Development of a Rapid and High-Throughput Multiplex Real-Time PCR Assay for Mycoplasma hominis and Ureaplasma Species.

Bacterial commensals of the human genitourinary tract, Mycoplasma hominis and Ureaplasma species (parvum and urealyticum) can be sexually transmitted, with the potential to cause nongonococcal urethritis, pelvic inflammatory disease, and infertility. Mycoplasma hominis and Ureaplasma species may also cause severe invasive infections in immunocompromised patients. Current culture-based methods for Mycoplasma/Ureaplasma identification are costly and laborious, with a turnaround time between 1 and 2 weeks. We developed a high-throughput, real-time multiplex PCR assay for the rapid detection of M. hominis and Ureaplasma species in urine, genital swab, body fluid, and tissue. In total, 282 specimens were tested by PCR and compared with historic culture results; a molecular reference method was used to moderate discrepancies. Overall result agreement was 99% for M. hominis (97% positive percentage agreement and 100% negative percentage agreement) and 96% for Ureaplasma species (96% positive percentage agreement and 97% negative percentage agreement). Specimen stability was validated for up to 7 days at room temperature. This multiplex molecular assay was designed for implementation in a high-complexity clinical microbiology laboratory. With this method, >90 samples can be tested in one run, with a turnaround time of 4 to 5 hours from specimen extraction to reporting of results. This PCR test is also more labor effective and cost-effective than the conventional culture-based test, thus improving laboratory efficiency and alleviating strains because of labor shortages.

African-American and Caucasian participation in postmortem human brain donation for neuropsychiatric research.

Increased African-American research participation is critical to the applicability and generalizability of biomedical research, as population diversity continues to increase both domestically and abroad. Yet numerous studies document historical origins of mistrust, as well as other barriers that may contribute to resistance in the African-American community towards participation in biomedical research. However, a growing body of more recent scientific evidence suggests that African-Americans value research and are willing to participate when asked. In the present study, we set out to determine factors associated with research participation of African-American families in postmortem human brain tissue donation for neuropsychiatric disorders as compared with Caucasian families, from same-day medical examiner autopsy referrals. We retrospectively reviewed brain donation rates, as well as demographic and clinical factors associated with donation in 1,421 consecutive referrals to three medical examiner's offices from 2010-2015. Overall, 69.7% of all next-of-kin contacted agreed to brain donation. While Caucasian families consented to donate brain tissue at a significantly higher rate (74.1%) than African-American families (57.0%) (p<0.001), African-American brain donation rates were as high as 60.5% in referrals from Maryland. Neither African-American nor Caucasian donors differed significantly from non-donors on any demographic or clinical factors ascertained, including age, sex, diagnosis of the donor, or in the relationship of the next-of-kin being contacted (p>0.05). However, Caucasian donors were significantly older, had more years of education, were more likely to be referred for study due to a psychiatric diagnosis, more likely to have comorbid substance abuse, and more likely to have died via suicide, as compared with African-American donors (p<0.05). When African-American participants are identified and approached, African-American families as well as Caucasian families are indeed willing to donate brain tissue on the spot for neuropsychiatric research, which supports the belief that African-American attitudes towards biomedical research may be more favorable than previously thought.

Contemporary epidemiology of infective endocarditis in patients with congenital heart disease: A UK prospective study.

OBJECTIVES: Infective endocarditis is a life-threatening complication of congenital heart disease (CHD), but there are few studies concerning the contemporary risk profile, preceding invasive procedures and outcomes in this patient population. The aim of this study was to investigate the epidemiology of infective endocarditis (IE) in patients with CHD. METHODS: Cases of IE in children and adults with CHD were prospectively recorded as part of the UK National Institute for Cardiovascular Outcomes Research (NICOR) National Congenital Heart Disease Audit. Patients were entered into the database between April 2008 and March 2016. RESULTS: Eight hundred episodes of IE were recorded in 736 patients with CHD. Sixty-five patients (9%) were infants (aged <1 year), 235 (32%) were children (aged 1-15 years), and 436 (59%) were adults (aged >15 years). The most common diagnoses were Tetralogy of Fallot (n = 150, 22.8%), ventricular septal defect (n = 129, 19.6%) and bicuspid aortic valve (n = 70, 10.7%). Dental procedures preceded 67 of 635 episodes (11%) of IE, and non-dental invasive procedures preceded 177 of 644 episodes (27.4%). The most common causative organisms were streptococci, accounting for 40% of cases. Overall in-hospital mortality was 6.7%. On multivariable analysis, adverse factors associated with in-hospital mortality were staphylococcal infection and presence of an underlying atrioventricular septal defect. CONCLUSIONS: Infective endocarditis in patients with CHD is an ongoing clinical challenge. In contemporary practice in tertiary congenital centers, 1 of 15 patients do not survive to hospital discharge. Streptococci remain the most common causative organism, and antecedent dental or medical procedures were undertaken in a significant minority in the 3 months before diagnosis. The presence of an atrioventricular septal defect or staphylococcal infection is associated with significantly increased risk of early mortality.

Reporting of vertebral fragility fractures: can radiologists help reduce the number of hip fractures?

Patients with osteoporotic vertebral fractures are at increased risk of hip fracture. In a cohort of hip fracture patients, many had previous imaging studies showing incidental vertebral fractures. Fifty-four percent of fractures were not reported by the radiologist, highlighting a missed opportunity for diagnosing and treating osteoporosis, thereby preventing further fractures. PURPOSE: Patients with osteoporotic vertebral fragility fractures (VFFs) are at increased risk of future fractures, including hip fractures. Treating osteoporosis in these patients has the potential to reduce the risk of subsequent hip fractures, which are associated with high morbidity, mortality and cost. In this retrospective cohort study, we investigated the reporting and follow-up of VFFs evident on imaging by radiologists at the John Radcliffe Hospital, Oxford. MATERIALS AND METHODS: Data from the local Fracture Liaison Service was used to case-find all incident hip fractures from 2013 presenting to the trust. We then identified patients who had also undergone a radiological procedure that included the thoracic and/or lumbar spine in the previous 6 years. All identified radiological images were re-examined for the presence of VFFs using the Genant semi-quantitative method. RESULTS: Seven hundred and thirty-two patients over the age of 50 with a hip fracture in 2013 were identified. One hundred and fifty-seven patients had previously undergone a radiological procedure involving the spine, and VFFs were identified in 65/157 (41%). Of these, only 30/65 (46%) were reported by a radiologist when the fracture was first visible. 32/35 (91%) of unreported VFFs were from imaging reported by non-musculoskeletal radiologists. Only 16/65 (25%) of patients with a VFF were documented as being on bone-specific therapy at the time of hip fracture. CONCLUSIONS: Our study highlights the under-reporting of osteoporotic vertebral fractures, particularly by non-musculoskeletal radiologists. Better systems for reporting and referring osteoporotic VFFs are necessary to increase the number of patients receiving appropriate osteoporosis treatment.

Cloud Computing for Pharmacometrics: Using AWS, NONMEM, PsN, Grid Engine, and Sonic.

Cloud computing allows pharmacometricians to access advanced hardware, network, and security resources available to expedite analysis and reporting. Cloud-based computing environments are available at a fraction of the time and effort when compared to traditional local datacenter-based solutions. This tutorial explains how to get started with building your own personal cloud computer cluster using Amazon Web Services (AWS), NONMEM, PsN, Grid Engine, and Sonic.

Fourier phase analysis of first-pass data: noninvasive detection of pulmonary sequestration.

Nuclear medicine Fourier phase analysis of first-pass data was used to evaluate blood flow to cystic lung masses in two children suspected of having pulmonary sequestrations. In both cases, the Fourier images provided a rapid, noninvasive, inexpensive analysis of the blood supply to the masses and permitted identification of the masses as pulmonary sequestrations. The analysis does not depend on the location of the mass and demonstrates the location of systemic rather than pulmonic arterial blood supply to the mass. In one case, Fourier analysis identified a second source of systemic blood supply not visible with other imaging modalities. Preoperative assessment of a cystic lung mass using Fourier analysis enables noninvasive classification of the mass as a pulmonary sequestration with systemic blood supply and aids the surgeon in resection.

A technique for preservation of the ileocecal valve in the neonatal short intestine.

We report a technique, appendiceal interposition, which permitted preservation of the ileocecal valve in an infant with a congenitally short intestine (jejunum, 12 cm; ileum, 1 cm). The procedure was performed on the first day of life in conjunction with jejunal lengthening by the Bianchi technique. The result was a small intestine of 21 cm in length with an intact ileocecal valve.

HCO emission from H II-molecular cloud interface regions.

A survey of well-known molecular clouds in the four strongest HCO NK-,K+ = 1(01)-0(00) hyperfine transitions has been carried out to determine the prevalence of HCO and to study its chemistry. HCO emission was observed in seven molecular clouds. Three of these, NGC 2264, W49, and NGC 7538, were not previously known sources of HCO. In addition, NGC 2024 and Sgr B2 were mapped and shown to have extensive HCO emission. The survey results show the HCO abundance to be enhanced in H II-molecular cloud interface regions and support a correlation between C+ and HCO emission. The strength of the HCO emission in NGC 2024 is interpreted in terms of this enhancement and the source structure and proximity to Earth.

Crucial bronchoscopic findings in esophageal atresia and tracheoesophageal fistula.

We performed a brief bronchoscopy in 42 newborn infants with esophageal atresia and tracheoesophageal fistula (TEF). The procedure was carried out in the operating room, usually just prior to thoracotomy for repair of the anomaly. Most infants had a gastrostomy tube placed initially. Observations included: (1) level of fistula, (2) presence of unusual variants, eg, double fistula, trifurcation fistula, (3) presence and severity of tracheobronchitis, and (4) position of the aortic arch. The endoscopic findings influenced the operative technique or management of 24 of the 42 infants (57%), including 13 infants (31%) with crucial findings which dictated a change in operative technique or management. Examples of the latter were an unsuspected cervical fistula associated with esophageal atresia, repaired by a cervical approach; other unusual variants of TEF (proximal fistula, trifurcation or quadrifurcation TEF), requiring specific dissection at thoracotomy; congenital stenosis of the right mainstem bronchus, requiring postoperative dilatation; or severe tracheobronchitis, which contraindicated thoracotomy. Observations at bronchoscopy accurately predicted the position of the aortic arch in ten recent infants, although the side of dominant pulsation was indeterminate in three other infants. Only one minor complication was attributable to the bronchoscopy. The procedure appeared to be safe and beneficial, and should be considered for all infants with esophageal atresia and TEF.

Multiple mating and siring success during natural oestrus in the ewe.

Ewes were each mated on four separate occasions, at 3, 9, 15 and 21 h after the start of oestrus and at each time by a different ram. The progeny were assigned to sires by blood typing, supplemented by resemblance between lambs and rams. The paternity of 64 lambs, born to 41 ewes, was established: 2 were conceived at a 3-h mating, 27 at 9h, 23 at 15 h and 12 at 21 h. The optimum time for a ram to inseminate, when in competition with others, is therefore 9-15 h after onset of oestrus, and this finding accords with behavioural observations. Ewes tended to lamb during the same half of the day as that when they had come into oestrus.

Balloon catheter dilatation of focal intestinal strictures following necrotizing enterocolitis.

Dilatation with a balloon catheter was successfully employed for 9 focal intestinal strictures which occurred in 5 infants following necrotizing enterocolitis. Eight of the 9 strictures were located in defunctionalized colon distal to an enterostomy; no infant had clinical intestinal obstruction. Because the dilatation achieved distal patency, subsequent closure of the enterostomy was accomplished without a formal laparotomy. The balloon dilatation technique may be valuable in the management of focal strictures that are not causing clinical intestinal obstruction.

Congenital primitive neuroectodermal tumor (neuroepithelioma) of the chest wall.

Primitive neuroectodermal tumor (neuroepithelioma) is a relatively common central nervous system tumor in children. Those arising from a peripheral nerve are extremely rare in childhood. There is only one reported case in 6-year-old where the tumor arose from the sciatic nerve. A case of neuroectodermal tumor of the chest wall, arising from the intercostal nerve, in a newborn is presented. The tumor metastasized to the brain. Prominent Homer-Wright rosettes, with central eosinophilic fibrillar substance similar to that seen in neuroepithelioma of the central nervous system, were present in the primary tumor and brain metastases. Ultrastructure, as revealed by transmission electron microscopy, is also described.

Nonrefluxing central venous catheters in children.

A central venous catheter occasionally is necessary in infants to maintain adequate nutritional support. Our experience shows that routine dressing changes are not necessary. Clinical studies of a slit-valve central venous catheter in which intraluminal thrombosis is eliminated are described. The combination of minimal dressing manipulation, adequate dressing protection, and the nonrefluxing slit-valve catheter have markedly reduced sepsis rates.

Studies with a slit-valve catheter for parenteral nutrition.

A new catheter for central venous total parenteral nutrition has a blunt occluded distal end and a longitudinally oriented slit one centimeter proximal. Studies in rats show that this slit-valve prevents both reflux of blood and intracatheter thrombosis. This may be a factor in reducing sepsis associated with central venous total parenteral nutrition.