NNT mediates redox-dependent pigmentation via a UVB- and MITF-independent mechanism.

Ultraviolet (UV) light and incompletely understood genetic and epigenetic variations determine skin color. Here we describe an UV- and microphthalmia-associated transcription factor (MITF)-independent mechanism of skin pigmentation. Targeting the mitochondrial redox-regulating enzyme nicotinamide nucleotide transhydrogenase (NNT) resulted in cellular redox changes that affect tyrosinase degradation. These changes regulate melanosome maturation and, consequently, eumelanin levels and pigmentation. Topical application of small-molecule inhibitors yielded skin darkening in human skin, and mice with decreased NNT function displayed increased pigmentation. Additionally, genetic modification of NNT in zebrafish alters melanocytic pigmentation. Analysis of four diverse human cohorts revealed significant associations of skin color, tanning, and sun protection use with various single-nucleotide polymorphisms within NNT. NNT levels were independent of UVB irradiation and redox modulation. Individuals with postinflammatory hyperpigmentation or lentigines displayed decreased skin NNT levels, suggesting an NNT-driven, redox-dependent pigmentation mechanism that can be targeted with NNT-modifying topical drugs for medical and cosmetic purposes.

Covid-19 Interstitial Pneumonia: Histological and Immunohistochemical Features on Cryobiopsies.

BACKGROUND: The pathogenetic steps leading to Covid-19 interstitial pneumonia remain to be clarified. Most postmortem studies to date reveal diffuse alveolar damage as the most relevant histologic pattern. Antemortem lung biopsy may however provide more precise data regarding the earlier stages of the disease, providing a basis for novel treatment approaches. OBJECTIVES: To ascertain the morphological and immunohistochemical features of lung samples obtained in patients with moderate Covid-19 pneumonia. METHODS: Transbronchial lung cryobiopsy was carried out in 12 Covid-19 patients within 20 days of symptom onset. RESULTS: Histopathologic changes included spots of patchy acute lung injury with alveolar type II cell hyperplasia, with no evidence of hyaline membranes. Strong nuclear expression of phosphorylated STAT3 was observed in >50% of AECII. Interalveolar capillaries showed enlarged lumen and were in part arranged in superposed rows. Pulmonary venules were characterized by luminal enlargement, thickened walls, and perivascular CD4+ T-cell infiltration. A strong nuclear expression of phosphorylated STAT3, associated with PD-L1 and IDO expression, was observed in endothelial cells of venules and interstitial capillaries. Alveolar spaces macrophages exhibited a peculiar phenotype (CD68, CD11c, CD14, CD205, CD206, CD123/IL3AR, and PD-L1). CONCLUSIONS: Morphologically distinct features were identified in early stages of Covid-19 pneumonia, with epithelial and endothelial cell abnormalities different from either classical interstitial lung diseases or diffuse alveolar damage. Alveolar type II cell hyperplasia was a prominent event in the majority of cases. Inflammatory cells expressed peculiar phenotypes. No evidence of hyaline membranes and endothelial changes characterized by IDO expression might in part explain the compliance and the characteristic pulmonary vasoplegia observed in less-advanced Covid-19 pneumonia.

Novel Insights in Anti-CD38 Therapy Based on CD38-Receptor Expression and Function: The Multiple Myeloma Model.

Multiple myeloma (MM) is a hematological disease characterized by the proliferation and accumulation of malignant plasmacells (PCs) in the bone marrow (BM). Despite widespread use of high-dose chemotherapy in combination with autologous stem cell transplantation (ASCT) and the introduction of novel agents (immunomodulatory drugs, IMiDs, and proteasome inhibitors, PIs), the prognosis of MM patients is still poor. CD38 is a multifunctional cell-surface glycoprotein with receptor and ectoenzymatic activities. The very high and homogeneous expression of CD38 on myeloma PCs makes it an attractive target for novel therapeutic strategies. Several anti-CD38 monoclonal antibodies have been, or are being, developed for the treatment of MM, including daratumumab and isatuximab. Here we provide an in-depth look atCD38 biology, the role of CD38 in MM progression and its complex interactions with the BM microenvironment, the importance of anti-CD38 monoclonal antibodies, and the main mechanisms of antibody resistance. We then review a number of multiparametric flow cytometry techniques exploiting CD38 antigen expression on PCs to diagnose and monitor the response to treatment in MM patients.

Features, reason for testing, and changes with time of 583 paroxysmal nocturnal hemoglobinuria clones from 529 patients: a multicenter Italian study.

In this study, we aimed at disclosing the main features of paroxysmal nocturnal hemoglobinuria (PNH) clones, their association with presentation syndromes, and their changes during follow-up. A large-scale, cooperative collection (583 clones from 529 patients) of flow cytometric and clinical data was entered into a national repository. Reason for testing guidelines were provided to the 41 participating laboratories, which followed the 2010 technical recommendations for PNH testing by Borowitz. Subsequently, the 30 second-level laboratories adopted the 2012 guidelines for high-resolution PNH testing, both upon order by the local clinicians and as an independent laboratory initiative in selected cases. Type3 and Type2 PNH clones (total and partial absence of glycosyl-phosphatidyl-inositol-anchor, respectively) were simultaneously present in 54 patients. In these patients, Type3 component was sevenfold larger than Type2 (p < 0.001). Frequency distribution analysis of solitary Type3 clone size (N = 442) evidenced two discrete patterns: small (20% of peripheral neutrophils) and large (> 70%) clones. The first pattern was significantly associated with bone marrow failure and myelodysplastic syndromes, the second one with hemolysis, hemoglobinuria, and thrombosis. Pediatric patients (N = 34) showed significant preponderance of small clones and bone marrow failure. The majority of PNH clones involved neutrophils, monocytes, and erythrocytes. Nevertheless, we found clones made exclusively by white cells (N = 13) or erythrocytes (N = 3). Rare cases showed clonal white cells restricted only to monocytes (6 cases) or neutrophils (3 cases). Retesting over 1-year follow-up in 151 cases showed a marked clone size increase in 4 cases and a decrease in 13, demonstrating that early breaking-down of PNH clones is not a rare event (8.6% of cases). This collaborative nationwide study demonstrates a clear-cut difference in size between Type2 and Type3 clones, emphasizes the existence of just two classes of PNH presentations based on Type3 clone size, depicts an asymmetric cellular composition of PNH clones, and documents the possible occurrence of changes in clone size during the follow-up.

Multiethnic GWAS Reveals Polygenic Architecture of Earlobe Attachment.

The genetic basis of earlobe attachment has been a matter of debate since the early 20th century, such that geneticists argue both for and against polygenic inheritance. Recent genetic studies have identified a few loci associated with the trait, but large-scale analyses are still lacking. Here, we performed a genome-wide association study of lobe attachment in a multiethnic sample of 74,660 individuals from four cohorts (three with the trait scored by an expert rater and one with the trait self-reported). Meta-analysis of the three expert-rater-scored cohorts revealed six associated loci harboring numerous candidate genes, including EDAR, SP5, MRPS22, ADGRG6 (GPR126), KIAA1217, and PAX9. The large self-reported 23andMe cohort recapitulated each of these six loci. Moreover, meta-analysis across all four cohorts revealed a total of 49 significant (p < 5 × 10-8) loci. Annotation and enrichment analyses of these 49 loci showed strong evidence of genes involved in ear development and syndromes with auricular phenotypes. RNA sequencing data from both human fetal ear and mouse second branchial arch tissue confirmed that genes located among associated loci showed evidence of expression. These results provide strong evidence for the polygenic nature of earlobe attachment and offer insights into the biological basis of normal and abnormal ear development.

A genome-wide association scan implicates DCHS2, RUNX2, GLI3, PAX1 and EDAR in human facial variation.

We report a genome-wide association scan for facial features in ∼6,000 Latin Americans. We evaluated 14 traits on an ordinal scale and found significant association (P values<5 × 10(-8)) at single-nucleotide polymorphisms (SNPs) in four genomic regions for three nose-related traits: columella inclination (4q31), nose bridge breadth (6p21) and nose wing breadth (7p13 and 20p11). In a subsample of ∼3,000 individuals we obtained quantitative traits related to 9 of the ordinal phenotypes and, also, a measure of nasion position. Quantitative analyses confirmed the ordinal-based associations, identified SNPs in 2q12 associated to chin protrusion, and replicated the reported association of nasion position with SNPs in PAX3. Strongest association in 2q12, 4q31, 6p21 and 7p13 was observed for SNPs in the EDAR, DCHS2, RUNX2 and GLI3 genes, respectively. Associated SNPs in 20p11 extend to PAX1. Consistent with the effect of EDAR on chin protrusion, we documented alterations of mandible length in mice with modified Edar funtion.

A genome-wide association study identifies multiple loci for variation in human ear morphology.

Here we report a genome-wide association study for non-pathological pinna morphology in over 5,000 Latin Americans. We find genome-wide significant association at seven genomic regions affecting: lobe size and attachment, folding of antihelix, helix rolling, ear protrusion and antitragus size (linear regression P values 2 × 10(-8) to 3 × 10(-14)). Four traits are associated with a functional variant in the Ectodysplasin A receptor (EDAR) gene, a key regulator of embryonic skin appendage development. We confirm expression of Edar in the developing mouse ear and that Edar-deficient mice have an abnormally shaped pinna. Two traits are associated with SNPs in a region overlapping the T-Box Protein 15 (TBX15) gene, a major determinant of mouse skeletal development. Strongest association in this region is observed for SNP rs17023457 located in an evolutionarily conserved binding site for the transcription factor Cartilage paired-class homeoprotein 1 (CART1), and we confirm that rs17023457 alters in vitro binding of CART1.

Investigación en salud mental en América Latina y el Caribe: identificando mecanismos para el fortalecimiento de capacidades / Mental health research in Latin America and the Caribbean: devising mechanisms for capacity strengthening

Objetivo: Identificar factores asociados al éxito de la investigación en salud mental en América Latina y el Caribe, como base para el fortalecimiento de las capacidades de investigación en la región. Material y métodos: Se envió un cuestionario a 792 investigadores y se recibió respuesta de 216. Los probandos fueron clasificados en base a los montos de financiamiento de sus investigaciones y número de publicaciones, en: [1] investigadores con financiamiento superior a USD 5,000 en el último año y con al menos 2 publicaciones internacionales en los últimos 5 años (n=50), [2] investigadores con menor financiamiento/publicaciones (n=88). Setenta y ocho investigadores fueron excluidos de la clasificación por tener información incompleta en estos parámetros. Resultados: Tanto los investigadores en el grupo 1 (IG1) como aquellos en el grupo 2 (IG2) declararon como filiación principal una institución universitaria (66% y 50%, respectivamente); la segunda filiación fue con institutos de investigación en el caso de los de IG1 (48%), y con hospitales (32%) o el sector privado (30%) para los de IG2. Comparativamente, los miembros de IG1 mostraron mayor participación en la formación de recursos humanos (76 vs 47%), consultorías (58 vs 36%), redes colaborativas (78 vs 51%), edición (86 vs 57%) y revisión científica (80 vs 43%); en cuanto a disponibilidad de recursos para investigación, los de IG1 indicaron tener mayor acceso a recursos de internet (66vs.33%), a revistas especializadas (64 vs 43%); a apoyo en epidemiología/bioestadística (82 vs 67%) y ciencias básicas (80 vs 41%), y un mayor número de estudiantes graduados (84 vs 41%). Los grupos difirieron en su percepción de los retos para lograr una eficiente implementación de la investigación en salud mental: IG1 identificaron la falta de recursos humanos capacitados como el principal reto, mientras que IG2, la falta de cultura de investigación en sus instituciones.

Objective: To identify factors associated to success in Mental Health (MH) research in the Latin America and Caribbean region, in order to use them as a basis for the strengthening of research capacities. Material and methods: A questionnaire was sent to 792 researchers in the region. Respondents (n=216) were classified according to their research success in: Group 1 researchers (G1Rs): funded with more than USD 5,000 in the last year, authors of at least 2 international publications in the past 5 years (n=50). Group 2 researchers (G2Rs): those reporting less funding or publications (n=88). Researchers who showed missing data in either parameter were excluded (n=78). Results: The main affiliation of both G1Rs and G2Rs were universities (66% and 50%, respectively); the second affiliation of G1Rs were research institutes (48%) while G2Rs were affiliated to hospitals (32%) or the private sector (30%). Compared to G2Rs, G1Rs had higher involvement in human resource training (76 vs 47%), consultantions (58 vs 36%), networking with colleagues (78 vs 51%), participation as scientific editors (86 vs 57%) or reviewers (80 vs 43%), more access to paid internet resources (66 vs 33%), specialized journals (64 vs 43%). In terms of research resources, GR1s declared having more support in epidemiology/biostatistics (82 vs 67%) and basic sciences (80 vs 41%); and more graduate students (84 vs 41%). G1Rs and G2Rs also differed in prioritizing the challenges they envisage for implementing mental health research: G1Rs ranked the lack of trained resources as the main challenge, while G2Rs remarked on the lack of research culture in their institutions. Conclusions: Our results identify mental health care institutions as the main focus for strengthening of research capacity . This task could be achieved through an intensive interactive work with already stronger universities and research institutes in the region.

Investigación en salud mental en América Latina y el Caribe: enfoque en las Ciencias Básicas / Mental Health Research in Latin America and Caribbean: focus on basic science

Objetivo: evaluar el estado de la investigación en ciencias básicas (CB) dentro del ámbito de la salud mental (SM), en países de América Latina y el Caribe (pALC). Material y métodos: Se recopiló información de bases de datos de publicaciones (PubMed/PsycINFO), mediante buscadores de internet y muestreo bola de nieve, para identificar investigadores y otros actores involucrados en la gestión de la salud mental. Los 2555 actores identificados recibieron un cuestionario dirigido a evaluar su capacidad personal/ institucional para realizar investigación en SM. Tanto la base de datos de publicaciones como las respuestas al cuestionario fueron utilizadas para el análisis. Resultados: Se identificó 2397 publicaciones en SM, de las cuales 222 estuvieron vinculadas a las CB. Sólo 9 de los 30 ALC tenían publicaciones en esta área. Uno de cada cuatro investigadores en Sm de la región la mayoría psiquiatras y neurólogos declaró tener entrenamiento formal en Cb. El 41% de los investigadores declaró contar con apoyo técnico para la investigación en CB en sus instituciones. Sólo 10% de los proyectos de investigación realizados en os últimos 5 años estuvieron relacionados a las CB. De estos, un 62% se realizó con colaboración local, y 30% con colaboradores de países desarrollados. La financiación provino principalmente de fundaciones, ONGs y universidades. Las principales motivaciones de los investigadores de SM en el área de las CB fueron el interés personal (77%), y la carga de enfermedad (63%). La investigación en CB no fue considerada una prioridad por la mayor parte de los investigadores (55%) y tomadores de decisión (56%). Los investigadores con entrenamiento formal en CB tuvieron más publicaciones y accedieron a mayores fondos de investigación que el resto de investigadores. Conclusiones: La investigación en CB en el área de SM permanece relegada en la región.

Objective: To evaluate the status of basic science (BSci) research in mental health (MH) in Latin American and the Caribbean (LAC) countries. Material and methods: A questionnaire was sent to 2555 MH researchers and stakeholders identified by a mapping process through publication databases (PubMed/PsycINFO), internet searches of institutions, and snowball sampling. The Questionnaire was designed to obtain information about their capacity for MH research. Both, the publication database and the questionnaire responses were the basis for all analyses. Results: Two hundred twenty-two out of 2397 MH publications were related to BSci. Only 9 LAC countries had MH publications in this area. One out of four researchers in MH in the region most of them psychiatrists and neurologists had formal training in BSci research methods. Forty one percent of them declared to have technical support for BSci research from their institutions. Barely 10% of the research projects in the past 5 year had a BSci approach. From them, 62% were local projects, and% were done in collaborations with developed countries, being funded mainly by NGOs or foundations. The main motivations for research in BSci were personal interest (77%) and burden of disease (63%). BSci research was not considered a priority by most researchers (55%) or stakeholders (56%). BSci researchers published more and had better financing than non-BSci is an integral component of MH research and cannot de set aside when establishing agendas and setting priorities in this health area. Given their characteristics, BSci researchers could effectively contribute to improve MH research capacity in the region.

What challenges does mental and neurological health research face in Latin American countries? / ¿Qué desafíos enfrenta la investigación en salud mental y neurológica en los países latinoamericanos?

OBJECTIVE: The World Health Organization Atlas Project identified important deficiencies in world mental and neurological health resources. These deficiencies, especially evident in low and middle-income countries, can be overcome by improving research capacity. The objective of this study is to assess the status of mental and neurological research in Latin American countries and identify the main difficulties encountered in conducting research, publishing results, and shaping health policies, interventions, and programs. METHOD: Semi-structured interviews were conducted with 34 key informants from 13 Latin American countries. RESULTS: Participants reported that production of mental and neurological research in Latin American countries is low. Lack of financial and human resources, including lack of support from government agencies, were identified as the main factors contributing to the dearth of local research. The few research projects that do take place in Latin American countries are often funded at researchers' personal expense. Few policies, interventions, or programs are generated from research results. To address these deficiencies, participants called for training in research methodology, mechanisms for identifying funding opportunities, and greater recognition of their research products. CONCLUSIONS: Researchers and stakeholders recognize the need to mobilize local and international efforts aimed at strengthening research capacity and results implementation. This will lead to an overall optimization of mental and neurological research in the region.

OBJETIVO: El proyecto Atlas de la Organización Mundial de la Salud identifica importantes deficiencias en salud mental y neurológica. Estas deficiencias, especialmente evidentes en países de medianos y bajos ingresos, pueden resolverse mejorando las capacidades en investigación. El objetivo de este estudio es evaluar el estado de la investigación en salud mental y neurológica en países Latinoamericanos, e identificar las principales dificultades encontradas al hacer investigación, publicar resultados, y generar políticas, intervenciones, y programas. MÉTODO: Entrevistas semi-estructuradas fueron realizadas a 34 informantes de 13 países Latinoamericanos. RESULTADOS: La producción de investigación en salud mental y neurológica en países Latinoamericanos es escasa, debido principalmente a la carencia de recursos financieros y humanos, incluyendo el casi ausente apoyo de agencias gubernamentales. Los pocos proyectos de investigación que se llevan a cabo son financiados mayormente con recursos propios de los investigadores. Pocas políticas, intervenciones o programas son generados a partir de resultados de investigación. Resolver estas deficiencias requerirá entrenar profesionales en metodología de la investigación, identificar oportunidades de financiación y lograr un mayor reconocimiento de los productos de la investigación. CONCLUSIONES: Hay necesidad de movilizar esfuerzos locales e internacionales orientados a fortalecer las capacidades en investigación y la implementación de resultados. Esto llevará a una optimización general de la investigación.

Bronchoalveolar lavage in malignancy.

Bronchoalveolar lavage is a useful diagnostic tool in diffuse or disseminated lung malignancies that do not involve the bronchial structures visible by endoscopy. The neoplastic histotype and the intraparenchymal neoplastic growth pattern are good predictors for diagnostic yield; adenocarcinoma, and tumors with lymphangitic or lepidic growth patterns are more easily diagnosed by bronchoalveolar lavage; in these cases the diagnostic yield reported is higher than 80%. In hematologic malignancies the diagnostic yield is quite good in secondary diffuse indolent B cell lymphomas and in primary B cell lymphomas of mucosa-associated lymphoid tissue (MALT) type but low in Hodgkin disease. Morphological analysis may be implemented by immunocytochemical or molecular tests to identify the cell lineage and the presence of monoclonality. Disorders in which bronchioloalveolar cell hyperplasia/dysplasia is a significant morphological component may have cytological features in bronchoalveolar lavage fluid that mimic lung neoplasms: acute respiratory distress syndrome (ARDS), acute interstitial pneumonitis (AIP), and acute exacerbation of idiopathic pulmonary fibrosis are the most important clinical entities in this group.

The ultrastructural basis of renal pathology in monoclonal gammopathies.

The kidney is frequently involved in the course of monoclonal gammopathies (MG). Renal involvement presents different clinical-morphological patterns, which can occur either at the onset or in a late phase of the hematological disease, as well as after chemotherapy. The reasons for the organ tropism of monoclonal immunoglobulins (Igs) are still unknown. Currently, it is well known that some primary structure alterations in monoclonal Igs and/or in their segments correlate to nephrotoxicity. On the other hand, it is impossible to predict the pathogenicity and the clinical manifestations induced by a specific monoclonal Ig based on its specific conformational modifications. Pathogenicity and organ tropism are probably complex phenomena, which involve specific protein factors, patient factors, target organ characteristics and monoclonal plasmacellular mass entities. However, aminoacidic sequence analysis of nephrotoxic Igs and some recent in vitro studies have allowed two different monoclonal light chain (LC) types to be distinguished. Glomerulopathic LCs (G-LCs) in the mesangium recognize their target structure and induce two distinct mesangiopathies, monoclonal Ig deposition disease (MIDD) and AL-amyloidosis (AL). Tubulopathic LCs (T-LCs) act on the proximal or on the distal tubule and cause, respectively, Fanconi syndrome (FS) and cast nephropathy. Pathogenic monoclonal Igs have the propensity to deposit in different renal parenchymal structures in extracellular sites, because of the transformation of soluble precursors in insoluble products. Evidence suggests that somatic mutations can destabilize the normal LCs globular soluble structure and this could be the major driving force for precipitation. Based on these features, MG can be classified as conformational and depositional diseases. Electronmicroscopy (EM) analysis of renal biopsies in MG patients with glomerular diseases distinguishes two morphological aspects. MIDD and a recently identified entity named proliferative glomerulonephritis (GN) with monoclonal IgG deposits are both characterized by non-organized granular electrondense deposits. AL, immunotactoid (IT) glomerulopathy and monoclonal cryoglobulinemia are, instead, characterized by organized deposits such as fibrils or microtubules. Tubular diseases in MG patients produce two different histological patterns. In FS, monoclonal Igs form crystals in the renal interstitium able to induce a local intense flogosis, while in cast nephropathy monoclonal Igs precipitate with Tamm-Horsfall protein (THP) in the proximal tubular lumen and induce tubular obstruction. The different morphological aspects are unrelated to specific clinical manifestations, while renal biopsy can diagnose different entities that can respond to different therapeutical schedules. This reveals the importance of the renal biopsy in the clinical management of the renal pathology in plasma cells dyscrasias, mainly when supported by the most advanced techniques of immunoelectronmicroscopy and polymerase chain reaction (PCR)-mediated analysis. Further elucidation of the molecular events involved in the pathogenesis of the different forms of renal damage is needed to design new and more effective therapeutical strategies. In particular, urinary proteomics seem to be promising in this setting.

Tasa de transporte de litio en enfermedad maníaco-depresiva e hipertensión arterial esencial / Lithium transport rate in maniac depressive illness and essential hipertension

Se presenta el estudio de la tasa de transporte de Litio (TTLi) en cinco grupos de personas: Control, pacientes con Enfermedad maníaco-depresiva (EMD), pacientes de Hipertensión Arterial Esencial (HAE) y parientes de EMD y HAE. Se estudia también la TTLi en 13 familias de EMD (dos a tres generaciones). Los resultados obtenidos nos llevan a la conclusión que la trasmisión genética de la EMD, en los casos en que el cromosoma X es el comprometido, es de carácter recesivo, mientras que en la HAE es de carácter dominante, y sugerimos que el estudio de la TTLi podría ser un elemento valioso para el diagnóstico del rasgo genético de la presencia potencial de ambas enfermedades, dentro de las limitaciones que son discutidas en la sección pertinente