Phenotypic characterization and predictive analysis of p.Asp47Asn LDL receptor mutation associated with Familial Hypercholesterolemia in a Chilean population

BackgroundFamilial hypercholesterolemia (FH) is an inherited disorder mainly caused by mutations in the LDL receptor (LDL-R) and characterized by elevation of low-density lipoprotein cholesterol (LDL-C) levels and premature cardiovascular disease.ObjectiveIn this study, we evaluated the clinical phenotype of the p.Asp47Asn, described as an uncertain pathogenic variant, and its effect on the structure of LDL-R and ligand interactions with apolipoproteins.Methods27 children and adolescents with suspected FH diagnosis were recruited from a pediatric endocrinology outpatient clinic. Blood samples were collected after 12 h fasting for lipid profile analysis. DNA sequencing was performed for six FH-related genes by Ion Torrent PGM platform and copy number variation by MLPA. For index cases, a familial cascade screening was done restricted to the same mutation found in the index case. In silico analysis were developed to evaluate the binding capacity of LDL-R to apolipoproteins B100 and E.ResultsLipid profile in children and adolescents demonstrated higher LDL-C levels in p.Asp47Asn carriers compared to the wild type genotype. In silico analysis predicted a reduction in the binding capacity of the ligand-binding modules LA1-2 of p.Asp47Asn LDL-R for ApoB100 and ApoE, which was not produced by local structural changes or folding defects but as a consequence of a decreased apparent affinity for both apolipoproteins.ConclusionThe clinical phenotype and the structural effects of p.Asp47Asn LDL-R mutation suggest that this variant associates to FH.     

DNA Methylation Variants at HIF3A Locus,B-Vitamin Intake,and Long-term Weight Change: Gene-Diet Interactions in Two U.S. Cohorts

The first epigenome-wide association study of BMI identified DNA methylation at an HIF3A locus associated with BMI. We tested the hypothesis that DNA methylation variants are associated with BMI according to intake of B vitamins. In two large cohorts, we found significant interactions between the DNA methylation–associated HIF3A single nucleotide polymorphism (SNP) rs3826795 and intake of B vitamins on 10-year changes in BMI. The association between rs3826795 and BMI changes consistently increased across the tertiles of total vitamin B₂ and B₁₂ intake (all P for interaction <0.01). The differences in the BMI changes per increment of minor allele were −0.10 (SE 0.06), −0.01 (SE 0.06), and 0.12 (SE 0.07) within subgroups defined by increasing tertiles of total vitamin B₂ intake and −0.10 (SE 0.06), −0.01 (SE 0.06), and 0.10 (SE 0.07) within subgroups defined by increasing tertiles of total vitamin B₁₂ intake. In two independent cohorts, a DNA methylation variant in HIF3A was associated with BMI changes through interactions with total or supplemental vitamin B₂, vitamin B₁₂, and folate. These findings suggest a potential causal relation between DNA methylation and adiposity.     

Diagnostic accuracy of a rapid urine-screening test (Multistix8SG) in cirrhotic patients with spontaneous bacterial peritonitis

OBJECTIVE: To assess the diagnostic accuracy of a rapid urine-screening test (Multistix8SG) for spontaneous bacterial peritonitis (SBP) in cirrhotic patients. METHODS: Seventy-two consecutive patients (44 males, 28 females; mean age 61.6 years) with cirrhosis and ascites were included in the study. A diagnostic paracentesis was performed on hospital admission in all patients and 2 days after antibiotic treatment in the case of SBP (polymorphonuclear [PMN] count over 250/mm in ascitic fluid). Each fresh sample of ascitic fluid was also tested using the Multistix8SG urine test, and the results were scored as negative, trace or positive. RESULTS: Nine of the 72 patients had SBP and the Multistix8SG urine test was positive. After 48 h of antibiotic therapy, the PMN count of three of these nine patients was still above 250/mm and the Multistix8SG test remained positive. In three other patients with SBP, the PMN count dropped below 250/mm and the Multistix8SG test result had become negative. Two of the nine SBP patients died before 48 h, and paracentesis was not performed in the ninth case. In the other 63 patients, the PMN count in ascitic fluid was below 250/mm; the Multistix8SG test revealed 17 trace results and 46 negative results. At the threshold of 250 PMN/mm in ascitic fluid, this test had a sensitivity and a specificity of 100%. CONCLUSION: A positive Multistix8SG urine test result in ascitic fluid appears to be an indication for antibiotic treatment.     

Pain and Bloody Ear Discharge in a Returning Traveler

Cochliomyia hominivorax, the New World screwworm, was a serious livestock pest in the southern United States until the 1960s, when it was successfully eradicated by the release of sterile male flies. It remains endemic in parts of the Caribbean and South America, and there is concern that climate change may extend its geographic distribution. Cochliomyia hominivorax is voracious and can cause extensive damage to soft tissue and bone. We describe the case of a 26-year-old traveler who presented with otalgia and bloody otorrhea after returning from a vacation in the Dominican Republic, where exposure to screwworm flies most likely occurred during a nap on the beach. The causative agent was recognized by its characteristic larval anatomy, which includes pigmented dorsal tracheal trunks and posterior spiracles with an open peritreme.A 26-year-old female presented with a 2-day history of otalgia and bloody otorrhea after returning from a week of vacation in the Dominican Republic, where she stayed in a beachside resort with screened windows, swam in the ocean, hiked near an estuary, and sunbathed on the beach. She admitted to falling asleep on the beach one evening after drinking alcohol. The next day she had right ear discomfort with the sensation of movement, and removed a fly from her ear. One day later during the flight home, she had acute onset of ear pain, which she initially attributed to pressure changes associated with her flight, until she began noting discharge from the ear. The discharge was initially clear but became bloody within 1–2 hours. She sought medical care the next day, when an otoscopic examination revealed multiple motile larvae (Figure 1, Panel A). She was immediately referred to an otolaryngologist who performed excision and debridement of her external auditory canal with atticotomy.Open in a separate windowFigure 1.Panel A: Excised tissue displaying multiple motile larvae that were removed from the external auditory canal of a returning traveler vacationing in the Dominican Republic with auricular myiasis. Panel B: The characteristic pigmented dorsal tracheal trunks (white open arrow) and posterior spiracles with an open peritreme (gray closed arrow and inset) of Cochliomyia hominivorax.Intraoperative findings revealed soft tissue larval infiltration extending to the temporal bone and tympanic membrane perforation without middle ear involvement. The patient tolerated the debridement well and was prescribed amoxicillin-clavulanic acid for potential secondary soft-tissue infection. She subsequently underwent tympanoplasty with a split-thickness skin graft. The removed larvae were preliminarily identified as Cochliomyia hominivorax by the University of Washington Microbiology Laboratory on the basis of characteristic anatomic findings. Definitive identification was confirmed by the U.S. Department of Agriculture Screwworm Research Unit.Cochliomyia hominivorax (“human eater”), the New World screwworm, has smooth larvae with pigmented dorsal tracheal trunks (white open arrow) and posterior spiracles with an open peritreme (gray closed arrow, and inset) (Figure 1, Panel B).¹ The larvae feed on the living tissue of warm-blooded mammals using sharp hook-like mandibles (Supplemental Video). Spines found in concentric rings along the larvae resemble a screw and aid in anchoring the larvae within tissue, hence the name “screwworm.”² A female adult can lay hundreds of eggs at one time, often within existing open wounds.³ The voracious larvae emerge within 24 hours and can cause extensive soft tissue damage, and in some cases, destruction of bone. After 5–7 days the pupae mature and attempt to leave the wound to eventually burrow into the ground to pupate.² Adult flies can live for 2–3 weeks. Female flies are attracted to the scent of normal secretions of the orifices of mammals, with case reports describing infestation of the ears, eyes, nose, mouth, vagina, and rectum,⁴ and are able to fly great distances (∼50 km/week) to find a suitable host in which to deposit their eggs.² The scent of tissue infested with C. hominivorax can attract additional adult female flies that deposit their eggs within the same site. Individuals who are immobile, developmentally delayed, mentally ill, or alcoholic are at a higher risk of infestation.³ Orbital myiasis can be particularly devastating, requiring extensive debridement and sometimes enucleation.⁴ Several case reports have described secondary myiasis of ulcerative cutaneous malignancies.⁵ Auricular myiasis, as observed in our patient, is extremely rare and primarily occurs in children.³ Treatment of New World screwworm infestation consists of debridement, antibiotics if evidence of secondary infection is present, and adjunctive ivermectin in severe cases.⁴Cochliomyia hominivorax was formerly endemic in the southern United States where it was responsible for a serious economic burden caused by livestock destruction, until the 1960s when it became the first pest to be successfully eradicated by the release of sterile male flies.⁶ The New World screwworm remains endemic in parts of the Caribbean and South America, and there is concern that climate change may extend its geographic distribution.⁷ Since the New World screwworm has been eradicated from the United States for decades, clinicians may be unaware of this potentially devastating infestation resulting from foreign travel to endemic areas. As a result of the risk for significant tissue destruction, particularly if diagnosis is delayed, it is important for clinicians to be able to promptly recognize and appropriately treat this form of myiasis.