TREX tetramer disruption alters RNA processing necessary for corticogenesis in THOC6 Intellectual Disability Syndrome.

THOC6 variants are the genetic basis of autosomal recessive THOC6 Intellectual Disability Syndrome (TIDS). THOC6 is critical for mammalian Transcription Export complex (TREX) tetramer formation, which is composed of four six-subunit THO monomers. The TREX tetramer facilitates mammalian RNA processing, in addition to the nuclear mRNA export functions of the TREX dimer conserved through yeast. Human and mouse TIDS model systems revealed novel THOC6-dependent, species-specific TREX tetramer functions. Germline biallelic Thoc6 loss-of-function (LOF) variants result in mouse embryonic lethality. Biallelic THOC6 LOF variants reduce the binding affinity of ALYREF to THOC5 without affecting the protein expression of TREX members, implicating impaired TREX tetramer formation. Defects in RNA nuclear export functions were not detected in biallelic THOC6 LOF human neural cells. Instead, mis-splicing was detected in human and mouse neural tissue, revealing novel THOC6-mediated TREX coordination of mRNA processing. We demonstrate that THOC6 is required for key signaling pathways known to regulate the transition from proliferative to neurogenic divisions during human corticogenesis. Together, these findings implicate altered RNA processing in the developmental biology of TIDS neuropathology.

GIMAP5 deficiency reveals a mammalian ceramide-driven longevity assurance pathway.

Preserving cells in a functional, non-senescent state is a major goal for extending human healthspans. Model organisms reveal that longevity and senescence are genetically controlled, but how genes control longevity in different mammalian tissues is unknown. Here, we report a new human genetic disease that causes cell senescence, liver and immune dysfunction, and early mortality that results from deficiency of GIMAP5, an evolutionarily conserved GTPase selectively expressed in lymphocytes and endothelial cells. We show that GIMAP5 restricts the pathological accumulation of long-chain ceramides (CERs), thereby regulating longevity. GIMAP5 controls CER abundance by interacting with protein kinase CK2 (CK2), attenuating its ability to activate CER synthases. Inhibition of CK2 and CER synthase rescues GIMAP5-deficient T cells by preventing CER overaccumulation and cell deterioration. Thus, GIMAP5 controls longevity assurance pathways crucial for immune function and healthspan in mammals.

Inocuidad de los productos lácteos y su influencia en la salud

Introducción: Las enfermedades transmitidas por los alimentos es un fenómeno a nivel internacional que provoca cada año miles de enfermos o muertos, con la inclusión de niños y ancianos. La inocuidad de los alimentos es el proceso que garantiza la calidad de estos durante el proceso productivo, almacenamiento y distribución. Evitar afectaciones en este proceso beneficia directamente la calidad de vida sin que represente un riesgo para la salud. Objetivo: Ofrecer acciones en el proceso industrial de la Unidad Empresarial de Base Planta de Leche y sus Derivados, perteneciente a la Empresa Productos Lácteos Guantánamo, que permitan la inocuidad de los productos lácteos a partir de insuficiencias identificadas. Método: Se realizó un estudio cualitativo y cuantitativo en dicha planta en el periodo 2021-2022, donde participaron 43 obreros (N=43) directamente vinculados con el proceso productivo de los productos lácteos. Se aplicaron el cuestionario y la observación participante para la obtención de la información. Se contó, además, con los criterios y opiniones de cuatros directivos y seis especialistas de la entidad. Resultados: Se hallaron limitaciones en la concepción de alternativas que constituyen acciones novedosas para la aplicación en el proceso industrial de resultados de calidad en la inocuidad de los productos lácteos a partir de la obsolescencia de los equipos y los niveles de conocimiento de Regular y Mal en el 100 % de los obreros. Se aplicaron acciones que permitan asegurar la inocuidad. Conclusiones: Las acciones que se presentan satisfacen las limitaciones diagnosticadas, corroborada durante el desempeño laboral para satisfacer las demandas sociales de inocuidad.

Introduction: Foodborne diseases are an international phenomenon that causes thousands of illnesses or deaths every year, including children and the elderly. Food safety is the process that guarantees food quality during the production process, storage and distribution. Avoiding effects in this process directly benefits the quality of life without representing a risk to health. Objective: To offer actions in the industrial process of the Milk and its Derivatives Plant Base Business Unit, belonging to the Productos Lácteos Guantánamo Company, that allow the safety of dairy products based on identified insufficiencies. Method: A qualitative and quantitative study was carried out in said plant in the period 2021-2022, where 43 workers (N=43) directly linked to the production process of dairy products participated. The questionnaire and participant observation were applied to obtain the information. In addition, the criteria and opinions of four directors and six specialists of the entity were included. Results: Limitations were found in the conception of alternatives that constitute novel actions for the application in the industrial process of quality results in the safety of dairy products based on the obsolescence of equipment and the levels of knowledge of Regular and Poor in 100% of the workers. Actions were applied to ensure safety. Conclusions: The actions presented satisfy the diagnosed limitations, corroborated during work performance to satisfy social demands for safety.

Introdução: As doenças transmitidas por alimentos são um fenómeno internacional que causa milhares de doenças ou mortes todos os anos, incluindo crianças e idosos. A segurança alimentar é o processo que garante a qualidade dos alimentos durante o processo de produção, armazenamento e distribuição. Evitar efeitos nesse processo beneficia diretamente a qualidade de vida sem representar risco à saúde. Objetivo: Oferecer ações no processo industrial da Unidad Empresarial de Base Planta de Leche y sus Derivados, pertencente à Empresa Produtos Lácteos Guantánamo, que permitam a segurança de produtos lácteos com base nas insuficiências identificadas. Método: Foi realizado um estudo qualitativo e quantitativo na referida fábrica no período 2021-2022, onde participaram 43 trabalhadores (N=43) ligados diretamente ao processo produtivo de laticínios. O questionário e a observação participante foram aplicados para obtenção das informações. Além disso, foram incluídos os critérios e opiniões de quatro diretores e seis especialistas da entidade. Resultados: Foram encontradas limitações na concepção de alternativas que constituam ações inéditas para a aplicação no processo industrial de resultados de qualidade na segurança de produtos lácteos com base na obsolescência dos equipamentos e nos níveis de conhecimento Regular e Ruim em 100% dos trabalhadores. Ações foram aplicadas para garantir a segurança. Conclusões: As ações apresentadas satisfazem as limitações diagnosticadas, corroboradas durante o desempenho do trabalho para satisfazer as demandas sociais de segurança.

Novel CARMIL2 (RLTPR) Mutation Presenting with Hyper-IgE and Eosinophilia: A Case Report.

BACKGROUND: Inborn errors of immunity are a growing group of disorders with a wide spectrum of genotypic and phenotypic profiles. CARMIL2 (previously named RLTPR) deficiency is a recently described cause of immune dysregulation, mainly presenting with allergy, mucocutaneous infections, and inflammatory bowel disease. CARMIL2 deficiency is categorized under diseases of immune dysregulation with susceptibility to lymphoproliferative conditions. CASE PRESENTATION: Here we describe a 29-years-old male from a consanguineous family, with food and sting allergy, allergic rhinitis, facial molluscum contagiosum (viral infection of the skin in the form of umbilicated papules), eosinophilia and highly elevated serum IgE level. Whole exome sequencing revealed numerous homozygous variants, including a CARMIL2 nonsense mutation, a gene regulating actin polymerization, and promoting cell protrusion formation. CONCLUSION: The selective role of CARMIL2 in T cell activation and maturation through cyto-skeletal organization is proposed to be the cause of immune dysregulation in individuals with CARMIL2 deficiency. CARMIL2 has an important role in immune pathways regulation, through cell maturation and differentiation, giving rise to a balance between Th1, Th2, and Th17 immune response. This case can improve the understanding of the different impacts of CARMIL2 mutations on immune pathways and further guide the diagnosis of patients with similar phenotypes.

Mechanisms of mRNA processing defects in inherited THOC6 intellectual disability syndrome.

THOC6 is the genetic basis of autosomal recessive THOC6 Intellectual Disability Syndrome (TIDS). THOC6 facilitates the formation of the Transcription Export complex (TREX) tetramer, composed of four THO monomers. The TREX tetramer supports mammalian mRNA processing that is distinct from yeast TREX dimer functions. Human and mouse TIDS model systems allow novel THOC6-dependent TREX tetramer functions to be investigated. Biallelic loss-of-functon(LOF) THOC6 variants do not influence the expression and localization of TREX members in human cells, but our data suggests reduced binding affinity of ALYREF. Impairment of TREX nuclear export functions were not detected in cells with biallelic THOC6 LOF. Instead, mRNA mis-splicing was observed in human and mouse neural tissue, revealing novel insights into THOC6-mediated TREX coordination of mRNA processing. We demonstrate that THOC6 is required for regulation of key signaling pathways in human corticogenesis that dictate the transition from proliferative to neurogenic divisions that may inform TIDS neuropathology.

Evans syndrome caused by a deleterious mutation affecting the adaptor protein SASH3.

Increasing evidence suggests multilineage cytopenias (also known as Evans syndrome) may be caused by inborn errors of immunity (IEI) with immune dysregulation. We studied a patient with autoimmune haemolytic anaemia and immune thrombocytopenia and identified a germline mutation in SASH3 (c.862C>T;p.Arg288Ter), indicating a recently identified IEI. Immunohistochemistry performed after clinically indicated splenectomy revealed severe hypoplasia/absence of germinal centres. The autoimmune phenotype was associated with an increased CD21low T-bet+ CD11c+ subset along with decreased regulatory T cells, impaired T-cell proliferation and T-cell exhaustion. The younger brother carries the same SASH3 mutation and shares immunophenotypic features but is currently clinical asymptomatic, indicating heterogeneity of SASH3 deficiency.

Systemic Inflammation and Normocytic Anemia in DOCK11 Deficiency.

BACKGROUND: Increasing evidence links genetic defects affecting actin-regulatory proteins to diseases with severe autoimmunity and autoinflammation, yet the underlying molecular mechanisms are poorly understood. Dedicator of cytokinesis 11 (DOCK11) activates the small Rho guanosine triphosphatase (GTPase) cell division cycle 42 (CDC42), a central regulator of actin cytoskeleton dynamics. The role of DOCK11 in human immune-cell function and disease remains unknown. METHODS: We conducted genetic, immunologic, and molecular assays in four patients from four unrelated families who presented with infections, early-onset severe immune dysregulation, normocytic anemia of variable severity associated with anisopoikilocytosis, and developmental delay. Functional assays were performed in patient-derived cells, as well as in mouse and zebrafish models. RESULTS: We identified rare, X-linked germline mutations in DOCK11 in the patients, leading to a loss of protein expression in two patients and impaired CDC42 activation in all four patients. Patient-derived T cells did not form filopodia and showed abnormal migration. In addition, the patient-derived T cells, as well as the T cells from Dock11-knockout mice, showed overt activation and production of proinflammatory cytokines that were associated with an increased degree of nuclear translocation of nuclear factor of activated T cell 1 (NFATc1). Anemia and aberrant erythrocyte morphologic features were recapitulated in a newly generated dock11-knockout zebrafish model, and anemia was amenable to rescue on ectopic expression of constitutively active CDC42. CONCLUSIONS: Germline hemizygous loss-of-function mutations affecting the actin regulator DOCK11 were shown to cause a previously unknown inborn error of hematopoiesis and immunity characterized by severe immune dysregulation and systemic inflammation, recurrent infections, and anemia. (Funded by the European Research Council and others.).

Phosphomevalonate kinase deficiency expands the genetic spectrum of systemic autoinflammatory diseases.

BACKGROUND: In the isoprenoid biosynthesis pathway, mevalonate is phosphorylated in 2 subsequent enzyme steps by MVK and PMVK to generate mevalonate pyrophosphate that is further metabolized to produce sterol and nonsterol isoprenoids. Biallelic pathogenic variants in MVK result in the autoinflammatory metabolic disorder MVK deficiency. So far, however, no patients with proven PMVK deficiency due to biallelic pathogenic variants in PMVK have been reported. OBJECTIVES: This study reports the first patient with functionally confirmed PMVK deficiency, including the clinical, biochemical, and immunological consequences of a homozygous missense variant in PMVK. METHODS: The investigators performed whole-exome sequencing and functional studies in cells from a patient who, on clinical and immunological evaluation, was suspected of an autoinflammatory disease. RESULTS: The investigators identified a homozygous PMVK p.Val131Ala (NM_006556.4: c.392T>C) missense variant in the index patient. Pathogenicity was supported by genetic algorithms and modeling analysis and confirmed in patient cells that revealed markedly reduced PMVK enzyme activity due to a virtually complete absence of PMVK protein. Clinically, the patient showed various similarities as well as distinct features compared to patients with MVK deficiency and responded well to therapeutic IL-1 inhibition. CONCLUSIONS: This study reported the first patient with proven PMVK deficiency due to a homozygous missense variant in PMVK, leading to an autoinflammatory disease. PMVK deficiency expands the genetic spectrum of systemic autoinflammatory diseases, characterized by recurrent fevers, arthritis, and cytopenia and thus should be included in the differential diagnosis and genetic testing for systemic autoinflammatory diseases.

Severe congenital neutropenia due to G6PC3 deficiency: early and delayed phenotype of a patient.

BACKGROUND: Severe Congenital Neutropenia type 4 (SCN4), is a rare autosomal recessive condition, due to mutations in the G6PC3 gene. The phenotype comprises neutropenia of variable severity and accompanying anomalies. CASE PRESENTATION: We report a male patient with confirmed G6PC3 deficiency presented with recurrent bacterial infections and multi-systemic complications. Our case was the first with a novel homozygous frameshift mutation in G6PC3. The patient demonstrated large platelets on his peripheral blood smear which is a rare presentation of this disease. CONCLUSION: As SCN4 patients could be easily missed, it is recommended to consider G6PC3 mutation for any case of congenital, unexplained neutropenia.

Biallelic NFATC1 mutations cause an inborn error of immunity with impaired CD8+ T-cell function and perturbed glycolysis.

The nuclear factor of activated T cells (NFAT) family of transcription factors plays central roles in adaptive immunity in murine models; however, their contribution to human immune homeostasis remains poorly defined. In a multigenerational pedigree, we identified 3 patients who carry germ line biallelic missense variants in NFATC1, presenting with recurrent infections, hypogammaglobulinemia, and decreased antibody responses. The compound heterozygous NFATC1 variants identified in these patients caused decreased stability and reduced the binding of DNA and interacting proteins. We observed defects in early activation and proliferation of T and B cells from these patients, amenable to rescue upon genetic reconstitution. Stimulation induced early T-cell activation and proliferation responses were delayed but not lost, reaching that of healthy controls at day 7, indicative of an adaptive capacity of the cells. Assessment of the metabolic capacity of patient T cells revealed that NFATc1 dysfunction rendered T cells unable to engage in glycolysis after stimulation, although oxidative metabolic processes were intact. We hypothesized that NFATc1-mutant T cells could compensate for the energy deficit due to defective glycolysis by using enhanced lipid metabolism as an adaptation, leading to a delayed, but not lost, activation responses. Indeed, we observed increased 13C-labeled palmitate incorporation into citrate, indicating higher fatty acid oxidation, and we demonstrated that metformin and rosiglitazone improved patient T-cell effector functions. Collectively, enabled by our molecular dissection of the consequences of loss-of-function NFATC1 mutations and extending the role of NFATc1 in human immunity beyond receptor signaling, we provide evidence of metabolic plasticity in the context of impaired glycolysis observed in patient T cells, alleviating delayed effector responses.

Use of a Biostimulant to Mitigate the Effects of Excess Salinity in Soil and Irrigation Water in Tomato Plants.

Global warming is linked to progressive soil salinisation, which reduces crop yields, especially in irrigated farmland on arid and semiarid regions. Therefore, it is necessary to apply sustainable and effective solutions that contribute to enhanced crop salt tolerance. In the present study, we tested the effects of a commercial biostimulant (BALOX®) containing glycine betaine (GB) and polyphenols on the activation of salinity defense mechanisms in tomato. The evaluation of different biometric parameters and the quantification of biochemical markers related to particular stress responses (osmolytes, cations, anions, oxidative stress indicators, and antioxidant enzymes and compounds) was carried out at two phenological stages (vegetative growth and the beginning of reproductive development) and under different salinity conditions (saline and non-saline soil, and irrigation water), using two formulations (different GB concentrations) and two doses of the biostimulant. Once the experiments were completed, the statistical analysis revealed that both formulations and doses of the biostimulant produced very similar effects. The application of BALOX® improved plant growth and photosynthesis and assisted osmotic adjustment in root and leaf cells. The biostimulant effects are mediated by the control of ion transport, reducing the uptake of toxic Na+ and Cl- ions and favoring the accumulation of beneficial K+ and Ca2+ cations, and a significant increase in leaf sugar and GB contents. BALOX® significantly reduced salt-induced oxidative stress and its harmful effects, as evidenced by a decrease in the concentration of oxidative stress biomarkers, such as malondialdehyde and oxygen peroxide, which was accompanied by the reduction of proline and antioxidant compound contents and the specific activity of antioxidant enzymes with respect to the non-treated plants.

Case Report: Refractory Cytopenia With a Switch From a Transient Monosomy 7 to a Disease-Ameliorating del(20q) in a NHEJ1-Deficient Long-term Survivor.

We report the case of a male Pakistani patient with a pathogenic homozygous loss of function variant in the non-homologous end-joining factor 1 (NHEJ1) gene. The growth retarded and microcephalic boy with clinodactyly of both hands and hyperpigmentation of the skin suffered from recurrent respiratory infections. He was five and a half years old when he came to our attention with refractory cytopenia and monosomy 7. Hematopoietic stem cell transplantation was considered but not feasible because there was no suitable donor available. Monosomy 7 was not detected anymore in subsequent bone marrow biopsies that were repeated in yearly intervals. Instead, seven and a half years later, a novel clone with a del(20q) appeared and steadily increased thereafter. In parallel, the patient's blood count, which had remained stable for over 20 years without necessitating any specific therapeutic interventions, improved gradually and the erythropoiesis-associated dysplasia resolved.

Calidad de las conferencias: un instrumento para su evaluación / Quality of lectures: an instrument for their evaluation

RESUMEN Introducción: desde la evaluación de las conferencias se puede contribuir a elevar la calidad de las clases. Objetivo: diseñar un instrumento para la evaluación de las conferencias en la carrera de Medicina. Métodos: se realizó una investigación con dos fases: en la primera se diseñó el instrumento a partir de la utilización de métodos téoricos y empíricos. En la segunda, mediante un estudio descriptivo transversal, se evaluó la validez de la guía. Se utilizó en una muestra de 90 conferencias escogidas al azar de las carreras de Medicina, Estomatología y Enfermería impartidas en el hospital Dr. Agostinho Neto durante el año 2017. Resultados: se diseña una guía de observación compuesta por indicadores que estructuran las dimensiones: condiciones organizativas e higiénico-sanitarias del aula, calidad de la introducción, el desarrollo y las conclusiones de la clase. Se demuestra pertinencia del instrumento para evaluar la calidad de las conferencias: insuficiencias en el uso de la pizarra (68,6 %); diapositivas con deficiente calidad (62,8 %); no orientación de trabajo independiente (54,3 %). Conclusiones: el instrumento puede contextualizarse en todos los perfiles de la educación médica y contribuye a la objetividad en la comparación de resultados de las diferentes comprobaciones, facilita la realización de estudios para evaluar la evolución de la calidad del proceso enseñanza-aprendizaje, permite asignar prioridades por el departamento de docencia del centro para la superación profesoral, así como evaluar el impacto de los programas de superación pedagógica del claustro.

ABSTRACT Introduction: the evaluation of lectures can contribute to improve the quality of lessons. Objective: to design an instrument for the evaluation of lectures in Medicine major. Methods: a two-phase research was carried out: in the first phase, the instrument was designed based on the use of theoretical and empirical methods. In the second, by means of a cross-sectional descriptive study, the validity of the guide was evaluated. It was used in a sample of 90 lectures randomly chosen from the Medicine, Dentistry and Nursing majors taught at Dr. Agostinho Neto Hospital during 2017. Results: an observation guide is designed comprising indicators that structure the dimensions: organizational and hygienic-sanitary conditions of the classroom, quality of the introduction, development and conclusions of the lectures. The relevance of the instrument to evaluate the quality of the lectures is demonstrated: insufficiencies in the use of blackboard 68,6 %; slides with deficient quality 62,8 %; no orientation of independent work 54,3 %. Conclusions: the instrument can be contextualized in all the profiles of medical education and contributes to the objectivity in the comparison of results of the different verifications, it facilitates the understanding of studies to evaluate the evolution of the quality of the teaching-learning process, it also allows assigning priorities by the teaching department of the center for the training of professors, as well as evaluating the impact of the programs of pedagogical training for the teaching staff.

Case Report of a Novel NFkB Mutation in a Lymphoproliferative Disorder Patient.

BACKGROUND: Lymphoproliferative disorders include a heterogeneous list of conditions that commonly involve dysregulation of lymphocyte proliferation resulting in lymphadenopathy and bone marrow infiltration. These disorders have various presentations, most notably autoimmune manifestations, organomegaly, lymphadenopathy, dysgammaglobulinemia, and increased risk of chronic infections. CASE PRESENTATION: A young boy presented with symptoms overlapping different lymphoproliferative disorders, including episodes of chronic respiratory tract infections, dysgammaglobulinemia, lymphadenopathy-associated with splenomegaly as well as skin rashes. Genetic studies revealed multiple heterozygous variants, including a novel mutation in the NFκB1 gene. CONCLUSION: This novel mutation can reveal new aspects in the pathogenesis of lymphoproliferative disorders and propose new treatments for them.

Synonymous mutation in adenosine triphosphatase copper-transporting beta causes enhanced exon skipping in Wilson disease.

Wilson disease (WD) is caused by biallelic pathogenic variants in adenosine triphosphatase copper-transporting beta (ATP7B); however, genetic testing identifies only one or no pathogenic ATP7B variant in a number of patients with WD. Synonymous single-nucleotide sequence variants have been recognized as pathogenic in individual families. The aim of the present study was to evaluate the prevalence and disease mechanism of the synonymous variant c.2292C>T (p.Phe764=) in WD. A cohort of 280 patients with WD heterozygous for a single ATP7B variant was investigated for the presence of c.2292C>T (p.Phe764=). In this cohort of otherwise genetically unexplained WD, the allele frequency of c.2292C>T (p.Phe764=) was 2.5% (14 of 560) compared to 7.1 × 10-6 in the general population (2 of 280,964 in the Genome Aggregation Database; p < 10-5 ; Fisher exact test). In an independent United Kingdom (UK) cohort, 2 patients with WD homozygous for p.Phe764= were identified. RNA analysis of ATP7B transcripts from patients homozygous or heterozygous for c.2292C>T and control fibroblasts showed that this variant caused high expression of an ATP7B transcript variant lacking exon 8. Conclusion: The synonymous ATP7B variant c.2292C>T (p.Phe764=) causes abnormal messenger RNA processing of ATP7B transcripts and is associated with WD in compound heterozygotes and homozygotes.

Low Density Granulocytes and Dysregulated Neutrophils Driving Autoinflammatory Manifestations in NEMO Deficiency.

NF-κB essential modulator (NEMO, IKK-γ) deficiency is a rare combined immunodeficiency caused by mutations in the IKBKG gene. Conventionally, patients are afflicted with life threatening recurrent microbial infections. Paradoxically, the spectrum of clinical manifestations includes severe inflammatory disorders. The mechanisms leading to autoinflammation in NEMO deficiency are currently unknown. Herein, we sought to investigate the underlying mechanisms of clinical autoinflammatory manifestations in a 12-years old male NEMO deficiency (EDA-ID, OMIM #300,291) patient by comparing the immune profile of the patient before and after hematopoietic stem cell transplantation (HSCT). Response to NF-kB activators were measured by cytokine ELISA. Neutrophil and low-density granulocyte (LDG) populations were analyzed by flow cytometry. Peripheral blood mononuclear cells (PBMC) transcriptome before and after HSCT and transcriptome of sorted normal-density neutrophils and LDGs were determined using the NanoString nCounter gene expression panels. ISG15 expression and protein ISGylation was based on Immunoblotting. Consistent with the immune deficiency, PBMCs of the patient were unresponsive to toll-like and T cell receptor-activators. Paradoxically, LDGs comprised 35% of patient PBMCs and elevated expression of genes such as MMP9, LTF, and LCN2 in the granulocytic lineage, high levels of IP-10 in the patient's plasma, spontaneous ISG15 expression and protein ISGylation indicative of a spontaneous type I interferon (IFN) signature were observed, all of which normalized after HSCT. Collectively, our results suggest that type I IFN signature observed in the patient, dysregulated LDGs and spontaneously activated neutrophils, potentially contribute to tissue damage in NEMO deficiency.