Not enough by half: NFAT5 haploinsufficiency in two patients with Epstein-Barr virus susceptibility.

Introduction: The transcription factor Nuclear factor of activated T cells 5 (NFAT5), pivotal in immune regulation and function, can be induced by osmotic stress and tonicity-independent signals. Objective: We aimed to investigate and characterize two unrelated patients with Epstein-Barr virus susceptibility and no known genetic etiology. Methods: After informed consent, we reviewed the electronic charts, extracted genomic DNA, performed whole-exome sequencing, filtered, and prioritized their variants, and confirmed through Sanger sequencing, family segregation analysis, and some functional assays, including lymphoproliferation, cytotoxicity, and characterization of natural killer cells. Results: We describe two cases of pediatric Mexican patients with rare heterozygous missense variants in NFAT5 and EBV susceptibility, a school-age girl with chronic-active infection of the liver and bowel, and a teenage boy who died of hemophagocytic lymphohistiocytosis. Discussion: NFAT5 is an important regulator of the immune response. NFAT5 haploinsufficiency has been described as an immunodeficiency syndrome affecting both innate and adaptive immunity. EBV susceptibility might be another manifestation in the spectrum of this disease.

Severe congenital neutropenia due to G6PC3 deficiency: Case series of five patients and literature review.

BACKGROUND AND OBJECTIVES: Glucose-6-phosphate catalytic subunit 3 (G6PC3) deficiency is characterized by severe congenital neutropenia with recurrent pyogenic infections, a prominent superficial venous pattern and cardiovascular and urogenital malformations caused by an alteration of glucose homeostasis, with increased endoplasmic reticulum stress and cell apoptosis. METHODS: We reviewed our patients with G6PC3 deficiency diagnosed along the last decade in Mexico; we also searched the PubMed/Medline database for the terms ('G6PC3 deficiency' OR 'Dursun syndrome' OR 'Severe congenital neutropenia type 4'), and selected articles published in English from 2009 to 2020. RESULTS: We found 89 patients reported from at least 14 countries in 4 continents. We describe five new cases from Mexico. Of the 94 patients, 56% are male, 48% from Middle East countries and none of them had adverse reactions to live vaccines; all presented with at least 1 severe infection prior to age 2. Seventy-five per cent had syndromic features, mainly atrial septal defect in 55% and prominent superficial veins in 62%. CONCLUSIONS: With a total of 94 patients reported in the past decade, we delineate the most frequent laboratory and genetic features, their treatment and outcomes, and to expand the knowledge of syndromic and non-syndromic phenotypes in these patients.

COVID-19 in the Context of Inborn Errors of Immunity: a Case Series of 31 Patients from Mexico.

INTRODUCTION: Patients with inborn errors of immunity (IEI) have a compromised or inappropriate immune response. Although they might be considered a high-risk group for severe SARS-CoV-2 infection, the reported impact of COVID-19 in these patients has been reassuring, while the differential susceptibility of distinct types of IEI remains unclear. OBJECTIVE: We aimed to describe the findings and outcomes of our known patients with IEI who were diagnosed with COVID-19. METHODS: In a retrospective study from March 2020 to February 2021, four centers in Mexico collected clinical, laboratory, and genetic data from pediatric and adult patients with known diagnoses of IEI who presented with COVID-19, based on compatible symptoms and positive SARS-CoV-2 testing or known household exposure. RESULTS: We report 31 patients with known IEI from Mexico who presented with SARS-CoV-2 infection. Seventy-four percent were male, 52% were pediatric, and 81% survived. Their ages ranged from 5 months to 56 years, with a median of 17 years. Sixty-five percent had predominant antibody deficiencies, 48% were hospitalized, and 26% required ICU. Pediatric patients had a higher hospital admission rate than adults. Inpatient mortality was 40%, and ICU mortality rate was 63%. Forty-eight percent developed pneumonia, while 36% had evidence of hyperinflammation (4 adults and 7 children). Predominant laboratory features were lymphopenia and thrombocytopenia, seen in 70 and 44% of patients, respectively. The serum D-dimer median value was 2.6 (0.5-20.6) µg/mL, and the median highest ferritin value was 1015 (32-10,303) ng/mL. Intravenous immunoglobulin was used in 80% of patients. Other treatments included macrolides (39%) and corticosteroids (29%). Six patients died from secondary infection or uncontrolled systemic inflammation. DISCUSSION: Although impaired immunity due to IEI may be a predisposing factor for severe COVID-19, most of our patients with IEI who acquired the SARS-CoV-2 infection developed a well-tolerated infection and survived, as have more than 80% of worldwide reported patients to date. An impaired immune or inflammatory response may be a predisposing factor for some and a protective factor for others. A systematic review of the literature could help identify those patients at risk of severe disease and complications. Healthcare-associated infections should be aggressively prevented.

Diagnostic and therapeutic caveats in Griscelli syndrome.

Griscelli syndrome (GS) is a rare autosomal recessive disease with characteristic pigment distribution, and there are currently 3 types according to the underlying genetic defect and clinical features. We present the case of a girl born from consanguineous parents who presented with predominant neurologic symptoms, silvery hair and granulomatous skin lesions. Cerebral magnetic resonance revealed diffuse white matter lesions, and central nervous system (CNS) lymphocytic infiltration was suspected. The patient underwent haematopoietic stem cell transplantation with graft failure and autologous reconstitution. She developed elevated liver enzyme with a cholestatic pattern. Multiple liver biopsies revealed centrilobular cholestasis and unspecific portal inflammation that improved with immunomodulatory treatment. She was revealed to have an impaired cytotoxicity in NK cells and a decreased expression of RAB27A. However, no variants were found in the gene. All types of GS present with pigment dilution and irregular pigment clumps that can be seen through light microscopy in hair and skin biopsy. Dermic granulomas and immunodeficiency with infectious and HLH predisposition have been described in GS type 2 (GS2). Neurologic alterations might be seen in GS type 1 (GS1) and GS type 2 (GS2), due to different mechanisms. GS1 presents with neurologic impairment secondary to myosin Va role in neuronal development and synapsis. Meanwhile, GS2 can present with neurologic impairment secondary to SNC HLH. Clinical features and cytotoxicity might aid in differentiating GS1 and GS2, especially since treatment differs.

Genetic, Immunological, and Clinical Features of the First Mexican Cohort of Patients with Chronic Granulomatous Disease.

PURPOSE: Chronic granulomatous disease (CGD) is a primary immunodeficiency characterized by an inability of phagocytes to produce reactive oxygen species, impairing their killing of various bacteria and fungi. We summarize here the 93 cases of CGD diagnosed in Mexico from 2011 to 2019. METHODS: Thirteen Mexican hospitals participated in this study. We describe the genetic, immunological, and clinical features of the 93 CGD patients from 78 unrelated kindreds. RESULTS: Eighty-two of the patients (88%) were male. All patients developed bacterial infections and 30% suffered from some kind of fungal infection. Fifty-four BCG-vaccinated patients (58%) presented infectious complications of BCG vaccine. Tuberculosis occurred in 29%. Granulomas were found in 56% of the patients. Autoimmune and inflammatory diseases were present in 15% of patients. A biological diagnosis of CGD was made in 89/93 patients, on the basis of NBT assay (n = 6), DHR (n = 27), and NBT plus DHR (n = 56). The deficiency was complete in all patients. The median age of biological diagnosis was 17 months (range, 0-186 months). A genetic diagnosis was made in 83/93 patients (when material was available), corresponding to CYBB (n = 64), NCF1 (n = 7), NCF2 (n = 7), and CYBA (n = 5) mutations. CONCLUSIONS: The clinical manifestations in these Mexican CGD patients were similar to those in patients elsewhere. This cohort is the largest in Latin America. Mycobacterial infections are an important cause of morbidity in Mexico, as in other countries in which tuberculosis is endemic and infants are vaccinated with BCG. X-linked CGD accounted for most of the cases in Mexico, as in other Latin American countries. However, a significant number of CYBA and NCF2 mutations were identified, expanding the spectrum of known causal mutations.

Cognitive performance after ischaemic stroke / Desempenho cognitivo após acidente vascular cerebral isquêmico

Cognitive impairment after stroke affects the patient recovery process. Therefore, the identification of factors associated with cognitive outcomes is important since it allows risk profiles of stroke survivors to be determined. OBJECTIVE: To assess cognitive outcome of stroke outpatients and investigate associations among clinical and demographic variables, vascular risk factors, depression symptoms and functional ability; and to describe the neuropsychological profile of these patients. METHODS: A cross-sectional design study was conducted. Subjects who suffered a first-ever ischaemic stroke 6 to 10 months prior to data collection underwent neuropsychological assessment and screening for depressive symptoms and functional ability. The outcome "cognitive performance" was analyzed considering two groups: "cognitive impairment" and "no cognitive impairment". RESULTS: There was a statistically significant association between cognitive impairment and female gender, age, stroke severity and functional ability. Regarding neuropsychological profile, the cognitive impairment group exhibited more generalized deficits in attention, visuospatial organization, verbal functions and verbal memory domains compared to the community control group. CONCLUSION: The occurrence of cognitive impairment among patients was high, especially in women, older participants, individuals with more severe stroke, and greater impairment in functional ability. Multiple cognitive domains are affected and this may hamper recovery and negatively impact independence and quality of life after stroke. .

O prejuízo na cognição após evento de Acidente Vascular Cerebral Isquêmico (AVCI) afeta a recuperação dos pacientes. Dessa forma, a identificação de fatores associados ao desfecho cognitivo torna-se relevante ao permitir traçar perfis de risco para os pacientes acometidos. OBJETIVO: Avaliar o desfecho cognitivo dos pacientes vitimados por AVCI em seguimento ambulatorial, verificar associação com variáveis clínicas e demográficas dos pacientes, sintomas de depressão e capacidade funcional, e descrever seu perfil neuropsicológico. MÉTODOS: Foi realizado um estudo transversal, cuja amostra foi composta por 45 pacientes que apresentaram primeiro evento de AVCI nos 6 a 10 meses anteriores à coleta de dados. Os sujeitos foram entrevistados e submetidos à avaliação neuropsicológica e da capacidade funcional e à verificação da presença de sintomas de depressão. O desfecho "desempenho cognitivo" foi analisado considerando dois grupos: "com prejuízo cognitivo" e "sem prejuízo cognitivo". RESULTADOS: Identificou-se associação estatisticamente significativa entre prejuízo cognitivo e sexo feminino, idade, gravidade do AVC e capacidade funcional. Considerando o perfil neuropsicológico, o grupo de sujeitos que apresentou prejuízo cognitivo exibiu déficits mais generalizados, nos domínios de atenção, organização visuoespacial, funções verbais e memória verbal, quando seu desempenho foi comparado aos controles da comunidade. CONCLUSÃO: A ocorrência de prejuízo cognitivo entre os pacientes é elevada, especialmente em mulheres, pessoas com maior idade, naqueles que sofreram AVC mais graves e naqueles que mostraram maior prejuízo na capacidade funcional. Diversos domínios cognitivos são afetados. Isto pode dificultar a recuperação, a independência e a qualidade de vida após o AVC. .

Cognitive performance after ischaemic stroke.

Cognitive impairment after stroke affects the patient recovery process. Therefore, the identification of factors associated with cognitive outcomes is important since it allows risk profiles of stroke survivors to be determined. OBJECTIVE: To assess cognitive outcome of stroke outpatients and investigate associations among clinical and demographic variables, vascular risk factors, depression symptoms and functional ability; and to describe the neuropsychological profile of these patients. METHODS: A cross-sectional design study was conducted. Subjects who suffered a first-ever ischaemic stroke 6 to 10 months prior to data collection underwent neuropsychological assessment and screening for depressive symptoms and functional ability. The outcome "cognitive performance" was analyzed considering two groups: "cognitive impairment" and "no cognitive impairment". RESULTS: There was a statistically significant association between cognitive impairment and female gender, age, stroke severity and functional ability. Regarding neuropsychological profile, the cognitive impairment group exhibited more generalized deficits in attention, visuospatial organization, verbal functions and verbal memory domains compared to the community control group. CONCLUSION: The occurrence of cognitive impairment among patients was high, especially in women, older participants, individuals with more severe stroke, and greater impairment in functional ability. Multiple cognitive domains are affected and this may hamper recovery and negatively impact independence and quality of life after stroke.

O prejuízo na cognição após evento de Acidente Vascular Cerebral Isquêmico (AVCI) afeta a recuperação dos pacientes. Dessa forma, a identificação de fatores associados ao desfecho cognitivo torna-se relevante ao permitir traçar perfis de risco para os pacientes acometidos. OBJETIVO: Avaliar o desfecho cognitivo dos pacientes vitimados por AVCI em seguimento ambulatorial, verificar associação com variáveis clínicas e demográficas dos pacientes, sintomas de depressão e capacidade funcional, e descrever seu perfil neuropsicológico. MÉTODOS: Foi realizado um estudo transversal, cuja amostra foi composta por 45 pacientes que apresentaram primeiro evento de AVCI nos 6 a 10 meses anteriores à coleta de dados. Os sujeitos foram entrevistados e submetidos à avaliação neuropsicológica e da capacidade funcional e à verificação da presença de sintomas de depressão. O desfecho "desempenho cognitivo" foi analisado considerando dois grupos: "com prejuízo cognitivo" e "sem prejuízo cognitivo". RESULTADOS: Identificou-se associação estatisticamente significativa entre prejuízo cognitivo e sexo feminino, idade, gravidade do AVC e capacidade funcional. Considerando o perfil neuropsicológico, o grupo de sujeitos que apresentou prejuízo cognitivo exibiu déficits mais generalizados, nos domínios de atenção, organização visuoespacial, funções verbais e memória verbal, quando seu desempenho foi comparado aos controles da comunidade. CONCLUSÃO: A ocorrência de prejuízo cognitivo entre os pacientes é elevada, especialmente em mulheres, pessoas com maior idade, naqueles que sofreram AVC mais graves e naqueles que mostraram maior prejuízo na capacidade funcional. Diversos domínios cognitivos são afetados. Isto pode dificultar a recuperação, a independência e a qualidade de vida após o AVC.

Implants of polyanionic collagen matrix in bone defects of ovariectomized rats.

In recent years, there has been a great interest in the development of biomaterials that could be used in the repair of bone defects. Collagen matrix (CM) has the advantage that it can be modified chemically to improve its mechanical properties. The aim of the present study was to evaluate the effect of three-dimensional membranes of native or anionic (submitted to alkaline treatment for 48 or 96 h) collagen matrix on the consolidation of osteoporosis bone fractures resulting from the gonadal hormone alterations caused by ovariectomy in rats subjected to hormone replacement therapy. The animals received the implants 4 months after ovariectomy and were sacrificed 8 weeks after implantation of the membranes into 4-mm wide bone defects created in the distal third of the femur with a surgical bur. Macroscopic analysis revealed the absence of pathological alterations in the implanted areas, suggesting that the material was biocompatible. Microscopic analysis showed a lower amount of bone ingrowth in the areas receiving the native membrane compared to the bone defects filled with the anionic membranes. In ovariectomized animals receiving anionic membranes, a delay in bone regeneration was observed mainly in animals not subjected to hormone replacement therapy. We conclude that anionic membranes treated with alkaline solution for 48 and 96 h presented better results in terms of bone ingrowth.

In vivo effects of Buthus occitanus tunetanus and Androctonus australis garzoni scorpion venoms on pregnant and non-pregnant rats

Scorpion envenomation in pregnant victims has been scarcely studied. Accidents with venomous animals can induce serious injuries for both mothers and embryos. In the current work, the lethality of Buthus occitanus tunetanus (Bot) and Androctonus australis garzoni (Aag) venoms was assessed in pregnant and non-pregnant murine rat models. The median lethal dose (LD50) was determined following the Spearman-Karber method. Our results showed great similarities of envenomation symptoms between term-pregnant and nonmated rats. An unusual vaginal bleeding was also seen in pregnant rats envenomed with Bot and Aag venoms. Our findings suggest that gestation may increase the venoms toxicity in rats.(AU)

Morphological characterization of a human glioma cell l ine.

A human malignant continuous cell line, named NG97, was recently established in our laboratory. This cell line has been serially subcultured over 100 times in standard culture media presenting no sign of cell senescence. The NG97 cell line has a doubling time of about 24 h. Immunocytochemical analysis of glial markers demonstrated that cells are positive for glial fibrillary acidic protein (GFAP) and S-100 protein, and negative for vimentin. Under phase-contrast microscope, cultures of NG97 showed cells with variable morphological features, such as small rounded cells, fusiform cells (fibroblastic-like cells), and dendritic-like cells. However, at confluence just small rounded and fusiform cells can be observed. At scanning electron microscopy (SEM) small rounded cells showed heterogeneous microextentions, including blebs and filopodia. Dendritic-like cells were flat and presented extensive prolongations, making several contacts with small rounded cells, while fusiform cells presented their surfaces dominated by microvilli.We believe that the knowledge about NG97 cell line may be useful for a deeper understanding of biological and immunological characteristics of gliomas.

Selection of tannase-producing Aspergillus niger strains

The aim of this work was to select strains of Aspergillus niger for tannase production. Growth of colonies in plates with tannic acid-containing medium indicated their ability to synthesize tannase. Tannase activity was also measured in solid-state fermentation. A. niger 11T25A5 was the best tannase producer (67.5 U.g-1/72 hours of fermentation.

Efecto de la nutrición sobre el desarrollo vegetativo y reproductivo del agente de la malformación floral y vegetativa del mango / Effect of nutrition on the vegetative and reproductive grown of the agent of floral and vegetative malformation of mango tree

Los aspectos fisiológicos del agente causal (fusarium subglutinans) de la malformación floral y vegetativa del mango (mangifera indica l.) referentes a la optimización de pruebas de patogenecidad, estudios epidemiológicos y de control, permite obtener datos consistentes en ambientes controlados. Debido a que entre los aspectos fisiológicos, el factor nutricional se señala como uno de los más influyentes en el crecimiento vegetativo y reproductivo de los hongos, el presente trabajo tiene como objetivo, evaluar la influencia de seis diferentes medios de cultivo sólidos y líquidos (pda, avena, v-8, armstrong, czapeck y extracto de mango), en el crecimiento micelial, esporulación y peso seco de dos aislamientos de f. subglutinas (iso-gv de la gema vegetativa e iso-gf de la gema floral). Los medios que favorecieron mayores pesos secos para las 2 cepas, fueron avena y v-8. En el medio de avena sólido, a pesar de haberse inducido un mayor crecimiento vegetativo para iso-gv y buen crecimiento para iso-gf, presentó baja esporulación en ambas cepas. El medio armstrong, sólido y líquido, favoreció la mayor esporulación. El efecto de diferentes combinaciones de fuentes de carbono (amida, fructosa, maltosa, sacarosa y nitrógeno (asparagina, peptona, nitrato de potasio, nitrato de sodio) sobre el comportamiento de fitopatógeno, demostró de manera general, que ambas cepas crecieron bien vegetativamente en todas las combinaciones amida-nitrato de sodio. En cuanto a la esporulación, fructosa - asparagina fue la que más estimuló la producción de conidios para gv, mientras que para gf, fue maltosa - nitrato de sodio

Acesso a ambulatório pediátrico de um hospital universitário / Evaluation of access to the pediatric service of a university hospital

No Brasil, verifica-se um descompasso entre o aumento das necessidades de atençäo á saúde e de sua oferta. O Sistema Unico de Saúde, cujas diretrizes preconizam a atençäol universal e eqüânime, determina a relavância desta temática dentro do campo da avaliaçäo dos serviços de saúde. Assim, foram estudados dois ambulatórios de pediatria de um hospital universitário, um geral e outro de uma subespecialidade (pneumologia), comparando os usuários quanto ao acesso. Foram aplicados 221 questionários entre clientes de ambos os ambulatórios de pediatria com o objetivo de se estudar e comparar variáveis socioeconômicas, procedência, acesso aos referidos ambulatórios e outros serviços de saúde.Evidenciou-se grande dificuldade de locomoçäo dos pacientes, a maioria dos quais säo encaminhados por serviços de saúde locais. Dos pacientes atendidos, 40 por cento näo receberam nenhum atendimento anteriormente à sua chegada ao hospital, fato que decorre principalmente de seu baixo nível socioeconômico, que os torna dependentes exclusivamente dos serviços públicos de saúde. A comparaçäo entre os dois ambulatórios mostra que os pacientes do ambulatório de especialidade têm melhor nível socioeconômico e säo menos dependentes dos serviços públicos, configurando desta forma uma situaçäo de ineqüidade. É apontado o melhor nível socioeconômico dos usuários da especialidade bem como questöes organizacionais do próprio serviço como os responsáveis pela iniqüidade verificada

Biocontrole pós-colheita da podridäo de Lasiodiplodia em frutos de manga por leveduras saprofíticas / Postharvest biocontrol of Lasiodiploidia rot of mango fruits by saprophytic yeasts

Among eigtheen yeast isolates studied as potential agents for biocontrol of stem-end rot on mango fruits caused by Lasiodiplodia theobromae, five were selected for testing in relation to three pathogen concentrations (1x10ü, 1x10 elevada à quarta and 1x10 elevada à quinta conidia/ml). The isolate LM-5 (Candida maritima) showed high control level at all pathogen concentrations. When tested against five pathogen isolates, LM-5 again demonstrated high control efficiency, and at in vitro test, highlyinhibited L.theobromae spore germination. The isolate LM-5 demonstrated great potential as agent for postharvest biocontrol of mango rot