Abstract
Objectives
We present the results of our experience in the diagnosis and follow up of the positive cases for propionic, methylmalonic acidemias and cobalamin deficiencies (PA/MMA/MMAHC) ...since the Expanded Newborn Screening was implemented in Madrid Region.
Methods
Dried blood samples were collected 48 h after birth. Amino acids and acylcarnitines were quantitated by MS/MS. Newborns with alterations were referred to the clinical centers for follow-up. Biochemical and molecular genetic studies for confirmation of a disease were performed.
Results
In the period 2011–2020, 588,793 children were screened, being 953 of them were referred to clinical units for abnormal result (192 for elevated C3 levels). Among them, 88 were false positive cases, 85 maternal vitamin B12 deficiencies and 19 were confirmed to suffer an IEM (8 PA, 4 MMA, 7 MMAHC). Ten out 19 cases displayed symptoms before the NBS results (6 PA, 1 MMA, 3 MMAHC). C3, C16:1OH+C17 levels and C3/C2 and C3/Met ratios were higher in newborns with PA/MMA/MMAHC. Cases diagnosed with B12 deficiency had mean B12 levels of 187.6 ± 76.9 pg/mL and their mothers 213.7 ± 95.0; 5% of the mothers were vegetarian or had poor eating while 15% were diagnosed of pernicious anemia. Newborns and their mothers received treatment with B12 with different posology, normalizing their levels and the secondary alterations disappeared.
Conclusions
Elevated C3 are a frequent cause for abnormal result in newborn screening with a high rate of false positive cases. Presymptomatic diagnosis of most of PA and some MMA/MMAHC is difficult. Vitamin B12 deficiency secondary to maternal deprivation is frequent with an heterogenous clinical and biochemical spectrum.
We present the results of our experience in the diagnosis of inborn errors of metabolism (IEM) since the Expanded Newborn Screening was implemented in our Region. Dried blood samples were collected ...48 h after birth. Amino acids and acylcarnitines were quantitated by mass spectrometry (MS)/MS. Newborns with alterations were referred to the clinical centers for follow‐up. Biochemical and molecular genetic studies for confirmation of a disease were performed. In the period 2011 to 2019, 592 822 children were screened: 902 of them were referred for abnormal results. An IEM was confirmed in 222 (1/2670): aminoacidopathies: 89 hyperphenylalaninemia (HPA) (51 benign HPA, 32 phenylketonuria, 4 DNAJC12 defect, and 2 primapterinuria), 6 hypermethioninemia, 3 tyrosinemia type 1 (TYR‐1), 1 TYR‐3, 4 maple syrup urine disease (MSUD), 2 branched‐chain amino acid transferase 2 deficiency, 2 homocystinuria, 1 cystinuria, 2 ornithine transcarbamylase (OTC) deficiency, 2 citrullinemia type I (CTLN1); FAO defects: 43 medium‐chain acyl‐CoA dehydrogenase deficiency (MCADD), 13 very long‐chain acyl‐CoA dehydrogenase deficiency, 2 long‐chain 3‐hydroxyacyl‐CoA dehydrogenase deficiency (LCHADD), 1 multiple acyl‐coA dehydrogenation deficiency, 11 systemic primary carnitine deficiency, 2 carnitine palmitoyltransferase type 2 (CPT‐II) deficiency, 1 CPT‐I deficiency; organic acidurias: 12 glutaric aciduria type 1 (GA‐1), 4 methylmalonic acidemia (MMA), 7 MMA including combined cases with homocystinuria (MMAHC), 6 propionic acidemia (PA), 7 3‐methylcrotonyl‐CoA carboxylase, 1 3‐hydroxy‐3‐methylglutaryl‐CoA lyase deficiency lyase deficiency. Only 19 infants (8.5%) were symptomatic at newborn screening result (1 LCHADD, 5 PA, 1 CPT‐II deficiency, 1 MMA, 3 MMAHC, 2 MSUD, 2 OTC deficiency, 1 CTLN1, 1 MCADD, 2 TYR‐1). No false negative cases were identified. Genetic diagnosis was conclusive in all biochemically confirmed cases, except for two infants with HPA, identifying pathogenic variants in 32 different genes. The conditions with the highest incidence were HPA (1/6661) and MCAD deficiencies (1/13 787).
Newborn Screening Programs (NSP) in Spain were born in the
city of Granada in 1968. Till the 1980s, they were developed around
the so-called “National Plan for Preventing Subnormality”, covering
up ...to 30% of the Spanish newborns. From 1982, when the health
system management was transferred to the different autonomous regions,
the NSP began to expand, and the bases to transform them into
an organized and multidisciplinary activity, integrated and coordinated
from the National Health System were settled. Despite this expansion,
it is not until the 1990s when their coverage reaches almost
100% newborns in Spain.
NSP grew up asymmetrically across the different autonomous
regions. In 2005 and 2006 the scientific societies SEQC (Spanish
Society of Clinical Chemistry) and AECNE (Spanish Society of
Newborn Screening), coordinated by the Health Promotion Area of
the General Directorate of Public Health, gathered together the necessary
information to elaborate a report on the NSP in Spain addressed
to the Interterritorial Council of the National Health System. In July
2013, that Council approved the seven diseases that should be part of
each region newborn screening panel, being the first step towards the
NSP harmonization in Spain. Currently, the NSP include between 8
and 29 diseases in their panels, thus more still more efforts are needed
in order to achieve a higher uniformity.
Los Programas de Cribado Neonatal (PCN) nacen en España en
Granada en el año 1968. Posteriormente, y hasta los años 80, se fueron
desarrollando en torno al llamado “Plan Nacional de Prevención
de la Subnormalidad” con una cobertura cercana al 30% de los recién
nacidos españoles. A partir de 1982, con el inicio de la gestión de
la sanidad a las comunidades autónomas (CCAA), los PCN se expandieron
y se comenzaron a sentar las bases para que éstos se convirtieran
en una actividad organizada y multidisciplinar, integrados y
coordinados desde el Sistema de Salud. A pesar de dicha expansión
no es hasta el inicio de la década de los 90 cuando se consigue una
cobertura próxima al 100% de los RN en España.
Los PCN fueron creciendo de forma muy asimétrica en las diferentes
CCAA y en los años 2005 y 2006 las Sociedades Científicas
SEQC (Sociedad Española de Química Clínica) y AECNE
(Asociación Española de Cribado Neonatal), con la coordinación
del Área de Promoción de la Salud de la Dirección General de Salud
Pública, recopilaron la información y elaboraron un informe, sobre
los PCN en España para el Consejo Interterritorial del sistema
Nacional de Salud (CISNS). En julio de 2013 este Consejo aprobó
las siete enfermedades que debían formar parte del panel de detección
de los PCN territoriales, primer paso hacia la armonización de
estos programas. Actualmente, los PCN incluyen entre 8 y 29 enfermedades
por lo que es necesario seguir trabajando para conseguir una
mayor uniformidad.
Abstract CD8-positive γδ T lymphocytes (GDCD8+ ) are specifically increased in peripheral blood of Behçet’s disease (BD) patients. GDCD8+ have shown a T regulatory (Treg ) function in autoimmune ...experimental models, human tumor infiltrates and intestinal intraepithelial lymphocytes from celiac patients. The aim of this study was to evaluate the Treg function of GDCD8+ and GDCD8− , freshly isolated from peripheral blood, in comparison to CD4+ CD25high naturally occurring Treg cells (nTreg ) in BD and healthy controls (HC). We tested their suppressive activity on CD4+ CD25− T effector cells (Teff ) proliferation by a CFSE dilution protocol, after suboptimal activation with anti-CD3, in the absence or presence of IL-2. Furthermore, secreted cytokines and suppressive latency associated peptide (LAP)-TGFβ surface upregulation were determined after GD activation. We found that Vδ1 chains contribution to GDCD8+ was higher in BD than in HC, but neither GDCD8+ nor GDCD8− ; (i) suppressed Teff proliferation, (ii) expressed LAP-TGFβ (iii) nor secreted IL-10, in either group. Moreover, GD presented a proinflammatory cytokine profile, mainly producing IFNγ and TNFα, in contrast to nTregs. In conclusion, peripheral GD could contribute more to the dysregulation of TH1 type of cytokines than to exerting a Treg function in BD.
CD8-positive gamma delta T lymphocytes (GDCD8+) are specifically increased in peripheral blood of Behcet's disease (BD) patients. GDCD8+ have shown a T regulatory (Treg) function in autoimmune ...experimental models, human tumor infiltrates and intestinal intraepithelial lymphocytes from celiac patients. The aim of this study was to evaluate the Treg function of GDCD8+ and GDCD8-, freshly isolated from peripheral blood, in comparison to CD4+CD25high naturally occurring Treg cells (nTreg) in BD and healthy controls (HC). We tested their suppressive activity on CD4+CD25- T effector cells (Teff) proliferation by a CFSE dilution protocol, after suboptimal activation with anti-CD3, in the absence or presence of IL-2. Furthermore, secreted cytokines and suppressive latency associated peptide (LAP)-TGF beta surface upregulation were determined after GD activation. We found that V delta 1 chains contribution to GDCD8+ was higher in BD than in HC, but neither GDCD8+ nor GDCD8-; (i) suppressed Teff proliferation, (ii) expressed LAP-TGF beta (iii) nor secreted IL-10, in either group. Moreover, GD presented a proinflammatory cytokine profile, mainly producing IFN gamma and TNF alpha , in contrast to nTregs. In conclusion, peripheral GD could contribute more to the dysregulation of TH1 type of cytokines than to exerting a Treg function in BD.
Tandem mass spectrometry (MS/MS) is being used for newborn screening since this laboratory testing technology increases the number of metabolic disorders that can be detected from dried blood-spot ...specimens. In the Community of Madrid, it was implemented in March 2011 and it includes 13 aminoacidopathies, fatty acid oxidation disorders and organic acidemias. The aim of this study was to describe our experience and evaluate the screening positive cases in a period of 9 years (2011-2019).
During the period of the study, a total of 592.822 neonates were screened with this expanded program by MS/MS in the Community of Madrid. Amino acids, acylcarnitines, and succinylacetone were quantified in all samples that met the quality criteria. Means, medians, percentiles and standard deviation of the analytes and ratios of interest were calculated.
901 patients (0,15 %) with a positive screening test were referred to clinical evaluation. 230 patients were diagnosed of 30 different inborn errors of metabolism (prevalence 1:2577), 11 of which were not included as a target in the Community of Madrid newborn screening program. The global positive predictive value was 25,6 %. During this period of time, two false negative cases were detected. The most prevalent disorders were phenylketonuria/hyperphenylalaninemia and medium chain acyl-CoA dehydrogenase deficiency (1:6444 and 1:13174 respectively). 93 % of the patients were detected in the presymptomatic stage.
During the last 9 years a large number of cases of IEM have been detected with an acceptable global positive predictive value. These results confirm the utility of inborn errors of metabolism newborn screening as a public health program.
Newborn Screening Programs (NSP) in Spain were born in the city of Granada in 1968. Till the 1980s, they were developed around the so-called "National Plan for Preventing Subnormality", covering up ...to 30% of the Spanish newborns. From 1982, when the health system management was transferred to the different autonomous regions, the NSP began to expand, and the bases to transform them into an organized and multidisciplinary activity, integrated and coordinated from the National Health System were settled. Despite this expansion, it is not until the 1990s when their coverage reaches almost 100% newborns in Spain. NSP grew up asymmetrically across the different autonomous regions. In 2005 and 2006 the scientific societies SEQC (Spanish Society of Clinical Chemistry) and AECNE (Spanish Society of Newborn Screening), coordinated by the Health Promotion Area of the General Directorate of Public Health, gathered together the necessary information to elaborate a report on the NSP in Spain addressed to the Interterritorial Council of the National Health System. In July 2013, that Council approved the seven diseases that should be part of each region newborn screening panel, being the first step towards the NSP harmonization in Spain. Currently, the NSP include between 8 and 29 diseases in their panels, thus more still more efforts are needed in order to achieve a higher uniformity.
