Preclinical and randomized phase I studies of plitidepsin in adults hospitalized with COVID-19.

Plitidepsin, a marine-derived cyclic-peptide, inhibits SARS-CoV-2 replication at nanomolar concentrations by targeting the host protein eukaryotic translation elongation factor 1A. Here, we show that plitidepsin distributes preferentially to lung over plasma, with similar potency against across several SARS-CoV-2 variants in preclinical studies. Simultaneously, in this randomized, parallel, open-label, proof-of-concept study (NCT04382066) conducted in 10 Spanish hospitals between May and November 2020, 46 adult hospitalized patients with confirmed SARS-CoV-2 infection received either 1.5 mg (n = 15), 2.0 mg (n = 16), or 2.5 mg (n = 15) plitidepsin once daily for 3 d. The primary objective was safety; viral load kinetics, mortality, need for increased respiratory support, and dose selection were secondary end points. One patient withdrew consent before starting procedures; 45 initiated treatment; one withdrew because of hypersensitivity. Two Grade 3 treatment-related adverse events were observed (hypersensitivity and diarrhea). Treatment-related adverse events affecting more than 5% of patients were nausea (42.2%), vomiting (15.6%), and diarrhea (6.7%). Mean viral load reductions from baseline were 1.35, 2.35, 3.25, and 3.85 log10 at days 4, 7, 15, and 31. Nonmechanical invasive ventilation was required in 8 of 44 evaluable patients (16.0%); six patients required intensive care support (13.6%), and three patients (6.7%) died (COVID-19-related). Plitidepsin has a favorable safety profile in patients with COVID-19.

La eficacia de rol, la claridad de rol y el rendimiento en jóvenes futbolistas / Role efficacy, role clarity and performance in youth footballers / Eficácia e clareza de papel e o rendimento em jovens futebolistas / Eficácia e clareza de papel e o rendimento em jovens futebolistas

El objetivo del trabajo fue doble: 1) Analizar las relaciones entre la eficacia de rol, la claridad de rol y el desempeño efectivo del rol como un indicador subjetivo del rendimiento; y 2) Examinar el papel de la claridad de rol como variable moduladora de la relación entre la eficacia de rol y el rendimiento, ambos objetivos en situaciones de ataque y de defensa en el fútbol. Doscientos noventa y cinco futbolistas varones de entre 12 y 16 años (M = 14.01, DE = 1.09) completaron los cuestionarios que evaluaban las variables del estudio. Los resultados mostraron que aquéllos que percibían una claridad de rol alta también percibían una eficacia de rol alta y un desempeño del rol alto. Además se confirmó el papel modulador de la claridad de rol en las funciones defensivas, concretamente, la eficacia de rol resultó ser un predictor positivo del rendimiento siempre y cuando la claridad de rol fuera alta

O objectivo do trabalho foi duplo: 1) Analisar as relações entre a eficácia de papel, a clareza de papel e o desempenho efectivo do papelcomo indicador subjectivo do rendimento; e 2) Examinar a função da clareza de papel como variável moderadora da relação entre a eficácia do papel eo rendimento, ambos objectivos em situações de ataque e defesa no futebol. Duzentos e noventa e cinco futebolistas rapazes com idades compreendidasentre os 12 e os 16 anos (M= 14.01, DP= 1.09) preencheram os questionários que avaliavam as variáveis do estudo. Os resultados revelam que aquelesque percebiam uma elevada clareza de papel também percebiam uma elevada eficácia de papel e um elevado desempenho de papel. Adicionalmente,confirmou-se o papel moderador da clareza de papel nas funções defensivas, concretamente, a eficácia de papel revelou ser um preditor positivo dorendimento sempre e quando a clareza de papel era elevada

The aim of this study was twofold: 1) to analyze the relationship between role efficacy, role clarity, and role performance effectiveness as a subjective measure of performance, and 2) to examine role clarity as a moderator of the role efficacy-performance relationship, in both an offensive and defensive context in football. Two hundred and ninety-five male soccer players between 12 and 16 years old (M = 14.01, SD = 1.09) completed the questionnaires assessing the variables of the study. Results showed that the players who reported higher role clarity also reported higher role efficacy and performed better than those with lower role clarity. Also, role clarity was a moderator for defensive role functions; specifically, role efficacy was a positive predictor of performance only when role clarity was high

O objectivo do trabalho foi duplo: 1) Analisar as relações entre a eficácia de papel, a clareza de papel e o desempenho efectivo do papelcomo indicador subjectivo do rendimento; e 2) Examinar a função da clareza de papel como variável moderadora da relação entre a eficácia do papel eo rendimento, ambos objectivos em situações de ataque e defesa no futebol. Duzentos e noventa e cinco futebolistas rapazes com idades compreendidasentre os 12 e os 16 anos (M= 14.01, DP= 1.09) preencheram os questionários que avaliavam as variáveis do estudo. Os resultados revelam que aquelesque percebiam uma elevada clareza de papel também percebiam uma elevada eficácia de papel e um elevado desempenho de papel. Adicionalmente,confirmou-se o papel moderador da clareza de papel nas funções defensivas, concretamente, a eficácia de papel revelou ser um preditor positivo dorendimento sempre e quando a clareza de papel era elevada

Use of virus vectors for the expression in plants of active full-length and single chain anti-coronavirus antibodies.

To extend the potential of antibodies and their derivatives to provide passive protection against enteric infections when supplied orally in crude plant extracts, we have expressed both a small immune protein (SIP) and a full-length antibody in plants using two different plant virus vectors based on potato virus X (PVX) and cowpea mosaic virus (CPMV). The alphaSIP molecule consisted of a single chain antibody (scFv) specific for the porcine coronavirus, transmissible gastroenteritis virus (TGEV) linked to the alpha-CH3 domain from human IgA. To express the full-length IgA, the individual light and heavy chains from the TGEV-specific mAb 6A.C3 were inserted into separate PVX constructs and plants were co-infected with both constructs. Western blot analysis revealed the efficient expression of both the SIP and IgA molecules. Analysis of crude plant extracts revealed that both the plant-expressed alphaSIP and IgA molecules could bind to and neutralize TGEV in tissue culture, indicating that active molecules were produced. Oral administration of crude extracts from antibody-expressing plant tissue to 2-day-old piglets showed that both the alphaSIP and full-length IgA molecules can provide in vivo protection against TGEV.

An antibody derivative expressed from viral vectors passively immunizes pigs against transmissible gastroenteritis virus infection when supplied orally in crude plant extracts.

To investigate the potential of antibody derivatives to provide passive protection against enteric infections when supplied orally in crude plant extracts, we have expressed a small immune protein (SIP) in plants using two different plant virus vectors based on potato virus X (PVX) and cowpea mosaic virus (CPMV). The epsilonSIP molecule consisted of a single-chain antibody (scFv) specific for the porcine coronavirus transmissible gastroenteritis virus (TGEV) linked to the epsilon-CH4 domain from human immunoglobulin E (IgE). In some constructs, the sequence encoding the epsilonSIP molecule was flanked by the leader peptide from the original murine antibody at its N-terminus and an endoplasmic reticulum retention signal (HDEL) at its C-terminus to allow the expressed protein to be directed to, and retained within, the endoplasmic reticulum. Western blot analysis of samples from Nicotiana clevelandii or cowpea tissue infected with constructs revealed the presence of SIP molecules which retained their ability to dimerize. The analysis of crude plant extracts revealed that the plant-expressed epsilonSIP molecules could bind to and neutralize TGEV in tissue culture, the levels of binding and neutralization reflecting the level of expression. Oral administration of crude extracts from SIP-expressing plant tissue to 2-day-old piglets demonstrated that the extracts which showed the highest levels of in vitro neutralization could also provide in vivo protection against challenge with TGEV.

Engineering the transmissible gastroenteritis virus genome as an expression vector inducing lactogenic immunity.

The genome of the coronavirus transmissible gastroenteritis virus (TGEV) has been engineered as an expression vector with an infectious cDNA. The vector led to the efficient (>40 micro g/10(6) cells) and stable (>20 passages) expression of a heterologous gene (green fluorescent protein [GFP]), driven by the transcription-regulating sequences (TRS) of open reading frame (ORF) 3a inserted in the site previously occupied by the nonessential ORFs 3a and 3b. Expression levels driven by this TRS were higher than those of an expression cassette under the control of regulating sequences engineered with the N gene TRS. The recombinant TGEV including the GFP gene was still enteropathogenic, albeit with a 10- to 10(2)-fold reduction in enteric tissue growth. Interestingly, a specific lactogenic immune response against the heterologous protein has been elicited in sows and their progeny. The engineering of an additional insertion site for the heterologous gene between viral genes N and 7 led to instability and to a new genetic organization of the 3' end of the recombinant viruses. As a consequence, a major species of subgenomic mRNA was generated from a TRS with the noncanonical core sequence 5'-CUAAAA-3'. Extension of the complementarity between the TRS and sequences at the 3' end of the viral leader was associated with transcriptional activation of noncanonical core sequences. The engineered vector led to expression levels as high as those of well-established vectors and seems very promising for the development of vaccines and, possibly, for gene therapy.