In vitro induction of hair follicle signatures using human dermal papilla cells encapsulated in fibrin microgels.

Cellular spheroids have been described as an appropriate culture system to restore human follicle dermal papilla cells (hFDPc) intrinsic properties; however, they show a low and variable efficiency to promote complete hair follicle formation in in vivo experiments. In this work, a conscientious analysis revealed a 25% cell viability in the surface of the dermal papilla spheroid (DPS) for all culture conditions, questioning whether it is an appropriate culture system for hFDPc. To overcome this problem, we propose the use of human blood plasma for the generation of fibrin microgels (FM) with encapsulated hFDPc to restore its inductive signature, either in the presence or in the absence of blood platelets. FM showed a morphology and extracellular matrix composition similar to the native dermal papilla, including Versican and Collagen IV and increasing cell viability up to 85%. While both systems induce epidermal invaginations expressing hair-specific keratins K14, K15, K71, and K75 in in vitro skin cultures, the number of generated structures increases from 17% to 49% when DPS and FM were used, respectively. These data show the potential of our experimental setting for in vitro hair follicle neogenesis with wild adult hFDPc using FM, being a crucial step in the pursuit of human hair follicle regeneration therapies.

Nimotuzumab Increases the Recovery Rate of Severe and Critical COVID-19 Patients: Evaluation in the Real-World Scenario.

EGFR signaling is an important regulator of SARS-CoV induced lung damage, inflammation and fibrosis. Nimotuzumab is a humanized anti-EGFR antibody registered for several cancer indications. An expanded access study was conducted to evaluate the safety and recovery rate of severe and critical patients with confirmed SARS-CoV-2 infection, treated with nimotuzumab in combination with the standard of care in the real-world scenario. The antibody was administered as an intravenous infusions every 72 h, up to 5 doses. In order to assess the impact of nimotuzumab, the recovery rate was compared with a paired retrospective cohort. Control patients received standard treatment according the national protocol but not nimotuzumab. Overall, 1,151 severe or critical patients received nimotuzumab in 21 hospitals of Cuba. Median age was 65 and 773 patients had at least one comorbidity. Nimotuzumab was very well-tolerated and mild or moderate adverse events were detected in 19 patients. 1,009 controls matching with the nimotuzumab patients, were selected using a "propensity score" method. The 14-day recovery rate of the nimotuzumab cohort was 72 vs. 42% in the control group. Controls had a higher mortality risk (RR 2.08, 95% CI: 1.79, 2.38) than the nimotuzumab treated patients. The attributable fraction was 0.52 (95% CI: 0.44%; 0.58), and indicates the proportion of deaths that were prevented with nimotuzumab. Our preliminary results suggest that nimotuzumab is a safe antibody that can reduce the mortality of severe and critical COVID-19 patients.

Overcoming Paradoxical Kinase Priming by a Novel MNK1 Inhibitor.

Targeting the kinases MNK1 and MNK2 has emerged as a valuable strategy in oncology. However, most of the advanced inhibitors are acting in an adenosine triphosphate (ATP)-competitive mode, precluding the evaluation of different binding modes in preclinical settings. Using rational design, we identified and validated the 4,6-diaryl-pyrazolo[3,4-b]pyridin-3-amine scaffold as the core for MNK inhibitors. Signaling pathway analysis confirmed a direct effect of the hit compound EB1 on MNKs, and in line with the reported function of these kinases, EB1 only affects the growth of tumor but not normal cells. Molecular modeling revealed the binding of EB1 to the inactive conformation of MNK1 and the interaction with the specific DFD motif. This novel mode of action appears to be superior to the ATP-competitive inhibitors, which render the protein in a pseudo-active state. Overcoming this paradoxical activation of MNKs by EB1 represents therefore a promising starting point for the development of a novel generation of MNK inhibitors.

Treatment of COVID-19 patients with the anti-CD6 antibody itolizumab.

OBJECTIVES: COVID-19 can lead to a hyperinflammatory state. CD6 is a glycoprotein expressed on mature T lymphocytes which is a crucial regulator of the T-cell activation. Itolizumab is a humanised antibody targeting CD6. Nonclinical and clinical data in autoimmune diseases indicate that it lowers multiple cytokines primarily involving the Th1/Th17 pathway. The primary objective of this study was to assess the impact of itolizumab in arresting the lung function deterioration of COVID-19 patients. Secondary objectives included safety, duration of ventilation, 14-day mortality and evaluation of interleukin 6 concentration. METHODS: Patients with confirmed SARS-CoV-2 received itolizumab in combination with other therapies included in the national protocol for COVID-19. RESULTS: Seventy critical, severe or moderate patients were treated with itolizumab in 10 Cuban hospitals. Median age was 68, and 94% had comorbidities. After 72 h, most patients improved the PO2/FiO2 ratio and reduced FiO2 requirements. Ventilation time was 8 days for critical and 1 day for severe cases. Ten patients had related adverse events while 3 subjects developed related serious events. In 30 patients, interleukin 6 decreased in individuals with high level and did not change in those with lower concentration. Fourteen-day lethality rate was 4% and 18% for moderate and severe patients, respectively. The proportion of moderate or severe patients with ventilation or death at day 14 was 9.8%. Time to treatment, neurological manifestations and biomarkers such as NLR were significantly associated with higher lethality. CONCLUSIONS: The opportune administration of itolizumab might interrupt the hyperinflammatory cascade and prevent COVID-19 morbidity and mortality.

A Novel Saliva-Based miRNA Signature for Colorectal Cancer Diagnosis.

Salivary microRNAs (miRNAs) are of high interest as diagnostic biomarkers for non-oral cancer. However, little is known about their value for colorectal cancer (CRC) detection. Our study aims to characterize salivary miRNAs in order to identify non-invasive markers for CRC diagnosis. The screening of 754 miRNAs was performed in saliva samples from 14 CRC and 10 healthy controls. The differential expressed miRNAs were validated by RT-qPCR in 51 CRC, 19 adenomas and 37 healthy controls. Receiver operating characteristic (ROC) curves and logistic regression models were performed to analyze the clinical value of these miRNAs. Twenty-two salivary miRNAs were significantly deregulated in CRC patients vs. healthy individuals (P < 0.05) in the discovery phase. From those, five upregulated miRNAs (miR-186-5p, miR-29a-3p, miR-29c-3p, miR-766-3p, and miR-491-5p) were confirmed to be significantly higher in the CRC vs. healthy group (P < 0.05). This five-miRNA signature showed diagnostic value (72% sensitivity, 66.67% specificity, AUC = 0.754) to detect CRC, which was even higher in combination with carcinoembryonic antigen (CEA) levels. Overall, after the first global characterization of salivary miRNAs in CRC, a five-miRNA panel was identified as a promising tool to diagnose this malignancy, representing a novel approach to detect cancer-associated epigenetic alterations using a non-invasive strategy.

MicroRNA-654-5p suppresses ovarian cancer development impacting on MYC, WNT and AKT pathways.

Ovarian cancer is the most lethal gynecological malignancy due to the silent nature on its early onset and the rapid acquisition of drug resistance. Histologically heterogeneous, it includes several subtypes with different mutational landscapes, hampering the development of effective targeted therapies. Non-coding RNAs are emerging as potential new therapeutic targets in cancer. To search for a microRNA signature related to ovarian carcinomas and study its potential as effective targeted therapy, we examined the expression of 768 miRNA in a large collection of tumor samples and found miR-654-5p to be infraexpressed in ovarian serous carcinomas, the most common and aggressive type. Restoration of miR-654-5p levels reduced tumor cell viability in vitro and in vivo and impaired sphere formation capacity and viability of ovarian cancer patient-derived ascitic cells ex vivo. CDCP1 and PLAGL2 oncogenes were found to be the most relevant direct miR-654-5p targets and both genes convey in a molecular signature associated with key cancer pathways relevant to ovarian tumorigenesis, such as MYC, WNT and AKT pathways. Together, we unveiled the tumor suppressor function of miR-654-5p, suggesting that its restoration or co-targeting of CDCP1 and PLAGL2 may be an effective therapeutic approach for ovarian cancer.

Superior Efficacy and Safety of a Nonemulsive Variant of the NGcGM3/VSSP Vaccine in Advanced Breast Cancer Patients.

NGcGM3 ganglioside is a tumor-specific antigen expressed in human breast tumors. The NGcGM3/VSSP vaccine, consisting in very small-sized proteoliposomes (VSSP) obtained by the incorporation of NGcGM3 into the outer membrane protein complex of Neisseria meningitidis, has been previously tested in a Phase II trial in patients with metastatic breast cancer (MBC) but emulsified with Montanide ISA 51. An Expanded Access study was carried out in MBC patients aiming to find if a nonemulsive formulation of NGcGM3/VSSP, without Montanide ISA 51, could be more safe and effective. A total of 104 patients were vaccinated with the nonemulsive formulation (900 µg), subcutaneously (SC), or with the emulsive formulation (200 µg), intramuscularly (IM). An intent-to-treat analysis of efficacy was performed with all patients, and 93 patients were split off according to the site of metastases (visceral/nonvisceral). Of note, SC-treated patients exhibited a superior median overall survival (OS) than IM-treated patients (23.6 vs. 8.2 months; log rank P = 0.001). Even though in the subset of patients with nonvisceral metastases SC vaccination duplicated the median OS compared to the alternative option (31.6 vs. 16.5 months), this difference did not reach statistical significance (log rank P = 0.118). Curiously, in patients with visceral metastases, the advantage of the nonemulsive formulation was more apparent (median OS 21.0 vs. 6.2 months; log rank P = 0.005). The vaccine was safe for both formulations.

Experiencias de la pesquisa activa estudiantil. Policlínico docente Fabio D´Celmo. Caonao. Cienfuegos / Experiences of student active research. Fabio D´Celmo Teaching Polyclinic. Caonao. Cienfuegos

RESUMEN Fundamento: ante la COVID 19, la pesquisa activa estudiantil en la Atención Primaria de Salud cobra vital importancia para la detección precoz de posibles pacientes infectados. Objetivo: describir las experiencias de la pesquisa activa estudiantil desarrollada en el Policlínico Fabio D´Celmo, de Cienfuegos. Métodos: estudio descriptivo, de corte transversal, realizado entre el 19 de marzo y el 20 de mayo de 2020, en un universo de 18 369 habitantes y 83 personas que participaron en la pesquisa. Los datos se obtuvieron de los informes diarios sobre la actividad, entregados al Puesto de Dirección del Área de Salud. Resultados: se describe la forma en que se organizó la pesquisa en el área. Fueron visitadas 66 493 viviendas y pesquisadas 161 772 personas. Se detectaron 854 pacientes con sintomatología respiratoria (0,52 %). En el Consultorio Médico Nro. 15, del Consejo Popular Guaos, una paciente resultó positiva al virus SARS-CoV-2, único caso positivo de COVID 19 en esta área de salud; los estudiantes contribuyeron a su detección y a la pesquisa de los 39 casos sospechosos que generó este foco. Se visitaron 25 983 personas mayores de 60 años, de ellas, 7734 ancianos que viven solos. No se ha reportado la COVID-19 entre los estudiantes. Conclusiones: el trabajo de pesquisa activa desarrollado fue positivo y contribuyó a detectar personas con sintomatología respiratoria, dentro de ellas un caso positivo a la COVID 19, sin que se afectara la salud de los pesquisadores, pues se siguieron las normas de bioseguridad.

ABSTRACT Background: in the face of COVID 19, the active investigation of students in Primary Health Care is of vital importance for the early detection of possible infected patients. Objective: to describe the experiences of the active student research carried out at the Fabio D´Celmo Polyclinic, in Cienfuegos. Methods: descriptive, cross-sectional study, carried out between March 19 and May 20, 2020, in a universe of 18,369 inhabitants and 83 people who participated in the research. The data were obtained from the daily reports on the activity, delivered to the Health Area Management Post. Results: the way in which the research was organized in the area is described; 66 493 homes were visited and 161 772 people were searched. 854 patients with respiratory symptoms (0.52%) were detected. In the Medical Office No. 15, of the Popular Council Guaos, a patient tested positive for the SARS-CoV-2 virus, the only positive case of COVID 19 in this health area. The students contributed to its detection and to the investigation of the 39 suspected cases generated by this outbreak; 25,983 people over 60 years of age were visited, of them 7,734 elderly living alone. COVID-19 has not been reported among students. Conclusions: the active research work carried out was positive and contributed to detect people with respiratory symptoms, among them a positive case for COVID 19, without affecting the health of the researchers, since the biosafety regulations were followed.

Malnutrición por defecto de un recién nacido por psicosis puerperal materna: presentación de un caso / Malnutrition per defect in a new born caused by maternal puerperal psychosis: presentation of a case

En el siguiente trabajo se realizó el seguimiento de un niño desde la etapa de recién nacido hasta el año de edad, por malnutrición por defecto, de causa materna por psicosis puerperal. En este caso, la madre rechazaba a su hijo, negándose a alimentarlo, privando al bebé de algo tan importante en esta etapa de la vida como la lactancia materna exclusiva, requiriendo de alimentación complementaria en edad temprana para lograr un aumento de peso adecuado y un desarrollo psicomotor que se igualara, en un período corto de tiempo, al de niños sanos de su misma edad, sin que aparecieran secuelas que dieran al traste con su desarrollo posterior. Se necesitó darle atención diferenciada por un equipo multidisciplinario formado por el médico general integral de su consultorio, pediatra del grupo básico de trabajo, un nutricionista, rehabilitadores del Centro de Neurodesarrollo, y especialista en psicología/psiquiatría para el tratamiento de la madre, logrando a los 5 meses de edad llevarlo al estado de normopeso; no obstante, se le dio seguimiento especializado hasta que cumplió el año de edad.

In the following work we carried out the follow up of a child since the stage of newborn up to the age of 1 year by malnutrition per defect, caused by maternal puerperal psychosis. In this case, the mother rejected the child, refusing to feed him, depriving the child of something so important in this life stage as the exclusive maternal breastfeeding. The child received a complementary feeding to achieve an adequate weight increase and a psychomotor development equal to the health children of the same age in a short period of time, without sequels that could damage his posterior development. It was necessary to give the child a differentiated care by a multidisciplinary team formed by the integral general physician working in his family consultation, the podiatrist of the main working group, a nutritionist, rehabilitators of the Neurodevelopment Center, and an specialist in psychology/psychiatry for treating his mother. When the child was 5 month old he reached the normal weight, but he received an specialized follow up until he was one year.

Malnutrición por defecto de un recién nacido por psicosis puerperal materna: presentación de un caso / Malnutrition by defect of a newborn because of the puerperal maternal psychosis: presentation of a case

En el siguiente trabajo se realizó el seguimiento de un niño desde la etapa de recién nacido hasta el año de edad por malnutrición por defecto, de causa materna por psicosis puerperal. En este caso, la madre rechazaba a su hijo, negándose a alimentarlo, privando al bebé de algo tan importante en esta etapa de la vida como la lactancia materna exclusiva, requiriendo de alimentación complementaria en edad temprana para lograr un aumento de peso adecuado y un desarrollo psicomotor que se igualara, en un período corto de tiempo, al de niños sanos de su misma edad, sin que aparecieran secuelas que dieran al traste con su desarrollo posterior. Se necesitó darle atención diferenciada por un equipo multidisciplinario formado por el médico general integral de su consultorio, pediatra del Grupo Básico de Trabajo, nutricionista, rehabilitadores del Centro de Neurodesarrollo, y por psicología y psiquiatría para el tratamiento a la madre, logrando a los 5 meses de edad llevarlo al estado de normopeso; no obstante, se le dio seguimiento especializado hasta que cumplió el año de edad.

In the following work we followed up a child since he was a newborn until he was 1 year old for malnutrition by defect, caused by the puerperal maternal psychosis. In this case the mother rejected her son, refusing to feed him, depriving him of such important thing in this stage of life like the exclusive maternal breastfeeding, requiring of complementary nourishing in such early age to get an adequate weight gain and a psychomotor development that would equalize, in a short period of time, the ones got by healthy children of the same age, without sequels affecting his subsequent development. It was necessary a differentiated attention by a multidisciplinary team formed by the integral general physician of the family consultation, a pediatrician of the Working Basic Group, a nutritionist, rehabilitators from the Centre of Neurodevelopment, and a psychologist and a psychiatrist for treating the mother. When he was 5 months old he reached the normal weight; nevertheless, he was given a specialized follow up until he was 1 year old.