An update to Hippocampome.org by integrating single-cell phenotypes with circuit function in vivo.

Understanding brain operation demands linking basic behavioral traits to cell-type specific dynamics of different brain-wide subcircuits. This requires a system to classify the basic operational modes of neurons and circuits. Single-cell phenotyping of firing behavior during ongoing oscillations in vivo has provided a large body of evidence on entorhinal-hippocampal function, but data are dispersed and diverse. Here, we mined literature to search for information regarding the phase-timing dynamics of over 100 hippocampal/entorhinal neuron types defined in Hippocampome.org. We identified missing and unresolved pieces of knowledge (e.g., the preferred theta phase for a specific neuron type) and complemented the dataset with our own new data. By confronting the effect of brain state and recording methods, we highlight the equivalences and differences across conditions and offer a number of novel observations. We show how a heuristic approach based on oscillatory features of morphologically identified neurons can aid in classifying extracellular recordings of single cells and discuss future opportunities and challenges towards integrating single-cell phenotypes with circuit function.

Endocannabinoid signalling in stem cells and cerebral organoids drives differentiation to deep layer projection neurons via CB1 receptors.

The endocannabinoid (eCB) system, via the cannabinoid CB1 receptor, regulates neurodevelopment by controlling neural progenitor proliferation and neurogenesis. CB1 receptor signalling in vivo drives corticofugal deep layer projection neuron development through the regulation of BCL11B and SATB2 transcription factors. Here, we investigated the role of eCB signalling in mouse pluripotent embryonic stem cell-derived neuronal differentiation. Characterization of the eCB system revealed increased expression of eCB-metabolizing enzymes, eCB ligands and CB1 receptors during neuronal differentiation. CB1 receptor knockdown inhibited neuronal differentiation of deep layer neurons and increased upper layer neuron generation, and this phenotype was rescued by CB1 re-expression. Pharmacological regulation with CB1 receptor agonists or elevation of eCB tone with a monoacylglycerol lipase inhibitor promoted neuronal differentiation of deep layer neurons at the expense of upper layer neurons. Patch-clamp analyses revealed that enhancing cannabinoid signalling facilitated neuronal differentiation and functionality. Noteworthy, incubation with CB1 receptor agonists during human iPSC-derived cerebral organoid formation also promoted the expansion of BCL11B+ neurons. These findings unveil a cell-autonomous role of eCB signalling that, via the CB1 receptor, promotes mouse and human deep layer cortical neuron development.

Intensification of the O3/TiO2/UV advanced oxidation process using a modified flotation cell.

The present work reports the use of a flotation cell as a prospective reactor for ozonation and the intensification of ozonation (catalytic ozonation and photocatalytic ozonation). The effect of the pH, ozone concentration and loading catalyst was investigated. The performance of the flotation cell was compared with that of conventional reactors used in ozonation through the ozone utilized index (OUI), which was proposed in this work and relates the amount of ozone supplied to the system per milligram of degraded pollutant. The flotation cell has the lowest OUI, which indicates that the ozone supplied is highly consumed. It was found that the modified flotation cell is an efficient reactor for ozonation, catalytic ozonation and photocatalytic ozonation processes because total diclofenac degradation was achieved in a short time, mass transfer limitations were not found (Ha = 7.26), and it presented a relatively low energy consumption (1.15 kW h m-3).

Loss of Cannabinoid CB1 Receptors Induces Cortical Migration Malformations and Increases Seizure Susceptibility.

Neuronal migration is a fundamental process of brain development, and its disruption underlies devastating neurodevelopmental disorders. The transcriptional programs governing this process are relatively well characterized. However, how environmental cues instruct neuronal migration remains poorly understood. Here, we demonstrate that the cannabinoid CB1 receptor is strictly required for appropriate pyramidal neuron migration in the developing cortex. Acute silencing of the CB1 receptor alters neuronal morphology and impairs radial migration. Consequently, CB1 siRNA-electroporated mice display cortical malformations mimicking subcortical band heterotopias and increased seizure susceptibility in adulthood. Importantly, rescuing the CB1 deficiency-induced radial migration arrest by knockdown of the GTPase protein RhoA restored the hyperexcitable neuronal network and seizure susceptibility. Our findings show that CB1 receptor/RhoA signaling regulates pyramidal neuron migration, and that deficient CB1 receptor signaling may contribute to cortical development malformations leading to refractory epilepsy independently of its canonical neuromodulatory role in the adult brain.

Prenatal exposure to cannabinoids evokes long-lasting functional alterations by targeting CB1 receptors on developing cortical neurons.

The CB1 cannabinoid receptor, the main target of Δ(9)-tetrahydrocannabinol (THC), the most prominent psychoactive compound of marijuana, plays a crucial regulatory role in brain development as evidenced by the neurodevelopmental consequences of its manipulation in animal models. Likewise, recreational cannabis use during pregnancy affects brain structure and function of the progeny. However, the precise neurobiological substrates underlying the consequences of prenatal THC exposure remain unknown. As CB1 signaling is known to modulate long-range corticofugal connectivity, we analyzed the impact of THC exposure on cortical projection neuron development. THC administration to pregnant mice in a restricted time window interfered with subcerebral projection neuron generation, thereby altering corticospinal connectivity, and produced long-lasting alterations in the fine motor performance of the adult offspring. Consequences of THC exposure were reminiscent of those elicited by CB1 receptor genetic ablation, and CB1-null mice were resistant to THC-induced alterations. The identity of embryonic THC neuronal targets was determined by a Cre-mediated, lineage-specific, CB1 expression-rescue strategy in a CB1-null background. Early and selective CB1 reexpression in dorsal telencephalic glutamatergic neurons but not forebrain GABAergic neurons rescued the deficits in corticospinal motor neuron development of CB1-null mice and restored susceptibility to THC-induced motor alterations. In addition, THC administration induced an increase in seizure susceptibility that was mediated by its interference with CB1-dependent regulation of both glutamatergic and GABAergic neuron development. These findings demonstrate that prenatal exposure to THC has long-lasting deleterious consequences in the adult offspring solely mediated by its ability to disrupt the neurodevelopmental role of CB1 signaling.

Low and High-Normal FMR1 Triplet Cytosine, Guanine Guanine Repeats Affect Ovarian Reserve and Fertility in Women Who Underwent In Vitro Fertilization Treatment? Results from a Cross-Sectional Study.

Dynamic mutations in the 5' untranslated region of FMR1 are associated with infertility. Premutation alleles interfere with prenatal development and increase infertility risks. The number of CGG repeats that causes the highest decrease in ovarian reserves remains unclear. We evaluated the effect of FMR1 CGG repeat lengths on ovarian reserves and in vitro fertilization (IVF) treatment outcomes in 272 women with alleles within the normal range. FMR1 CGG repeat length was investigated via PCR and capillary electrophoresis. Alleles were classified as low-normal, normal, and high-normal. Serum levels of follicle-stimulating hormone and anti-Mullerian hormone (AMH) in the follicular phase of the menstrual cycle were measured, and antral follicles (AFC) were counted. IVF outcomes were collected from medical records. Regarding FMR1 CGG repeat length alleles, 63.2% of women presented at least one low-normal allele. Those carrying low-normal alleles had significantly lower AMH levels than women carrying normal or high-normal alleles. Low-normal/low-normal genotype was the most frequent, followed by low-normal/normal and normal/normal. A comparison of ovarian reserve markers and reproductive outcomes of the three most frequent genotypes revealed that AFC in the low-normal/normal genotype was significantly lower than the low-normal/low-normal genotype. The low number of FMR1 CGG repeats affected AMH levels and AFC but not IVF outcomes per cycle of treatment.

Factors influencing NiTi endodontic file separation: A thematic review.

Nickel-titanium (NiTi) file separation during endodontic treatment is an undesirable event. This phenomenon needs to be understood by knowing the factors influencing fracture in endodontic files. There is a large amount of literature where these factors and their influence have been studied, increasing the knowledge about the mechanisms involved, mainly related to wire technology, file shapes and geometry, operator manipulation, the anatomy of the root canal, and the irrigation and sterilization processes. As many factors are involved, the complexity of the fracture phenomena increases and the isolated correlation of one factor with the file fracture becomes a small part of comprehending the separation phenomena. This thematic review aims to compile important reports from 2014 to 2022 on the factors influencing NiTi file separation. The information obtained was classified into wire technology, file geometry, operational aspects, irrigation and sterilization, and anatomy. For this purpose, the Scopus, Web of Science and ScienceDirect databases were consulted using a search string. Filters were applied to consolidate the final set of relevant papers covering the subject of factors influencing endodontic file separation. It was found that the fracture of NiTi files incorporates different mechanisms that operate simultaneously during the endodontic procedure and strongly affect the instrument performance. The collected information promotes good practices to prevent file separation.

NK cells from Men Who Have Sex with Men at high risk for HIV-1 infection exhibit higher effector capacity.

Despite being under constant exposure to HIV-1, some individuals do not show serological or clinical evidence of infection and are known as HESN (HIV-Exposed Seronegative). Multiple studies in different HESN cohorts have linked the NK cells as a correlate of resistance; however, little is known about the role of these cells in Men Who Have Sex with Men (MSM) with high risk sexual behaviors. We evaluated a general overview of activation and effector features of NK cells of MSM co-cultured with LT CD4+ HIV+ in which MSM at high risk of HIV-1 infection (HR-MSM) exhibit higher capacity to eliminate infected cells, reduced percentages of CD69+ cells when compared to MSM at low risk of infection (LR-MSM). In addition, we found that, despite the lower levels of CD69+ NK cells on HR-MSM group, within this population, higher percentages of CD69+ IFN-γ+ and CD69+ NKG2D+ NK cells were found together with higher levels of RANTES and Granzyme B production with higher antiviral capacity, resulting in a lower concentration of p24 protein and p24+ CD4+ T cells. Altogether, this information suggests that NK cells of MSM could impact the capacity to face the viral infection.

The search of a genetic basis for noise-induced hearing loss (NIHL).

BACKGROUND AND AIM: Knowledge about the genetic factors responsible for noise-induced hearing loss (NIHL) is still limited. This study investigated whether genetic factors are associated or not to susceptibility to NIHL. SUBJECTS AND METHODS: The family history and genotypes were studied for candidate genes in 107 individuals with NIHL, 44 with other causes of hearing impairment and 104 controls. Mutations frequently found among deaf individuals were investigated (35delG, 167delT in GJB2, Δ(GJB6- D13S1830), Δ(GJB6- D13S1854) in GJB6 and A1555G in MT-RNR1 genes); allelic and genotypic frequencies were also determined at the SNP rs877098 in DFNB1, of deletions of GSTM1 and GSTT1 and sequence variants in both MTRNR1 and MTTS1 genes, as well as mitochondrial haplogroups. RESULTS: When those with NIHL were compared with the control group, a significant increase was detected in the number of relatives affected by hearing impairment, of the genotype corresponding to the presence of both GSTM1 and GSTT1 enzymes and of cases with mitochondrial haplogroup L1. CONCLUSION: The findings suggest effects of familial history of hearing loss, of GSTT1 and GSTM1 enzymes and of mitochondrial haplogroup L1 on the risk of NIHL. This study also described novel sequence variants of MTRNR1 and MTTS1 genes.

Immune characterization of a Colombian family cluster with SARS-CoV-2 infection. / Caracterización inmunológica de un grupo familiar colombiano con infección por SARS-CoV-2

INTRODUCTION: Immunological markers have been described during COVID-19 and persist after recovery. These immune markers are associated with clinical features among SARSCoV-2 infected individuals. Nevertheless, studies reporting a comprehensive analysis of the immune changes occurring during SARS-CoV-2 infection are still limited. OBJECTIVE: To evaluate the production of proinflammatory cytokines, the antibody response, and the phenotype and function of NK cells and T cells in a Colombian family cluster with SARS-CoV-2 infection. MATERIALS AND METHODS: Proinflammatory cytokines were evaluated by RT-PCR and ELISA. The frequency, phenotype, and function of NK cells (cocultures with K562 cells) and T-cells (stimulated with spike/RdRp peptides) were assessed by flow cytometry. Anti-SARS-CoV-2 antibodies were determined using indirect immunofluorescence and plaque reduction neutralization assay. RESULTS: During COVID-19, we observed a high proinflammatory-cytokine production and a reduced CD56bright-NK cell and cytotoxic response. Compared with healthy controls, infected individuals had a higher frequency of dysfunctional CD8+ T cells CD38+HLA-DR-. During the acute phase, CD8+ T cells stimulated with viral peptides exhibited a monofunctional response characterized by high IL-10 production. However, during recovery, we observed a bifunctional response characterized by the co-expression of CD107a and granzyme B or perforin. CONCLUSION: Although the proinflammatory response is a hallmark of SARS-CoV-2 infection, other phenotypic and functional alterations in NK cells and CD8+ T cells could be associated with the outcome of COVID-19. However, additional studies are required to understand these alterations and to guide future immunotherapy strategies.

Introducción. Se han descrito diferentes marcadores inmunológicos durante la COVID-19, los cuales persisten incluso después de la convalecencia y se asocian con los estadios clínicos de la infección. Sin embargo, aún son pocos los estudios orientados al análisis exhaustivo de las alteraciones del sistema inmunológico en el curso de la infección. Objetivo. Evaluar la producción de citocinas proinflamatorias, la reacción de anticuerpos, y el fenotipo y la función de las células NK y los linfocitos T en una familia colombiana con infección por SARS-CoV-2. Materiales y métodos. Se evaluaron las citocinas proinflamatorias mediante RT-PCR y ELISA; la frecuencia, el fenotipo y la función de las células NK (en cocultivos con células K562) y linfocitos T CD8+ (estimulados con péptidos spike/RdRp) mediante citometría de flujo, y los anticuerpos anti-SARS-CoV-2, mediante inmunofluorescencia indirecta y prueba de neutralización por reducción de placa. Resultados. Durante la COVID-19 hubo una producción elevada de citocinas proinflamatorias, con disminución de las células NK CD56bright y reacción citotóxica. Comparados con los controles sanos, los individuos infectados presentaron con gran frecuencia linfocitos T CD8+ disfuncionales CD38+HLA-DR-. Además, en los linfocitos T CD8+ estimulados con péptidos virales, predominó una reacción monofuncional con gran producción de IL-10 durante la fase aguda y una reacción bifuncional caracterizada por la coexpresión de CD107a y granzima B o perforina durante la convalecencia. Conclusión. Aunque la reacción inflamatoria caracteriza la infección por SARS-CoV-2, hay otras alteraciones fenotípicas y funcionales en células NK y linfocitos T CD8+ que podrían asociarse con la progresión de la infección. Se requieren estudios adicionales para entender estas alteraciones y guiar futuras estrategias de inmunoterapia.

Effect of land cover and landscape fragmentation on anopheline mosquito abundance and diversity in an important Colombian malaria endemic region.

Landscape structure influences the distribution and abundance of anopheline mosquitoes and has an indirect impact on malaria transmission. This work aimed to determine the effect of land cover and landscape fragmentation on anopheline mosquito abundance and diversity in an important Colombian malaria endemic area, the Bajo Cauca region. Diversity indices were calculated for Anopheles mosquitoes collected in various localities of the region. Land cover types were characterized using orthorectified aerial photographs to estimate landscape metrics. The relationship between landscape fragmentation and species diversity was evaluated by regression analysis. The correlation between species abundance and land cover types was determined using canonical correspondence analyses. Results showed a statistically significant tendency for a lower diversity of the Anopheles community in landscapes with higher patch number, patch density and effective mesh size. For most species, there was evidence of a significant relationship between species abundance and land covers modified by anthropic activities which generate forest loss. These results indicate that activities that modify the landscape structure and land cover composition generate changes that affect the spatial distribution and composition of epidemiologically-important Anopheles species, which may impact malaria distribution in a region. This information is useful to guide control interventions that promote unfavorable landscapes for malaria vector propagation.

Long-term hippocampal interneuronopathy drives sex-dimorphic spatial memory impairment induced by prenatal THC exposure.

Prenatal exposure to Δ9-tetrahydrocannabinol (THC), the most prominent active constituent of cannabis, alters neurodevelopmental plasticity with a long-term functional impact on adult offspring. Specifically, THC affects the development of pyramidal neurons and GABAergic interneurons via cannabinoid CB1 receptors (CB1R). However, the particular contribution of these two neuronal lineages to the behavioral alterations and functional deficits induced by THC is still unclear. Here, by using conditional CB1R knockout mice, we investigated the neurodevelopmental consequences of prenatal THC exposure in adulthood, as well as their potential sex differences. Adult mice that had been exposed to THC during embryonic development showed altered hippocampal oscillations, brain hyperexcitability, and spatial memory impairment. Remarkably, we found a clear sexual dimorphism in these effects, with males being selectively affected. At the neuronal level, we found a striking interneuronopathy of CCK-containing interneurons in the hippocampus, which was restricted to male progeny. This THC-induced CCK-interneuron reduction was not evident in mice lacking CB1R selectively in GABAergic interneurons, thus pointing to a cell-autonomous THC action. In vivo electrophysiological recordings of hippocampal LFPs revealed alterations in hippocampal oscillations confined to the stratum pyramidale of CA1 in male offspring. In addition, sharp-wave ripples, a major high-frequency oscillation crucial for learning and memory consolidation, were also altered, pointing to aberrant circuitries caused by persistent reduction of CCK+ basket cells. Taken together, these findings provide a mechanistic explanation for the long-term interneuronopathy responsible for the sex-dimorphic cognitive impairment induced by prenatal THC.

Oral administration of the cannabigerol derivative VCE-003.2 promotes subventricular zone neurogenesis and protects against mutant huntingtin-induced neurodegeneration.

BACKGROUND: The administration of certain cannabinoids provides neuroprotection in models of neurodegenerative diseases by acting through various cellular and molecular mechanisms. Many cannabinoid actions in the nervous system are mediated by CB1 receptors, which can elicit psychotropic effects, but other targets devoid of psychotropic activity, including CB2 and nuclear PPARγ receptors, can also be the target of specific cannabinoids. METHODS: We investigated the pro-neurogenic potential of the synthetic cannabigerol derivative, VCE-003.2, in striatal neurodegeneration by using adeno-associated viral expression of mutant huntingtin in vivo and mouse embryonic stem cell differentiation in vitro. RESULTS: Oral administration of VCE-003.2 protected striatal medium spiny neurons from mutant huntingtin-induced damage, attenuated neuroinflammation and improved motor performance. VCE-003.2 bioavailability was characterized and the potential undesired side effects were evaluated by analyzing hepatotoxicity after chronic treatment. VCE-003.2 promoted subventricular zone progenitor mobilization, increased doublecortin-positive migrating neuroblasts towards the injured area, and enhanced effective neurogenesis. Moreover, we demonstrated the proneurogenic activity of VCE-003.2 in embryonic stem cells. VCE-003.2 was able to increase neuroblast formation and striatal-like CTIP2-mediated neurogenesis. CONCLUSIONS: The cannabigerol derivative VCE-003.2 improves subventricular zone-derived neurogenesis in response to mutant huntingtin-induced neurodegeneration, and is neuroprotective by oral administration.

Contribution of Altered Endocannabinoid System to Overactive mTORC1 Signaling in Focal Cortical Dysplasia.

Alterations of the PI3K/Akt/mammalian target of rapamycin complex 1 (mTORC1) signaling pathway are causally involved in a subset of malformations of cortical development (MCDs) ranging from focal cortical dysplasia (FCD) to hemimegalencephaly and megalencephaly. These MCDs represent a frequent cause of refractory pediatric epilepsy. The endocannabinoid system -especially cannabinoid CB1 receptor- exerts a neurodevelopmental regulatory role at least in part via activation of mTORC1 signaling. Therefore, we sought to characterize the possible contribution of endocannabinoid system signaling to FCD. Confocal microscopy characterization of the CB1 receptor expression and mTORC1 activation was conducted in FCD Type II resection samples. FCD samples were subjected to single nucleotide polymorphism screening for endocannabinoid system elements, as well as CB1 receptor gene sequencing. Cannabinoid CB1 receptor levels were increased in FCD with overactive mTORC1 signaling. CB1 receptors were enriched in phospho-S6-positive cells including balloon cells (BCs) that co-express aberrant markers of undifferentiated cells and dysplastic neurons. Pharmacological regulation of CB1 receptors and the mTORC1 pathway was performed in fresh FCD-derived organotypic cultures. HU-210-evoked activation of CB1 receptors was unable to further activate mTORC1 signaling, whereas CB1 receptor blockade with rimonabant attenuated mTORC1 overactivation. Alterations of the endocannabinoid system may thus contribute to FCD pathological features, and blockade of cannabinoid signaling might be a new therapeutic intervention in FCD.

Síndrome de Peutz-Jeghers: reporte de caso / Peutz-Jeghers syndrome: Case report

Resumen El síndrome de Peutz-Jeghers es una enfermedad hereditaria, autosómica dominante, caracterizada por la presencia de múltiples pólipos gastrointestinales de tipo hamartomatoso y se asocia con hiperpigmentación mucocutánea. A continuación, se reporta un caso de un paciente de 25 años con historia de hemicolectomía derecha por una intususcepción ileocolónica secundaria a un pólipo gigante en el íleon terminal. Se trata de un paciente que consultó por rectorragia, con evidencia en el examen físico de lesiones hipercromáticas color café oscuro en la mucosa yugal. Se realizó una colonoscopia total, en la que se observaron múltiples pólipos. Se practicó una mucosectomía endoscópica a algunos de ellos, histopatológicamente compatibles con pólipos hamartomatosos.

Abstract Peutz-Jeghers syndrome is an autosomal dominant hereditary disease characterized by multiple hamartomatous-type gastrointestinal polyps associated with mucocutaneous hyperpigmentation. A case of a 25-year-old male patient with a history of right hemicolectomy due to ileocolonic intussusception secondary to a giant polyp in the terminal ileum is reported. This patient consulted for rectal bleeding, with evidence on physical examination of dark brown hyperchromatic lesions on the buccal mucosa. A total colonoscopy was performed, noting multiple polyps. Endoscopic mucosectomy was conducted on some of them, being histopathologically compatible with hamartomatous polyps.

Fatigue failure of NiTi endodontic files: scoping review / Falla por fatiga de limas NiTi para endodoncia: revisión exploratoria de cobertura temática

Abstract Introduction: Nickel-Titanium (NiTi) endodontic files are made of hyperelastic material with shape memory. However, these files suffer a sudden fracture during the endodontic treatment, which is considered an unfavorable prognosis. Many studies have been conducted to establish fatigue resistance focused on file brands and determine which is better. Although the most common failure mechanisms have been established for motorized endodontic files, the information is scattered, making it difficult to develop clear research trends. Methods: a scoping review was carried out using Scopus, Dimensions.ai, Web of Science, and Science Direct databases to answer screening questions related to the predominant fracture mechanism in NiTi files, test types, and equipment used for experimentation and to identify the most active authors. Results: using the general search terms, 432 research papers were found, of which 75 were finally selected after eliminating duplicates and applying exclusion criteria. Conclusions: typical failure mechanisms for rotatory and reciprocating files were identified based on the panoramic review and bibliometric indicators. Also, the standard mechanical tests for endodontic files and the characteristics of their assemblies were summarized. The most active authors in the area and their nationality were tagged. Finally, gaps for future research are proposed to generate a comprehensive knowledge of NiTi file failure.

Resumen Introducción: las limas de Níquel-Titanio (NiTi) utilizadas en endodoncia están hechas de un material hiperelástico con memoria de forma. Sin embargo, estas limas sufren fractura repentina durante el tratamiento, lo cual se considera un pronóstico desfavorable. Se han realizado diversos estudios para establecer la resistencia a la fatiga de limas, y determinar cuál marca es mejor. Aunque se han establecido los mecanismos de falla más comunes para las limas de endodoncia motorizadas, la información se encuentra dispersa, dificultando la definición de tendencias claras de investigación. Métodos: se realizó una revisión de cobertura temática utilizando las bases de datos Scopus, Dimensions.ai, Web of Science y Science Direct, para responder a preguntas orientadoras relacionadas con el mecanismo de fractura predominante en las limas NiTi, tipos de pruebas y equipos utilizados para la experimentación e identificar los autores más activos en el área. Resultados: utilizando términos generales de búsqueda, se encontraron 435 trabajos de investigación. Finalmente se seleccionaron 75, tras eliminar duplicados y aplicar criterios de exclusión. Conclusiones: a partir de la revisión panorámica de literatura y empleando algunos indicadores bibliométricos, se identificaron los mecanismos de falla más comunes para las limas rotatorias y reciprocantes. Se obtuvo información sobre ensayos mecánicos y los montajes más utilizados para las limas de endodoncia. Se identificaron los autores más activos en el área y su nacionalidad. Por último, se sugieren oportunidades de investigación para generar un conocimiento exhaustivo sobre la falla de las limas NiTi.

Immune characterization of a Colombian family cluster with SARS-CoV-2 infection / Caracterización inmunológica de un grupo familiar colombiano con infección por SARS-CoV-2

Abstract | Introduction: Immunological markers have been described during COVID-19 and persist after recovery. These immune markers are associated with clinical features among SARS- CoV-2 infected individuals. Nevertheless, studies reporting a comprehensive analysis of the immune changes occurring during SARS-CoV-2 infection are still limited. Objective: To evaluate the production of proinflammatory cytokines, the antibody response, and the phenotype and function of NK cells and T cells in a Colombian family cluster with SARS-CoV-2 infection. Materials and methods: Proinflammatory cytokines were evaluated by RT-PCR and ELISA. The frequency, phenotype, and function of NK cells (cocultures with K562 cells) and T-cells (stimulated with spike/RdRp peptides) were assessed by flow cytometry. Anti-SARS-CoV-2 antibodies were determined using indirect immunofluorescence and plaque reduction neutralization assay. Results: During COVID-19, we observed a high proinflammatory-cytokine production and a reduced CD56bright-NK cell and cytotoxic response. Compared with healthy controls, infected individuals had a higher frequency of dysfunctional CD8+ T cells CD38+HLA-DR-. During the acute phase, CD8+ T cells stimulated with viral peptides exhibited a monofunctional response characterized by high IL-10 production. However, during recovery, we observed a bifunctional response characterized by the co-expression of CD107a and granzyme B or perforin. Conclusion: Although the proinflammatory response is a hallmark of SARS-CoV-2 infection, other phenotypic and functional alterations in NK cells and CD8+ T cells could be associated with the outcome of COVID-19. However, additional studies are required to understand these alterations and to guide future immunotherapy strategies.

Resumen | Introducción. Se han descrito diferentes marcadores inmunológicos durante la COVID-19, los cuales persisten incluso después de la convalecencia y se asocian con los estadios clínicos de la infección. Sin embargo, aún son pocos los estudios orientados al análisis exhaustivo de las alteraciones del sistema inmunológico en el curso de la infección. Objetivo. Evaluar la producción de citocinas proinflamatorias, la reacción de anticuerpos, y el fenotipo y la función de las células NK y los linfocitos T en una familia colombiana con infección por SARS-CoV-2. Materiales y métodos. Se evaluaron las citocinas proinflamatorias mediante RT-PCR y ELISA; la frecuencia, el fenotipo y la función de las células NK (en cocultivos con células K562) y linfocitos T CD8+ (estimulados con péptidos spike/RdRp) mediante citometría de flujo, y los anticuerpos anti-SARS-CoV-2, mediante inmunofluorescencia indirecta y prueba de neutralización por reducción de placa. Resultados. Durante la COVID-19 hubo una producción elevada de citocinas proinflamatorias, con disminución de las células NK CD56 bright y reacción citotóxica. Comparados con los controles sanos, los individuos infectados presentaron con gran frecuencia linfocitos T CD8+ disfuncionales CD38+HLA-DR-. Además, en los linfocitos T CD8+ estimulados con péptidos virales, predominó una reacción monofuncional con gran producción de IL-10 durante la fase aguda y una reacción bifuncional caracterizada por la coexpresión de CD107a y granzima B o perforina durante la convalecencia. Conclusión. Aunque la reacción inflamatoria caracteriza la infección por SARS-CoV-2, hay otras alteraciones fenotípicas y funcionales en células NK y linfocitos T CD8+ que podrían asociarse con la progresión de la infección. Se requieren estudios adicionales para entender estas alteraciones y guiar futuras estrategias de inmunoterapia.

Carcinoma ex-pleomorphic adenoma of upper lip showing copy number loss of tumor suppressor genes.

BACKGROUND: Carcinoma ex-pleomorphic adenoma (CXPA) is a malignant salivary gland tumor that arises rarely in the minor salivary glands. Although the etiology of CXPA remains unclear, the role of some tumor suppressor genes and oncogenes in CXPA is documented; however, other genes still need to be studied. STUDY DESIGN AND OBJECTIVE: An uncommon case of CXPA involving the upper lip is presented, which was analyzed by a panel of tumor suppressor genes by multiplex ligation-dependent probe amplification. RESULTS: The genes investigated in this study, a loss of copy number was detected for CASP8, CD44, CDH1, DAPK1, ESR1, RASSF1, and TP73. Immunohistochemical reactions for the validation of some of these results showed negativity for CD44, RASSF1, and p73. CONCLUSION: A loss of copy number of the genes CD44, RASSF1, and TP73 may contribute to the carcinogenesis of CXPAs.

Role of the mitochondrial mutations, m.827A>G and the novel m.7462C>T, in the origin of hearing loss.

Samples from 30 deaf probands exhibiting features suggestive of syndromic mitochondrial deafness or from families with maternal transmission of deafness were selected for investigation of mutations in the mitochondrial genes MT-RNR1 and MT-TS1. Patients with mutation m.1555A>G had been previously excluded from this sample. In the MT-RNR1 gene, five probands presented the m.827A>G sequence variant, of uncertain pathogenicity. This change was also detected in 66 subjects of an unaffected control sample of 306 Brazilian individuals from various ethnic backgrounds. Given its high frequency, we consider it unlikely to have a pathogenic role on hereditary deafness. As to the MT-TS1 gene, one proband presented the previously known pathogenic m.7472insC mutation and three probands presented a novel variant, m.7462C>T, which was absent from the same control sample of 306 individuals. Because of its absence in control samples and association with a family history of hearing impairment, we suggest it might be a novel pathogenic mutation.

Epidemiología de la enfermedad ácido péptica

En el presente estudio se analizan diferentes factores epidemiológicos con la enfermedad acido-péptica y se trata de establecer además el grado de incapacidad laboral generado por la enfermedad. Las conclusiones son similares a las establecidad previamente en otros estudios