ObjectiveNonsense mutations account for 5–70% of all genetic disorders. In the United States, nonsense mutations in the CLN1/PPT1 gene underlie >40% of the patients with infantile neuronal ceroid lipofuscinosis (INCL), a devastating neurodegenerative lysosomal storage disease. We sought to generate a reliable mouse model of INCL carrying the most common Ppt1 nonsense mutation (c.451C>T) found in the United States patient population to provide a platform for evaluating nonsense suppressors in vivo.MethodsWe knocked-in c.451C>T nonsense mutation in the Ppt1 gene in C57 embryonic stem (ES) cells using a targeting vector in which LoxP flanked the Neo cassette, which was removed from targeted ES cells by electroporating Cre. Two independently targeted ES clones were injected into blastocysts to generate syngenic C57 knock-in mice, obviating the necessity for extensive backcrossing.ResultsGeneration of Ppt1-KI mice was confirmed by DNA sequencing, which showed the presence of c.451C>T mutation in the Ppt1 gene. These mice are viable and fertile, although they developed spasticity (a “clasping” phenotype) at a median age of 6 months. Autofluorescent storage materials accumulated throughout the brain regions and in visceral organs. Electron microscopic analysis of the brain and the spleen showed granular osmiophilic deposits. Increased neuronal apoptosis was particularly evident in cerebral cortex and abnormal histopathological and electroretinographic (ERG) analyses attested striking retinal degeneration. Progressive deterioration of motor coordination and behavioral parameters continued until eventual death.InterpretationOur findings show that Ppt1-KI mice reliably recapitulate INCL phenotype providing a platform for testing the efficacy of existing and novel nonsense suppressors in vivo.

INTRODUCTION: Adnexal masses are common in pregnancy. Our objective is to evaluate outcomes of oophorectomy in pregnant women. METHODS: This is a retrospective cohort study using data from the National Inpatient Sample between 2007 and 2013. Reproductive-aged women undergoing oophorectomy were identified using International Classification of Diseases – 9 diagnosis and procedure codes. Ectopic pregnancies were excluded. The risk of ovarian cancer was compared between pregnant and non-pregnant women undergoing oophorectomy. Secondary outcomes included surgical and pregnancy complications. Univariable analyses and multivariable regression were performed with adjustment for confounding factors. RESULTS: 203,075 women aged 15–45 years undergoing oophorectomy were identified, of whom 17,285 (8.5%) were pregnant. Pregnancy was associated with a lower frequency of ovarian cancer diagnosis (0.9% vs 2.5%, P<.001, OR 0.4, 95% CI 0.3 – 0.4). This relationship persisted with multivariable regression (P<.001, aOR 0.3, 95% CI 0.3 – 0.4). Pregnant women were significantly less likely to undergo a laparoscopic procedure or to undergo hysterectomy, omentectomy, or lymph node dissection compared with non-pregnant women. Pregnant women were also significantly more likely to die during hospitalization; this relationship persisted in multivariable regression (0.16% vs 0.05%, P<.001, aOR 4.1, 95% CI 2.5 – 6.6). There were no significant differences in frequency of transfusion or venous thromboembolism. Of pregnant women, 82% delivered during hospitalization for oophorectomy, 78% of these delivered by cesarean. CONCLUSION: Oophorectomy in pregnancy is associated with a lower frequency of ovarian cancer diagnosis, though is associated with higher frequency of laparotomy and death during hospitalization.

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