Identification of suitable reference microRNA for qPCR analysis in pediatric inflammatory bowel disease.

Pediatric inflammatory bowel disease (IBD) accounts for 10-15% of IBD and is associated with considerable morbidity for patients. Dysregulated microRNAs (miRNA, miR), small noncoding RNA molecules that modulate gene expression, have been the target of research in IBD diagnosis, surveillance, and therapy. Proper selection of reference genes, which are a prerequisite for accurate measurement of miRNA expression, is currently lacking. We hypothesize that appropriate normalization requires unique reference genes for different tissue and disease types. Through the study of 28 pediatric intestinal samples, we sought to create a protocol for selection of suitable endogenous reference genes. Candidate reference genes (miR-16, 193a, 27a, 103a, 191) were analyzed by RT-quantitative (q)PCR. Criteria used for designation of suitable reference genes were as follows: 1) ubiquitous: present in all tissue samples with quantification cycle value 15-35; 2) uniform expression: no differential expression between control and disease samples (P > 0.05); 3) stability: stability value <0.5 by NormFinder. Our results suggest the use of miR-27a/191 for Crohn's disease small bowel, none of the five candidate genes for Crohn's disease colon, and miR-16/27a for ulcerative colitis. Additionally, target miR-874 had differential expression when normalized with different reference genes. Our results demonstrate that reference gene choice for qPCR analysis has a significant effect on study results and that proper data normalization is imperative.

Alvarado Scores Predict Additive Value of Magnetic Resonance Imaging in Workup of Suspected Appendicitis in Children.

BACKGROUND: The presentation of appendicitis in pediatrics is variable, and diagnostic imaging is often used. Magnetic resonance imaging (MRI) is replacing computed tomography in some centers, particularly after a nondiagnostic ultrasound (NDUS). Nonetheless, MRI is not widely used in this setting because of cost, procedure time, institutional capacity, and high rates of negative scans. We hypothesized that the Alvarado Score (AS) could be used to determine the additive diagnostic value of MRI after an NDUS. MATERIALS AND METHODS: Retrospective review of patients aged ≤18 y at a single tertiary care children's hospital who received an ultrasound for suspected appendicitis during 10 consecutive months in 2017. NDUS were defined as nonvisualization of the appendix or secondary signs without radiologic diagnosis. AS were retrospectively calculated from the electronic medical record. Primary outcomes were pathology-confirmed appendicitis, appendectomy, and perforation. RESULTS: AS was determined for 352 patients out of 463 who met inclusion criteria (76%). Sixty-two percent had an NDUS, and 45% of these patients received MRI. Patients with high-risk AS were significantly more likely to have MRI diagnostic of appendicitis (P = 0.0015), and low-risk AS patients were more likely to have a negative or equivocal MRI (P = 0.0169). Twenty-one MRI scans were required per each additional diagnosis of appendicitis in patients with low AS after NDUS versus 4.2 in intermediate-risk AS patients and 2.1 in high-risk AS patients. CONCLUSIONS: Risk stratification with AS can help assess the additive diagnostic utility of MRI after NDUS. MRI may be overutilized for diagnosing acute appendicitis in pediatric patients with low-risk AS.

Impact of Decolonization Protocols and Recurrence in Pediatric MRSA Skin and Soft-Tissue Infections.

BACKGROUND: Methicillin-resistant staphylococcus aureus (MRSA) colonization is associated with the development of skin and soft-tissue infection in children. Although MRSA decolonization protocols are effective in eradicating MRSA colonization, they have not been shown to prevent recurrent MRSA infections. This study analyzed the prescription of decolonization protocols, rates of MRSA abscess recurrence, and factors associated with recurrence. MATERIALS AND METHODS: This study is a single-institution retrospective review of patients ≤18 y of age diagnosed with MRSA culture-positive abscesses who underwent incision and drainage (I&D) at a tertiary-care children's hospital. The prescription of an MRSA decolonization protocol was recorded. The primary outcome was MRSA abscess recurrence. RESULTS: Three hundred ninety-nine patients with MRSA culture-positive abscesses who underwent I&D were identified. Patients with previous history of abscesses, previous MRSA infection groin/genital region abscesses, higher number of family members with a history of abscess/cellulitis or MRSA infection, and I&D by a pediatric surgeon were more likely to be prescribed decolonization. Decolonized patients did not have lower rates of recurrence. Recurrence was more likely to occur in patients with previous abscesses, previous MRSA infection, family history of abscesses, family history of MRSA infection, Hispanic ethnicity, and those with fever on admission. CONCLUSIONS: MRSA decolonization did not decrease the rate of recurrence of MRSA abscesses in our patient cohort. Patients at high risk for MRSA recurrence such as personal or family history of abscess or MRSA infection, Hispanic ethnicity, or fever on admission did not benefit from decolonization.

To wrap or not to wrap: A retrospective review of circumcision dressing and post-procedural bleeding.

INTRODUCTION: Bleeding is an infrequent, but important, complication after circumcision. Our aim was to examine postoperative bleeding events after circumcision comparing patients managed with a circumferential wrap to ointment alone. METHODS: Boys ≤18 years of age who underwent circumcision at a tertiary children's hospital were retrospectively reviewed between 2017 and 2018. Postoperative bleeding was defined by phone calls, clinic or Emergency Department visits, or return to the operating room. Outcomes were examined by univariate association and multivariable modeling. RESULTS: Of 681 boys undergoing circumcision, 503 (74%) patients received a wrap dressing and 178 (26%) only ointment. There were 28 (4%) patients who had a postoperative bleeding event: 14/503 (2.7%) among wrap dressings and 14/178 (7.8%) among ointment alone (p < 0.01). The majority of events were phone calls related to bleeding (75%). Univariate analysis demonstrated no association between postoperative bleeding and surgeon specialty (p = 0.72), age at circumcision (p = 0.44) or technique type (p = 0.09). After controlling for age, technique type, and surgeon specialty, dressing type remained significantly associated with postoperative bleeding (OR = 2.81, p < 0.01). CONCLUSION: This single-center, retrospective review found circumferential wrap dressings are associated with a decrease in bleeding events after circumcision. LEVEL OF EVIDENCE: III - retrospective case-control study.