Patients with coronavirus disease‐2019 may be discharged based on clinical resolution of symptoms, and evidence for viral RNA clearance from the upper respiratory tract. Understanding the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) viral clearance profile is crucial to establish a re‐testing plan on discharge and ending isolation of patients. We aimed to evaluate the number of days that a patient needed to achieve undetectable levels of SARS‐CoV‐2 in upper respiratory tract specimens (nasopharyngeal swab and/or an oropharyngeal swab). The clearance and persistence of viral RNA was evaluated in two groups of positive patients: those who achieved two negative reverse transcription‐polymerase chain reaction (RT‐PCR) tests and those who kept testing positive. Patients were organized thereafter in two subgroups, mild illness patients discharged home and inpatients who had moderate to severe illness. Results from RT‐PCR tests were then correlated with results from the evaluation of the immune response. The study evidenced that most patients tested positive for more than 2 weeks and that persistence of viral RNA is not necessarily associated with severe disease but may result from a weaker immune response instead.

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SummaryThe considerable clinical heterogeneity of patients with common variable immunodeficiency disorders (CVID) shares some similarity with bonemarrow failure disorders such as Diamond-Blackfan anaemia (DBA) and Shwachman-Diamond syndrome (SDS), now recognized as defects in ribosome biogenesis or ribosomopathies. The recognition of a patient with DBA who subsequently developed CVID lends support to our previous finding of a heterozygous mutation in the SBDS gene of SBDS in another CVID patient, suggesting that ribosome biogenesis defects are responsible for a subset of CVID. Genetic defects in the ribosomal translational machinery responsible for various bone marrow failure syndromes are recognized readily when they manifest in children, but diagnosing these in adults presenting with complex phenotypes and hypogammaglobulinaemia can be a challenge. In this perspective paper, we discuss our clinical experience in CVID patients with ribosomopathies, and review the immunological abnormalities in other conditions associated with ribosomal dysfunction. With genetic testing available for various bone marrow failure syndromes, our hypothesis that ribosomal abnormalities may be present in patients with CVID could be proved in future studies by testing for mutations in specific ribosomal genes. New knowledge might then be translated into novel therapeutic strategies for patients in this group of immunodeficiency disorders.

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We report the first RhD variant associated with a 3-bp in-frame duplication in the RHD gene, predicted to be located within the RhD protein transmembrane domain that might be expected to result in a weak-D-like phenotype, concordant with serologic findings.

A 12-year-old Spanish male was referred to our department to elucidate the etiology of a congenital macrocytic anemia (MCV 104 fL; Hb 9.0 g/dL) with dyserythropoiesis, needing blood transfusions during the first 4 months of life, and mild thrombocytopenia (73 × 109/L) with large platelets. At the age of 3 months, he presented: Hb 5.9 g/dL, reticulocytes 105 × 1012/L, peripheral blood smear with anisocytosis, poikilocytosis, and basophilic stippling. He has a low PK activity (45 % of normal). The bone marrow was suggestive of congenital dyserythropoietic anemia (CDA), with erythroid hyperplasia and dyserythropoiesis and orthochromatic erythroblasts showing an irregular nuclear contour and bi-or multinucleated erythroblasts; electronic microscopy identified vacuoles in the cytoplasm of erythroblasts.CDAs are rare forms of bone marrow failure syndromes characterized by ineffective erythropoiesis [1,2]. There are three classical forms of CDA presentation due to mutations in CDANI, C15orf41, SEC23B, and KIF23 [3-5] and rarer forms associated with pathological variants in KLF1 and GATA1. GATA1 variants are associated with deregulation of erythroid and megakaryocytic lineages differentiation [6][7][8].

Glucose-6-phosphate isomerase (GPI) deficiency cause hereditary nonspherocytic hemolytic anemia (HNSHA) of variable severity in individuals homozygous or compound heterozygous for mutations in GPI gene. This work presents clinical features and genotypic results of two patients of Portuguese origin with GPI deficiency. The patients suffer from a mild hemolytic anemia (Hb levels ranging from 10 to 12.7g/mL) associated with macrocytosis, reticulocytosis, hyperbilirubinemia, hyperferritinemia and slight splenomegaly. Genomic DNA sequencing revealed in one patient homozygosity for a new missense mutation in exon 3, c.260G>C (p.Gly87Ala), and in the second patient compound heterozygosity for the same missense mutation (p.Gly87Ala), along with a frameshift mutation resulting from a single nucleotide deletion in exon 14, c.1238delA (p.Gln413Arg fs*24). Mutation p.Gln413Arg fs*24 is the first frameshift null mutation to be described in GPI deficiency. Molecular modeling suggests that the structural change induced by the p.Gly87Ala pathogenic variant has direct impact in the structural arrangement of the region close to the active site of the enzyme.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency, an X-linked disorder, is usually observed in hemizygote males and very rarely in females. The G6PD class 1 variants, very uncommon, are associated with chronic hemolytic anemia. Here we report a Portuguese woman who suffered in her sixties from a chronic hemolytic anemia due to G6PD deficiency. Molecular studies revealed heterozygosity for an in-frame 18-bp deletion, mapping to exon 10 leading to a deletion of 6 residues, 362-367 (LNERKA), which is a novel G6PD class 1 variant, G6PD Tondela. Two of her three daughters, asymptomatic, with G6PD activity within the normal range, are heterozygous for the same deletion. The patient's leukocyte and reticulocyte mRNA studies revealed an almost exclusive expression of the mutant allele, explaining the chronic hemolytic anemia. Patient whole blood genomic DNA HUMARA assay showed a balanced pattern of X chromosome inactivation (XCI), but granulocyte DNA showed extensive skewing, harboring the mutated allele, implying that in whole blood, lymphocyte DNA, with a very long lifetime, may cover up the current high XCI skewing. This observation indicates that HUMARA assay in women should be assessed in granulocytes and not in total leukocytes. © 2011 Elsevier Inc. All rights reserved. IntroductionX chromosome inactivation (XCI) is an epigenetic process, unique in mammals, by which one of the two X chromosomes in each cell is inactivated in females early in embryogenesis [1]. Therefore, women are a mosaic of paternal and maternal active X chromosome and a theoretical 1:1 ratio of two cell lines with inactive maternal to paternal X chromosome could be expected. The XCI ratio is usually assessed analyzing protein variants directly in heterozygous females' cells [2], transcribed mRNA expression [3] or DNA methylation status of polymorphic X-linked genes, such as the human androgen receptor (HUMARA) gene [4]. Deviation from the theoretical 1:1 ratio between the 2 parental alleles is called skewing. Several reports using the HUMARA assay show that the incidence of skewing is relatively low at birth and in adult non-hematopoietic tissues, but higher and agedependent in hematopoietic cells, with 30-40% of healthy elderly women reported to have skewing (greater than 75% expression of one parental X chromosome) [5][6][7][8][9][10][11][12]. T lymphocytes show less evidence of skewing with age than neutrophils, presumably reflecting the neutrophils' short half-life, whereas T lymphocytes are long-living cells, and in consequence, some T cells are produced near the time of study but others are derived from earlier stem cells [11][12][13]. This agerelated skewing in blood cells has been attributed to clonality as a consequence of hematopoietic stem cell senescence [5,7,9]. Swierczek et al. report a discrepancy between the skewed XCI observed in blood cells of 45 elderly women (ages 65-92 years; mean, 81.3 years) using the methylation-based HUMARA assay and a transcriptional based assay: with HUMARA assay in granulocyte DNA they found an ag...