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Newborn screening (NBS) for cystic fibrosis (CF) based on pancreatitis-associated protein (PAP) has been performed for several years. While some influencing factors are known, there is currently a lack of information on the influence of seasonal temperature on PAP determination or on the course of PAP blood concentration in infants during the first year of life. Using data from two PAP studies at the Heidelberg NBS centre and storage experiments, we compared PAP determinations in summer and winter and determined the direct influence of temperature. In addition, PAP concentrations measured in CF-NBS, between days 21–35 and 36–365, were compared. Over a 7-year period, we found no significant differences between PAP concentrations determined in summer or winter. We also found no differences in PAP determination after 8 days of storage at 4 °C, room temperature or 37 °C. When stored for up to 3 months, PAP samples remained stable at 4 °C, but not at room temperature (p = 0.007). After birth, PAP in neonatal blood showed a significant increasing trend up to the 96th hour of life (p < 0.0001). During the first year of life, blood PAP concentrations continued to increase in both CF- (36–72 h vs. 36–365 d p < 0.0001) and non-CF infants (36–72 h vs. 36–365 d p < 0.0001). Seasonal effects in central Europe appear to have a limited impact on PAP determination. The impact of the increase in blood PAP during the critical period for CF-NBS and beyond on the applicability and performance of PAP-based CF-NBS algorithms needs to be re-discussed.

Early diagnosis of severe combined immunodeficiency (SCID), spinal muscular atrophy (SMA), and sickle cell disease (SCD) improves health outcomes by providing a specific treatment before the onset of symptoms. A high-throughput nucleic acid-based method in newborn screening (NBS) has been shown to be fast and cost-effective in the early detection of these diseases. Screening for SCD has been included in Germany's NBS Program since Fall 2021 and typically requires high-throughput NBS laboratories to adopt analytical platforms that are demanding in terms of instrumentation and personnel. Thus, we developed a combined approach applying a multiplexed quantitative real-time PCR (qPCR) assay for simultaneous SCID, SMA, and 1st-tier SCD screening, followed by a tandem mass spectrometry (MS/MS) assay for 2nd-tier SCD screening. DNA is extracted from a 3.2-mm dried blood spot from which we simultaneously quantify T-cell receptor excision circles for SCID screening, identify the homozygous SMN1 exon 7 deletion for SMA screening, and determine the integrity of the DNA extraction through the quantification of a housekeeping gene. In our two-tier SCD screening strategy, our multiplex qPCR identifies samples carrying the HBB: c.20A>T allele that is coding for sickle cell hemoglobin (HbS). Subsequently, the 2nd tier MS/MS assay is used to distinguish heterozygous HbS/A carriers from samples of patients with homozygous or compound heterozygous SCD. Between July 2021 and March 2022, 96,015 samples were screened by applying the newly implemented assay. The screening revealed two positive SCID cases, while 14 newborns with SMA were detected. Concurrently, the qPCR assay registered HbS in 431 samples which were submitted to 2nd-tier SCD screening, resulting in 17 HbS/S, five HbS/C, and two HbS/β thalassemia patients. The results of our quadruplex qPCR assay demonstrate a cost-effective and fast approach for a combined screening of three diseases that benefit from nucleic-acid based methods in high-throughput NBS laboratories.

Newborn screening (NBS) programs are effective measures of secondary prevention and have been successively extended. We aimed to evaluate NBS for methylmalonic acidurias, propionic acidemia, homocystinuria, remethylation disorders and neonatal vitamin B12 deficiency, and report on the identification of cofactor-responsive disease variants. This evaluation of the previously established combined multiple-tier NBS algorithm is part of the prospective pilot study “NGS2025” from August 2016 to September 2022. In 548,707 newborns, the combined algorithm was applied and led to positive NBS results in 458 of them. Overall, 166 newborns (prevalence 1: 3305) were confirmed (positive predictive value: 0.36); specifically, methylmalonic acidurias (N = 5), propionic acidemia (N = 4), remethylation disorders (N = 4), cystathionine beta-synthase (CBS) deficiency (N = 1) and neonatal vitamin B12 deficiency (N = 153). The majority of the identified newborns were asymptomatic at the time of the first NBS report (total: 161/166, inherited metabolic diseases: 9/14, vitamin B12 deficiency: 153/153). Three individuals were cofactor-responsive (methylmalonic acidurias: 2, CBS deficiency: 1), and could be treated by vitamin B12, vitamin B6 respectively, only. In conclusion, the combined NBS algorithm is technically feasible, allows the identification of attenuated and severe disease courses and can be considered to be evaluated for inclusion in national NBS panels.

Community urban gardens are a type of urban agriculture. They provide spaces for social interaction and increase in life quality of maintainers and consumers. This study implements a community garden in Vila Buritis II, Planaltina, Brasília, Federal District, Brazil. The population of Planaltina has low purchasing power and high indices of social vulnerability. The implementation of the vegetable garden resulted from a partnership between agroecology students from the Federal Institute of Brasília, the local community, and the government. The work relied on donations, reuse of materials, and volunteer labor. Educational actions in the neighborhood approached the relationship between production in the garden and food choices, with a focus on healthy eating, consumption of fresh food, and reduction of industrialized items. The building of the vegetable garden converted an idle area, with a history of accumulating garbage and rubble, into a productive space for social coexistence and leisure. Twenty-seven edible species and an undefined number of ornamental plants were planted. Between 30 and 40 residents visit the space regularly and collect food for their daily meals. However, the maintenance and increase of garden productivity still need technical assistance and resources.

Este artigo trata da construção de Livros de Artista com os meios estruturais da imagem lenticular, considerando aspectos formais e visuais, sequência e serialidade. São analisadas algumas possibilidades de produção com diferentes linguagens, passando pela desconstrução e ressignificação de paisagens capturadas por câmera digital. As imagens lenticulares, configuradas como livros de artista, propõem uma outra relação perceptiva e estética a partir do deslocamento do espectador diante da obra. O estudo busca aproximações conceituais nas obras de Gaspar Antoine Bois-Clair, Yaacov Agam e Carlos Vergara; o embasamento teórico nos autores Anne Cauquelin, Maria Celeste de Almeida Wanner, Annateresa Fabris, e Paulo Silveira, entre outros.

Isovaleric aciduria (IVA) is a rare disorder of leucine metabolism and part of newborn screening (NBS) programs worldwide. However, NBS for IVA is hampered by, first, the increased birth prevalence due to the identification of individuals with an attenuated disease variant (so-called “mild” IVA) and, second, an increasing number of false positive screening results due to the use of pivmecillinam contained in the medication. Recently, machine learning (ML) methods have been analyzed, analogous to new biomarkers or second-tier methods, in the context of NBS. In this study, we investigated the application of machine learning classification methods to improve IVA classification using an NBS data set containing 2,106,090 newborns screened in Heidelberg, Germany. Therefore, we propose to combine two methods, linear discriminant analysis, and ridge logistic regression as an additional step, a digital-tier, to traditional NBS. Our results show that this reduces the false positive rate by 69.9% from 103 to 31 while maintaining 100% sensitivity in cross-validation. The ML methods were able to classify mild and classic IVA from normal newborns solely based on the NBS data and revealed that besides isovalerylcarnitine (C5), the metabolite concentration of tryptophan (Trp) is important for improved classification. Overall, applying ML methods to improve the specificity of IVA could have a major impact on newborns, as it could reduce the newborns’ and families’ burden of false positives or over-treatment.

Vitamin B12 deficiency, mostly of maternal origin in newborns, is a well-treatable condition but can cause severe neurologic sequelae in infants. Early detection of vitamin B12 deficiency allows the pre-symptomatic treatment of affected children. This evaluation assesses the characteristics of maternal vitamin B12 deficiency detected by newborn screening. In a prospective single-center study, a systematic screening strategy for vitamin B12 deficiency using a combination of two second-tier strategies was applied. In addition to confirmatory diagnostics in children, the systematic work-up of vitamin B12 status was also performed for their mothers. Maternal characteristics were assessed including ethnic origin, diet, and vitamin supplementation during pregnancy. For affected mothers, a work-up by internal medicine was recommended. In total, 121 mother–infant couples were analyzed. 66% of mothers adhered to a balanced diet including meat. The cause of maternal vitamin B12 deficiency was unknown in 56% of cases, followed by dietary causes in 32%, and organic causes in 8%. All mothers following a vegan diet and most mothers with a vegetarian diet took vitamin preparations during pregnancy, whereas only 55.8% of mothers with a balanced diet took folic acid or other vitamins. Maternal vitamin B12, folic acid, and homocysteine levels were significantly correlated with the child’s folic acid levels, and with homocysteine, methylmalonic, and methylcitric acid levels in first and second NBS dried blood spots. Most children had normal blood counts and showed normocytosis. Although 36.7% of mothers showed anemia, only one presented with macrocytosis. Adherence to vitamin supplementation in pregnancy is low despite the recommendation for supplementation of folic acid. Ideally, the evaluation of mothers for vitamin B12 levels and appropriate therapy should be initiated in early pregnancy. In infants detected through newborn screening, the multidisciplinary assessment and therapy of both children and mothers should be performed.

O presente artigo pretende examinar a trajetória política e as táticas e estratégias empregadas pelo cacique Kaingang Orides Belino Correia da Silva, que, entre os anos 2000 a 2003, foi vice-prefeito e prefeito, por curto período, em Ipuaçu-SC. Oriundo de uma família de líderes indígenas e funcionário da Funai, Orides assumiu o cacicado da Terra Indígena Xapecó em 1999, implementando mudanças estruturais na sua comunidade. Em 2000, elegeu-se vice-prefeito e, em 2002, assumiu a Prefeitura de Ipuaçu, tornando-se o primeiro prefeito indígena da região Sul do Brasil. Investigando a atuação de Orides no circuito político-institucional, intenta-se observar os benefícios conquistados para sua comunidade a partir da sua atuação política. As fontes utilizadas neste artigo são publicações de jornais, com as quais se organizam cronologicamente os episódios das relações políticas do cacique Orides.

Endonucleolytic removal of 5’-leader sequences from tRNA precursor transcripts (pre-tRNAs) by ribonuclease P (RNase P) is essential for protein synthesis. Beyond RNA-based RNase P enzymes, protein-only versions of the enzyme exert this function in various eukarya (there termed PRORPs) and in some bacteria (Aquifex aeolicus and close relatives); both enzyme types belong to distinct subgroups of the PIN domain metallonuclease superfamily. Homologs of Aquifex RNase P (HARPs) are also expressed in some other bacteria and many archaea, where they coexist with RNA-based RNase P and do not represent the main RNase P activity. Here, we solved the structure of the bacterial HARP from Halorhodospira halophila by cryo-electron microscopy, revealing a novel screw-like dodecameric assembly. Biochemical experiments demonstrate that oligomerization is required for RNase P activity of HARPs. We propose that the tRNA substrate binds to an extended spike-helix (SH) domain that protrudes from the screw-like assembly to position the 5’-end in close proximity to the active site of the neighboring dimer. The structure suggests that eukaryotic PRORPs and prokaryotic HARPs recognize the same structural elements of pre-tRNAs (tRNA elbow region and cleavage site). Our analysis thus delivers the structural and mechanistic basis for pre-tRNA processing by the prokaryotic HARP system.

ABSTRACT Introduction: The involvement of the immune system in the process of neoplasms has been increasingly studied due to its potential in antitumor therapy. Objectives: To evaluate the expression of heat shock protein 27 (Hsp27) in tissue samples from invasive ductal carcinomas with and without the presence of axillary sentinel lymph node metastasis, as well as to study the relationship of cell proliferation rate (Ki-76) and apoptosis (caspase 3) compared with samples of fibrocystic change in breast tissue. Methods: Cross-sectional study with tissue samples collected at the Hospital São Lucas, between September 2001 and October 2009, divided into three groups: 1. fibrocystic breast change (control group); 2. invasive ductal carcinoma of the breast with presence of lymph node metastasis; and 3. absence of lymph node metastasis. Results: Increased Hsp27 expression was observed in the group of non-metastasis carcinomas when compared to the other groups. There was difference in Ki-67 protein expression and in the ratio between cell proliferation and apoptosis among the carcinoma groups compared to the control group. However, there was no significant difference between the carcinoma groups or between the expressions of caspase 3 in the three groups, when compared to each other. Conclusion: Quantitative analysis of Hsp27 demonstrated increased protein expression in primary neoplasm tissues and in lymph nodes in carcinomas without axillary sentinel lymph node metastasis. Although the methodology and the number of cases do not allow concluding that this behavior in metastatic disease is a constant finding, this justifies the need to evaluate this finding in future studies.

O presente artigo pretende examinar a trajetória política e as táticas e estratégias empregadas pelo cacique Kaingang Orides Belino Correia da Silva, que, entre os anos 2000 a 2003, foi vice-prefeito e prefeito, por curto período, em Ipuaçu-SC. Oriundo de uma família de líderes indígenas e funcionário da Funai, Orides assumiu o cacicado da Terra Indígena Xapecó em 1999, implementando mudanças estruturais na sua comunidade. Em 2000, elegeu-se vice-prefeito e, em 2002, assumiu a Prefeitura de Ipuaçu, tornando-se o primeiro prefeito indígena da região Sul do Brasil. Investigando a atuação de Orides no circuito político-institucional, intenta-se observar os benefícios conquistados para sua comunidade a partir da sua atuação política. As fontes utilizadas neste artigo são publicações de jornais, com as quais se organizam cronologicamente os episódios das relações políticas do cacique Orides.

Bebidas à base de Camellia sinensis tem sido utilizadas por muitos séculos na medicina tradicional, sendo a infusão preparada com as suas folhas. Um dos produtos à base de C. sinensis amplamente consumido é o chá verde, sendo que seus benefícios vem sendo associados a sua composição química. Por este motivo o objetivo deste trabalho foi avaliar a possível atividade genotóxica do extrato de chá verde através da avaliação de danos no DNA pelo Teste Cometa. Desta forma, foi conduzido um ensaio in vitro a partir de leucócitos isolados de sangue periférico de indivíduos saudáveis. Os leucócitos isolados foram tratados com diferentes concentrações de extrato de C. sinensis, posteriormente sendo avaliado o dano no DNA pelo Teste Cometa. Os resultados encontrados neste trabalho indicaram que nas concentrações 3μg/mL, 10μg/mL, 30μg/mL, 100μg/mL e 300μg/mL o extrato de C. sinensis comportou-se igual ao controle não apresentando diferença significativa no índice de danos do DNA, o que indica que nas concentrações utilizadas o extrato não apresentou efeito genotóxico. O uso do chá verde na dieta pode ter muitos benefícios para o organismo, principalmente pelos seus compostos bioativos e, por este motivo, fazem-se necessários estudos para avaliar sua toxicidade, para melhor avaliar a segurança em seu uso.

Uma das etapas imprescindíveis para a implantação de um Sistema de Gestão Ambiental (SGA) em acordo com a Norma ISO 14001 é o levantamento e a análise dos aspectos e impactos ambientais. Sendo assim, este trabalho tem como objetivo a identificação dos aspectos e impactos ambientais de uma Cooperativa de Energia Elétrica do Sul Catarinense, assim como a elaboração de uma matriz de avaliação riscos ambientais, a fim de possibilitar oportunidades de melhoria para uma futura implantação de um SGA. A metodologia utilizada foi de uma pesquisa descritiva e estudo de caso, tendo uma abordagem qualitativa. Os resultados evidenciaram que a empresa possui um total de 55 aspectos e impactos ambientais, sendo que 26 destes não possuem medida de controle. Neste sentido, foram propostas ações de melhoria às 26 ocorrências, juntamente com um plano de gerenciamento de resíduos sólidos e a reformulação do programa de educação ambiental da Cooperativa.

Este artigo apresenta e valida um modelo de mobilidade veicular urbano resultante da análise de traços reais de mobilidade de táxis da cidade de Roma (Itália). O modelo de mobilidade é aplicado em um simulador de tráfego, possibilitando-se gerar traços sintéticos de mobilidade para simuladores de redes. A principal vantagem de se gerar traços sintéticos é que, além de manter aspectos do padrão de mobilidade real, permite-se um maior controle das variáveis do ambiente de simulação. Apesar do modelo proposto não conseguir capturar todos os aspectos reais da mobilidade dos táxis, mostrou-se efetivo em reproduzir características observadas como relevantes na movimentação veicular urbana.

O presente trabalho apresenta um estudo sobre a estruturação de indicadores de desempenho para a medição da sustentabilidade de um a organização do segmento da construção civil pesada. O objetivo do estudo foi desenvolver uma estrutura de indicadores de sustentabilidade, que envolve-se as áreas de meio ambiente, segurança do trabalho, saúde ocupacional e gestão de talentos, a fim de demonstrar a maturidade da organização em tornar-se uma empresa referente em sustentabilidade no seu segmento. Os procedimentos metodológicos adotados foram preponderantemente da pesquisa bibliográfica e documental. Por meio deste estudo foi possível estruturar vinte e três novas métricas, com indicadores focados na sustentabilidade, parametrizados e implantados nas obras.

Introduction: It presents the development of a proposed conceptual model for digital educational games in the context of the classroom in which the analysis focused on learning for sustainable habits production and consumption in the residential environment. The existing conceptual models are more linked to the creation and recently with evaluation of the games. In the context of the implementation, most models focus more on the description of the games validation processes than in mediating the pedagogical practice itself. Therefore, the research question is about how to create, implement and evaluate digital educational games for teaching and learning process in the classroom, with the focus on learning the culture of sustainability in the residential environment. Methodology: As far as the methodological characterization is concerned, the investigation was supported by bibliographical and field research for data collection. It was emphasized the use of Design Science Research Methodology as methodological procedure for the development of the conceptual model. Results: The system has been tested in a pilot study in two groups of Young Apprentice Program at SENAC/SC to check its consistency. Conclusions: It was concluded based on the results produced by the application of the pilot that the proposed conceptual model contributes to the process of teaching and learning in the classroom.

O trabalho tem como objetivo identificar a percepção da opinião pública internacional diante das principais alternativas de resposta às mudanças climáticas: inovação tecnológica versus mudanças no estilo de vida individual; e os determinantes por trás dessa posição. A investigação utiliza dados em painel para quinze países, coletados em três unidades de tempo. Os resultados mostram um predomínio da escolha por soluções tecnológicas para resolver o problema, especialmente nos países emergentes, mas tal percepção vem enfrentando queda nos últimos anos, cedendo espaço para aceitação da opção por mudanças no comportamento individual. São as nações que possuem condições ambientais menos favoráveis, maior desenvolvimento humano e forte desenvolvimento institucional as que tendem a ver como menos efetivas as soluções tecnológicas para o problema das mudanças climáticas.

Introduction Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have become common in hospitals and the community environment, and this wide resistance has limited patient treatment. Clindamycin (CL) represents an important alternative therapy for infections caused by S. aureus. Antimicrobial susceptibility testing using standard methods may not detect inducible CL resistance. This study was performed to detect the phenotypes of resistance to macrolides-lincosamides-streptogramin B (MLSB) antibiotics, including CL, in clinical samples of S. aureus from patients at a tertiary hospital in Santa Maria, State of Rio Grande do Sul, Brazil. Methods One hundred and forty clinical isolates were submitted to the disk diffusion induction test (D-test) with an erythromycin (ER) disk positioned at a distance of 20mm from a CL disk. The results were interpreted according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI). Results In this study, 29 (20.7%) of the 140 S. aureus samples were resistant to methicillin (MRSA), and 111 (79.3%) were susceptible to methicillin (MSSA). The constitutive resistance phenotype (cMLSB) was observed in 20 (14.3%) MRSA samples and in 5 (3.6%) MSSA samples, whereas the inducible resistance phenotype (iMLSB) was observed in 3 (2.1%) MRSA samples and in 8 (5.8%) MSSA samples. Conclusions The D-test is essential for detecting the iMLSB phenotype because the early identification of this phenotype allows clinicians to choose an appropriate treatment for patients. Furthermore, this test is simple, easy to perform and inexpensive.

The polyphenol contents and antioxidant capacity of Brazilian red grape juices and wine vinegars were analyzed. Additionally, it was analyzed the human polyphenol absorption and acute effect in plasmatic oxidative metabolism biomarkers after juice ingestion. The organic Bordo grape juice (GBO) presented a higher level of trans-resveratrol, quercitin, rutin, gallic acid, caffeic acid and total flavonoids then other juices and vinegars as well as antioxidant capacity. The plasmatic polyphenol increased 27.2% after GBO juice ingestion. The results showed that juices and vinegars from Brazilian crops present similar chemical and functional properties described in studies performed in other countries.

Abstract: A new low-cost process for mid resolution FIR-Arrays for thermal imaging in the LWIR range (8- 14 μm) has been developed. Integrated array devices with 1k and 5k pixel array size have been designed and manufactured using this process which is fully compatible with semiconductor production. The devices are suitable to support detection of humans e.g. in security and automotive safety applications. [ABSTRACT FROM AUTHOR]

Objective: To evaluate a systematic newborn screening (NBS) strategy for vitamin B12 deficiency.Study Design: In a prospective single-center NBS study, a systematic screening strategy for vitamin B12 deficiency was developed and evaluated. Tandem-mass spectrometry screening was complemented by 2 second-tier strategies, measuring methylmalonic/3-OH-propionic/methylcitric acid, and homocysteine from dried blood spots.Results: In a cohort of 176 702 children screened over 27 months, 33 children were detected by NBS in whom (maternal) vitamin B12 deficiency was confirmed. Homocysteine was the most sensitive marker for vitamin B12 deficiency, but only combination with a second-tier strategy evaluating methylmalonic acid allowed for detection of all 33 children. Mothers were of various ethnic origins, and 89% adhered to a balanced diet. Treatment in children was performed predominantly by oral vitamin B12 supplementation (84%), and all children remained without clinical symptoms at short-term follow-up.Conclusions: Vitamin B12 deficiency is a treatable condition but can cause severe neurologic sequelae in infants if untreated. The proposed screening strategy is feasible and effective to identify moderate and severe cases of vitamin B12 deficiency. With an incidence of 1:5355 newborns, vitamin B12 deficiency is more frequent than inborn errors of metabolism included in NBS panels. Treatment of vitamin B12 deficiency is easy, and additional benefits can be achieved for previously undiagnosed affected mothers. This supports inclusion of vitamin B12 deficiency into NBS but also stresses the need for increased awareness of vitamin B12 deficiency in caregivers of pregnant women. [ABSTRACT FROM AUTHOR]

Background: Aromatic l-amino acid decarboxylase deficiency (AADCD) is a rare, autosomal-recessive neurometabolic disorder caused by variants in dopa decarboxylase (DDC) gene, resulting in a severe combined deficiency of serotonin, dopamine, norepinephrine, and epinephrine. Birth prevalence of AADCD varies by population. In pilot studies, 3-O-methyldopa (3-OMD) was shown to be a reliable biomarker for AADCD in high-throughput newborn screening (NBS) allowing an early diagnosis and access to gene therapy. To evaluate the usefulness of this method for routine NBS, 3-OMD screening results from the largest three German NBS centers were analyzed., Methods: A prospective, multicenter (n = 3) NBS pilot study evaluated screening for AADCD by quantifying 3-OMD in dried blood spots (DBS) using tandem mass spectrometry (MS/MS)., Results: In total, 766,660 neonates were screened from January 2021 until June 2023 with 766,647 with unremarkable AADCD NBS (766,443 by 1st-tier analysis and 204 by 2nd-tier analysis) and 13 with positive NBS result recalled for confirmatory diagnostics (recall-rate about 1:59,000). Molecular genetic analysis confirmed AADCD (c.79C > T p.[Arg27Cys] in Exon 2 und c.215 A > C p.[His72Pro] in Exon 3) in one infant. Another individual was highly suspected with AADCD but died before confirmation (overall positive predictive value 0.15). False-positive results were caused by maternal L-Dopa use (n = 2) and prematurity (30th and 36th week of gestation, n = 2). However, in 63% (n = 7) the underlying etiology for false positive results remained unexplained. Estimated birth prevalence (95% confidence interval) was 1:766,660 (95% CI 1:775,194; 1:769,231) to 1:383,330 (95% CI 1:384,615; 1:383,142). The identified child remained asymptomatic until last follow up at the age of 9 months., Conclusions: The proposed screening strategy with 3-OMD detection in DBS is feasible and effective to identify individuals with AADCD. The estimated birth prevalence supports earlier estimations and confirms AADCD as a very rare disorder. Pre-symptomatic identification by NBS allows a disease severity adapted drug support to diminish clinical complications until individuals are old enough for the application of the gene therapy., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Newborn screening (NBS) for cystic fibrosis (CF) based on pancreatitis-associated protein (PAP) has been performed for several years. While some influencing factors are known, there is currently a lack of information on the influence of seasonal temperature on PAP determination or on the course of PAP blood concentration in infants during the first year of life. Using data from two PAP studies at the Heidelberg NBS centre and storage experiments, we compared PAP determinations in summer and winter and determined the direct influence of temperature. In addition, PAP concentrations measured in CF-NBS, between days 21-35 and 36-365, were compared. Over a 7-year period, we found no significant differences between PAP concentrations determined in summer or winter. We also found no differences in PAP determination after 8 days of storage at 4 °C, room temperature or 37 °C. When stored for up to 3 months, PAP samples remained stable at 4 °C, but not at room temperature ( p = 0.007). After birth, PAP in neonatal blood showed a significant increasing trend up to the 96th hour of life ( p < 0.0001). During the first year of life, blood PAP concentrations continued to increase in both CF- (36-72 h vs. 36-365 d p < 0.0001) and non-CF infants (36-72 h vs. 36-365 d p < 0.0001). Seasonal effects in central Europe appear to have a limited impact on PAP determination. The impact of the increase in blood PAP during the critical period for CF-NBS and beyond on the applicability and performance of PAP-based CF-NBS algorithms needs to be re-discussed.

Newborn screening (NBS) programs are effective measures of secondary prevention and have been successively extended. We aimed to evaluate NBS for methylmalonic acidurias, propionic acidemia, homocystinuria, remethylation disorders and neonatal vitamin B 12 deficiency, and report on the identification of cofactor-responsive disease variants. This evaluation of the previously established combined multiple-tier NBS algorithm is part of the prospective pilot study "NGS2025" from August 2016 to September 2022. In 548,707 newborns, the combined algorithm was applied and led to positive NBS results in 458 of them. Overall, 166 newborns (prevalence 1: 3305) were confirmed (positive predictive value: 0.36); specifically, methylmalonic acidurias (N = 5), propionic acidemia (N = 4), remethylation disorders (N = 4), cystathionine beta-synthase (CBS) deficiency (N = 1) and neonatal vitamin B 12 deficiency (N = 153). The majority of the identified newborns were asymptomatic at the time of the first NBS report (total: 161/166, inherited metabolic diseases: 9/14, vitamin B 12 deficiency: 153/153). Three individuals were cofactor-responsive (methylmalonic acidurias: 2, CBS deficiency: 1), and could be treated by vitamin B 12 , vitamin B 6 respectively, only. In conclusion, the combined NBS algorithm is technically feasible, allows the identification of attenuated and severe disease courses and can be considered to be evaluated for inclusion in national NBS panels.