查看更多>>摘要：Background: Alkaptonuria (AKU) is a rare tyrosine metabolism disorder caused by homogentisate 1,2-dioxygenase (HGD) mutations and homogentisic acid (HGA) accumulation. In this study, we investigated the genotype-phenotype relationship in AKU patients with a novel HGD gene mutation from a Chinese Hani family. Methods: Routine clinical examination and laboratory evaluation were performed, urine alkalinization test and urinary gas chromatography-mass spectrometry were used to assess HGA. Gene sequencing was utilized to study the defining features of AKU. NetGene2-2.42 and BDGP software was used to predict protein structure online. Flow cytometry and RT-PCR were used to analyze HGD proteins and HGD mRNA, respectively. Results: Two pediatric patients fulfilled diagnostic criteria for AKU with eddish-brown or black diapers and urine HGA testing. Sequencing testing revealed that all members of this family had a novel samesense mutation c.15G > A at the edge of exon 1 of the HGD. By flow cytometry, the expression of HGD protein in the pediatric patients' peripheral blood mononuclear cells was barely expressed. NetGene2-2.42 and BDGP software showed that the mutation reduced the score of the 5 & PRIME; splice donor site and disrupted its normal splicing, and the RT-PCR product also demonstrated that the defect in the HGD protein was due to the lack of the first exon containing the start codon ATG after the mutation. Conclusions: The novel mutation c.15G > A in HGD is associated with the AKU phenotype. It may affect the splicing of exon 1, leading to exon skipping, which impairs the structure and function of the protein.
查看更多>>摘要：Background: Short chain fatty acids (SCFAs) and branched chain amino acids (BCAAs) are frequently determined in faeces, and widely used as biomarkers of gut-microbiota activity. However, collection of faeces samples from neonates is not straightforward, and to date levels of these metabolites in newborn's faeces and urine samples have not been described. Methods: A targeted gas chromatography - mass spectrometry (GC-MS) method for the determination of SCFAs and BCAAs in both faeces and urine samples has been validated. The analysis of 210 urine and 137 faeces samples collected from preterm (PI), term infants (TI) and their mothers was used to report faecal and urinary SCFA and BCAA levels in adult and neonatal populations. Results: A significant correlation among five SCFAs and BCAAs in faeces and urine samples was observed. Reference ranges of SCFAs and BCAAs in mothers, PI and TI were reported showing infant's lower concentrations in faeces and higher concentrations in urine. Conclusion: This method presents a non-invasive approach for the simultaneous assessment of SCFAs and BCAAs in faecal and urine samples and the results will serve as a knowledge base for future experiments that will focus on the study of the impact of nutrition on the microbiome of lactating mothers and their infants.
查看更多>>摘要：Background: SARS-CoV-2 is an RNA virus that primarily causes respiratory disease; however, infection of other tissue has been reported. Evaluation of SARS-CoV-2 in tissue specimens may increase understanding of SARSCoV-2 pathobiology. Materials and Methods: A qualitative test for detection of SARS-CoV-2 in formalin-fixed paraffin-embedded (FFPE) tissues was developed and validated using droplet digital PCR (ddPCR), which has a lower limit of detection than reverse transcription (RT)-qPCR. After extraction of total RNA from unstained FFPE tissue, SARS-CoV-2 nucleocapsid (N1, N2) target sequences were amplified and quantified, along with human RPP30 as a control using the Bio-Rad SARS-CoV-2 ddPCR kit. Results: SARS-CoV-2 was detected in all 21 known positive samples and none of the 16 negative samples. As few as approximately 5 viral copies were reliably detected. Since January 2021, many tissue types have been clinically tested. Of the 195 clinical specimens, the positivity rate was 35% with placenta and fetal tissue showing the highest percentage of positive cases. Conclusion: This sensitive FFPE-based assay has broad clinical utility with applications as diverse as pregnancy loss and evaluation of liver transplant rejection. This assay will aid in understanding atypical presentations of COVID-19 as well as long-term sequelae.
Valle, AndreaRodriguez, JavierCamina, FelixMartinez-Olmos, Miguel A....
5页
查看更多>>摘要：Background: To examine glycaemic status, and the impact of at-admission HbA1c levels on outcome, in a large group of participants hospitalized for COVID-19. Methods: We inclued 515 participants with confirmed COVID-19 infection, with or without known diabetes, who met the following additional criteria: 1) age > 18 years, 2) HbA1c was determined at admission; 3) fasting plasma glucose was determined in the week of admission, and 4) discharge or death was reached before the end of the study. We examined attributes of participants at admission and 3-6 months post-discharge. To assess the associations of pre-admission attributes with in-hospital mortality, logistic regression analyses were performed. Results: Mean age was 70 years, 98.8% were of white race, 49% were female, 31% had known diabetes (KD), an additional 7% met the HbA1c criterion for diabetes, and 13.6% died. In participants with KD, FPG and HbA1c levels were not associated with mortality in adjusted analyses; however, in participants without KD, whereas FPG showed direct association with mortality, HbA1c showed slight inverse association. Conclusions: There was a very high prevalence of people without KD with HbA1c levels above normal atadmission. This alteration does not seem to have been related to blood glucose levels.
NSTL
Elsevier