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Objective:The aim of this study was to investigate the ADAM 10 rs.653765 SNP genetic polymorphism in the hepatocellular carcinoma occurrence (de novo and post DAAs). Methods: This study was conducted on 360 participants divided to 4 groups. Group 1: 90 chronic adult patients infected with HCV received DAAs regimens and evolved HCC during the period of follow up. Group 2: Another 90 HCV patients received the same DAAs regimens and did not show HCC manifestations during the same follow up period. Group 3 included 90 de novo HCC patients (did not receive any DAAs). Finally, 90 apparently healthy participants as group 4. Clinical and laboratory data were evaluated, and ADAM 10 genotyping were performed using qPCR. Results: The study showed statistically significant between HCC de novo and HCC deterioration on top of DAAs according to three scoring systems (Child Pugh, BCLC and HKLC) with p-value <0.05. Regarding ADAM10 gene polymorphism, the study showed a significant difference between CC versus CT+TT genotypes of HCC groups according to Child Bugh, BCLC and HKLC staging systems. Yet, no significant difference was found when ADAM10 genotypes and allele frequencies were compared between the four different studied groups. No difference in the survival rate between HCC de novo and on the top of DAAs but more aggressive stages with HCC on top of DAAs. Conclusion: ADAM10 genotypes did not show any significant association with HCC. Also, no differences in the death rate recorded between the de novo HCC and HCC post DAAs treatment with statistical significant worse staging of HCC post DAAs and were noted. the study showed a significant difference between CC versus CT+TT genotypes of HCC groups according to Child Bugh, BCLC and HKLC staging systems.

Background: Tetrahydrobiopterin is a coenzyme of phenylalanine hydroxylase and other enzymes essential for the synthesis of tyrosine and other neurotransmitters. Despite proper nutritional regulation of blood phenylalanine levels, tetrahydrobiopterin deficiency results in progressive neurologic illness. Objective:The study aims to optimize a mass spec-dependent assay for quantitative measurement of biopterin and neopterin as distinctive markers of the atypical phenylketonuria due to tetrahydrobiopterin deficiency. Patients and methods:The study enrolled 46 patients with typical hyperphenylalaninaemia, 14 atypical cases, and 50 healthy children as a control group. Quantitative measurements of biopterin, neopterin, phenylalanine, and tyrosine were performed by ultra-performance liquid chromatography-mass spectrometry in the dried blood spots of patients and control samples. Results: Validation of the analytical protocol was performed at a biopterin and neopterin concentration range from 0 to 100 nmol/l. The regression coefficients for the linearity of the calibration curves exceeded 0.98. The lower limit of detection of biopterin and neopterin ranged from 1.5 to 2.5 nmol/l. The intra-day and inter-day precision and accuracy ranged from 96% -105%, and from 97% -110%. The mean recoveries were 105 ± 7% for biopterin and 106 ± 9% for neopterin. The short and long-term stability of the stored samples was seven days at room temperature and 12 weeks at -20 °C. Biopterin and neopterin were significantly higher in the classic group compared to the atypical and control group (p < 0.05), with their level being in the order classic > control > atypical group. Phenylalanine levels had a significant positive correlation with biopterin and neopterin levels in classic phenylketonuria (p < 0.05). Conclusions:The performance of the developed assay for biopterin and neopterin in the dried blood spot by ultraperformance liquid chromatography-mass spectrometry was accurate and precise thus, provide a legitimate diagnostic tool for cases of atypical phenylketonuria.