Combination therapy of Epidermal Growth Factor and Growth Hormone-Releasing Hexapeptide in acute ischemic stroke: a phase I/II non-blinded, randomized clinical trial

ObjectiveThis study tested the hypothesis that a neuroprotective combined therapy based on epidermal growth factor (EGF) and growth hormone-releasing hexapeptide (GHRP6) could be safe for acute ischemic stroke patients, admitting up to 30% of serious adverse events (SAE) with proven causality.MethodsA multi-centric, randomized, open-label, controlled, phase I-II clinical trial with parallel groups was conducted (July 2017 to January 2018). Patients aged 18–80 years with a computed tomography-confirmed ischemic stroke and less than 12 h from the onset of symptoms were randomly assigned to the study groups I (75 μg rEGF + 3.5 mg GHRP6 i.v., n=10), II (75 μg rEGF + 5 mg GHRP6 i.v., n=10), or III (standard care control, n=16). Combined therapy was given BID for 7 days. The primary endpoint was safety over 6 months. Secondary endpoints included neurological (NIHSS) and functional [Barthel index and modified Rankin scale (mRS)] outcomes.ResultsThe study population had a mean age of 66 ± 11 years, with 21 men (58.3%), a baseline median NIHSS score of 9 (95% CI: 8–11), and a mean time to treatment of 7.3 ± 2.8 h. Analyses were conducted on an intention-to-treat basis. SAEs were reported in 9 of 16 (56.2%) patients in the control group, 3 of 10 (30%) patients in Group I (odds ratio (OR): 0.33; 95% CI: 0.06–1.78), and 2 of 10 (20%) patients in Group II (OR: 0.19; 95% CI: 0.03–1.22); only two events in one patient in Group I were attributed to the intervention treatment. Compliance with the study hypothesis was greater than 0.90 in each group. Patients treated with EGF + GHRP6 had a favorable neurological and functional evolution at both 90 and 180 days, as evidenced by the inferential analysis of NIHSS, Barthel, and mRS and by their moderate to strong effect size. At 6 months, proportion analysis evidenced a higher survival rate for patients treated with the combined therapy. Ancillary analysis including merged treated groups and utility-weighted mRS also showed a benefit of this combined therapy.ConclusionEGF + GHRP6 therapy was safe. The functional benefits of treatment in this study supported a Phase III study.Clinical Trial RegistrationRPCEC00000214 of the Cuban Public Registry of Clinical Trials, Unique identifier: IG/CIGB-845I/IC/1601

Intralesional infiltrations of arteriosclerotic tissue cells-free filtrate reproduce vascular pathology in healthy recipient rats

Lower-extremity arterial disease is a major health problem with increasing prevalence, often leading to non-traumatic amputation, disability and mortality. The molecular mechanisms underpinning abnormal vascular wall remodeling are not fully understood. We hypothesized on the existence of a vascular tissue memory that may be transmitted through soluble signaling messengers, transferred from humans to healthy recipient animals, and consequently drive the recapitulation of arterial wall thickening and other vascular pathologies. We examined the effects of the intralesional infiltration for 6 days of arteriosclerotic popliteal artery-derived homogenates (100 µg of protein) into rats’ full-thickness wounds granulation tissue. Animals infiltrated with normal saline solution or healthy brachial arterial tissue homogenate obtained from traumatic amputation served as controls. The significant thickening of arteriolar walls was the constant outcome in two independent experiments for animals receiving arteriosclerotic tissue homogenates. This material induced other vascular morphological changes including an endothelial cell phenotypic reprogramming that mirrored the donor’s vascular histopathology. The immunohistochemical expression pattern of relevant vascular markers appeared to match between the human tissue and the corresponding recipient rats. These changes occurred within days of administration, and with no cross-species limitation. The identification of these “vascular disease drivers” may pave novel research avenues for atherosclerosis pathobiology

Efectos sobre la fertilidad del cerdo de dos candidatos vacunales recombinantes basados en la GnRH

This paper describes for the first time the generation of the K88ab-GnRH hybrid fimbriae, the fusion of N. meningitidis P64k protein (P64k-GnRH), and its evaluation as vaccine candidates to control fertility in mammals. Twenty hybrid male pigs were randomly distributed in four groups: placebos and immunized with K88ab-GnRH, P64k-GnRH and a GnRH analogue (GnRHm1), linked to a tetanus toxoid (TT) T-helper epitope (positive control), respectively. The pigs were immunized at 9-10 weeks of age, using a two-dose scheme, and were sacrificed sixteen weeks later. K88ab-GnRH, P64k-GnRH, and GnRHm1-TT induced higher, similar, and lower testosterone levels in the serum, compared to the placebo, respectively. In the K88ab-GnRH group, the pigs underwent a reduction in testicle size and weight (P < 0.01), and the weight of epididymes compared to the placebo; none of them was able to ejaculate. In the P64k-GnRH group, the pigs had a reduction in testicle weight (P < 0.05), and only one of them was able to ejaculate. The testicles of the pigs immunized with K88ab-GnRH and P64k-GnRH showed structural and functional damage; spermatogenesis was also affected. The accessory sexual glands of the P64k-GnRH group were normal, in contrast to K88ab-GnRH, where interstitial fibrosis was observed. The damage caused by K88ab-GnRH and P64k-GnRH in the target organs evaluated were in all cases lower than the affectations caused by the GnRHm1-TT peptide.En este trabajo se describe, por primera vez, la obtención de la fimbria híbrida K88ab-GnRH, la fusión de la GnRH a la proteína P64k de N. meningitidis (P64k-GnRH) y su evaluación como candidatos vacunales para controlar la fertilidad en mamíferos. Veinte cerdos machos híbridos fueron asignados al azar a cuatro grupos: placebo e inmunizados con K88ab-GnRH, P64k-GnRH y con un péptido análogo de GnRH (GnRHm1), unido a un epítopo T cooperador del toxoide tetánico (TT) (control positivo), respectivamente. Los cerdos se inmunizaron con 9-10 semanas de edad, en un esquema de 2 dosis y se sacrificaron 16 semanas después. K88ab-GnRH, P64k-GnRH y GnRHm1-TT indujeron niveles de testosterona en suero, mayor, similar y menor, comparados con el placebo, respectivamente. En el grupo K88ab-GnRH los cerdos disminuyeron (P < 0,01) el largo y peso de los testículos y el peso de los epidídimos, comparado con el placebo y ninguno llegó a eyacular. En el grupo P64k-GnRH los cerdos disminuyeron el peso de los testículos (P < 0,05), y sólo uno llegó a eyacular. Los testículos de los cerdos inmunizados con K88ab-GnRH y P64k-GnRH mostraron daños estructurales, funcionales y afectación de la espermatogénesis. Las glándulas sexuales accesorias del grupo P64k-GnRH estaban normales a diferencia de K88ab-GnRH, en las que se observó fibrosis intersticial. Los daños provocados por K88ab-GnRH y P64k-GnRH en los órganos diana evaluados, resultaron inferiores en todos los casos, a las afectaciones que generó el péptido GnRHm1-TT

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Comparison between nasopharyngeal swabs and saliva as reliable specimens for the diagnosis of SARS-CoV-2

Introduction: The SARS-CoV-2 virus is a positive-strand RNA virus. The virus can also be detected in many different specimens as throat swabs, nasal swabs, sputum, saliva, blood, etc.Objective: The aim of this paper is to compare the reliability of different types of specimen collection, saliva and swabs samples for the detection of SARS-CoV-2.Material and Methods: A sample of 22 COVID-19 positive patients was selected. Paired samples from saliva, nasopharyngeal, oropharyngeal and nasopharyngeal + oropharyngeal swabs were collected on the 7th day after diagnosis. The hyssops and medium employed was IMPROSWAB and IMPROVIRAL NAT Medium, Germany. The sample evaluation was conducted through RT-PCR. The results were compared using Fisher’s exact test and ROC curve. The gold standard proposed in this paper was the nasopharyngeal + oropharyngeal swabs specimen.Results: The gold standard method detected 10 true positive cases, of which oropharyngeal swabs, nasopharyngeal swabs and saliva only detected three positive cases. Significant differences (Fisher’s exact test p = 0.003) were detected in the comparison between saliva and the gold standart proposed. The ROC curve analysis showed that saliva had an area under the curve of 0.650, with a 30% of sensibility. However, the nasopharyngeal and nasopharyngeal + oropharyngeal samples had an area under curve of 0.950 and 1.000, respectively, with a sensibility of 90% and 100%, respectively.Conclusion: Saliva samples are not a reliable specimen for SARS-CoV-2 RNA detection. In turn, the most reliable specimens are nasopharyngeal and nasopharyngeal + oropharyngeal samples collected by swabbing.Introducción: El SARS-CoV-2 es un virus ARN positivo. Este virus puede ser detectado en diferentes tipos de secreción como hisopada bucal, nasal, esputo, saliva, sangre, etc.Objetivo: El objetivo de este estudio es comparar la confiabilidad de diferentes tipos de muestras, saliva y exudado, en la detección de SARS-CoV-2.Material y Métodos: Una muestra de 22 pacientes con diagnóstico de Covid-19 fue estudiada. Se tomaron muestras pareadas de saliva y exudado nasofaríngeo y orofaríngeo en cada paciente. Se emplearon los hisopos y medios de la firma alemana IMPROVE®. Los resultados de las determinaciones por RT-PCR se compararon mediante test de Fisher (test de la probabilidad exacta de Fisher) y cada sets de muestras fue evaluada individualmente y luego comparadas por curvas ROC. El estándar de oro propuesto fue el doble hisopado nasofaríngeo/orofaríngeo.Resultados: El método de oro propuesto detectó 10 casos positivos. La coincidencia de detección entre todos los sets de muestras fue de 3 casos (30%). Se obtuvieron diferencias significativas (Fisher p = 0.003) en la comparación de los casos detectados en saliva vs el estándar de oro. El análisis de curvas ROC mostró un área bajo la curva de 0.650 (30% de sensibilidad) para la saliva. En el caso del hisopado nasofaríngeo y el estándar de oro mostraron un área bajo la curva de 0.95 y 1.00, respectivamente, con una sensibilidad del 90% y 100%, respectivamente.Conclusiones: La saliva no es una muestra confiable para la detección de SARS-CoV-2. La muestra más confiable para el diagnóstico fue el hisopado nasofaríngeo y el doble hisopado

The Nucleocapsid Protein of SARS-CoV-2, Combined with ODN-39M, Is a Potential Component for an Intranasal Bivalent Vaccine with Broader Functionality

Despite the rapid development of vaccines against COVID-19, they have important limitations, such as safety issues, the scope of their efficacy, and the induction of mucosal immunity. The present study proposes a potential component for a new generation of vaccines. The recombinant nucleocapsid (N) protein from the SARS-CoV-2 Delta variant was combined with the ODN-39M, a synthetic 39 mer unmethylated cytosine-phosphate-guanine oligodeoxynucleotide (CpG ODN), used as an adjuvant. The evaluation of its immunogenicity in Balb/C mice revealed that only administration by intranasal route induced a systemic cross-reactive, cell-mediated immunity (CMI). In turn, this combination was able to induce anti-N IgA in the lungs, which, along with the specific IgG in sera and CMI in the spleen, was cross-reactive against the nucleocapsid protein of SARS-CoV-1. Furthermore, the nasal administration of the N + ODN-39M preparation, combined with RBD Delta protein, enhanced the local and systemic immune response against RBD, with a neutralizing capacity. Results make the N + ODN-39M preparation a suitable component for a future intranasal vaccine with broader functionality against Sarbecoviruses

Synthetic Growth Hormone-Releasing Peptides (GHRPs): A Historical Appraisal of the Evidences Supporting Their Cytoprotective Effects

Background: Growth hormone-releasing peptides (GHRPs) constitute a group of small synthetic peptides that stimulate the growth hormone secretion and the downstream axis activity. Mounting evidences since the early 1980s delineated unexpected pharmacological cardioprotective and cytoprotective properties for the GHRPs. However, despite intense basic pharmacological research, alternatives to prevent cell and tissue demise before lethal insults have remained as an empty niche in the clinical armamentarium. Here, we have rigorously reviewed the investigational development of GHRPs and their clinical niching perspectives. Methodology: PubMed/MEDLINE databases, including original research and review articles, were explored. The search design was date escalated from 1980 and included articles in English only. Results and Conclusions: GHRPs bind to two different receptors (GHS-R1a and CD36), which redundantly or independently exert relevant biological effects. GHRPs’ binding to CD36 activates prosurvival pathways such as PI-3K/AKT1, thus reducing cellular death. Furthermore, GHRPs decrease reactive oxygen species (ROS) spillover, enhance the antioxidant defenses, and reduce inflammation. These cytoprotective abilities have been revealed in cardiac, neuronal, gastrointestinal, and hepatic cells, representing a comprehensive spectrum of protection of parenchymal organs. Antifibrotic effects have been attributed to some of the GHRPs by counteracting fibrogenic cytokines. In addition, GHRP family members have shown a potent myotropic effect by promoting anabolia and inhibiting catabolia. Finally, GHRPs exhibit a broad safety profile in preclinical and clinical settings. Despite these fragmented lines incite to envision multiple pharmacological uses for GHRPs, especially as a myocardial reperfusion damage-attenuating candidate, this family of “drugable” peptides awaits for a definitive clinical niche

Effects of Two GnRH-based Recombinant Vaccine Candidates on Pig Fertility

This paper describes for the first time the generation of the K88ab-GnRH hybrid fimbria, the fusion of N. meningitidis P64k protein (P64k-GnRH), and its evaluation as vaccine candidates to control fertility in mammals. Twenty hybrid male pigs were randomly distributed in four groups: placebos and immunized with K88ab-GnRH, P64k-GnRH and a GnRH analogue (GnRHm1), linked to a tetanus toxoid (TT) T-helper epitope (positive control), respectively. The pigs were immunized at 9-10 weeks of age, using a two-dose scheme, and were sacrificed sixteen weeks later. K88ab-GnRH, P64k-GnRH, and GnRHm1-TT induced higher, similar, and lower testosterone levels in the serum, compared to the placebo, respectively. In the K88ab-GnRH group, the pigs underwent a reduction in testicle size and weight (P < 0.01), and a reduction in the weight of epididymes compared to the placebo; none of them was able to ejaculate. In the P64k-GnRH group, the pigs had a reduction in testicle weight (P < 0.05), and only one of them was able to ejaculate. The testicles of the pigs immunized with K88ab-GnRH and P64k-GnRH showed structural and functional damage; spermatogenesis was also affected. The accessory sexual glands of the P64k-GnRH group were normal, in contrast to K88ab-GnRH, where interstitial fibrosis was observed. The damage caused by K88ab-GnRH and P64k-GnRH in the target organs evaluated were in all cases lower than the affectations caused by the GnRHm1-TT peptide