Rare autosomal dominant hereditary hemochromatosis associated with SLC40A1 gene: ferroportin disease or type 4 hereditary hemochromatosis?

Ferroportin (FPN1), encoded by the SLC40A1 gene, is the unique cellular iron exporter identified in mammals. FPN1 transfers iron from the intestine and macrophages into the bloodstream. This function is negatively regulated by hepcidin. Mutations in SLC40A1 may affect FPN1 function, originating distinct autosomal dominant diseases: (i) the Ferroportin Disease (FD), due to loss-of-function mutations, is characterized by decreased iron export from enterocytes and severely affected iron transfer in macrophages, giving rise to a marked iron accumulation in spleen and liver; and (ii) the Type 4 Hereditary Hemochromatosis (HH), resulting from gain-of-function mutations conferring resistance to hepcidin-mediated FPN1 degradation and consequently high cellular iron export. In this study, 335 individuals suspected of having non-classic HH were enrolled. Six genes related with iron metabolism were analysed by SSCP, dHPLC or NGS. The latter used TruSeq or Nextera XT libraries and a MiSeq platform (Illumina). Genetic variants found were validated by Sanger sequencing. Predictive consequences at protein level were evaluated using Polyphen-2 and SIFT softwares. From all patients analysed, three SLC40A1 pathogenic variants were detected in heterozygosity in three women: two missense, c.238G>A, p.Gly80Ser and c.610G>A, p.Gly204Ser; and one deletion, c.485_487delTTG; p.Val162del. These variants had been reported in public databases, but they were not known to be present in the Portuguese population. The p.Gly80Ser and the p.Val162del are FPN1 loss-of-function mutations and were found associated with hyperferritinemia and low transferrin saturation (FD). In contrast, the p.Gly204Ser induced a gain of FPN1 function with a full iron export capacity giving the patient a type 4-HH phenotype, which includes iron overload, hyperferritinemia and high transferrin saturation. Detailed clinical evaluation of the suspected patients are useful to unravel the effect of different mutations in FPN1 function, expression and regulation.This work was partially supported by INSA_2013DGH910 and GenomePT (POCI-01-0145-FEDER-022184).info:eu-repo/semantics/publishedVersio

Novel mutation in addition to functional TMPRSS6 gene polymorphisms originate an IRIDA-like phenotype in an African child

Iron-refractory iron deficiency anemia (IRIDA) is a rare autosomal recessive anemia often unresponsive to oral iron intake and partially responsive to parenteral iron treatment. The disease originates from mutations in TMPRSS6 gene, encoding Matriptase 2, a transmembrane serine protease that plays an essential role in down-regulating hepcidin. Once TMPRSS6 is mutated, the corresponding protein is absent or inactive at the hepatocyte membrane leading to uncontrolled high levels of hepcidin and impaired iron absorption. This study aimed to investigate a 4-year-old boy of sub-Saharan ancestry (Mozambique/Angola), presenting with microcytic hypochromic anemia, low transferrin saturation, normal ferritin, and having a partial response to intravenous iron treatment. He is a -α3.7-thalassemia carrier. TMPRSS6 was screened for variants by Next-Generation Sequencing using Nextera XT libraries in a MiSeq platform (Illumina). Genetic variants found were validated by Sanger sequencing. In silico analyses were performed in HSF, SIFT, Poly-Phen2 and Missense3D softwares. A novel missense mutation (c.871G>A) was found in heterozygosity, in TMPRSS6 exon 8. In silico analysis indicates the conserved amino acid change (G291S) may be damaging to the protein stability. Due to its location in the CUB1 domain, it may also affect the enzyme activation and substrate recognition. Additionally, 3 SNPs previously associated with a greater risk of developing iron deficiency anemia (K253E, V736A and Y739Y) were also identified in TMPRSS6. Although IRIDA is known as an autosomal recessive disease, we conclude that, in this case, the result of a digenic inheritance of the novel damaging mutation (c.871G>A; G291S) and the 3 common modulating SNPs in the same gene and a co-inheritance of the α-thalassemia HBA deletion may lead to an IRIDA-like phenotype. Further functional studies of the mutated protein as well as family studies should be conducted.This work was partially supported by INSA_2013DGH910 and GenomePT (POCI-01-0145-FEDER-022184).info:eu-repo/semantics/publishedVersio

Spontaneous Neonatal Humeral Artery Thromboembolism: a Case Report

info:eu-repo/semantics/publishedVersio

Hypophosphatemia As a Possible Biomarker for Epileptic Seizures at the Emergency Department

Introduction: Hypophosphatemia seems to be temporally associated with seizures, despite not being considered a trigger. We aimed to evaluate hypophosphatemia as a biomarker for seizures. Methods: Retrospective study, including all consecutive patients admitted at our central hospital's emergency department from 01/01-31/03/2021, screened as "altered consciousness/syncope" or "seizures", with available phosphate levels. Results: 277 patients included, mostly male (61.7%), mean age 64.3 years. Final diagnosis was "seizure" in 34.7% and "other diagnosis" in 65.3%. Patients with seizures were younger (p200U/L) was more frequent in the seizure group (p=0.04). Odds ratio (OR) of hypophosphatemia for seizures was 4.330 (CI 95% 2.170-8.640, p<0.001), persisting after correction for confounders. OR of hyperCK was 1.890 (CI 95% 1.060-3.371, p=0.03), losing significance when adjusted. Sensitivity was low for both. Hypophosphatemia was more specific (91.2% vs 79.9%). Conclusions: Our findings support hypophosphatemia as a seizure biomarker. More studies are needed.info:eu-repo/semantics/publishedVersio

Eficiência do processo de biodigestão em fossa séptica biodigestora inoculada com esterco de ovino.

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Comparative genomics of two pseudomonas aeruginosa clinical isolates to elucidate the composition of their mobilomes

Recently, we have set a collaboration with Hospital de Braga, located in the North of Portugal, that handles over 600 P. aeruginosa isolates per year, aiming to rouse a holistic research approach to provide relevant information and tools to the clinicians to circumvent the multi-resistance phenomena in P. aeruginosa. Since then, we have set procedures aiming a systematic phenotypic characterization of the clinical isolates and developed strategies for the identification of pathogenicity islands and SNPs among the clinical isolates via comparative genomics. In this context, we have determined the full genome sequence of two clinical isolates using the high-throughput system Illumina Genome Analyzer IIx. These two clinical isolates, named 138244 and 152504, are representatives of allelic sequence types ST175 (widely disseminated and associated with multidrug-resistance) and ST560 (rare allele), respectively. Importantly, under standardized experimental procedures, isolate 138244 did not produce pigments and evidenced an antibiotic pan-resistant phenotype whereas 152504 produced a high amount of pyocyanin pigment and was susceptible to all antibiotics tested. A comparative genomic analysis using the genome sequences of both isolates and of all P. aeruginosa strains deposited in Genbank so far, allowed the identification of the accessory genome content of both isolates. Apparently, isolate 152504 harbors in its genome 243 unique genes, often clustered together in the same locus. Based on the genome annotation information, the pool of unique genes mainly encode several virulence factors, chemical stress resistance systems as well as 106 hypothetical proteins, some of which predicted members of the secretome of P. aeruginosa 152504. The accessory genome of 138244 mainly includes genes associated with mobile elements (phages, transposases, integrons) and genes encoding for 190 hypothetical proteins. Currently, research approaches are focused on the functional elucidation of sets of genes encoding hypothetical proteins of both isolates and in the description and characterization of their secretomes.Fundação para a Ciência e a Tecnologia (FCT

Insights into the accessory genome of two pseudomonas aeruginosa clinical isolates using comparative genomics

Recently, we have set a collaboration with Hospital de Braga, located in the North of Portugal, that handles over 600 P. aeruginosa isolates per year, aiming to rouse a holistic research approach to provide relevant information and tools to the clinicians to circumvent the multi-resistance phenomena in P. aeruginosa. Since then, we have set procedures aiming a systematic phenotypic characterization of the clinical isolates and developed strategies for the identification of pathogenicity islands and SNPs among the clinical isolates via comparative genomics. In this context, we have determined the full genome sequence of two clinical isolates using the high-throughput system Illumina Genome Analyzer IIx. These two clinical isolates, named 138244 and 152504, are representatives of allelic sequence types ST175 (widely disseminated and associated with multidrug-resistance) and ST560 (rare allele), respectively. Importantly, under standardized experimental procedures, isolate 138244 did not produce pigments and evidenced an antibiotic pan-resistant phenotype whereas 152504 produced a high amount of pyocyanin pigment and was susceptible to all antibiotics tested. A comparative genomic analysis using the genome sequences of both isolates and of all P. aeruginosa strains deposited in Genbank so far, allowed the identification of the accessory genome content of both isolates. Apparently, isolate 152504 harbors in its genome 254 unique genes, often clustered together in the same locus. Based on the genome annotation information, the pool of unique genes mainly encode several virulence factors, chemical stress resistance systems as well as 183 hypothetical proteins, 45 of which predicted members of the secretome of P. aeruginosa 152504. The accessory genome of 138244 mainly includes genes associated with mobile elements (phages, transposases, integrons) and genes encoding for 160 hypothetical proteins. Currently, research approaches are focused on the functional elucidation of sets of genes encoding hypothetical proteins of both isolates and in the description and characterization of their secretomes.Fundação para a Ciência e a Tecnologia (FCT

In situ modification of nanostructure configuration through the manipulation of hydrogen bonded amphiphile self-association

Herein, we report the synthesis of a novel amphiphilic salt containing a number of hydrogen bond donating (HBD) and accepting (HBA) functionalities. This amphiphile has been shown to self-associate via hydrogen bond formation in a DMSO solution, confirmed through a combination of NMR, UV-Vis and dynamic light scattering and supported by X-ray diffraction studies. The combination of different HBD and HBA functionalities within the amphiphile structure gives rise to a variety of competitive, self-associative hydrogen bonding modes that result in the formation of ‘frustrated’ hydrogen bonded nanostructures. These nanostructures can be altered through the addition of competitive HBD arrays and/or HBA anionic guests. The addition of these competitive species modifies the type of self-associative hydrogen bonding modes present between the amphiphilic molecules, triggering the in situ formation of novel hydrogen bonded nanostructures