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Electronic cigarettes (e-cigarettes) are handheld, battery-powered vaporizing devices. It is estimated that more than 25% of youth have used these devices recreationally. While vaping-associated lung injury is an increasingly recognized risk, little is known about the risk of traumatic injuries associated with e-cigarette malfunction.A multi-institutional retrospective study was performed by querying the electronic health records at nine children's hospitals. Patients who sustained traumatic injuries while vaping from January 2016 through December 2019 were identified. Patient demographics, injury characteristics, and the details of trauma management were reviewed.15 children sustained traumatic injuries due to e-cigarette explosion. The median age was 17 y (range 13-18). The median injury severity score was 2 (range 1-5). Three patients reported that their injury coincided with their first vaping experience. Ten patients required hospital admission, three of whom required intensive care unit admission. Admitted patients had a median length of stay of 3 d (range 1-6). The injuries sustained were: facial burns (6), loss of multiple teeth (5), thigh and groin burns (5), hand burns (4), ocular burns (4), a radial nerve injury, a facial laceration, and a mandible fracture. Six children required operative intervention, one of whom required multiple operations for a severe hand injury.In addition to vaping-associated lung injury, vaping-associated traumatic injuries are an emerging and worrisome injury pattern sustained by adolescents in the United States. This report highlights another means by which e-cigarettes pose an increasing risk to a vulnerable youth population.

Studies are urgently needed to characterize immunogenicity, efficacy, and safety of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mRNA vaccines in kidney transplant (KT) recipients, excluded from major clinical trials. Complex ELISPOT and other cellular response techniques have been applied, but simpler tools are needed. An easy-to-use real-world monitoring of SARS-CoV-2 IgG antibodies against the Spike protein and QuantiFERON® SARS-CoV-2 IFNI³ release assay (IGRA) were performed at baseline and 28 days after the second dose in KT recipients and controls (dialysis patients and healthy ones). All healthy controls and >95% dialysis controls became positive for anti-S IgG antibodies, while only 63.3% of KT patients seroconverted with a very low antibody level. A positive IGRA was documented in 96.9% of controls, 89.3% peritoneal dialysis, 77.6% hemodialysis, 61.3% of KT patients transplanted more than 1 year ago and only 36% of those transplanted within the previous 12 months. Overall, 100% of healthy controls, 95.4% of dialysis patients and 78.8% KT recipients developed any immune response (humoral and/or cellular) against SARS-CoV-2. KT patients showed low rates of immune responses to mRNA Coronavirus infectious disease 2019 vaccines, especially those with recent transplantations. Simple humoral and cellular monitoring is advisable, so that repeated doses may be scheduled according to the results.

PURPOSE Several preoperation prognosis models used on the treatment of metastatic bone disease on appendicular skeleton have been devised. The purpose of this study was to compare the performance of different survival prognostic models on patients with metastatic bone disease in long bones in a Chilean population. METHODS This is a multicentric retrospective study. We retrospectively reviewed the medical records of 136 patients who were confirmed with metastatic bone disease of the appendicular skeleton and who were treated surgically from 2016 to 2019. The minimum follow-up time was 12 months. All patients were assessed using four appendicular metastatic bone disease scoring systems. A preoperative predicted survival time for all 136 patients was retrospectively calculated making use of the revised Katagiri, PathFx, Optimodel and IOR score model. RESULTS The PathFx model demonstrated an accuracy at predicting 3 (area under the curve [AUC] = 0.61) and 6-month (AUC = 0.65) survival time after surgical management. IOR score model demonstrated an accuracy at predicting 12-month survival time (AUC = 0.64). The survival rate reached the 44% in a year. The median survival time to death or last follow-up time was 14.9 months (SD ± 15). CONCLUSION PathFx score model demonstrated the highest accuracy at predicting a survival time of 3 and 6 months. IOR score model was the most accurate measure at predicting a survival time of 12-months. To our knowledge, this is the first study reporting a comparative analysis of metastatic bone disease with predicting models in a country located in Latin America. PathFx's and IOR score models are the ones to be used in the Chilean population as the predictive models in metastatic bone disease of the appendicular skeleton.

Surgical site infection (SSI) remains a significant source of patient morbidity and resource utilization in children undergoing colorectal surgery. We examined the utility of a protocolized perioperative care bundle in reducing SSI in pediatric patients undergoing colorectal surgery.We conducted a prospective cohort study of patients ≤18 years of age undergoing colorectal surgery at ten United States children's hospitals. Using a perioperative care protocol comprising eight elements, or "colon bundle", we divided patients into low (1-4 elements) or high (5-8 elements) compliance cohorts. Procedures involving colorectal repair or anastomosis with abdominal closure were included. Demographics and clinical outcomes were compared between low and high compliance cohorts. Compliance was compared with a retrospective cohort. The primary outcome was superficial SSI incidence at 30 days.Three hundred and thirty-six patients were included in our analysis: 138 from the low compliance cohort and 198 from the high compliance cohort. Age and gender were similar between groups. Preoperative diagnosis was similar except for more patients in the high compliance cohort having inflammatory bowel disease (18.2% versus 5.8%, p0.01). The most common procedure performed was small bowel to colorectal anastomosis. Wound classification and procedure acuity were similar between groups. Superficial SSI at 30 days occurred less frequently among the high compliance compared to the low compliance cohort (4% versus 9.7%, p = 0.036). Median postoperative length of stay and 30-day rates of readmission, reoperation, intra-abdominal abscess and anastomotic leak requiring operation were not significantly different between groups. None of the individual colon bundle elements were independently protective against superficial SSI.Standardization of perioperative care is associated with a reduction in superficial SSI in pediatric colorectal surgery. Expansion of standardized protocols for children undergoing colorectal surgery may improve outcomes and decrease perioperative morbidity.Clinical Research Paper LEVEL OF EVIDENCE: Level II.

Cardiac organoids are 3-dimensional (3D) structures composed of tissue or niche-specific cells, obtained from diverse sources, encapsulated in either a naturally derived or synthetic, extracellular matrix scaffold, and include exogenous biochemical signals such as essential growth factors. The overarching goal of developing cardiac organoid models is to establish a functional integration of cardiomyocytes with physiologically relevant cells, tissues, and structures like capillary-like networks composed of endothelial cells. These organoids used to model human heart anatomy, physiology, and disease pathologies in vitro have the potential to solve many issues related to cardiovascular drug discovery and fundamental research. The advent of patient-specific human-induced pluripotent stem cell-derived cardiovascular cells provide a unique, single-source approach to study the complex process of cardiovascular disease progression through organoid formation and incorporation into relevant, controlled microenvironments such as microfluidic devices. Strategies that aim to accomplish such a feat include microfluidic technology-based approaches, microphysiological systems, microwells, microarray-based platforms, 3D bioprinted models, and electrospun fiber mat-based scaffolds. This article discusses the engineering or technology-driven practices for making cardiac organoid models in comparison with self-assembled or scaffold-free methods to generate organoids. We further discuss emerging strategies for characterization of the bio-assembled cardiac organoids including electrophysiology and machine-learning and conclude with prospective points of interest for engineering cardiac tissues in vitro.

PURPOSE The purpose of this study was to investigate factors impacting transplant-free survival among infants with biliary atresia. METHODS A multi-institutional, retrospective cohort study was performed at nine tertiary-level children's hospitals in the United States. Infants who underwent Kasai portoenterostomy (KP) from January 2009 to May 2017 were identified. Clinical characteristics included age at time of KP, steroid use, surgical approach, liver pathology, and surgeon experience. Likelihood of transplant-free survival (TFS) was evaluated using logistic regression, adjusting for patient and surgeon-level factors. Secondary outcomes at 1 year included readmission, cholangitis, reoperation, mortality, and biliary clearance. RESULTS Overall, 223 infants underwent KP, and 91 (40.8%) survived with their native liver. Mean age at surgery was 63.9 days (± 24.7 days). At 1 year, 78.5% experienced readmission, 56.9% developed cholangitis, 3.8% had a surgical revision, and 5 died. Biliary clearance at 3 months was achieved in 76.6%. Controlling for patient and surgeon-level factors, each additional day of age toward operation was associated with a 2% decrease in likelihood of TFS (OR 0.98, 95% CI 0.97-0.99). CONCLUSION Earlier surgical intervention by Kasai portoenterostomy at tertiary-level centers significantly increases likelihood for TFS. Policy-level interventions to facilitate early screening and surgical referral for infants with biliary atresia are warranted to improve outcomes.

Background and purpose Innovative delivery of research education and training in pharmacy education improves student perception of, and involvement in, professional pharmacy research. The purpose of this project was to utilize a flipped-classroom video technology approach to introduce our Capstone research experience and to assess the impact of the video technology on student's perceptions of pharmacy research. Educational activity and setting Faculty recorded one-min videos that provided an overview of current research. Each video included a research introduction, explanation of student incorporation into research, and impact of research on the field of expertise. Students were assigned to review faculty videos. To assess impact of the videos on students' perceptions of research, t-tests compared five variables before and after students watched the videos. These variables included research curiosity, research interest, research inspiration, research motivation, and pharmacy research interest. Findings Across all students, no differences were detected. A statistically significant interaction emerged that suggested prior pharmacy experience may impact research curiosity, interest, and inspiration. Summary: Findings demonstrated that the impact of research videos on pharmacy students' attitudes and perceptions towards research may depend on students' past experiences in a pharmacy setting. Further research is needed to identify factors that impact students' perceptions of pharmacy research. The information from this study provided our faculty members with valuable insights that can be used to better prepare students in the laboratory, classroom, and beyond.

To measure changes in student professionalism from co-curricular activities.A prospective cohort study followed first-year pharmacy students at The University of Texas at El Paso School of Pharmacy. Students completed one co-curricular activity each semester. Learning was assessed using the RxPro VALUE rubric and Professionalism Assessment Tool (PAT) at the beginning and end of the first professional year. The RxPro VALUE rubric included six domains (connections to experience, lifelong learning, diversity of communities and cultures, civic identity and commitment, empathy, and reflection and self-assessment) and was rated from 0 (not yet applied) to 4 (exemplary). The PAT contained 33-items in five domains (reliability, responsibility, and accountability; lifelong learning and adaptability; relationships with others; upholding principles of integrity and respect; and citizenship and professional engagement) and was rated from 1 (knows) to 5 (teaches) (maximum score = 165).Mean scores on the RxPro VALUE rubric increased (p 0.05) from baseline to end of year in five domains: connections to experience (1.44 vs. 2.07), diversity of communities and cultures (1.75 vs. 2.3), civic identity and commitment (1.34 vs. 2.33), empathy (1.44 vs. 2.37), and reflection and self-assessment (1.53 vs 2.19). No significant changes were seen in PAT scores from baseline (M = 114.19, standard deviation = 32.832) to end of year (M = 119.56, standard deviation = 29.63, p = 0.35).The RxPro VALUE rubric detected growth in professional behaviors among first-year doctor of pharmacy students, while no significant change was observed in the PAT.

Introduction The primary objective of the study was to assess factors that predict pursuit of an oncology post-graduate year 2 (PGY-2) residency. Additional objectives included identifying the role of and exploring opportunities for PharmD curricula to increase student interest in the oncology field. Methods An anonymous 15-minute survey was developed and administered to oncology pharmacists and residents. Study participants were included if they were actively practicing as clinical pharmacists in an oncology setting in the US, maintained an active pharmacist license, and >18 years of age. Responses were analyzed using descriptive and inferential statistics. All results are reported in aggregate, with the exception of quotes obtained from open-ended responses. Results Eighty participants were included in analyses. After controlling for variance due to age, sex, race, years of practice, the number of oncology advanced pharmacy practice experiences (APPEs) correlated with pursuit of a PGY-2 residency in oncology (p = 0.047). Additionally, participants' perceived level of preparedness from didactic oncology training predicted pursuit of an oncology PGY-2 residency (p = 0.002). Emerging themes in pursuing oncology from open-ended items revealed that inclusion of supportive care in the didactic curriculum and having a family member or friend diagnosed with cancer were important factors. Additionally, participants' responses regarding PharmD curricula included recommendations to inform didactic and experiential education. Conclusions The results support the need for reevaluation of oncology education in PharmD curricula. Further studies could explore specific aspects of didactic curriculum that impact the level of student preparedness, and which elements encourage a student pharmacist to seek further training in the oncology field.

Intercostal nerve cryoblation during the Nuss procedure for pectus excavatum decreases pain, opiate requirement, and hospital length of stay (LOS) compared to thoracic epidural analgesia. However, long-term complications of cryoablation, including neuropathic pain development, are not well studied. We conducted a multi-institutional retrospective review of patients following intercostal nerve cryoablation during Nuss bar insertion (11/2015–7/2018). Patients completed the Leeds Assessment of Neuropathic Symptoms and Signs, a validated questionnaire for detecting neuropathic symptoms. Primary outcome was neuropathic pain development. Secondary outcomes included duration of chest numbness and LOS. T test was performed; p

PurposeMinimally-invasive repair of pectus excavatum by the Nuss procedure is associated with significant postoperative pain, prolonged hospital stay, and high opiate requirement. We hypothesized that intercostal nerve cryoablation during the Nuss procedure reduces hospital length of stay (LOS) compared to thoracic epidural analgesia.DesignThis randomized clinical trial evaluated 20 consecutive patients undergoing the Nuss procedure for pectus excavatum between May 2016 and March 2018. Patients were randomized evenly via closed-envelope method to receive either cryoanalgesia or thoracic epidural analgesia. Patients and physicians were blinded to study arm until immediately preoperatively.SettingSingle institution, UCSF-Benioff Children's Hospital.Participants20 consecutive patients were recruited from those scheduled for the Nuss procedure. Exclusion criteria were age

Ceftolozane/tazobactam (C/T) has emerged as a potential agent for the treatment of extensively drug-resistant (XDR) Pseudomonas aeruginosa infections. As it is a time-dependent antimicrobial, prolonged infusion may help achieve pharmacokinetic/pharmacodynamic (PK/PD) targets. To compare alternative steady-state concentrations (Css) of C/T in continuous infusion (CI) against three XDR P. aeruginosa ST175 isolates with C/T minimum inhibitory concentration (MIC) values of 2 to 16 mg/L in a hollow-fiber infection model (HFIM). Duplicate 10-day HFIM assays were performed to evaluate Css of C/T in CI: one compared 20 and 45 mg/L against the C/T-susceptible isolate while the other compared 45 and 80 mg/L against the two C/T-non-susceptible isolates. C/T resistance emerged when C/T-susceptible isolate was treated with C/T in CI at a Css of 20 mg/L; which showed a deletion in the gene encoding AmpC β-lactamase. The higher dosing regimen (80 mg/L) showed a slight advantage in effectiveness. The higher dosing regimen has the greatest bactericidal effect, regardless of C/T MIC. Exposure to the suboptimal Css of 20 mg/L led to the emergence of C/T resistance in the susceptible isolate. Antimicrobial regimens should be optimized through C/T levels monitoring and dose adjustments to improve clinical management.

Ceftazidime-avibactam (CZA) has emerged as a promising solution to the lack of new antibiotics against Pseudomonas aeruginosa infections. Data from in vitro assays of CZA combinations, however, are scarce. The objective of our study was to perform a time-kill analysis of the effectiveness of CZA alone and in combination with other antibiotics against a collection of extensively drug-resistant (XDR) P. aeruginosa isolates. Twenty-one previously characterized representative XDR P. aeruginosa isolates were selected. Antibiotic susceptibility was tested by broth microdilution, and results were interpreted using CLSI criteria. The time-kill experiments were performed in duplicate for each isolate. Antibiotics were tested at clinically achievable free-drug concentrations. Different treatment options, including CZA alone and combined with amikacin, aztreonam, meropenem, and colistin, were evaluated to identify the most effective combinations. Seven isolates were resistant to CZA (MIC ≥ 16/4 mg/liter), including four metallo-β-lactamase (MBL)-carrying isolates and two class A carbapenemases. Five of them were resistant or intermediate to aztreonam (MIC ≥ 16 mg/liter). Three isolates were resistant to amikacin (MIC ≥ 64 mg/liter) and one to colistin (MIC ≥ 4 mg/liter). CZA monotherapy had a bactericidal effect in 100% (14/14) of the CZA-susceptible isolates. Combination therapies achieved a greater overall reduction in bacterial load than monotherapy for the CZA-resistant isolates. CZA plus colistin was additive or synergistic in 100% (7/7) of the CZA-resistant isolates, while CZA plus amikacin and CZA plus aztreonam were additive or synergistic in 85%. CZA combined with colistin, amikacin, or aztreonam was more effective than monotherapy against XDR P. aeruginosa isolates. A CZA combination could be useful for treating XDR P. aeruginosa infections, including those caused by CZA-resistant isolates. IMPORTANCE The emergence of resistance to antibiotics is a serious public health problem worldwide and can be a cause of mortality. For this reason, antibiotic treatment is compromised, and we have few therapeutic options to treat infections. The main goal of our study is to search for new treatment options for infections caused by difficult-to-treat resistant germs. Pseudomonas aeruginosa is a Gram-negative bacterium distributed throughout the world with the ability to become resistant to most available antibiotics. Ceftazidime-avibactam (CZA) emerged as a promising solution to the lack of new antibiotics against infections caused by P. aeruginosa strains. This study intended to analyze the effect of CZA alone or in combination with other available antibiotics against P. aeruginosa strains. The combination of CZA with other antibiotics could be more effective than monotherapy against extensively drug-resistant P. aeruginosa strains.

In vivo diagnostic imaging of bacterial infections is currently reliant on targeting their metabolic pathways, an ineffective method to identify microbial species with low metabolic activity. Here, we establish HS-198 as a small-molecule fluorescent conjugate that selectively targets the highly conserved bacterial protein HtpG (high-temperature protein G), within Borrelia burgdorferi, the bacterium responsible for Lyme disease. We describe the use of HS-198 to target morphologic forms of B. burgdorferi in both the logarithmic growth phase and the metabolically dormant stationary phase as well as in inactivated spirochetes. Furthermore, in a murine infection model, systemically injected HS-198 identified B. burgdorferi as revealed by imaging in postnecropsy tissue sections. These findings demonstrate how small-molecule probes directed at conserved bacterial protein targets can function to identify the microbe using noninvasive imaging and potentially as scaffolds to deliver antimicrobial agents to the pathogen.

Background Perforated appendicitis is the most common cause of intraabdominal abscess (IAA) in children. The optimal postoperative antibiotic regimen to reduce IAA has evolved in the last decade from triple-drug to 2-drug therapy (CM). Recent retrospective studies show decreased infectious complications with monotherapy PT. To date prospective comparative data are lacking. Therefore, a prospective randomized trial comparing PT versus CM was conducted. Methods A multi-institutional prospective randomized trial was performed in children with perforated appendicitis comparing postoperative antibiotic regimens PT or CM. The primary outcome was 30-day postoperative IAA formation. Perforation was strictly defined as a hole in the appendix or fecalith in the abdomen, documented with intraoperative photographs. Results One hundred sixty-two patients were enrolled during the study period. No differences in age, weight, or duration of presenting symptoms were identified. In addition, length of stay, duration of intravenous antibiotic treatment, discharge oral antibiotic treatment, and antibiotic-related complications did not differ between groups. Compared to the CM group, the PT group had significantly lower IAA rate [6.1% vs 23.8%, odd ratio (OR) 4.80, P = 0.002], lower postoperative computed tomography imaging rate (13.9% vs 29.3%, OR 2.57, P = 0.030), and fewer emergency room visits (8.8% vs 26.4%, OR 3.73, P = 0.022). Multivariate logistic regression analysis found the use of CM versus PT (OR 9.21, P = 0.021) to be the most significant predictor for developing IAA. Conclusions In children with perforated appendicitis, postoperative monotherapy with PT is superior to standard 2-drug therapy with CM and does not increase antibiotic-related complications or antibiotic exposure duration.

Background and objectives Patients undergoing cement fixation for hip arthroplasty are at increased risk of developing bone cement implantation syndrome (BCIS). We sought to determine: what is the occurrence of BCIS in patients with cancer after hip arthroplasty? What are the risk factors in patients with cancer for the development of this syndrome? What is the outcome for patients with cancer having BCIS? Methods We identified 374 patients with cancer who underwent cemented hip arthroplasty between 2010 and 2014. Patient characteristics, operative variables, and outcomes were collected. Results BCIS occurred in 279 (75%) patients. A total of 353 (94%) patients had bone metastases and 179 (48%) patients had lung metastases at the time of surgery. Age greater than 60 (hazard ratio [HR] 2.09, P = .02) and the presence of lung metastases (HR 1.77, P = .019) were associated with increased risk of BCIS. Increased perioperative use of vasopressors (HR 1.72, P = .023) and increased hospital stay beyond 10 days (HR 2.67, P = .003) was associated with BCIS. Conclusions BCIS is a frequent clinical event in patients with cancer undergoing femoral cemented arthroplasty with increased risk for patients over age 60 and those with compromised lung function due to lung metastases and lung cancer. Patients who develop BCIS are more likely to require longer postoperative hospitalization. Careful preoperative assessment and intraoperative communication are crucial steps to reduce the consequences of BCIS.

Purpose This quality improvement project aimed to assess the elements of the current workflow process that meets requirements for transitions of care from a family medicine inpatient to outpatient service following the Transitional Care Management (TCM) program developed by the Centers for Medicare & Medicaid Services. The purpose of the study was to assess the current family medicine workflow and determine whether the current process meets the criteria for billing and calculate the potential loss of reimbursement. Methods Interviews with key personnel, review of practice policies, and a retrospective chart review were performed for clinic patients discharged from the inpatient to outpatient family medicine service. Results A total of 37 patients met inclusion criteria for the chart review. Of these, 8% of patients seen at the outpatient clinic met all criteria for TCM. Potential reimbursement for those who met TCM criteria was $293.14 USD; the estimated potential TCM reimbursement for patients not meeting criteria was $1997.76 USD. Conclusion Standardized, team-based TCM services have shown to decrease readmission rates for high-risk patient populations. Results from this project identified processes in place at the family medicine practice to facilitate the development of a standardized transitional care service which could meet both TCM and best practice models.

Background: Past research has focused on understanding influenza vaccine acceptance in non-Hispanic white populations; however, research on the social causes of influenza vaccine acceptance rates in Hispanic populations is slowly developing. Objective: The purpose of this study was to assess theoretically driven predictors (i.e. attitudes, perceptions, behaviors, etc.) on influenza and the intention to vaccinate. Methods: A survey was administered to assess predictors of intentions to receive the influenza vaccine. The survey included items adapted from the National Flu Survey. Results: Key constructs common in models of health behaviors emerged as predictors of behavioral intentions to receive the flu vaccine. Recent vaccination within the past year ( P < 0.001), perceived effectiveness of the flu vaccine ( P < 0.004), and perceived safety of the flu vaccine ( P = 0.009) were predictors of intentions to vaccinate. Exploratory analyses revealed that government distrust was a statistically significant predictor of intentions to vaccinate ( P = 0.044). Conclusion: The above results have important implications for health-care providers and public health educators. The better we understand the relationship between theoretically driven predictors and vaccine behaviors, the more educators and health-care providers can focus on meaningful, culturally sensitive, targeted-vaccine education.

Benign thyroid disease in children represents a wide spectrum of disease. While most benign thyroid disorders may be either monitored or managed medically, surgery is an important treatment, particularly for Graves’ disease or large multinodular goiters. Multidisciplinary teams including specialists in pediatric endocrinology, genetics, radiology, pathology and surgery at high volume centers offer the safest and most effective management.

Background Policing is a stressful occupation. Most research examining police stress focuses solely on patrol officers, and often focuses on black and white officers only. Further, organizational sources of stress tend to be more important for police officers generally. Aims To explore sources and severity of stress in a predominately Hispanic, mid-sized, Southwestern police department. Methods A cross-sectional study of 147 police officers of all ranks was conducted. A modified version of the Police Stress Survey (PSS) was administered during daily briefings. Logistical regression models were utilized to examine predictors of stress based on gender, race/ethnicity, rank and tenure. Results Overall, occupational stressors proved more prominent than organizational stressors, contrary to previous research. Additionally, race/ethnicity was predictive of both total stress and occupational stress; rank was predictive of organizational stress; and tenure in law enforcement was predictive of occupational stress. Conclusions The sources of stress that impact police officers may be shifting as the climate surrounding law enforcement changes. What is stressful for officers depends on the race/ethnicity, rank and tenure of the officer within the organizational structure. Potential interventions designed to alleviate stress should be mindful of the variety of needs within any given department.

BACKGROUND/PURPOSE: The differential diagnosis for prenatal suprarenal masses (SRM) is broad and includes neuroblastoma, adrenal hemorrhage and subdiaphragmatic extralobar pulmonary sequestration (SEPS). We sought to elucidate the appropriate postnatal management for fetuses found to have a SRM. METHODS: We conducted a retrospective review of patients prenatally diagnosed with SRM at our institution between 1998 and 2018. Prenatal characteristics, imaging, and neonatal outcomes were collected. We also performed a PubMed literature search and pooled analysis of all patients with a prenatally diagnosed SRM previously described in the literature. RESULTS: The literature review yielded 32 studies, of which 19 were single case reports. In our case series, twelve patients were included. Seven patients were delivered vaginally, one was terminated. Postnatal diagnoses included: SEPS (n=5), adrenal hemorrhage (n=3), polycystic kidney (n=2), splenic cyst (n=1), and unknown for one patient. All but two of the final diagnoses had been on the initial diagnostic differential. With the exception of the terminated fetus, all remain alive today. On pooled analysis, patients who underwent operative management were diagnosed later 32 weeks versus 24 weeks) and had a significant predominance of left-sided lesions (59.5% vs. 39.2%). The published literature demonstrates a trend towards observation versus resection over the past 30 years. CONCLUSIONS: Patients prenatally diagnosed with a SRM have an excellent prognosis. Our series demonstrates a high incidence of SEPS, which were all resected, and adrenal hemorrhage, which were observed with repeat imaging. These patients can be followed with serial postnatal ultrasounds to determine the diagnosis prior to deciding the appropriate treatment.

Our objective was to analyze in vitro the persistence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in the packaging material of the drugs dispensed to hospital wards. Additionally, to evaluate if the protection with a double plastic bag prevents the contamination of the medication dispensed to an intensive care unit (ICU).On the first part, different materials containing different drugs within an ICU were sampled to confirm the lack of contamination by SARS-CoV-2. The confirmation of the virus was performed using real time reverse transcription polymerase chain reaction. As a control group, in the microbiology laboratory we inoculated the virus into the different surfaces containing the same drugs included in the first part. Samples were obtained with a sterile swab at 3, 6, 8, 10, 14, 21, and 30 days after inoculation and analyzed through real time reverse transcription polymerase chain reaction.None of the studied materials containing the drugs within an ICU was contaminated by SARS-CoV-2. In the second part, SARS-CoV-2 was found in all surfaces for up to 30 days.The use of double-bag unit-dose system to deliver medication in a pandemic seems effective to prevent the potential transmission of SARS-CoV-2. A striking SARS-CoV-2 RNA stability of up to 30 days was found in the surfaces containing the drugs.

Fetal surgery corrects severe congenital anomalies in utero to prevent their severe consequences on fetal development. The significant risk of open fetal operations to the pregnant mother has driven innovation toward minimally invasive procedures that decrease the risks inherent to hysterotomy. In this article, we discuss the basic principles of minimally invasive fetal surgery, the general history of its development, specific conditions and procedures used to treat them, and the future of the field.

We describe our experience using a modified suture fistula technique for addressing tension in longer gap esophageal atresia (EA). Esophagoesophagopexy (EEP) is the tacking of the proximal and distal ends of esophageal pouches without formal anastomosis. In this retrospective cohort, we review the outcomes of patients with EA after EEP. We reviewed the operative reports of EA cases treated at our institution from 1997 to 2016 and identified all patients described as having EEP. Of 129 EA cases, five patients underwent EEP. Formal anastomosis was not done due to patient’s instability, prematurity, or long gap. Median birth weight was 1.4 kg (0.6–2.2 kg), and median gestational age at birth was 29 weeks (25–34 weeks). Age at time of EEP ranged 0–5 months. Esophagoesophageal fistula was confirmed in three patients. All three had strictures requiring weekly dilations. One of these patients died. The two surviving patients underwent fundoplication. We describe an alternative technique for esophageal anastomosis in patients for whom a standard anastomosis is not possible. EEP can lead to a functional anastomosis through fistulization and avoid the morbidity of multiple thoracotomies and lengthening procedures. Families should be educated on the potential need for dilations and antireflux procedures.

Anesthesiologists have a high prevalence of burnout with adverse effects on professionalism and safety. The objective of this study was to assess the impact of an interactive anesthesiology educational program on the wellness of anesthesia providers and their children, as assessed by a modified Professional Fulfillment Index.Prospective observational study.Perioperative area.Thirty clinicians participated in the program. Twenty respondents, representing 67% of participants and each corresponding to a parent and their child or children, completed the post-event survey.An interactive anesthesiology educational program incorporating children, between the ages of five and eighteen years old, of anesthesia providers was held in the perioperative area. The program was held over four hours and was comprised of four sessions including pediatric anesthesia, neuroanesthesia, airway, and ultrasound stations.Anesthesia providers and their children were administered a post-event assessment, including a modified Professional Fulfillment Index and satisfaction survey.All twenty (100%) of respondents indicated it was "very true" or "completely true" that their child was happy with the program, and that it was worthwhile and satisfying to both the anesthesia provider and their child. Nineteen (95%) of reporting participants indicated it was "very true" or "completely true" that it was meaningful to have the department host such a program and 17 (85%) respondents felt their child now better understands the anesthesia work of the parent. All clinician volunteers indicated it was "very true" or "completely true" that they were contributing professionally during the program in ways that they valued most.An interactive educational wellness initiative provides an effective and feasible method for increasing professional fulfillment and satisfaction among anesthesia providers while educating our youngest generation of learners. Implementation of such a program may also occur with modifications such as televideo to maintain COVID-19 precautions.

Background Extracorporeal membrane oxygenation (ECMO) artificially supports respiratory and cardiac function when conventional techniques fail. ECMO has been described as a treatment modality for acute pulmonary and cardiac failure following orthotopic liver transplantation (OLT). Here, we present a series of adult OLT recipients placed on ECMO after transplantation for both respiratory and cardiac indications and review the literature on the role of ECMO in the setting of OLT. Methods For the patient series, we cross-referenced all patients who underwent OLT at our institution between 2007 and 2018 with the ECMO database of our institution and described these cases. For the literature review, we identified cases and series that described the use of ECMO after liver transplantation in adult recipients. Results A total of 1792 patients underwent OLT. Eight patients were placed on ECMO (0.4%), 5 men and 3 women aged 28 to 68 years (4 venovenous and 4 venoarterial). Three of (38%) 8 patients survived to discharge and are alive today. In the literature, we identified 3 series and 12 case reports of ECMO following OLT, with the majority of the literature derived from the Asian OLT experience. Conclusions ECMO following liver transplantation should be considered as a viable rescue strategy in patients with severe cardiopulmonary failure. ECMO is particularly effective if the cause of cardiopulmonary failure is recognized promptly and is thought to be transient. This is the largest series in the United States and demonstrates a 38% survival rate, which is comparable to other reports in the literature from Asia.

Streptococcus pyogenes infections are common in the pediatric population in the form of tonsillopharyngitis or cutaneous disease. Less frequently, it presents with invasive forms such as pyomyositis or necrotizing fasciitis. Mortality in children is of 7 %. We present an 11-year-old patient with a subscapular venolymphatic malformation who consulted for febrile syndrome associated with pain and an acute increase in the size of the lesion. During the hospitalization, diagnosis of pyomyositis due to Streptococcus pyogenes was reached.Las infecciones por Streptococcus pyogenes son frecuentes en la población pediátrica en forma de faringoamigdalitis o cuadros cutáneos. Con menor frecuencia, se presenta con formas invasivas, tales como piomiositis o fascitis necrotizante. La mortalidad en niños reportada en estas últimas es de un 7,69 %. Se presenta a una paciente de 11 años con malformación venolinfática subescapular que consultó por síndrome febril asociado a dolor y aumento agudo del tamaño de la lesión. Durante la internación, se arribó al diagnóstico de piomiositis por Streptococcus pyogenes.

Background: Extensively drug-resistant (XDR) Pseudomonas aeruginosa (P. aeruginosa) and particularly P. aeruginosa high-risk clones, are of growing concern because treatment options are limited. For years, colistin monotherapy has been the only available treatment, but is well known that is not an optimal treatment. A combination of colistin with another antibiotic could be a possible therapeutic option. Objectives: This study aimed to investigate effective antibiotic combinations against 20 XDR P. aeruginosa isolates obtained in a Spanish multicentre study (2015). Methods: Forty-five checkerboards with six antipseudomonal antibiotics (amikacin, aztreonam, ceftazidime, meropenem, colistin, and ceftolozane/tazobactam) were performed to determine whether combinations were synergic or additive by fractional inhibitory concentration indices. On average, 15 different regimens were evaluated in duplicate against the three most prevalent high-risk clones (ST175, ST235, ST111) by time-kill analyses over 24 h. The combination showing synergism in the three high-risk clones was validated in all studied XDR isolates. Results: In time-kill curves, the untreated control failed, as did each study regimen when administered alone. Two combinations were synergistic in the three high-risk clones that were initially studied: amikacin plus ceftazidime and colistin plus meropenem, with the second being the most effective combination. The efficacy of colistin plus meropenem was then tested in all 20 isolates. A synergistic bacterial density reduction for the duration of the study occurred in 80% of the entire XDR collection. Conclusions: These data suggest that colistin plus meropenem may be a useful combination for the treatment of infections due to XDR P. aeruginosa, including high-risk clones, which warrants evaluation in a clinical trial. (C) 2019 International Society for Antimicrobial Chemotherapy. Published by Elsevier Ltd. All rights reserved.

OBJECTIVES: The aim of this study was to assess the indirect effect of paediatric 13-valent pneumococcal conjugate vaccine (PCV13) vaccination on people ≥65 years of age with invasive pneumococcal disease (IPD) in Catalonia and to determine factors predictive of mortality. METHODS: During 2014-2016, 1285 IPD cases were reported to the Public Health Agency of Catalonia. The indirect effect of paediatric PCV13 vaccination was calculated by comparing the incidence rate (IR) in 2016 (PCV13 year) with that in 2009 (pre-PCV13). Predictors of mortality were determined using multivariate logistic regression. RESULTS: Comparing 2016 and 2009, IPD decreased by 19% (IR 40.1 and 32.5 per 100 000 person-years, respectively). PCV13 serotypes decreased by 57% (IR 23.7 and 10.1), while non-PCV13 serotypes increased by 36% (IR 16.4 and 22.4). During 2014-2016, the mortality rate was 17.5%, and mortality was associated with age ≥85 years (adjusted odds ratio (aOR) 2.91, 95% confidence interval (CI) 1.89, 4.48), meningitis (aOR 2.29, 95% CI 1.25, 4.19), non-focal bacteraemia (aOR 3.73, 95% CI 2.00, 6.94), and ≥1 high-risk condition (aOR 1.89, 95% CI 1.08, 3.32). PPV23non13 serotypes were associated with lower mortality than PCV13 serotypes (aOR 0.54, 95% CI 0.34, 0.86). CONCLUSIONS: The incidence of IPD in people ≥65 years of age decreased after the introduction of paediatric PCV13, and this was due to a reduction in PCV13 serotypes, although an increase in non-PCV13 serotypes was observed. Mortality was associated with age, meningitis, non-focal bacteraemia, ≥1 high-risk condition, and PCV13 serotypes. This study was partially supported by SpIDnet (Assessing the impact of vaccination with conjugate vaccines on the epidemiology of invasive pneumococcal disease in Europe), a network funded by the European Centre for Disease Prevention and Control ( ECDC/2015/031 ); the Catalan Agency for the Management of Grants for University Research ( AGAUR Grant number 2017/SGR 1342 and 2017/SGR 0742 ); the Centro de Investigación Biomédica en Red de Epidemiología y Salud Pública ( CIBERESP CB06/02/0076 and CB15/00067 ); and the Centro de Investigación Biomédica en Red de Enfermedades Respiratorias ( CIBERES CB06/06/0037 ). Sí

Patients with peritoneal carcinomatosis (PC) of gastric origin have a poor prognosis of life with an average survival of 1-3 months. Systemic chemotherapy has improved the survival of those patients with gastric metastatic cancer at 7-10 months. However, this benefit could not be reproduced in those patients with PC. The current literature for the use of hyperthermic intraperitoneal chemotherapy (HIPEC) for gastric PC has significant variation related to patient selection, treatment intent (palliative vs. attempt at curative treatment), surgical technique, intraperitoneal chemotherapy agent utilized, and systemic chemotherapy administered adjuvantly. From the perspective of patient selection for cytoreduction and HIPEC, patients with extensive PC are not candidates. In addition, unresectable location would make a patient a poor candidate for cytoreduction and HIPEC. Optimally, those with positive peritoneal cytology alone could benefit most. However, the role of cytoreductive surgery and HIPEC in patients with PC of gastric origin has not yet been clarified.Los pacientes con carcinomatosis peritoneal (CP) de origen gástrico tienen un mal pronóstico de vida, con una supervivencia media de 1 a 3 meses. La quimioterapia sistémica ha mejorado la supervivencia de los pacientes con cáncer gástrico metastásico a los 7-10 meses. Sin embargo, este beneficio no se ha podido reproducir en los pacientes con CP. En cuanto a lo relacionado con la literatura actual para el uso de HIPEC (hyperthermic intraperitoneal chemotherapy) en la CP de origen gástrico, existe una variación significativa en la selección de pacientes, la intención de tratamiento (paliativo frente a intento de tratamiento curativo), la técnica quirúrgica, el agente quimioterapéutico intraperitoneal utilizado y la quimioterapia sistémica adyuvante administrada. Desde la perspectiva de la selección de pacientes para citorreducción y tratamiento con HIPEC, los pacientes con CP extensa no son candidatos. Además, lesiones irresecables por su localización harían al paciente un pobre candidato para citorreducción y tratamiento con HIPEC. De manera óptima, aquellos pacientes con citología peritoneal positiva en ausencia de CP son quienes más podrían beneficiarse. Sin embargo, el papel de la cirugía citorreductora y del tratamiento con HIPEC en los pacientes con CP de origen gástrico aún no ha sido esclarecido.

Objective To create culturally appropriate colonoscopy education services and handouts for the Hispanic population undergoing a colonoscopy procedure to ensure correct bowel preparation. Setting Project DIS-IMPACT was conducted at a federally qualified, nonprofit community health center along the United States–Mexico border. Practice description Patients identified in the study included those who were at least 50 years of age and had not received a colonoscopy between July 1, 2012, and February 28, 2013. Participants included uninsured Hispanics, able to speak English or Spanish, registered with the clinic, and referred by a provider working at the health center. Practice innovation The process of evaluating and developing colonoscopy screening education included creating a standing order, developing medication education handouts in both English and Spanish, and documenting areas of improvement at monthly pharmacy staff meetings. Main outcome measures Outcomes of the program included a descriptive statistics analysis of the served population and culturally appropriate counseling recommendations used in the medication (polyethylene glycol electrolyte solution) education handouts. Results A total of 143 participants received colonoscopy, including 49 men and 94 women. As the classes were conducted, the patient information handouts were adapted accordingly based on culturally appropriate recommendations from participants and providers. Conclusion Pharmacists played a unique role in providing culturally appropriate precolonoscopy education material to assist in successful colonoscopy preparation.

Objective To describe the development and implementation of a human papillomavirus (HPV) vaccine patient assistance program (PAP) for university students, and to acquire information on the number who accessed the program and completed the series. Setting University of Texas at El Paso University Student Health Clinic Pharmacy, Fall 2011–Spring 2014. Practice description A community pharmacy located within the university student health clinic providing services to an underinsured student population. Practice innovation Existing evidence shows the benefit of using PAP in community pharmacies but is nonspecific regarding the use of PAP for vaccines in an uninsured and underinsured Hispanic student population. The implementation of this unique HPV vaccine program in a community setting aims to increase awareness, access, and rates. Main outcome measures Primary measures included results from a needs-assessment questionnaire that were used to implement the HPV vaccine program. After implementation, utilization data were collected on the number of students who qualified and enrolled in the HPV PAP and the number of students who completed the HPV series. Results The preliminary data from a needs assessment indicated that a majority (72.1%, n = 80) of students did not understand how HPV is transmitted. A total of 89 students qualified for PAP. The majority were women (81%). A total of 71 students (79.8%) received their second dose and 43 (48.3%) completed the series. Conclusions Although pharmacists continue to provide vaccine services, minorities such as the Hispanic population continue to be underimmunized. Students may not be taking the proper precautions to prevent the acquisition of HPV. For these reasons services such as this HPV vaccine program are warranted. Pharmacists need to continue to educate and advocate on the importance of vaccines and how they prevent disease.

Cryoanalgesia prevents pain by freezing the affected peripheral nerve. We report the use of intraoperative cryoanalgesia during the Nuss procedure for pectus excavatum and describe our initial experience, modifications of technique, and lessons learned.We retrospectively reviewed the medical records of patients who received cryoanalgesia during the Nuss procedure between June 1, 2015, and April 30, 2016, at our institutions and analyzed modifications in surgical technique during this early adoption period.Eight male and two female patients underwent the Nuss procedure with cryoanalgesia. The mean postoperative length of stay (LOS) was 2days (range 1-3). Average inpatient pain scores were 3.4, 3.2, and 4.6 on postoperative days 1-3, respectively (N=10, 7, and 2). At a 1-week postoperative visit, mean pain score was 1.1 (N=6). Compared to the preceding 15 Nuss patients at our institution, who were treated with a thoracic epidural, postoperative LOS was significantly shorter with cryoanalgesia (2.0±0.82 vs. 6.3±1.3days, P0.001). We modified our technique for patient habitus and adopted single-lung ventilation for improved visualization.Cryoanalgesia may be the ideal pain management strategy for Nuss patients because it is effective and long lasting. Intraoperative application is easily integrated into the Nuss procedure.Treatment study: case series; Evidence level IV.

Parkinson's disease (PD) is the second most common neurodegenerative disorder. It is characterised by selective loss of dopaminergic neurons in the substantia nigra pars compacta, which results in dopamine depletion, leading to a number of motor and non-motor symptoms.In recent years, the development of new animal models using nuclease-based genome-editing technology (ZFN, TALEN, and CRISPR/Cas9 nucleases) has enabled the introduction of custom-made modifications into the genome to replicate key features of PD, leading to significant advances in our understanding of the pathophysiology of the disease.We review the most recent studies on this new generation of in vitro and in vivo PD models, which replicate the most relevant symptoms of the disease and enable better understanding of the aetiology and mechanisms of PD. This may be helpful in the future development of effective treatments to halt or slow disease progression.

Objectives Most national-level data regarding lesbian, gay, bisexual, transgender, queer/questioning, intersex, and allies (LGBTQIA) immunizations are limited. The primary objective of this study was to identify factors that influence behaviors, attitudes, and perceptions toward the uptake of the influenza vaccine within the Hispanic LGBTQIA community. Methods This was a prospective survey that assessed vaccine acceptability and practices regarding the influenza vaccine within the LGBTQIA community. Collection of data occurred through the use of social media platforms from July 2016 to May 2018. A total of 126 participants (mean age 32.03 ± 11.68 years) completed a 15-minute, 26-item, English/Spanish survey. A number of outcome measures assessed perceptions of vaccine effectiveness and safety. In addition, perceived severity of influenza symptoms and perceived susceptibility to contract influenza was assessed. Results A logistic regression model assessed the impact of several factors on influenza vaccine uptake. Perceived susceptibility of contracting influenza from the vaccine (P = 0.015) and perceived ease of receiving the influenza vaccine (P = 0.005) were the strongest predictors of vaccine uptake. Results showed no association between disclosure of sexual orientation and influenza immunization uptake (χ2 = 3.55; P = 0.17). Exploratory analyses revealed that non-Hispanic patients were more likely to perceive that their health care providers were aware of their sexual orientation compared with Hispanic patients (χ2 = 8.66; P = 0.013). Conclusion Several factors emerged as predictors of influenza vaccine uptake in the LGBTQIA population. Further studies are needed to explore additional factors such as disclosure of sexual orientation and variation of uptake based on vaccine type (STD vs. non-STD vaccines).

IMPORTANCE: Sutureless gastroschisis repair offers an alternative to the traditional sutured method and has been associated with decreased intubation time. Published study results are inconsistent regarding the advantages of sutureless closure. OBJECTIVE: To compare the clinical outcomes of sutureless and sutured gastroschisis repair. DESIGN, SETTING, AND PARTICIPANTS: A single-center cohort review was performed of all consecutive patients (n = 97) who underwent gastroschisis repair from February 1, 2007, to April 30, 2017, at the University of California, San Francisco. Patients’ medical records were evaluated for clinical characteristics and outcomes. Cases with incomplete data during initial hospitalization were excluded. MAIN OUTCOMES AND MEASURES: Length of hospital stay, time to full enteral feeds, total parenteral nutrition duration, days requiring intravenous analgesia, days intubated, wound infection rate, antibiotic treatment duration, rate of umbilical hernias that required an operation, and readmission rate. RESULTS: In total, 97 patients (47 [48%] were female and 50 [52%] were male with a mean [SD] age of 2.8 [2.8] days) underwent gastroschisis repair, of which 7 were excluded for incomplete medical record. Of the 90 patients included in the study, 50 (56%) underwent sutured closure and 40 (44%) underwent sutureless closure. No statistical difference was found between the sutured and sutureless groups in length of hospital stay (mean [SD] days, 43.9 [40.4] vs 36.7 [21.2]; P = .71), time to full enteral feeds (mean [SD] days, 31.4 [20.2] vs 27.9 [17.3]; P = .22), total parenteral nutrition duration (mean [SD] days, 33.5 [29.8] vs 27.4 [18.2]; P = .23), wound infection rates (14 [28%] vs 10 [25%]; P = .81), and readmission rates (5 [10%] vs 7 [18%]; P = .36). The sutureless group, compared with the sutured group, had substantially fewer days receiving antibiotics (mean [SD], 7.2 [6.4] vs 12.4 [13.2]; P = .003), fewer days intubated (mean [SD], 2.8 [3.3] vs 6.8 [1.3]; P = .001), fewer days receiving intravenous analgesia (mean [SD], 4.2 [4.0] vs 7.1 [4.5]; P = .003), and fewer patients that required silo reduction (25 [63%] vs 48 [96%]; P

The abrupt onset of respiratory failure secondary to asthma, known as acute asphyxial asthma (AAA) in adults, is uncommonly reported in children. Here, we report a case of a child with the acute onset of respiratory failure consistent with AAA complicated by the finding of a neck mass during resuscitation. This 11-year-old boy with a history of asthma initially presented in respiratory failure with altered mental status after the complaint of difficulty in breathing minutes before collapsing at home. Initially, his respiratory failure was thought to be secondary to status asthmaticus, and treatment was initiated accordingly. However, a neck mass noted during the resuscitation was cause for concern, and other etiologies for his respiratory failure were considered, including an airway obstructing neck mass. After pediatric surgery and anesthesia consultation for intubation and possible tracheostomy placement, general anesthesia was induced in the operating room with an inhaled anesthetic, with prompt resolution of the bronchspasm and decompression of the neck mass. Review of the imaging and clinical course ultimately yielded a diagnosis of cervical lung herniation as the etiology of his neck mass. We report this case of AAA and cervical lung herniation and a review of the literature of these 2 uncommon phenomena in children.

Parkinson's disease is a progressive neurodegenerative disorder characterised by a loss of dopaminergic neurons in the substantia nigra pars compacta, which results in a significant decrease in dopamine levels and consequent functional motor impairment.Although its aetiology is not fully understood, several pathogenic mechanisms, including oxidative stress, have been proposed. Current therapeutic approaches are based on dopamine replacement drugs; these agents, however, are not able to stop or even slow disease progression. Novel therapeutic approaches aimed at acting on the pathways leading to neuronal dysfunction and death are under investigation.In recent years, such natural molecules as polyphenols, alkaloids, and saponins have been shown to have a neuroprotective effect due to their antioxidant and anti-inflammatory properties. The aim of our review is to analyse the most relevant studies worldwide addressing the benefits of some phytochemicals used in in vitro models of Parkinson's disease.

Single-site umbilical incision laparoscopic surgery (SSULS) is increasingly being used to treat a variety of childhood surgical diseases. Existing SSULS approaches have inefficient triangulation and poor ergonomics. In an effort to overcome these shortcomings, magnet-assisted laparoscopy was developed. Specialized magnetic graspers are introduced through a standard 12-mm port and are controlled by a powerful external magnet. This study is a retrospective analysis of all magnet-assisted laparoscopic operations performed at the Fundacion Hospitalaria Private Children's Hospital from September 2009 to January 2011. Outcomes include demographics, diagnosis, operative time, intraoperative complications, and conversion rates. Forty-four magnet-assisted laparoscopic operations were performed. The operations included 23 appendectomies, 8 cholecystectomies, 3 Nissen fundoplications, 2 gastrojejunostomies, 2 splenectomies, 2 ovarian tumor/cyst resections, 1 retroperitoneal lymphangioma resection, 1 left adrenalectomy, 1 total abdominal colectomy and 1 pulmonary wedge resection. The mean operative times for the most commonly performed operations were 61 minutes for appendectomy and 93 minutes for cholecystectomy. The operations were classified as follows: Group I, adjunct to conventional laparoscopy (5 operations); Group II, adjunct to multiple-access umbilical laparoscopy (11 operations); and Group III, true single-port laparoscopy (28 operations). Among Group II/III operations, 6 operations required 1 additional port outside the umbilicus. No operations required more that 1 additional port, and no operations were converted to the open technique. There were no intraoperative complications. Magnet-assisted laparoscopic surgery is safe and effective in children. The use of magnetic graspers improves triangulation and ergonomics while reducing the number and size of abdominal incisions.

In situ hybridization methods enable the mapping of mRNA expression within intact biological samples. With current approaches, it is challenging to simultaneously map multiple target mRNAs within whole-mount vertebrate embryos, representing a significant limitation in attempting to study interacting regulatory elements in systems most relevant to human development and disease. Here, we report a multiplexed fluorescent in situ hybridization method based on orthogonal amplification with hybridization chain reactions (HCR). With this approach, RNA probes complementary to mRNA targets trigger chain reactions in which fluorophore-labeled RNA hairpins self-assemble into tethered fluorescent amplification polymers. The programmability and sequence specificity of these amplification cascades enable multiple HCR amplifiers to operate orthogonally at the same time in the same sample. Robust performance is achieved when imaging five target mRNAs simultaneously in fixed whole-mount and sectioned zebrafish embryos. HCR amplifiers exhibit deep sample penetration, high signal-to-background ratios and sharp signal localization.

Agonist-induced internalization of somatostatin receptors (ssts) determines subsequent cellular responsiveness to peptide agonists and influences sst receptor scintigraphy. To investigate sst2A trafficking, rat sst2A tagged with epitope was expressed in human embryonic kidney cells and tracked by antibody labeling. Confocal microscopical analysis revealed that stimulation with sst and octreotide induced internalization of sst2A. Internalized sst2A remained sequestrated within early endosomes, and 60 min after stimulation, internalized sst2A still colocalized with β-arrestin1-enhanced green fluorescence protein (EGFP), endothelin-converting enzyme-1 (ECE-1), and rab5a. Internalized 125I-Tyr11-SST-14 was rapidly hydrolyzed by endosomal endopeptidases, with radioactive metabolites being released from the cell. Internalized 125I-Tyr1-octreotide accumulated as an intact peptide and was released from the cell as an intact peptide ligand. We have identified ECE-1 as one of the endopeptidases responsible for inactivation of internalized SST-14. ECE-1-mediated cleavage of SST-14 was inhibited by the specific ECE-1 inhibitor, SM-19712, and by preventing acidification of endosomes using bafilomycin A1. ECE-1 cleaved SST-14 but not octreotide in an acidic environment. The metallopeptidases angiotensin-1 converting enzyme and ECE-2 did not hydrolyze SST-14 or octreotide. Our results show for the first time that stimulation with SST-14 and octreotide induced sequestration of sst2A into early endosomes and that endocytosed SST-14 is degraded by endopeptidases located in early endosomes. Furthermore, octreotide was not degraded by endosomal peptidases and was released as an intact peptide. This mechanism may explain functional differences between octreotide and SST-14 after sst2A stimulation. Moreover, further investigation of endopeptidase-regulated trafficking of neuropeptides may result in novel concepts of neuropeptide receptor inactivation in cancer diagnosis.

Although cell surface metalloendopeptidases degrade neuropeptides in the extracellular fluid to terminate signaling, the function of peptidases in endosomes is unclear. We report that isoforms of endothelin-converting enzyme-1 (ECE-1a-d) are present in early endosomes, where they degrade neuropeptides and regulate post-endocytic sorting of receptors. Calcitonin gene-related peptide (CGRP) co-internalizes with calcitonin receptor-like receptor (CLR), receptor activity-modifying protein 1 (RAMP1), beta-arrestin2, and ECE-1 to early endosomes, where ECE-1 degrades CGRP. CGRP degradation promotes CLR/RAMP1 recycling and beta-arrestin2 redistribution to the cytosol. ECE-1 inhibition or knockdown traps CLR/RAMP1 and beta-arrestin2 in endosomes and inhibits CLR/RAMP1 recycling and resensitization, whereas ECE-1 overexpression has the opposite effect. ECE-1 does not regulate either the resensitization of receptors for peptides that are not ECE-1 substrates (e.g., angiotensin II), or the recycling of the bradykinin B(2) receptor, which transiently interacts with beta-arrestins. We propose a mechanism by which endosomal ECE-1 degrades neuropeptides in endosomes to disrupt the peptide/receptor/beta-arrestin complex, freeing internalized receptors from beta-arrestins and promoting recycling and resensitization.

Calcitonin receptor-like receptor (CLR) and the receptor activity-modifying protein 1 (RAMP1) comprise a receptor for calcitonin gene-related peptide (CGRP). Although CGRP induces endocytosis of CLR/RAMP1, little is known about post-endocytic sorting of these proteins. We observed that the duration of stimulation with CGRP markedly affected post-endocytic sorting of CLR/RAMP1. In HEK and SK-N-MC cells, transient stimulation (10(-7) M CGRP, 1 h), induced CLR/RAMP1 recycling with similar kinetics (2-6 h), demonstrated by labeling receptors in living cells with antibodies to extracellular epitopes. Recycling of CLR/RAMP1 correlated with resensitization of CGRP-induced increases in [Ca(2+)](i). Cycloheximide did not affect resensitization, but bafilomycin A(1), an inhibitor of vacuolar H(+)-ATPases, abolished resensitization. Recycling CLR and RAMP1 were detected in endosomes containing Rab4a and Rab11a, and expression of GTPase-defective Rab4aS22N and Rab11aS25N inhibited resensitization. After sustained stimulation (10(-7) M CGRP, >2 h), CLR/RAMP1 trafficked to lysosomes. RAMP1 was degraded approximately 4-fold more rapidly than CLR (RAMP1, 45% degradation, 5 h; CLR, 54% degradation, 16 h), determined by Western blotting. Inhibitors of lysosomal, but not proteasomal, proteases prevented degradation. Sustained stimulation did not induce detectable mono- or polyubiquitination of CLR or RAMP1, determined by immunoprecipitation and Western blotting. Moreover, a RAMP1 mutant lacking the only intracellular lysine (RAMP1K142R) internalized and was degraded normally. Thus, after transient stimulation with CGRP, CLR and RAMP1 traffic from endosomes to the plasma membrane, which mediates resensitization. After sustained stimulation, CLR and RAMP1 traffic from endosomes to lysosomes by ubiquitin-independent mechanisms, where they are degraded at different rates.

Summary. Early virological response may predict outcome following treatment with peginterferon alpha-2a and ribavirin in patients chronically infected with hepatitis C virus (HCV). As total HCV core antigen may constitute an alternative direct marker to HCV RNA for assessing the levels of viraemia in such patients, we evaluated the correlation between HCV core antigen and HCV RNA, and whether HCV core antigen at baseline, 4 and 12 weeks after treatment could predict sustained virological response (SVR) to combined therapy, in comparison with HCV RNA. A total of 290 serum samples from 58 previously treatment naive chronic HCV patients were examined for HCV core antigen and HCV-RNA by means of quantitative HCV RNA when receiving combination therapy for the first time. SVR was significantly associated with basal HCV core antigen but not with HCV RNA. There was a good correlation between HCV core antigen and HCV RNA (r2 = 0.781). The negative predictive value of HCV core antigen testing in predicting nonresponse at weeks 4 and 12 were 75 and 100%, and for undetectable or a 2-log drop in HCV RNA were 69.6 and 75% respectively. HCV core antigen detection is quick, and easy to perform alternative to HCV RNA, and could be used as a marker of HCV viraemia for monitoring the progress of therapy.

Awareness with recall after general anesthesia is an infrequent, but well described, phenomenon that may result in posttraumatic stress disorder. There are no recent data on the incidence of this complication in the United States. We, therefore, undertook a prospective study to determine the incidence of awareness with recall during general anesthesia in the United States. This is a prospective, nonrandomized descriptive cohort study that was conducted at seven academic medical centers in the United States. Patients scheduled for surgery under general anesthesia were interviewed in the postoperative recovery room and at least a week after anesthesia and surgery by using a structured interview. Data from 19,575 patients are presented. A total of 25 awareness cases were identified (0.13% incidence). These occurred at a rate of 1-2 cases per 1000 patients at each site. Awareness was associated with increased ASA physical status (odds ratio, 2.41; 95% confidence interval, 1.04-5.60 for ASA status III-V compared with ASA status I-II). Age and sex did not influence the incidence of awareness. There were 46 additional cases (0.24%) of possible awareness and 1183 cases (6.04%) of possible intraoperative dreaming. The incidence of awareness during general anesthesia with recall in the United States is comparable to that described in other countries. Assuming that approximately 20 million anesthetics are administered in the United States annually, we can expect approximately 26,000 cases to occur each year.

The INNO-LiPA Mycobacteria Test (Innogenetics, N.V., Belgium) is a PCR-based reverse hybridization assay for the simultaneous identification of several mycobacterial species. We evaluated two simplified lysis methods for mycobacterial DNA release for application in the INNO-LiPA Mycobacteria Test. The two methods were based on either (i) heat treatment or (ii) sonication. Both methods were performed directly on 45 positive liquid cultures (MB-BacT, BioMérieux, Marcy l'Etoile, France) containing 17 different mycobacterial species. These two simple lysis procedures demonstrated similar effectiveness (100%) to that recommended by the manufacturer. They also significantly shortened the time required for mycobacterial DNA release.