Alagille syndrome (ALGS) is a multisystem condition characterized by cholestasis and bile duct paucity on liver biopsy and variable involvement of the heart, skeleton, eyes, kidneys, and face and caused by pathogenic variants in the JAG1 or NOTCH2 gene. The variable expressivity of the clinical phenotype and the lack of genotype–phenotype correlations lead to significant diagnostic difficulties. Here we present an analysis of 18 patients with cholestasis who were diagnosed with ALGS. We used an NGS panel targeting coding exons of 52 genes, including the JAG1 and NOTCH2 genes. Sanger sequencing was used to verify the mutation in the affected individuals and family members. The specific facial phenotype was seen in 16/18 (88.9%). Heart defects were seen in 8/18 (44.4%) patients (pulmonary stenosis in 7/8). Butterfly vertebrae were seen in 5/14 (35.7%) patients. Renal involvement was detected in 2/18 (11.1%) cases—one patient had renal cysts, and one had obstructive hydronephrosis. An ophthalmology examination was performed on 12 children, and only one had posterior embryotoxon (8.3%). A percutaneous liver biopsy was performed in nine cases. Bile duct paucity was detected in six/nine cases (66.7%). Two patients required liver transplantation because of cirrhosis. We identified nine novel variants in the JAG1 gene—eight frameshift variants (c.1619_1622dupGCTA (p.Tyr541X), c.1160delG (p.Gly387fs), c.964dupT (p.C322fs), c.120delG (p.L40fs), c.1984dupG (p.Ala662Glyfs), c.3168_3169delAG (p.R1056Sfs*51), c.2688delG (p.896CysfsTer49), c.164dupG (p.Cys55fs)) and one missense variant, c.2806T > G (p.Cys936Gly). None of the patients presented with NOTCH2 variants. In accordance with the classical criteria, only six patients could meet the diagnostic criteria in our cohort without genetic analysis. Genetic testing is important in the diagnosis of ALGS and can help differentiate it from other types of cholestasis.

After the sudden outbreak of the COVID-19 pandemic, scientists and clinicians around the world have significantly expanded understanding of the pathogenesis of the disease as well as the impact of SARS-CoV-2 on various organs and tissues. To date, it is accepted to consider the new coronavirus infection as a multisystem disease, but the data on the effect on fertility remains unclear. Previous works by other authors have presented controversial results, and there is no evidence of a direct effect of the new coronavirus on the male gonads. Thus, further studies are needed to verify the hypothesis that the testicles are the target organ for SARS-CoV-2. Groups were formed: Group I (n = 109; age from 25 to 75 years, Median (IQR) - 60 (23) years), cause of death - new coronavirus infection; Group II (n = 21, age from 25 to 75 years, Median (IQR) - 55 (29.5) years) - autopsy testicular material obtained outside the pandemic. We used RT-PCR to detect the presence of viral RNA in testicular tissue. In addition, we investigated the levels of proteins that provide viral invasion, such as ACE-2 and Furin. In the present study, we detected genetic material of a new coronavirus and increased proteins required for viral invasion in testicular tissue of patients with COVID-19 by RT-PCR. Thus, based on our findings, we assume that testicular tissue is potentially vulnerable to SARS-CoV-2.Communicated by Ramaswamy H. Sarma