Multimodal Analgesia Approach in Acute Low Back Pain Management: A Phase III Study of a Novel Analgesic Combination of Etoricoxib/Tramadol.

INTRODUCTION: Pain and disability management are crucial for a speedy recovery. Combining analgesics with different mechanisms of action provides greater pain relief with lower doses, promoting efficient multimodal analgesia. This study evaluated the efficacy and safety between two fixed-dose combinations (FDC): etoricoxib/tramadol compared to paracetamol/tramadol for the management of acute low back pain (LBP) in a 7-day treatment. METHODS: We conducted a phase IIIb, prospective, randomized, and multicenter study in patients with acute LBP treated with etoricoxib 90 mg/tramadol 50 mg (one packet of granules diluted in 100 ml of water, once a day [QD], for 7 days) or paracetamol 975 mg/tramadol 112.5 mg (one tablet of 325 mg/37.5 mg, three times a day [TID], for 7 days) to assess the efficacy (in terms of pain and disability improvement) and safety. RESULTS: One hundred and twenty-four patients were randomized to receive either etoricoxib/tramadol QD (n = 61) or paracetamol/tramadol TID (n = 63). From the magnitude of change in pain evaluations, differences were observed between the treatment groups at 3 [p = 0.054, CI 95% - 0.648 (- 0.010 to 1.306)] and 5 days (p = 0.041). The proportion of patients with a 30% reduction in Visual Analogue Scale (VAS) score was statistically significant when comparing the treatment groups on the third day of follow-up [p = 0.008, CI 95% 0.241 (0.061-0.421)]. An improvement in LBP's disability to perform activities of daily routine (Oswestry and Roland-Morris questionnaires) was observed in both treatment groups. A total of 79 adverse events (AEs) (38 [48.1%] with etoricoxib/tramadol and 41 [51.9%] with paracetamol/tramadol) were reported. The most frequent AEs were nausea (17.7%) and dizziness (16.4%). CONCLUSIONS: The results show the clinical benefits of etoricoxib/tramadol FDC, such as the sparing effect of tramadol dose per day, early therapeutic response rate compared with paracetamol/tramadol; which translates into faster pain relief, better adherence, less tramadol drug dependency, and a reduction of related AEs incidence. TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT04968158.

Lectin-type oxidized LDL receptor-1 as a potential therapeutic target for cerebral cavernous malformations treatment.

Introduction: Cerebral cavernous malformations (CCMs) are pathologic lesions comprised of clusters of thin-walled capillaries characterized by abnormal proliferation, angiogenesis, and bleeding secondary to somatic or germline mutations in endothelial cells. CCMs can cause headaches, seizures and/or neurological defects. There is a clinical need to develop better tools to detect CCMs and follow their progression in conjunction with the current use of neuroimaging techniques. Here we present data supporting the utility of LOX-1 (lectin-type oxidized LDL receptor 1), a 50 kDa transmembrane protein implicated in endothelial cell dysfunction and ischemia, as a putative biomarker for CCM. Methods: CCM urine samples (n = 23) were collected from pediatric CCM patients. Matched healthy controls (n = 24) were collected from pediatric patients with either Chiari I malformation or fatty filum terminale, and otherwise normal findings. All samples were collected with patient/family consent and institutional review board approval.Samples were analyzed with Olink Proteomic Proximity Extension Assay (PEA). Differences in expression for 2,925 unique proteins were quantified between healthy control urine samples and CCM urine samples. The results were normalized, validated, and analyzed for demographic bias. In addition to urine samples, CCM tissue from patients was harvested and used to create primary cell lines for in vitro analysis of LOX-1 expression, in addition to immunofluorescence of lesional tissue excised at surgery. Results: ANOVA analysis of the CCM urine samples showed a statistically significant increase in LOX-1 compared to the control samples, with CCM patients exhibiting a > 5-fold increase in urinary expression. Corroborating these elevated levels of circulating marker, analysis of source tissue from surgically resected CCMs revealed that LOX-1 is increased in both CCM patient cavernoma primary cell lines and operative specimens. Conclusion: LOX-1 is involved with pathways implicated in CCM pathogenesis and our data here reveals that LOX-1 expression is significantly elevated in CCM patients as compared to matched healthy control individuals, including both source tissue from surgically excised CCMs and in analysis of samples collected from outside of the central nervous system, particularly urine. This proof-of-principle data suggests that LOX-1 may have potential utility as a target for CCM treatment and supports further investigation related to its potential mechanistic impact on CCM pathogenesis.

Perceptions of Government-Assisted Payors: A Survey and Analysis of American Shoulder and Elbow Surgeons Current Medicare, Medicare Advantage and Medicaid Practice Allocation.

INTRODUCTION: Significant discrepancy and variance exist in the United States health care system with regards to patient access to medical care based on a patient's insurance type, whether that be government-assisted or a private insurer. There are currently three major government-assisted insurance programs: Medicare, Medicare Advantage, and Medicaid, each of which have their own patient mix and regulatory processes that govern care delivery. The purpose of this study was to evaluate the current perceptions of shoulder and elbow surgeons surrounding practice patterns and barriers to access for patients whose primary insurance is a government-assisted payor. METHODS: This was a national, observational study that surveyed the American Shoulder and Elbow Surgeons (ASES) society membership. This 15-question survey assessed surgeon demographics, practice types, reimbursement models, as well as rates and trends of their access to patients with government-assisted insurance. Subgroup analysis between practice type and barriers to access for patients with one of these three government-assisted insurance were also analyzed and compared. Pearson's Chi-squared test or Fisher's exact test was used to test association between categorical responses and categorical/binary respondent characteristics. A P value < .05 was deemed statistically significant. RESULTS: A total of 257 ASES members completed the survey. Mean years in practice for respondents was 14. For Medicare patients, the most common perceived barriers were reimbursement (49%) followed by administrative burden (33%) and then implant reimbursement at the surgeon's primary surgical facility (32%). For Medicare Advantage patients the most common barrier to access was administrative burden (52%), reimbursement (50%), and the patient's ability to access peri-operative services such as physical therapy, home health etc. (40%). The most common barriers for Medicaid patients were relatively evenly distributed between reimbursement (62%), low patient engagement in their care (61%), and patient's ability to access peri-operative services (60%). CONCLUSION: Amongst members of the ASES, barriers to patient access varied by government-assisted payor. For Medicare advantage, administrative burden was largest barrier to access. Whereas for Medicare and Medicaid, reimbursement was the most significant barrier. Further investigation and understanding of these barriers to patient access are necessary to improve availability of shoulder and elbow subspecialized care to a broader population of patients insured by government-assisted payors.

Standardized intrapulmonary lymph node dissection in lung cancer specimens: A national Colombian analysis.

Objective: In patients with non-small cell lung cancer, lymph node assessment is essential for appropriate staging. The intrapulmonary lymph nodes (IPLNs) should be considered when assigning the N stage but are infrequently evaluated in Colombian centers, resulting in understaging that may hinder optimal treatment. Methods: We conducted a prospective study of IPLN dissection in patients with clinical stage I or II non-small cell lung cancer who underwent surgical resection at 9 institutions in Colombia between 2021 and 2023. IPLN dissection was performed by trained surgeons who collected lymph nodes from fresh specimens after resection and before formalin fixation. Results: One hundred patients were eligible for the analysis. Their mean age was 67 ± 10.9 years, and 76% were women. Most (74%) had adenocarcinoma, 20% had neuroendocrine tumors, and 6% had squamous cell carcinoma. Successful sampling and histopathologic analysis of at least one IPLN station was obtained in 85% of patients, 9% had upstaging due to positive N2 lymph nodes, and 5% had upstaging due to positive N1 lymph nodes. Among the patients with pN0 or pN1 disease, 3.2% (3 out of 91) were upstaged exclusively due to positive IPLNs. Conclusions: Fresh-specimen dissection to collect IPLNs is appropriate and feasible to achieve more accurate pathological staging in Colombian lung cancer patients. In clinical N0 patients, IPLN dissection maximizes selection for adjuvant therapy.

Levels of Plasma Endothelin-1, Circulating Endothelial Cells, Endothelial Progenitor Cells, and Cytokines after Cardiopulmonary Bypass in Children with Congenital Heart Disease: Role of Endothelin-1 Regulation.

Congenital heart disease (CHD) can be complicated by pulmonary arterial hypertension (PAH). Cardiopulmonary bypass (CPB) for corrective surgery may cause endothelial dysfunction, involving endothelin-1 (ET-1), circulating endothelial cells (CECs), and endothelial progenitor cells (EPCs). These markers can gauge disease severity, but their levels in children's peripheral blood still lack consensus for prognostic value. The aim of our study was to investigate changes in ET-1, cytokines, and the absolute numbers (Ɲ) of CECs and EPCs in children 24 h before and 48 h after CPB surgery to identify high-risk patients of complications. A cohort of 56 children was included: 41 cases with CHD-PAH (22 with high pulmonary flow and 19 with low pulmonary flow) and 15 control cases. We observed that Ɲ-CECs increased in both CHD groups and that Ɲ-EPCs decreased in the immediate post-surgical period, and there was a strong negative correlation between ET-1 and CEC before surgery, along with significant changes in ET-1, IL8, IL6, and CEC levels. Our findings support the understanding of endothelial cell precursors' role in endogenous repair and contribute to knowledge about endothelial dysfunction in CHD.

Mild Hypoxia Accelerates Cerebral Cavernous Malformation Disease Through CX3CR1-CX3CL1 Signaling.

BACKGROUND: Heterogeneity in the severity of cerebral cavernous malformations (CCMs) disease, including brain bleedings and thrombosis that cause neurological disabilities in patients, suggests that environmental, genetic, or biological factors act as disease modifiers. Still, the underlying mechanisms are not entirely understood. Here, we report that mild hypoxia accelerates CCM disease by promoting angiogenesis, neuroinflammation, and vascular thrombosis in the brains of CCM mouse models. METHODS: We used genetic studies, RNA sequencing, spatial transcriptome, micro-computed tomography, fluorescence-activated cell sorting, multiplex immunofluorescence, coculture studies, and imaging techniques to reveal that sustained mild hypoxia via the CX3CR1-CX3CL1 (CX3C motif chemokine receptor 1/chemokine [CX3C motif] ligand 1) signaling pathway influences cell-specific neuroinflammatory interactions, contributing to heterogeneity in CCM severity. RESULTS: Histological and expression profiles of CCM neurovascular lesions (Slco1c1-iCreERT2;Pdcd10fl/fl; Pdcd10BECKO) in male and female mice found that sustained mild hypoxia (12% O2, 7 days) accelerates CCM disease. Our findings indicate that a small reduction in oxygen levels can significantly increase angiogenesis, neuroinflammation, and thrombosis in CCM disease by enhancing the interactions between endothelium, astrocytes, and immune cells. Our study indicates that the interactions between CX3CR1 and CX3CL1 are crucial in the maturation of CCM lesions and propensity to CCM immunothrombosis. In particular, this pathway regulates the recruitment and activation of microglia and other immune cells in CCM lesions, which leads to lesion growth and thrombosis. We found that human CX3CR1 variants are linked to lower lesion burden in familial CCMs, proving it is a genetic modifier in human disease and a potential marker for aggressiveness. Moreover, monoclonal blocking antibody against CX3CL1 or reducing 1 copy of the Cx3cr1 gene significantly reduces hypoxia-induced CCM immunothrombosis. CONCLUSIONS: Our study reveals that interactions between CX3CR1 and CX3CL1 can modify CCM neuropathology when lesions are accelerated by environmental hypoxia. Moreover, a hypoxic environment or hypoxia signaling caused by CCM disease influences the balance between neuroinflammation and neuroprotection mediated by CX3CR1-CX3CL1 signaling. These results establish CX3CR1 as a genetic marker for patient stratification and a potential predictor of CCM aggressiveness.

Transcriptomic signatures of individual cell types in cerebral cavernous malformation.

Cerebral cavernous malformation (CCM) is a hemorrhagic neurovascular disease with no currently available therapeutics. Prior evidence suggests that different cell types may play a role in CCM pathogenesis. The contribution of each cell type to the dysfunctional cellular crosstalk remains unclear. Herein, RNA-seq was performed on fluorescence-activated cell sorted endothelial cells (ECs), pericytes, and neuroglia from CCM lesions and non-lesional brain tissue controls. Differentially Expressed Gene (DEG), pathway and Ligand-Receptor (LR) analyses were performed to characterize the dysfunctional genes of respective cell types within CCMs. Common DEGs among all three cell types were related to inflammation and endothelial-to-mesenchymal transition (EndMT). DEG and pathway analyses supported a role of lesional ECs in dysregulated angiogenesis and increased permeability. VEGFA was particularly upregulated in pericytes. Further pathway and LR analyses identified vascular endothelial growth factor A/ vascular endothelial growth factor receptor 2 signaling in lesional ECs and pericytes that would result in increased angiogenesis. Moreover, lesional pericytes and neuroglia predominantly showed DEGs and pathways mediating the immune response. Further analyses of cell specific gene alterations in CCM endorsed potential contribution to EndMT, coagulation, and a hypoxic microenvironment. Taken together, these findings motivate mechanistic hypotheses regarding non-endothelial contributions to lesion pathobiology and may lead to novel therapeutic targets. Video Abstract.

Investigación científica en el campo de las ataxias espinocerebelosas: una mirada bibliométrica / Scientific Research in The Field of Spinocerebellar Ataxias: A Bibliometric View

El objetivo del presente estudio consistió en caracterizar la producción científica relacionada con el campo de las ataxias espinocerebelosas, las cuales constituyen enfermedades neurodegenerativas, manifestadas por cuadros clínicos progresivos e invalidantes. La investigación es de tipo censal-documental y recupera metadatos de Scopus, correspondientes a 5654 investigaciones relacionadas con este problema de salud, durante el período 1961-2020. El procesamiento explora las principales características bibliométricas de los documentos publicados, el ritmo de crecimiento, la paternidad de las obras, el impacto por índice de citas, así como las redes de colaboración y la estructura que sigue el flujo del conocimiento. Se observa un despegue notable de la producción científica desde inicios de los años 90 del siglo pasado, coincidiendo con el desarrollo de investigaciones afines en el campo de la genética. También se constata un predominio en el estudio de los tipos SCA1, SCA2, SCA3, SCA6, y SCA17, donde los cuatro primeros corresponden a las ataxias de mayor prevalencia a escala mundial. El corpus documental refleja la consolidación de grupos de investigación relativamente estables, encabezados por líderes científicos y caracterizados por la ampliación sostenida de la colaboración internacional y por el trabajo colectivo e interdisciplinario. También se aprecia la tendencia hacia el aumento del número de referencias dentro de cada documento. Los mayores volúmenes productivos se concentran en países desarrollados, junto a países en vías de desarrollo donde existen elevados niveles de prevalencia en esta enfermedad(AU)

The objective of the present study was to characterize the scientific production related to the field of spinocerebellar ataxias, which constitute neurodegenerative diseases, manifested by progressive and disabling clinical conditions. The research is census-documentary type and recovers metadata from Scopus, corresponding to 5,654 investigations related to this health problem, during the period 1961-2020. The processing explores the main bibliometric characteristics of the published documents, the pace of growth, the authorship of the works, the impact by citation index, as well as the collaboration networks and the structure that follows the flow of knowledge. A notable takeoff in scientific production has been observed since the beginning of the 90s of the last century, coinciding with the development of related research in the field of genetics. There is also a predominance in the study of the types SCA1, SCA2, SCA3, SCA6, and SCA17, where the first four correspond to the most prevalent ataxias worldwide. The documentary corpus reflects the consolidation of relatively stable research groups, headed by scientific leaders and characterized by the sustained expansion of international collaboration and collective and interdisciplinary work. There is also a trend towards increasing the number of references within each document. The largest productive volumes are concentrated in developed countries, along with developing countries where there are high levels of prevalence of this disease(AU)

Circulating Plasma miRNA Homologs in Mice and Humans Reflect Familial Cerebral Cavernous Malformation Disease.

Patients with familial cerebral cavernous malformation (CCM) inherit germline loss of function mutations and are susceptible to progressive development of brain lesions and neurological sequelae during their lifetime. To date, no homologous circulating molecules have been identified that can reflect the presence of germ line pathogenetic CCM mutations, either in animal models or patients. We hypothesize that homologous differentially expressed (DE) plasma miRNAs can reflect the CCM germline mutation in preclinical murine models and patients. Herein, homologous DE plasma miRNAs with mechanistic putative gene targets within the transcriptome of preclinical and human CCM lesions were identified. Several of these gene targets were additionally found to be associated with CCM-enriched pathways identified using the Kyoto Encyclopedia of Genes and Genomes. DE miRNAs were also identified in familial-CCM patients who developed new brain lesions within the year following blood sample collection. The miRNome results were then validated in an independent cohort of human subjects with real-time-qPCR quantification, a technique facilitating plasma assays. Finally, a Bayesian-informed machine learning approach showed that a combination of plasma levels of miRNAs and circulating proteins improves the association with familial-CCM disease in human subjects to 95% accuracy. These findings act as an important proof of concept for the future development of translatable circulating biomarkers to be tested in preclinical studies and human trials aimed at monitoring and restoring gene function in CCM and other diseases.

ß1 integrin monoclonal antibody treatment ameliorates cerebral cavernous malformations.

ß1 integrins are important in blood vessel formation and function, finely tuning the adhesion of endothelial cells to each other and to the extracellular matrix. The role of integrins in the vascular disease, cerebral cavernous malformation (CCM) has yet to be explored in vivo. Endothelial loss of the gene KRIT1 leads to brain microvascular defects, resulting in debilitating and often fatal consequences. We tested administration of a monoclonal antibody that enforces the active ß1 integrin conformation, (clone 9EG7), on a murine neonatal CCM mouse model, Krit1flox/flox ;Pdgfb-iCreERT2 (Krit1ECKO ), and on KRIT1-silenced human umbilical vein endothelial cells (HUVECs). In addition, endothelial deletion of the master regulator of integrin activation, Talin 1 (Tln1), in Krit1ECKO mice was performed to assess the effect of completely blocking endothelial integrin activation on CCM. Treatment with 9EG7 reduced lesion burden in the Krit1ECKO model and was accompanied by a strong reduction in the phosphorylation of the ROCK substrate, myosin light chain (pMLC), in both retina and brain endothelial cells. Treatment of KRIT1-silenced HUVECs with 9EG7 in vitro stabilized cell-cell junctions. Overnight treatment of HUVECs with 9EG7 resulted in significantly reduced total surface expression of ß1 integrin, which was associated with reduced pMLC levels, supporting our in vivo findings. Genetic blockade of integrin activation by Tln1ECKO enhanced bleeding and did not reduce CCM lesion burden in Krit1ECKO mice. In sum, targeting ß1 integrin with an activated-specific antibody reduces acute murine CCM lesion development, which we found to be associated with suppression of endothelial ROCK activity.

Neuroinflammation Plays a Critical Role in Cerebral Cavernous Malformation Disease.

BACKGROUND: Cerebral cavernous malformations (CCMs) are neurovascular lesions caused by loss of function mutations in 1 of 3 genes, including KRIT1 (CCM1), CCM2, and PDCD10 (CCM3). CCMs affect ≈1 out of 200 children and adults, and no pharmacologic therapy is available. CCM lesion count, size, and aggressiveness vary widely among patients of similar ages with the same mutation or even within members of the same family. However, what determines the transition from quiescent lesions into mature and active (aggressive) CCM lesions is unknown. METHODS: We use genetic, RNA-sequencing, histology, flow cytometry, and imaging techniques to report the interaction between CCM endothelium, astrocytes, leukocytes, microglia/macrophages, neutrophils (CCM endothelium, astrocytes, leukocytes, microglia/macrophages, neutrophils interaction) during the pathogenesis of CCMs in the brain tissue. RESULTS: Expression profile of astrocytes in adult mouse brains using translated mRNAs obtained from the purification of EGFP (enhanced green fluorescent protein)-tagged ribosomes (Aldh1l1-EGFP/Rpl10a) in the presence or absence of CCM lesions (Slco1c1-iCreERT2;Pdcd10fl/fl; Pdcd10BECKO) identifies a novel gene signature for neuroinflammatory astrocytes. CCM-induced reactive astrocytes have a neuroinflammatory capacity by expressing genes involved in angiogenesis, chemotaxis, hypoxia signaling, and inflammation. RNA-sequencing analysis on RNA isolated from brain endothelial cells in chronic Pdcd10BECKO mice (CCM endothelium), identified crucial genes involved in recruiting inflammatory cells and thrombus formation through chemotaxis and coagulation pathways. In addition, CCM endothelium was associated with increased expression of Nlrp3 and Il1b. Pharmacological inhibition of NLRP3 (NOD [nucleotide-binding oligomerization domain]-' LRR [leucine-rich repeat]- and pyrin domain-containing protein 3) significantly decreased inflammasome activity as assessed by quantification of a fluorescent indicator of caspase-1 activity (FAM-FLICA [carboxyfluorescein-fluorochrome-labeled inhibitors of caspases] caspase-1) in brain endothelial cells from Pdcd10BECKO in chronic stage. Importantly, our results support the hypothesis of the crosstalk between astrocytes and CCM endothelium that can trigger recruitment of inflammatory cells arising from brain parenchyma (microglia) and the peripheral immune system (leukocytes) into mature active CCM lesions that propagate lesion growth, immunothrombosis, and bleedings. Unexpectedly, partial or total loss of brain endothelial NF-κB (nuclear factor κB) activity (using Ikkbfl/fl mice) in chronic Pdcd10BECKO mice does not prevent lesion genesis or neuroinflammation. Instead, this resulted in a trend increase in the number of lesions and immunothrombosis, suggesting that therapeutic approaches designed to target inflammation through endothelial NF-κB inhibition may contribute to detrimental side effects. CONCLUSIONS: Our study reveals previously unknown links between neuroinflammatory astrocytes and inflamed CCM endothelium as contributors that trigger leukocyte recruitment and precipitate immunothrombosis in CCM lesions. However, therapeutic approaches targeting brain endothelial NF-κB activity may contribute to detrimental side effects.

Cryobanking European Mink (Mustela lutreola) Mesenchymal Stem Cells and Oocytes.

The European mink (Mustela lutreola) is one of Europe's most endangered species, and it is on the brink of extinction in the Iberian Peninsula. The species' precarious situation requires the application of new ex situ conservation methodologies that complement the existing ex situ and in situ conservation measures. Here, we report for the first time the establishment of a biobank for European mink mesenchymal stem cells (emMSC) and oocytes from specimens found dead in the Iberian Peninsula, either free or in captivity. New emMSC lines were isolated from different tissues: bone marrow (emBM-MSC), oral mucosa (emOM-MSc), dermal skin (emDS-MSC), oviduct (emO-MSc), endometrium (emE-MSC), testicular (emT-MSC), and adipose tissue from two different adipose depots: subcutaneous (emSCA-MSC) and ovarian (emOA-MSC). All eight emMSC lines showed plastic adhesion, a detectable expression of characteristic markers of MSCs, and, when cultured under osteogenic and adipogenic conditions, differentiation capacity to these lineages. Additionally, we were able to keep 227 Cumulus-oocyte complexes (COCs) in the biobank, 97 of which are grade I or II. The European mink MSC and oocyte biobank will allow for the conservation of the species' genetic variability, the application of assisted reproduction techniques, and the development of in vitro models for studying the molecular mechanisms of infectious diseases that threaten the species' precarious situation.

Mesenchymal Stem Cells in Embryo-Maternal Communication under Healthy Conditions or Viral Infections: Lessons from a Bovine Model.

Bovine mesenchymal stem cells are a relevant cell population found in the maternal reproductive tract that exhibits the immunomodulation capacity required to prevent embryo rejection. The phenotypic plasticity showed by both endometrial mesenchymal stem cells (eMSC) and embryonic trophoblast through mesenchymal to epithelial transition and epithelial to mesenchymal transition, respectively, is essential for embryo implantation. Embryonic trophoblast maintains active crosstalk via EVs and soluble proteins with eMSC and peripheral blood MSC (pbMSC) to ensure the retention of eMSC in case of pregnancy and induce the chemotaxis of pbMSC, critical for successful implantation. Early pregnancy-related proteins and angiogenic markers are detected as cargo in EVs and the soluble fraction of the embryonic trophectoderm secretome. The pattern of protein secretion in trophectoderm-EVs changes depending on their epithelial or mesenchymal phenotype and due to the uptake of MSC EVs. However, the changes in this EV-mediated communication between maternal and embryonic MSC populations infected by viruses that cause abortions in cattle are poorly understood. They are critical in the investigation of reproductive viral pathologies.

Comparison of Biological Features of Wild European Rabbit Mesenchymal Stem Cells Derived from Different Tissues.

Although the European rabbit is an "endangered" species and a notorious biological model, the analysis and comparative characterization of new tissue sources of rabbit mesenchymal stem cells (rMSCs) have not been well addressed. Here, we report for the first time the isolation and characterization of rMSCs derived from an animal belonging to a natural rabbit population within the native region of the species. New rMSC lines were isolated from different tissues: oral mucosa (rOM-MSC), dermal skin (rDS-MSC), subcutaneous adipose tissue (rSCA-MSC), ovarian adipose tissue (rOA-MSC), oviduct (rO-MSC), and mammary gland (rMG-MSC). The six rMSC lines showed plastic adhesion with fibroblast-like morphology and were all shown to be positive for CD44 and CD29 expression (characteristic markers of MSCs), and negative for CD34 or CD45 expression. In terms of pluripotency features, all rMSC lines expressed NANOG, OCT4, and SOX2. Furthermore, all rMSC lines cultured under osteogenic, chondrogenic, and adipogenic conditions showed differentiation capacity. In conclusion, this study describes the isolation and characterization of new rabbit cell lines from different tissue origins, with a clear mesenchymal pattern. We show that rMSC do not exhibit differences in terms of morphological features, expression of the cell surface, and intracellular markers of pluripotency and in vitro differentiation capacities, attributable to their tissue of origin.

Microneedle-mediated Intratumoral Delivery of Anti-CTLA-4 Promotes cDC1-dependent Eradication of Oral Squamous Cell Carcinoma with Limited irAEs.

Head and neck squamous cell carcinoma (HNSCC) ranks sixth in cancer incidence worldwide and has a 5-year survival rate of only 63%. Immunotherapies-principally immune checkpoint inhibitors (ICI), such as anti-PD-1 and anti-CTLA-4 antibodies that restore endogenous antitumor T-cell immunity-offer the greatest promise for HNSCC treatment. Anti-PD-1 has been recently approved for first-line treatment of recurrent and metastatic HNSCC; however, less than 20% of patients show clinical benefit and durable responses. In addition, the clinical application of ICI has been limited by immune-related adverse events (irAE) consequent to compromised peripheral immune tolerance. Although irAEs are often reversible, they can become severe, prompting premature therapy termination or becoming life threatening. To address the irAEs inherent to systemic ICI therapy, we developed a novel, local delivery strategy based upon an array of soluble microneedles (MN). Using our recently reported syngeneic, tobacco-signature murine HNSCC model, we found that both systemic and local-MN anti-CTLA-4 therapy lead to >90% tumor response, which is dependent on CD8 T cells and conventional dendritic cell type 1 (cDC1). However, local-MN delivery limited the distribution of anti-CTLA-4 antibody from areas distal to draining lymphatic basins. Employing Foxp3-GFPDTR transgenic mice to interrogate irAEs in vivo, we found that local-MN delivery of anti-CTLA-4 protects animals from irAEs observed with systemic therapy. Taken together, our findings support the exploration of MN-intratumoral ICI delivery as a viable strategy for HNSCC treatment with reduced irAEs, and the opportunity to target cDC1s as part of multimodal treatment options to boost ICI therapy.

Caracterización del equilibrio dinámico y la tipología de pie en futbolistas juveniles / Characterization of dynamic balance and standing typology in youth soccer players

Resumen Introducción: El equilibrio dinámico (ED) es la base para todas las actividades motoras deportivas. Algunas variables como el tipo de pie y el mecanismo de Windlass (MW) están asociadas a dicho equilibrio y al riesgo de lesión en la extremidad inferior. Se hace necesario el reconocimiento de estos elementos en la población con el fin de identificar potenciales factores de riesgo y promover las respectivas medidas preventivas. Objetivo: Caracterizar el equilibrio dinámico, algunas variables sociodemográficas, antropométricas y anatómico-funcionales del pie en jóvenes futbolistas del Club Deportivo Cañasgordas Latinos. Materiales y métodos: Estudio cuantitativo, observacional y de corte transversal. Se incluyeron 10 futbolistas. Se evaluó ED por medio del Y-Balance Test. Se evaluó el MW por medio del Test de Jack. Para determinar la tipología del pie se utilizó el método HERZCO. Resultados: Se identificaron aspectos susceptibles en el equipo y posibles factores de riesgo de lesión como la prevalencia de pies cavos y la inactivación del mecanismo de Windlass en el 70% de la muestra. Ningún deportista presento riesgo de lesión asociado a asimetrías en el equilibrio dinámico. Conclusiones: Se sugiere complementar la evaluación del ED con factores anatómico-funcionales en futuros estudios para determinar el riesgo de lesión.

Abstract Introduction: Dynamic balance (ED) is at the base of all sports motor activity. Some variables such as the type of foot and the Windlass Mechanism (WM) are associated with this balance and the risk of injury on the lower limb. The recognition of these elements in the population is necessary to identify potential risk factors and promote the respective preventive measures. Objective: To characterize the dynamic balance sociodemographic, anthropometric and some anatomical-functional variables of the foot in young soccer players from the Canasgordas Latinos club. Materials and methods: Quantitative, observational, and cross-sectional study. A total of 10 soccer players were included. The ED was evaluated using the Y-Balance Test. MW was evaluated through the Jack's Test. The HERZCO method was used to determine the type of the foot. Results: Susceptible aspects of the equipment and possible risk factors for injury, such as the prevalence of pes cavus and the inactivation of the Windlass Mechanism in 70% of the sample were identified. Any athlete presented an injury risk associated with asymmetries in dynamic balance. Conclusions: It is suggested to complement the evaluation of ED with anatomical-functional factors in athletes to determine the risk of injury.

Gastroschisis: A Successful, Prospectively Evaluated Treatment Model in a Middle-Income Country.

BACKGROUND: This research adopted a care protocol from high-income countries in a level II/III hospital in a middle-income country to decrease morbidity and mortality associated with gastroschisis. METHODS: We established a multidisciplinary protocol to treat patients with gastroschisis prospectively from November 2012 to November 2018. This included prenatal diagnosis, presence of a neonatologist and pediatric surgeon at birth, and either performing primary closure on the patients with an Apgar score of 8/9, mild serositis, and no breathing difficulty or placing a preformed silo, when unable to fulfill these criteria, under sedation and analgesia (no intubation) in the operating room or at the patients' bedside. The subsequent management took place in the neonatal intensive care unit. The data were analyzed through the Mann-Whitney and Student's t-distribution for the two independent samples; the categorical variables were analyzed through a chi-square distribution or Fisher's exact test. RESULTS: In total, 55 patients were included in the study: 33 patients (60%) were managed with a preformed silo, whereas 22 patients (40%) underwent primary closure. Prenatal diagnosis (P = 0.02), birth at the main hospital (P = 0.02), and the presence of a pediatric surgeon at birth (P = 0.04) were associated with successful primary closure. The primary closure group had fewer fasting days (P < 0.001) and a shorter neonatal intensive care unit length of stay (P = 0.025). The survival rate was 92.7% (51 patients). CONCLUSION: The treatment model modified to fit the means of our hospital proved successful.

El método de experto en tesis doctorales cubanas del ámbito educacional: un estudio bibliométrico y de contenido / The expert method in Cuban doctoral dissertation in the educational field: a bibliometric and content study

La presente investigación tiene como objetivo caracterizar la evolución y empleo del método de experto en investigaciones pedagógicas y educacionales cubanas. Para cumplir este propósito se desarrolló un estudio bibliométrico y de contenido, con una muestra conformada por 1000 tesis doctorales defendidas durante el período 1995-2019, lo cual representa el 31,96 por ciento de la producción nacional en estas ramas de la ciencia. Se identificaron 29 métodos de uso más frecuente, así como una red de términos que demarcan cinco conglomerados de campos de aplicación. Los métodos se clasifican, generalmente, como empíricos o teóricos, con un grado de normalización relativamente bajo. En particular, el método de experto ha sido aplicado de manera creciente y sistemática durante las dos últimas décadas, con énfasis marcado en el caso particular del método Delphi. Las terminologías más comunes son experto y especialista, sin que se aprecien diferencias conceptuales notables entre ellas. La búsqueda de validez ha sido sustituida paulatinamente por otras nociones cualitativas como viabilidad y el binomio pertinencia-factibilidad. La búsqueda de consenso ha perdido terreno frente a otros objetivos más flexibles, como la evaluación de la calidad de un resultado científico, principalmente metodologías, concepciones y modelos. También se estableció un modelo matemático basado en la regresión logística binaria, que resulta útil para estimar la probabilidad de que el método de experto se implemente durante una investigación doctoral. En este modelo intervienen siete variables, entre las cuales la encuesta y los métodos estadísticos resultan más influyentes e interrelacionados(AU)

The objective of this research is to characterize the evolution and use of the expert method in Cuban pedagogical and educational research. To fulfil this purpose, a bibliometric and content study was developed. A sample was made up of 1000 doctoral dissertations from 1995-2019, which represents 31.96percent of the national production in these branches of science. Twenty-nine most frequently used methods were identified, as well as a network of terms that define five conglomerates of application fields. Methods are generally classified as empirical or theoretical, with a relatively low degree of standardization. In particular, the expert method has been applied increasingly and systematically during the last two decades, with marked emphasis on the particular case of Delphi method. The most common terminologies are expert and specialist, with no notable conceptual differences between them. The search for validity has gradually been replaced by other qualitative notions, such as feasibility and pertinence-feasibility binomial. The search for consensus has lost ground compared to other more flexible objectives, such as the evaluation of the quality of a scientific result, mainly methodologies, conceptions and models. A mathematical model based on binary logistic regression was also established, which is useful to estimate the probability that the Expert Method is implemented during a doctoral research. Seven variables are involved in this model, among which the survey and statistical methods are more influential and interrelated(AU)

Mercurio total en bagre rayado y bocachico del río Meta, Colombia / Total mercury in striped catfish and bocachico from river Meta, Colombia

RESUMEN El consumo de pescado contaminado con mercurio representa un grave problema para la salud humana, en especial, en poblaciones, en las cuales, forma parte de su dieta diaria. Este problema requiere un seguimiento de la concentración de mercurio en peces depredadores y no depredadores de agua dulce, especialmente, en zonas poco estudiadas, como la región de la Orinoquia colombiana. Por lo tanto, el objetivo de este estudio fue determinar la concentración de mercurio total en muestras de bagre rayado (Pseudoplatystoma fasciatum metaense) y de bocachico (Prochilodus mariae), de la parte alta del río Meta, Colombia, peces muy consumidos por las poblaciones aledañas. Ocho muestras de cada especie de pez se analizaron por duplicado, por espectrofotometría de absorción atómica, con generador de hidruros. Se estableció, que el método aplicado es trazable al material de referencia certificado Dorm-4; también, se realizó la validación del método. El valor promedio de mercurio total en bagre rayado fue de 0,055 ± 0,0107 mg/kg y para el bocachico, 0,026 ± 0,0054 mg/kg de pescado fresco. Las muestras analizadas no sobrepasan el valor límite establecido por la Organización Mundial de la Salud y Environmental Protection Agency; sin embargo, el índice de riesgo por efectos no cancerígenos, en algunas muestras de bagre rayado, presentan un valor superior a uno, por lo que su consumo representa un riesgo para la salud, especialmente, durante la gestación y la primera infancia, así como de pescadores y aquellas comunidades de la región, que dependen del consumo de este alimento.

ABSTRACT The consumption of fish contaminated with mercury represents a serious problem for human health, especially in populations in which it is part of their daily diet. This problem requires the monitoring of mercury concentration in freshwater predatory and non-predatory fish, especially in poorly studied areas such as the Orinoquia region of Colombia. The aim of this study was to determine total mercury concentration in samples of catfish (Pseudoplatystoma fasciatum metaense) and bocachico (Prochilodus mariae) from the river Meta, Meta-Colombia. Fishes very much consumed by the adjacent populations. Eight samples of each fish species were analyzed in duplicate by atomic absorption spectrophotometry with a hydride generator. It was established that the applied method is traceable to the Dorm-4 certified reference material; the method was also validated. The average value of total mercury in striped catfish was 0.055 ± 0.0107 mg / kg, and for bocachico 0.026 ± 0.0054 mg / kg of fresh fish. These data obtained do not exceed the limit value established by the World Health Organization and Environmental Protection Agency. However, the risk index for non-carcinogenic effects in some samples of striped catfish have a value greater than one, therefore their consumption represents a health risk, especially during pregnancy and early childhood, as well as for fishermen and those communities in the area that depend on the consumption of this food.

Trivalent Subunit Vaccine Candidates for COVID-19 and Their Delivery Devices.

The COVID-19 pandemic highlights the need for platform technologies enabling rapid development of vaccines for emerging viral diseases. The current vaccines target the SARS-CoV-2 spike (S) protein and thus far have shown tremendous efficacy. However, the need for cold-chain distribution, a prime-boost administration schedule, and the emergence of variants of concern (VOCs) call for diligence in novel SARS-CoV-2 vaccine approaches. We studied 13 peptide epitopes from SARS-CoV-2 and identified three neutralizing epitopes that are highly conserved among the VOCs. Monovalent and trivalent COVID-19 vaccine candidates were formulated by chemical conjugation of the peptide epitopes to cowpea mosaic virus (CPMV) nanoparticles and virus-like particles (VLPs) derived from bacteriophage Qß. Efficacy of this approach was validated first using soluble vaccine candidates as solo or trivalent mixtures and subcutaneous prime-boost injection. The high thermal stability of our vaccine candidates allowed for formulation into single-dose injectable slow-release polymer implants, manufactured by melt extrusion, as well as microneedle (MN) patches, obtained through casting into micromolds, for prime-boost self-administration. Immunization of mice yielded high titers of antibodies against the target epitope and S protein, and data confirms that antibodies block receptor binding and neutralize SARS-CoV and SARS-CoV-2 against infection of human cells. We present a nanotechnology vaccine platform that is stable outside the cold-chain and can be formulated into delivery devices enabling single administration or self-administration. CPMV or Qß VLPs could be stockpiled, and epitopes exchanged to target new mutants or emergent diseases as the need arises.