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BACKGROUND: The incidence of systemic lupus in children with discoid lupus is unknown. OBJECTIVE: This study assessed the baseline characteristics of patients with pediatric discoid lupus erythematosus (pDLE). METHODS: Medical records at 17 sites were reviewed for pediatric dermatology and rheumatology patients with discoid lupus erythematosus. The inclusion criteria were clinical and/or histopathologic diagnosis of discoid lupus erythematosus with an age at onset of <18 years. Baseline data were collected at the first documented visit. Outcomes included diagnosis of systemic lupus erythematosus (SLE) at the baseline visit using the 1997 American College of Rheumatology (primary) and the 2012 Systemic Lupus International Collaborating Clinics (secondary) criteria. RESULTS: Of the >1500 charts reviewed, 438 patients met the inclusion criteria. The cohort was predominantly female (72%) and racially/ethnically diverse. A diagnosis of SLE at the baseline visit (pDLE + SLE) was rendered in 162 (37%) patients using the American College of Rheumatology and in 181 (41%) patients using the Systemic Lupus International Collaborating Clinics criteria. Patients with pDLE + SLE were older at the time of rash onset (median, 12.9 vs 8.9 years; P < .001), with shorter time from discoid lupus erythematosus onset to diagnosis, compared with patients with pDLE-only (median, 2 vs 7 months; P < .001). Patients with pDLE + SLE were more likely to be female (P = .004), with generalized discoid lupus erythematosus and clinically aggressive disease, including end-organ involvement, positive serologies, and higher- titer levels of antinuclear antibodies (P < .001). LIMITATIONS: Retrospective study. CONCLUSION: A diagnosis of discoid lupus erythematosus in adolescence should prompt thorough screening for SLE.
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Lupus Eritematoso Discoide , Lupus Eritematoso Sistémico , Adolescente , Niño , Estudios de Cohortes , Estudios Transversales , Femenino , Humanos , Lupus Eritematoso Discoide/diagnóstico , Lupus Eritematoso Discoide/epidemiología , Lupus Eritematoso Sistémico/epidemiología , Masculino , Estudios RetrospectivosRESUMEN
Identification of qualitative variants of von Willebrand disease (VWD) can be a diagnostic challenge because of discrepant results obtained in the multiple laboratory tests available for its appropriate classification. We report two cases of infrequent inherited variants of VWD with unclear preliminary results with the test panel available at the time of first consultation and that were finally diagnosed as a VWD type 2A/IID with a c.8318 G > C, p.Cys2773Ser mutation and a VWD type 2M with c.4225 T > G, p.Val1409Phe mutation, respectively. The description of these two cases highlights that despite the limited diagnostic panel for the evaluation of von Willebrand Factor (VWF) functionality, the multimeric analysis and genetic family studies were fundamental tools to achieve the final diagnosis.
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Enfermedades de von Willebrand/diagnóstico , Adulto , Femenino , Humanos , Persona de Mediana Edad , Adulto JovenRESUMEN
BACKGROUND: Epidermolysis bullosa (EB) is a group of rare and currently incurable genetic blistering disorders. As more pathogenic-driven therapies are being developed, there is an important need for EB-specific validated outcomes measures designed for use in clinical trials. OBJECTIVES: To test the reliability and construct validity of an instrument for scoring clinical outcomes of research for EB (iscorEB), a new combined clinician- and patient-reported outcomes tool. METHODS: We conducted an observational study consisting of independent 1-day assessments (six assessors) at two academic hospitals. The assessments consisted of iscorEB clinician (iscorEB-c), Birmingham Epidermolysis Bullosa Severity (BEBS) and global severity assessment for physicians; and iscorEB patient (iscorEB-p), Quality of Life evaluation in Epidermolysis Bullosa and Children's Dermatology Life Quality Index for patients. Construct validity and intraclass correlation coefficients (ICCs) for interobserver, intraobserver and test-retest reliability were calculated. RESULTS: Overall, 31 patients with a mean age of 19·5 years (1·8-45·2) were included. Disease severity was mild in 42% of cases, moderate in 29% and severe in 29%. The interobserver ICC was 0·96 for both the clinician-reported section of iscorEB-c and BEBS. The ICC for intraobserver reliability was 0·91 and 0·70 for the skin and mucosal domains of iscorEB-c, respectively. Cronbach's alpha for iscorEB-c was 0·89. The test-retest reliability of iscorEB-p was 0·97 and Cronbach's alpha was 0·84. The clinical score differentiated between subjects with mild, moderate and severe disease, and both clinical and patient subscores discriminated between recessive dystrophic EB and other EB subtypes. CONCLUSIONS: iscorEB has robust reliability and construct validity, including strong ability to distinguish EB types and severities. Further studies are planned to test its responsiveness to change.
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Epidermólisis Ampollosa/terapia , Índice de Severidad de la Enfermedad , Adolescente , Adulto , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Variaciones Dependientes del Observador , Evaluación de Resultado en la Atención de Salud , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Buschke-Ollendorff syndrome (BOS) is a rare, often benign, autosomal skin disorder. BOS commonly presents with nontender connective tissue naevi and sclerotic bony lesions (osteopoikilosis [OPK]). Herein, we summarize the presenting features of BOS and potential associations by conducting a systematic review of the literature and summarizing a cohort seen at the Hospital for Sick Children (HSC), Toronto, Canada. PubMed was searched using the following terms: 'BOS'; 'dermatofibrosis lenticularis'; 'OPK'; 'LEMD3'; 'elastoma'; 'collagenoma'. Only case reports were included, without date or language restrictions. Cases were further narrowed to those where patients or their families had a combination of skin and bony lesions, or a positive genetic test. Data were summarized using frequencies. In total, 594 reports were discovered, of which 546 (92%) were excluded. The remaining 48 accounted for 164 cases. Skin lesions were noted in 24% of cases and bony lesions in 20%, while 54% of patients had both. In 1% of cases the diagnosis was made on genetic testing alone. A family history was noted in 92% of cases. All patients with spinal stenosis (2%) or shortened status (7%) had OPK. Six per cent of patients had neurological problems. However, 50% of the cohort from HSC had cognitive delays, and only cases from 2007 onwards reported cognitive delays (the prevalence was 17% among those cases). This review confirms the classical diagnostic features of BOS. In addition, it highlights a previously unreported association between a shortened stature and OPK, as well as a possible association with cognitive delays.
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Osteopoiquilosis/diagnóstico , Enfermedades Cutáneas Genéticas/diagnóstico , Adolescente , Adulto , Distribución por Edad , Edad de Inicio , Anciano , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Osteopoiquilosis/complicaciones , Enfermedades Cutáneas Genéticas/complicaciones , Adulto JovenAsunto(s)
Vesícula , Piel , Administración Cutánea , Vesícula/diagnóstico , Vesícula/etiología , Niño , HumanosRESUMEN
In this study, we assessed the association between single-nucleotide polymorphisms (SNPs) in seven candidate genes involved in orchestrating the immune response against cytomegalovirus (CMV) and the 12-month incidence of CMV infection in 315 CMV-seropositive kidney transplant (KT) recipients. Patients were managed either by antiviral prophylaxis or preemptive therapy. CMV infection occurred in 140 patients (44.4%), including 13 episodes of disease. After adjusting for various clinical covariates, patients harboring T-allele genotypes of interleukin-28B (IL28B) (rs12979860) SNP had lower incidence of CMV infection (adjusted hazard ratio [aHR]: 0.66; 95% confidence interval [CI]: 0.46-0.96; p-value = 0.029). In the analysis restricted to patients not receiving prophylaxis, carriers of the TT genotype of toll-like receptor 9 (TLR9) (rs5743836) SNP had lower incidence of infection (aHR: 0.61; 95% CI: 0.38-0.96; p-value = 0.035), whereas the GG genotype of dendritic cell-specific ICAM 3-grabbing nonintegrin (DC-SIGN) (rs735240) SNP exerted the opposite effect (aHR: 1.86; 95% CI: 1.18-2.94; p-value = 0.008). An independent association was found between the number of unfavorable SNP genotypes carried by the patient and the incidence of CMV infection. In conclusion, specific SNPs in IL28B, TLR9 and DC-SIGN genes may play a role in modulating the susceptibility to CMV infection in CMV-seropositive KT recipients.
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Infecciones por Citomegalovirus/genética , Inmunidad Innata/genética , Fallo Renal Crónico/cirugía , Trasplante de Riñón , Polimorfismo de Nucleótido Simple , Adulto , Anciano , Alelos , Moléculas de Adhesión Celular/genética , Infecciones por Citomegalovirus/sangre , Femenino , Genotipo , Humanos , Incidencia , Interferones , Interleucinas/genética , Fallo Renal Crónico/sangre , Fallo Renal Crónico/genética , Lectinas Tipo C/genética , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Receptores de Superficie Celular/genética , Receptores de TrasplantesRESUMEN
Although 8% of reported FVIII gene (F8) mutations responsible for haemophilia A (HA) affect mRNA processing, very few have been fully characterized at the mRNA level and/or systematically predicted their biological consequences by in silico analysis. This study is aimed to elucidate the effect of potential splice site mutations (PSSM) on the F8 mRNA processing, investigate its correlation with disease severity, and assess their concordance with in silico predictions. We studied the F8 mRNA from 10 HA patient's leucocytes with PSSM by RT-PCR and compared the experimental results with those predicted in silico. The mRNA analysis could explain all the phenotypes observed and demonstrated exon skipping in six cases (c.222G>A, c.601+1delG, c.602-11T>G, c.671-3C>G, c.6115+9C>G and c.6116-1G>A) and activation of cryptic splicing sites, both donor (c.1009+1G>A and c.1009+3A>C) and acceptor sites (c.266-3delC and c.5587-1G>A). In contrast, the in silico analysis was able to predict the score variation of most of the affected splice site, but the precise mechanism could only be correctly determined in two of the 10 mutations analysed. In addition, we have detected aberrant F8 transcripts, even in healthy controls, so this must be taken into account as they could mask the actual contribution of some PSSM. We conclude that F8 mRNA analysis using leucocytes still constitutes an excellent approach to investigate the transcriptional effects of the PSSM in HA, whereas prediction in silico is not always reliable for diagnostic decision-making.
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Factor VIII/genética , Hemofilia A/diagnóstico , Hemofilia A/genética , Leucocitos/metabolismo , Mutación , Empalme del ARN , ARN Mensajero , Biología Computacional/métodos , Análisis Mutacional de ADN , Exones , Humanos , Intrones , Fenotipo , Sitios de Empalme de ARNAsunto(s)
Genotipo , Mosaicismo , Neurofibromatosis 1/genética , Neurofibromina 1/genética , Fenotipo , Adolescente , Adulto , Biopsia , Niño , Femenino , Estudios de Asociación Genética , Pruebas Genéticas , Humanos , Masculino , Neurofibromatosis 1/diagnóstico , Neurofibromatosis 1/patología , Piel/patología , Adulto JovenAsunto(s)
Manchas Café con Leche/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Proteínas de la Membrana/genética , Mosaicismo , Eliminación de Secuencia/genética , Proteínas Adaptadoras Transductoras de Señales , Adolescente , Manchas Café con Leche/diagnóstico , Diagnóstico Diferencial , Heterocigoto , Humanos , Masculino , Neurofibromatosis 1/diagnóstico , Neurofibromatosis 1/genéticaAsunto(s)
Antagonistas Adrenérgicos beta/administración & dosificación , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Hemangioma/tratamiento farmacológico , Nadolol/administración & dosificación , Propranolol/administración & dosificación , Administración Oral , Esquema de Medicación , Estudios de Factibilidad , Femenino , Humanos , Lactante , Masculino , Resultado del TratamientoRESUMEN
Essentials The Royal disease (RD) is a form of hemophilia B predicted to be caused by a splicing mutation. We generated an iPSC-based model of the disease allowing mechanistic studies at the RNA level. F9 mRNA analysis in iPSC-derived hepatocyte-like cells showed the predicted abnormal splicing. Mutated F9 mRNA level was very low but we also found traces of wild type transcripts. SUMMARY: Background The royal disease is a form of hemophilia B (HB) that affected many descendants of Queen Victoria in the 19th and 20th centuries. It was found to be caused by the mutation F9 c.278-3A>G. Objective To generate a physiological cell model of the disease and to study F9 expression at the RNA level. Methods Using fibroblasts from skin biopsies of a previously identified hemophilic patient bearing the F9 c.278-3A>G mutation and his mother, we generated induced pluripotent stem cells (iPSCs). Both the patient's and mother's iPSCs were differentiated into hepatocyte-like cells (HLCs) and their F9 mRNA was analyzed using next-generation sequencing (NGS). Results and Conclusion We demonstrated the previously predicted aberrant splicing of the F9 transcript as a result of an intronic nucleotide substitution leading to a frameshift and the generation of a premature termination codon (PTC). The F9 mRNA level in the patient's HLCs was significantly reduced compared with that of his mother, suggesting that mutated transcripts undergo nonsense-mediated decay (NMD), a cellular mechanism that degrades PTC-containing mRNAs. We also detected small proportions of correctly spliced transcripts in the patient's HLCs, which, combined with genetic variability in splicing and NMD machineries, could partially explain some clinical variability among affected members of the European royal families who had lifespans above the average. This work allowed the demonstration of the pathologic consequences of an intronic mutation in the F9 gene and represents the first bona fide cellular model of HB allowing the study of rare mutations at the RNA level.
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Factor IX/genética , Hemofilia B/genética , Hepatocitos/metabolismo , Células Madre Pluripotentes Inducidas/metabolismo , Mutación , ARN Mensajero/genética , Adolescente , Empalme Alternativo , Diferenciación Celular , Línea Celular , Factor IX/metabolismo , Femenino , Predisposición Genética a la Enfermedad , Hemofilia B/sangre , Hemofilia B/diagnóstico , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Fenotipo , ARN Mensajero/metabolismo , Análisis de Secuencia de ARNRESUMEN
OBJECTIVE: Our goal was to conduct the item generation and piloting phases of a new discriminative and evaluative tool for pediatric post-thrombotic syndrome. METHODS: We followed a formative model for the development of the tool, focusing on the signs/symptoms (items) that define post-thrombotic syndrome. For item generation, pediatric thrombosis experts and subjects diagnosed with extremity post-thrombotic syndrome during childhood nominated items. In the piloting phase, items were cross-sectionally measured in children with limb deep vein thrombosis to examine item performance. RESULT: Twenty-three experts and 16 subjects listed 34 items, which were then measured in 140 subjects with previous diagnosis of limb deep vein thrombosis (70 upper extremity and 70 lower extremity). The items with strongest correlation with post-thrombotic syndrome severity and largest area under the curve were pain (in older children), paresthesia, and swollen limb for the upper extremity group, and pain (in older children), tired limb, heaviness, tightness and paresthesia for the lower extremity group. CONCLUSION: The diagnostic properties of the items and their correlations with post-thrombotic syndrome severity varied according to the assessed venous territory. The information gathered in this study will help experts decide which item should be considered for inclusion in the new tool.
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Extremidad Inferior/irrigación sanguínea , Síndrome Postrombótico/diagnóstico , Extremidad Superior/irrigación sanguínea , Niño , Preescolar , Femenino , Humanos , Extremidad Inferior/patología , Masculino , Síndrome Postrombótico/etiología , Extremidad Superior/patología , Trombosis de la Vena/complicacionesRESUMEN
Root elongation, hematoxylin staining, and changes in the ultrastructure of root-tip cells of an Al-tolerant maize variety (Zea mays L. C 525 M) exposed to nutrient solutions with 20 &mgr;M Al (2.1 &mgr;M Al3+ activity) for 0, 4, and 24 h were investigated in relation to the subcellular distribution of Al using scanning transmission electron microscopy and energy-dispersive x-ray microanalysis on samples fixed by different methods. Inhibition of root-elongation rates, hematoxylin staining, cell wall thickening, and disturbance of the distribution of pyroantimoniate-stainable cations, mainly Ca, was observed only after 4 and not after 24 h of exposure to Al. The occurrence of these transient, toxic Al effects on root elongation and in cell walls was accompanied by the presence of solid Al-P deposits in the walls. Whereas no Al was detectable in cell walls after 24 h, an increase of vacuolar Al was observed after 4 h of exposure. After 24 h, a higher amount of electron-dense deposits containing Al and P or Si was observed in the vacuoles. These results indicate that in this tropical maize variety, tolerance mechanisms that cause a change in apoplastic Al must be active. Our data support the hypothesis that in Al-tolerant plants, Al can rapidly cross the plasma membrane; these data clearly contradict the former conclusions that Al mainly accumulates in the apoplast and enters the symplast only after severe cell damage has occurred.
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The present study was undertaken to determine the effects of clindamycin and tetracycline, both intravenously administered, on antibody response to thymus-dependent antigen (PC-KLH) in BALB/c mice. The immunological parameters evaluated were: DPFC/spleen (direct plaque forming-cells), antibody secretion median rate (PC50), heterogeneity index (Hi), number of total splenic lymphocytes and cellular viability. The results showed that clindamycin (i.v.) increased the humoral response; 28 mg/kg was the dose that showed the greatest enhancement (+73%). The PC50 was not affected by clindamycin but Hi decreased at 28 mg/kg and increased at 2.8 mg/kg doses, although neither result was statistically significant. When tetracycline was given i.v., a slight decrease in the anti-PC DPFC number was observed. Although the PC50 was greater at 10 mg/kg (p less than 0.05), Hi was smaller at the 1 mg/kg dose (p less than 0.05).
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Adyuvantes Inmunológicos , Clindamicina/farmacología , Inmunosupresores , Tetraciclina/farmacología , Animales , Formación de Anticuerpos/efectos de los fármacos , Femenino , Hemocianinas/inmunología , Técnica de Placa Hemolítica , Linfocitos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos BALB C , Fosfatidilcolinas/farmacología , Bazo/citología , Bazo/efectos de los fármacosRESUMEN
Penicillin G, cefotaxime and clavulanic acid administered intravenously were studied for their immunomodulating properties. BALBC/c mice were immunized using PC-KLH as thymus-dependent antigen at the same time as the antibiotic was injected. The effect on antibody response was evaluated 5 days after immunization. Critical immunological parameters such as direct antibody-producing cells, Ab-secretion median rate, secretion rate heterogeneity and cellular viability, were studied. The most stimulatory effects were with penicillin G at the 4 x 10(6) IU/kg dose (+73%), cefotaxime at the 366 mg/kg dose (+80%) and clavulanic acid at 1 mg/kg (+218%). The experiments using inhibition of plaque formation with free hapten demonstrate that all the drugs studied decreased the antibody secretion rate and this parameter appeared more heterogeneous when cefotaxime and clavulanic acid were given; however, with penicillin this parameter was more heterogeneous at 2 x 10(5) mg/kg and 1 x 10(3) IU/kg respectively. When clavulanic acid was injected, the number of lymphocytes per spleen, size and friability of the spleen were increased versus the control mice. The present data show that all of the drugs studied present immunostimulating effects on humoral response, against thymus-dependent antigen, but have differently pronounced enhancements.
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Antibacterianos/farmacología , Animales , Células Productoras de Anticuerpos/efectos de los fármacos , Cefotaxima/farmacología , Ácidos Clavulánicos/farmacología , Femenino , Técnica de Placa Hemolítica , Linfocitos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos BALB C , Penicilina G/farmacología , Fosfatidilcolinas/farmacología , Bazo/citologíaRESUMEN
BACKGROUND: In von Willebrand factor (VWF) the effect of mutations potentially affecting mRNA processing or splicing is less predictable than that of other mutations (e.g. nonsense or missense substitutions). Bioinformatic tools can provide a valuable means to determine the consequences of potential splice site mutations (PSSM), but functional studies are mandatory to elucidate the true effect of the variation detected. OBJECTIVES, PATIENTS AND METHODS: After identification of PSSM in VWD patients, we began a systematic study of their in vivo effect in RNA extracted from the patients' platelets and leukocytes. RESULTS AND CONCLUSIONS: Thirteen pairs of primers were designed for full amplification of VWF mRNA by RT-PCR that, after sequencing of aberrant products, enabled elucidation of the PSSM consequences for mRNA processing. This procedure was used to study seven different PSSM identified in four patients demonstrating diverse molecular mechanisms such as exon skipping (c.533-2A>G and c.8155+3G>C) and the activation of a cryptic splice site (c.7730-1G>C). No visible effect was evident for c.1533+15G>A and c.5170+10C>T and the consequence of c.[546G>A;7082-2A>G] was hidden by nonsense-mediated mRNA decay (NMD). Results were compared with in silico predictions of four splice-site analysis tools. We demonstrate selective degradation of VWF mRNA bearing PSSM by NMD for several mutations, which suggests that NMD represents a general mechanism for truncating mutations in VWF. Furthermore, because NMD efficiency varies between cell types, use of RNA from both platelets and leukocytes for in vivo study of VWF PSSM offers complementary results, particularly in cases in which NMD occurs in the allele carrying the mutation.
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Plaquetas/metabolismo , Leucocitos/metabolismo , Mutación , Empalme del ARN , Factor de von Willebrand/genética , Femenino , Humanos , Masculino , Linaje , ARN Mensajero/genéticaRESUMEN
Psoriasis is a common skin condition seen in pediatrics. Treatment modalities used to treat psoriasis in children are different from those prevailing in the adult population and require adequate testing in pediatric subjects. This article reviews the published evidence on the different treatment modalities for pediatric psoriasis over the past 5 years.