pm 0.006$ MeV$/c^2$, $m(\Xi_b^{*-}) - m(\Xi_b^0) - m(\pi^{-}) = 23.96 \pm 0.12 \pm 0.06$ MeV$/c^2$, $\Gamma(\Xi_b^{*-}) = 1.65 \pm 0.31 \pm 0.10$ MeV, where the first and second uncertainties are statistical and systematic, respectively. The width of the lighter state is consistent with zero, and we place an upper limit of $\Gamma(\Xi_b^{\prime -}) < 0.08$ MeV at 95% confidence level. Relative production rates of these states are also reported.Comment: 17 pages, 2 figure

Measurement of the CP-violating phase \phi s in Bs->J/\psi\pi+\pi- decays

Measurement of the mixing-induced CP-violating phase phi_s in Bs decays is of prime importance in probing new physics. Here 7421 +/- 105 signal events from the dominantly CP-odd final state J/\psi pi+ pi- are selected in 1/fb of pp collision data collected at sqrt{s} = 7 TeV with the LHCb detector. A time-dependent fit to the data yields a value of phi_s=-0.019^{+0.173+0.004}_{-0.174-0.003} rad, consistent with the Standard Model expectation. No evidence of direct CP violation is found.Comment: 15 pages, 10 figures; minor revisions on May 23, 201

Observation of associated production of a ZZ boson with a DD meson in the~forward region

A search for associated production of a $Z$ boson with an open charm meson is presented using a data sample, corresponding to an integrated luminosity of $1.0\,\mathrm{fb}^{-`}$ of proton--proton collisions at a centre-of-mass energy of 7\,TeV, collected by the LHCb experiment. %% Seven candidate events for associated production of a $Z$ boson with a $D^0$ meson and four candidate events for a $Z$ boson with a $D^+$ meson are observed with a combined significance of 5.1standard deviations. The production cross-sections in the forward region are measured to be $$\sigma_{Z\rightarrow\mu^+\mu^-\!,D^0} = 2.50\pm1.12\pm0.22pb$$ $$\sigma_{Z\rightarrow\mu^+\mu^-\!,D^+} = 0.44\pm0.23\pm0.03pb,$$ where the first uncertainty is statistical and the second systematic.Comment: 18 pages, 2 figure

Measurements of the B+B^+, B0B^0, Bs0B_s^0 meson and Λb0\Lambda_b^0 baryon lifetimes

Measurements of $b$-hadron lifetimes are reported using $pp$ collision data, corresponding to an integrated luminosity of 1.0fb$^{-1}$, collected by the LHCb detector at a centre-of-mass energy of $7$Tev. Using the exclusive decays $B^+\to J/\psi K^+$, $B^0\to J/\psi K^*(892)^0$, $B^0\to J/\psi K^0_{\rm S}$, $\Lambda_b^0\to J/\psi \Lambda$ and $B^0_s\to J/\psi \phi$ the average decay times in these modes are measured to be $\tau_{B^+\to J/\psi K^+}$ = $1.637 \pm$ 0.004 $\pm$ 0.003 ps, $\tau_{B^0\to J/\psi K^*(892)^0}$ = $1.524 \pm$ 0.006 $\pm$ 0.004 ps, $\tau_{B^0\to J/\psi K^0_{\rm S}}$ = $1.499 \pm$ 0.013 $\pm$ 0.005 ps, $\tau_{\Lambda_b^0\to J/\psi \Lambda}$ = $1.415 \pm$ 0.027 $\pm$ 0.006 ps and $\tau_{B^0_s\to J/\psi \phi}$ = $1.480 \pm$ 0.011 $\pm$ 0.005 ps, where the first uncertainty is statistical and the second is systematic. These represent the most precise lifetime measurements in these decay modes. In addition, ratios of these lifetimes, and the ratio of the decay-width difference, $\Delta\Gamma_d$, to the average width, $\Gamma_d$, in the $B^0$ system, $\Delta \Gamma_d/\Gamma_d = -0.044 \pm 0.025 \pm 0.011$, are reported. All quantities are found to be consistent with Standard Model expectations.Comment: 28 pages, 4 figures. Updated reference

Observation of an Excited Bc+ State

Using pp collision data corresponding to an integrated luminosity of 8.5 fb-1 recorded by the LHCb experiment at center-of-mass energies of s=7, 8, and 13 TeV, the observation of an excited Bc+ state in the Bc+π+π- invariant-mass spectrum is reported. The observed peak has a mass of 6841.2±0.6(stat)±0.1(syst)±0.8(Bc+) MeV/c2, where the last uncertainty is due to the limited knowledge of the Bc+ mass. It is consistent with expectations of the Bc∗(2S31)+ state reconstructed without the low-energy photon from the Bc∗(1S31)+→Bc+γ decay following Bc∗(2S31)+→Bc∗(1S31)+π+π-. A second state is seen with a global (local) statistical significance of 2.2σ (3.2σ) and a mass of 6872.1±1.3(stat)±0.1(syst)±0.8(Bc+) MeV/c2, and is consistent with the Bc(2S10)+ state. These mass measurements are the most precise to date

Neddylation inhibition prevents acetaminophen-induced liver damage by enhancing the anabolic cardiolipin pathway

\ua9 2024 The AuthorsDrug-induced liver injury (DILI) is a significant cause of acute liver failure (ALF) and liver transplantation in the Western world. Acetaminophen (APAP) overdose is a main contributor of DILI, leading to hepatocyte cell death through necrosis. Here, we identified that neddylation, an essential post-translational modification involved in the mitochondria function, was upregulated in liver biopsies from patients with APAP-induced liver injury (AILI) and in mice treated with an APAP overdose. MLN4924, an inhibitor of the neuronal precursor cell-expressed developmentally downregulated protein 8 (NEDD8)-activating enzyme (NAE-1), ameliorated necrosis and boosted liver regeneration in AILI. To understand how neddylation interferes in AILI, whole-body biotinylated NEDD8 (bioNEDD8) and ubiquitin (bioUB) transgenic mice were investigated under APAP overdose with and without MLN4924. The cytidine diphosphate diacylglycerol (CDP-DAG) synthase TAM41, responsible for producing cardiolipin essential for mitochondrial activity, was found modulated under AILI and restored its levels by inhibiting neddylation. Understanding this ubiquitin-like crosstalk in AILI is essential for developing promising targeted inhibitors for DILI treatment