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Traditional culture methods for identifying the plant fungal pathogens Sclerotinia sclerotiorum (Lib.) de Bary and Botrytis cinerea Pers.:Fr. are slow and laborious. The goal of this study was to develop a multiplex real-time PCR (qPCR) assay to detect and quantify DNA from S. sclerotiorum and B. cinerea. A primer set (SsIGS_5) for S. sclerotiorum was designed that targeted the intergenic spacer (IGS) regions of the ribosomal DNA. Addition of a probe to the assay increased its specificity: when the primer/probe set was tested against 21 fungal species (35 strains), amplification was detected from all S. sclerotiorum strains and no other species. For qPCR, the SsIGS_5 primer and probe set exhibited a linear range from 7·0 ng to 0·07 pg target DNA (R2 = 0·99). SsIGS_5 was then multiplexed with a previously published primer/probe set for B. cinerea to develop a high-throughput method for the detection and quantification of DNA from both pathogens. When multiplexed, the sensitivity and specificity of both assays were not different from individual qPCR reactions. The multiplex assay is currently being used to detect and quantify S. sclerotiorum and B. cinerea DNA from aerosol samples collected in commercial seed alfalfa fields. Significance and Impact of the Study A primer and probe set for the quantification of Sclerotinia sclerotiorum DNA in a PCR assay was developed. The probe-based nature of this assay signifies an improvement over previous assays for this species by allowing multiplex reactions while maintaining high sensitivity. The primer/probe set was used in a multiplex real-time PCR assay for the quantification of S. sclerotiorum and Botrytis cinerea DNA, enabling rapid analysis of environmental samples. In crops susceptible to both pathogens, this multiplex assay can be used to quickly quantify the presence of each pathogen.

Objective To evaluate whether pulse oximetry can be used as a screening test to detect congenital heart disease (CHD) in otherwise well newborns before nursery discharge. Methods Births at Lakeland Regional Medical Center (LRMC) who met inclusion criteria, had a single pulse oximeter reading before discharge. A normal reading was considered to be >94%. Abnormal readings were repeated on a different extremity and if still abnormal, an echocardiography was performed. The number of children with CHD and the number of echocardiograms were compared with the previous year at LRMC. Results There were 2114 infants enrolled, 88 echocardiograms (3.8%) were performed, and 43 were abnormal (yield = 48.9%), of which 12 required management (13.6%). The control group consisted of 2851 births, producing 108 echocardiograms (3.8%), 42 of which were abnormal (38.9%), and 13 required management (12.0%). No comparisons were significant. During both years, 3 children with cyanotic CHD were born. Routine pulse oximetry failed to detect a child with total anomalous pulmonary venous return. Not a single normal echocardiogram was generated by the study. Conclusion Routine pulse oximetry was nearly 100% specific for detecting cyanotic CHD; as a result there was no increase in the number of echocardiograms. Routine pulse oximetry did detect a child who might not have been detected otherwise but failed to detect one that should have been detected. (J Pediatr 2003;142:268-72)

Radiofrequency Ablation in Pediatric Ebstein's Anomaly. Introduction: Abnormal anatomy and complex electrophysiology in patients with Ebstein's anomaly of the tricuspid valve may confound attempts at radiofrequency ablation (RFA). Methods and Results: Data for 65 pediatric Ebstein's patients (9.8 ± 5.4 years, 4 months to 20 years; 39 ± 25 kg, 5.1 to 108 kg) were obtained from the Pediatric Radiofrequency Ablation Registry. The degree of tricuspid regurgitation (DOTR) and the degree of Ebstein's anomaly were assessed with echocardiography/Doppler. Leading indications were drug refractoriness (24 [37%] of 65 patients) and life-threatening arrhythmia (14 [22%] of 65 patients). For the 65 patients, 82 typical (nondecremental) accessory pathways (APs) (62% right free wall, 34% right septal, and 4% left sided), 17 other supraventricular tachycardias (1 ectopic atrial, 7 AV reentry, 5 Mahaim, and 4 intra-atrial reentry tachycardias), and 1 ventricular mechanism were mapped. Thirty-four (52%) of 65 patients had a single AP (21 right free wall, 10 septal, and 3 left); 19 (29%) of 65 patients multiple APs; 6 (9%) of 65 patients a single AP plus a non-AP mechanism; and 6 (9%) of 65 patients non-AP mechanism(s) only. REA acute success rates and recurrence rates for right free wall, right septal, and other mechanisms were 79%/32%, 89%/29%, and 75%/27%. Mild DOTR and a body surface area (BSA) ≤ 1.7 m2 independently predicted a better acute success rate. BSA ≤ 1.7 m2 also predicted long-term success. Conclusion: In this patient subset, life-threatening arrhythmias and multiple electrophysiologic mechanisms are commonly encountered during REA. Mild DOTR and a BSA ≥ 1.7 m2 predict a higher acute success rate. While acute success rates are relatively high, recurrence is frequent.

A member of the Tennessee state legislature recently proposed a bill that would mandate all newborn infants to undergo pulse oximetry screening for the purpose of identifying those with critical structural heart disease before discharge home. The Tennessee Task Force on Screening Newborn Infants for Critical Congenital Heart Defects was convened on September 29, 2005. This group reviewed the current medical literature on this topic, as well as data obtained from the Tennessee Department of Health, and debated the merits and potential detriments of a statewide screening program. The estimated incidence of critical congenital heart disease is 170 in 100000 live births, and of those, 60 in 100000 infants have ductal-dependent left-sided obstructive lesions with the potential of presentation by shock or death if the diagnosis is missed. Of the latter group, the diagnosis is missed in ∼9 in 100 000 by fetal ultrasound assessment and discharge examination and might be identified by a screening program. Identification of the missed diagnosis in these infants before discharge could spare many of them death or neurologic sequelae. Four major studies using pulse oximetry screening were analyzed, and when data were restricted to critical left-sided obstructive lesions, sensitivity values of 0% to 50% and false-positive rates of between 0.01% and 12% were found in asymptomatic populations. Because of this variability and other considerations, a meaningful cost/benefit analysis could not be performed. It was the consensus of the task force to provide a recommendation to the legislature that mandatory screening not be implemented at this time. In addition, we determined that a very large, prospective, perhaps multistate study is needed to define the sensitivity and false-positive rates of lower-limb pulse oximetry screening in the asymptomatic newborn population and that there needs to be continued partnering between the medical community, parents, and local, state, and national governments in decisions regarding mandated medical care.

There are many ways to express the problem of undiagnosed congenital heart disease (CHD) in infants. Vaidyanathan, et al. [1] describe the problem in a statistically detailed manner. I prefer this analysis: for every 700,000 live births, there will be 4,500 infants with CHD, 25% of these children will leave the hospital without a diagnosis, and roughly 30 infants will die from CHD before any cardiac diagnosis is made [2]. This exceeds the total operative mortality from the most complicated operation in CHD surgery, the Norwood [3,4]. If you search hypoplastic left heart syndrome on PubMed you’ll get 1856 articles, if you search for undiagnosed CHD you’ll get only 67 articles. This lack of research interest is despite, even in an era of extensive fetal diagnosis, the most common diagnosis made at autopsy for infants is CHD-nearly 40% [5].

The efficacy of CT scan in the diagnosis of pediatric appendicitis has hot been established.Every patient under the age of 18 who was diagnosed and treated for acute appendicitis in the 1 year period from March 1998-March 1999 at Lakeland Regional Medical Center were included. The presentation, laboratory evaluation, imaging evaluation, hospital course, and pathologic evaluation were reviewed. Selected imaging studies were reviewed by an independent radiologist.Forty-six patients were treated for appendicitis; 17 of them received CT scans (37%). The CT scans predicted appendicitis in 9 of 17 cases (sensitivity = 53%). False-negative studies resulted in some morbidity but no mortality. The radiology review indicated that three cases clearly did not demonstrate appendicitis. The other false-negative studies were secondary to either technical or professional factors.The efficacy of CT imaging in the diagnosis of acute appendicitis in children has still not been demonstrated. This limited series indicates that if CT scanning is to be used in pediatric patients, more attention to technical and professional factors may be required. Some of these factors, particularly the ingestion of oral contrast, are particularly problematic in small children and may limit the effectiveness of this modality.

To the Editor .— The Pediatrics article by Bernard et al,1 which purported to link infant swimming to the later development of asthma, had a number of epidemiologic and statistical problems but, more importantly, seems to be an article published for political rather than scientific purposes. The press release by the American Academy of Pediatrics (AAP) on June 4, 2007, trumpeting this very weak study as evidence against infant swimming was in poor taste.2 The AAP clearly opposes infant swimming.2–4 Its press release stated that …

As an outspoken critic of the current state of pre-sports screening in the U.S. ([1][1]), I applaud Corrado et al. ([2][2]) and Pelliccia et al. ([3][3]) for their scientific and thorough efforts to detect conditions responsible for sudden cardiac death in athletes and adopting standardized

Many cutaneous disorders are associated with activation or increased numbers of mast cells. Electrical stimulation has been shown to be effective in treating many of these disorders. This study is designed to examine the effect of electrical stimulation on mast cells in acute wounds. Four pathogen-free pigs received 20 wounds, each of which was subjected to biopsy at various times after wounding. Half of the wounds were treated with electrical stimulation and the other half were treated with a sham electrode. The biopsy specimens were fixed in Carnoy's medium and stained with alcian blue and Nuclear Fast Red. Mast cells from both sets of wounds were counted and analyzed. Highly significant reductions in the number of mast cells were seen with electrical stimulation on days 1 and 2 compared with nonstimulated control wounds. Electron microscopy was performed to compare the stimulated and control mast cells for characteristic features in morphology, location, and evidence of degranulation. Electrical stimulation did not appear to induce degranulation. The ability of electrical stimulation to decrease the number of mast cells may be related to a reduction of either proliferation or migration of these cells and may prove to be a valuable therapeutic technique.