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Cystic fibrosis (CF) is the most common lethal inherited disorder among Caucasians and results from mutation in the gene encoding the CF transmembrane conductance regulator. In addition to its multisystem clinical effects, the disease is characterized by increased proinflammatory mediators and oxidant stress, and systemic redox imbalance with reduced glutathione (GSH), together with alterations in circulating and tissue (n-6) and (n-3) fatty acids, particularly a decrease in docosahexaenoic acid. The metabolism of phospholipids and fatty acids is closely related to GSH through the methionine-homocysteine cycle, in which choline via betaine provides methyl groups to regenerate S-adenosylmethionine, important in generating phosphatidylcholine and amino acid precursors for GSH. Current research focuses both on fatty acid supplementations to normalize altered (n-6) to (n-3) fatty acid balance and decrease generation of (n-6) fatty acid-derived inflammatory mediators, and strategies to improve oxidant defenses and redox balance. However, further research is needed before such strategies can be included in clinical care of individuals with CF. [ABSTRACT FROM AUTHOR]

Illegal hunting is a major threat to the elephants of Africa, with more elephants killed by poachers than die from natural causes. DNA from tusks has been used to infer the source populations for confiscated ivory, relying on nuclear genetic markers. However, mitochondrial DNA (mtDNA) sequences can also provide information on the geographic origins of elephants due to female elephant philopatry. Here, we introduce the Loxodonta Localizer (LL; www.loxodontalocalizer.org), an interactive software tool that uses a database of mtDNA sequences compiled from previously published studies to provide information on the potential provenance of confiscated ivory. A 316 bp control region sequence, which can be readily generated from DNA extracted from ivory, is used as a query. The software generates a listing of haplotypes reported among 1917 African elephants in 24 range countries, sorted in order of similarity to the query sequence. The African locations from which haplotype sequences have been previously reported are shown on a map. We demonstrate examples of haplotypes reported from only a single locality or country, examine the utility of the program in identifying elephants from countries with varying degrees of sampling, and analyze batches of confiscated ivory. The LL allows for the source of confiscated ivory to be assessed within days, using widely available molecular methods that do not depend on a particular platform or laboratory. The program enables identification of potential regions or localities from which elephants are being poached, with capacity for rapid identification of populations newly or consistently targeted by poachers., (© The American Genetic Association 2019.)

The adequate preparation of cystic fibrosis (CF) youth for the transfer from pediatric to adult-based health care services is essential to meet the needs of this changing population. This paper describes the evolution of a transition clinic for patients with CF into a multidimensional quality improvement transition initiative. Three transition interventions (a patient transition clinical pathway; collaboration with the adult clinic; and a tool to measure transfer readiness) were sequentially implemented and evaluated. Each was found to be a valuable addition to a comprehensive transition protocol and today are endorsed as part of transition best practices., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The performance of brain-machine interfaces (BMIs) that continuously control upper limb neuroprostheses may benefit from distinguishing periods of posture and movement so as to prevent inappropriate movement of the prosthesis. Few studies, however, have investigated how decoding behavioral states and detecting the transitions between posture and movement could be used autonomously to trigger a kinematic decoder. We recorded simultaneous neuronal ensemble and local field potential (LFP) activity from microelectrode arrays in primary motor cortex (M1) and dorsal (PMd) and ventral (PMv) premotor areas of two male rhesus monkeys performing a center-out reach-and-grasp task, while upper limb kinematics were tracked with a motion capture system with markers on the dorsal aspect of the forearm, hand, and fingers. A state decoder was trained to distinguish four behavioral states (baseline, reaction, movement, hold), while a kinematic decoder was trained to continuously decode hand end point position and 18 joint angles of the wrist and fingers. LFP amplitude most accurately predicted transition into the reaction (62%) and movement (73%) states, while spikes most accurately decoded arm, hand, and finger kinematics during movement. Using an LFP-based state decoder to trigger a spike-based kinematic decoder [r = 0.72, root mean squared error (RMSE) = 0.15] significantly improved decoding of reach-to-grasp movements from baseline to final hold, compared with either a spike-based state decoder combined with a spike-based kinematic decoder (r = 0.70, RMSE = 0.17) or a spike-based kinematic decoder alone (r = 0.67, RMSE = 0.17). Combining LFP-based state decoding with spike-based kinematic decoding may be a valuable step toward the realization of BMI control of a multifingered neuroprosthesis performing dexterous manipulation.

Purpose of Review: This review is based upon the recent literature regarding eradication of newly acquired infection with Pseudomonas aeruginosa (Psa) in patients with cystic fibrosis (CF) and the economic and other effects of such an early eradication policy in a CF clinic., Recent Findings: Various Psa eradication protocols which utilize intravenous or aerosol anti-pseudomonal antibiotics, with or without oral antibiotics, have been reported. The recent ELITE trial reported successful eradication of 90% of Psa in selected Psa antibody negative patients after 28 days of tobramycin for inhalation. Another recent report of a protocol based on intravenous antibiotic use reported elimination of 'first growth' Psa in over 96% of all patients, accompanied by decreased chronic Psa infection, decreased anti-Psa treatment costs and decreased hospitalization costs., Summary: The effects of early eradication protocols for Psa have included decreased prevalence of chronic Psa infection, improved patient health and pulmonary function, and decreased hospital and antibiotic costs.

To examine the spatiotemporal distribution of discriminable information about reach-to-grasp movements in the primary motor cortex upper extremity representation, we implanted four microelectrode arrays in the anterior bank and lip of the central sulcus in each of two monkeys. We used linear discriminant analysis to compare information, quantified as decoding accuracy, contained in various neurophysiological signals. For all signal types, decoding accuracy increased immediately after the movement cue, peaked around movement onset, and declined during the static hold. Spike recordings and local field potential (LFP) time domain amplitude provided more discriminable information than LFP frequency domain power. Discriminable information on movement type was distributed evenly across recording sites by LFP amplitude and 1-4 Hz power but unevenly by 100-170 Hz power and spike recordings. These latter two signal types provided higher decoding accuracies closer to the hemispheric surface than deep in the anterior bank and also provided accuracies that varied along the central sulcus. This variation in the distribution of movement-type information may be related to differences in the rostral versus caudal regions of the primary motor cortex and to its underlying somatotopic organization. The even distribution of information by LFP amplitude and 1-4 Hz power compared with the more localized distribution by 100-170 Hz power and spikes suggest that these different neurophysiological signals reflect different underlying processes that distribute information through the motor cortex during reach-to-grasp movements.

Background: Acquisition of Pseudomonas aeruginosa (Psa) and infection with mucoid strains is associated with repeated pulmonary exacerbations which often require intravenous and long-term nebulised antibiotic treatments, repeated hospitalizations and leads to a more precipitous decline in lung function. Anti-Psa antibiotic therapy early in the course of Psa infection in patients with cystic fibrosis (CF) may result in eradication of Psa and prevention or delay of colonization with the organism. From January 1995 to December 2009 our paediatric CF clinic has followed an early eradication protocol for the first appearance of Psa. In this paper we report on the economic effects after 15 years as reflected in hospitalization and antibiotic usage and cost., Methods: The Psa-eradication protocol includes 2 weeks of IV piperacillin and tobramycin, followed by oral ciprofloxacin for 3 weeks, and nebulised colistimethate for 6 months. The same protocol is used for newly diagnosed CF patients who grow Psa on their first visit or who grow a mucoid strain, multiresistant strain of Psa or whose Psa co-cultured with Burkholderia cepacia complex, and for patients in whom Psa recurs after initial clearance., Results: 195 Psa eradication courses were completed from 1995 to 2009 with an overall Psa clearance rate of 90%. Patients that only cultured a Psa classic (non-mucoid) strain had a clearance rate was 96.5%. The percentage of children chronically infected with Psa has declined from 44% in 1994 to 15% in 2009.Total days spent in hospital for all reasons declined by 43%; chronic Psa hospital days declined by 75%; IV and nebulised anti-Psa antibiotic costs reduced by 44%., Conclusions: Results indicate that application of a Pseudomonas eradication protocol as described in this report has economic and resource utilization benefits in addition to clinical benefits., (Copyright © 2011 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.)

With increasing life expectancy and the need for lung transplantation in the cystic fibrosis (CF) population, there are increasing reports of chronic kidney disease (CKD). However, values for baseline or longitudinal glomerular filtration rate (GFR) as measured by exogenous clearance markers are lacking in this population. Retrospective cross-sectional study in 2 to 18-year-olds cared for at a single CF center who had a GFR measured by plasma disappearance of Technetium-99 m diethylenetriaminepentaacetic acid (mGFR). The primary outcome was evidence of renal dysfunction as defined by CKD stage II or below (mGFR <90 ml/min/1.73 m(2), persistent abnormalities in urinary sediment, abnormal renal imaging). Of 63 patients evaluated, four had apparent renal dysfunction, one demonstrated decreased mGFR, and three others had persistent microscopic hematuria. The mean mGFR was substantially higher (140 ± 24 ml/min/1.73 m(2)) than expected or previously reported for healthy children. We did not demonstrate the presence of significant renal impairment after limited aminoglycoside exposure in the first decade following diagnosis with CF. However, we did document the presence of glomerular hyperfiltration in a significant proportion of our CF patients.

Background: Choline is an important constituent of acetylcholine. Choline is needed for acetylcholine in the nonneuronal acetylcholine system that includes epithelial cells of the lung and intestine, endothelial cells, and immune cells. Plasma free choline concentrations are low in children with cystic fibrosis (CF), but the implications for acetylcholine are unknown., Objective: We determined the relation between plasma free choline and related metabolites and leukocyte acetylcholine in children with CF and in a control group of healthy children without CF., Design: This was a cross-sectional study in 34 children with CF who were pancreatic insufficient and taking pancreatic enzyme-replacement therapy and in 16 healthy children. Plasma free choline, betaine, dimethylglycine, methionine, homocysteine, and leukocyte acetylcholine concentrations were quantified by using isotope-dilution HPLC-tandem mass spectrometry., Results: Mean (±SE) plasma free choline was 9.30 ± 0.37 and 6.54 ± 0.38 μmol/L (P < 0.05) and leukocyte acetylcholine was 1.21 ± 0.016 and 0.077 ± 0.011 pmol leukocyte acetylcholine/10(6) cells (P < 0.05) in control children and children with CF, respectively. Leukocyte acetylcholine was positively correlated with plasma free choline concentration in children with CF (r = 0.412, P < 0.05) but not in control children. Plasma betaine, dimethylglycine, and methionine concentrations were also lower in children with CF than in control children (P < 0.05)., Conclusions: A low free choline and methyl status in children with CF is associated with reduced acetylcholine in leukocytes. Whether these changes are explained by a mutation in the CF transmembrane conductance regulator or disturbances in choline metabolism and the implications for immune cell dysfunction in CF are unknown. This trial was registered at clinicaltrials.gov as NCT01150136.

Rationale: infection with Burkholderia cepacia complex (BCC) bacteria in cystic fibrosis (CF) is associated with an unpredictable rate of pulmonary decline. Some BCC, but not others, elaborate copious mucoid exopolysaccharide, endowing them with a gross mucoid phenotype, the clinical significance of which has not been described., Objectives: to determine whether there was a correlation between bacterial mucoid phenotype, as assessed in a semiquantitative manner from plate culture, and severity of disease as assessed by the rate of decline in lung function., Methods: we performed a retrospective clinical review of 100 patients with CF attending the Vancouver clinics between 1981 and 2007 and analyzed the rate of lung function decline (% predicted FEV(1))., Measurements and Main Results: patients infected exclusively with nonmucoid BCC had a more rapid decline in lung function (annual FEV(1) change, -8.51 ± 2.41%) than those infected with mucoid bacteria (-3.01 ± 1.09%; P < 0.05). Linear mixed-effects data modeling revealed a statistically significant inverse association between semiquantitative mucoid exopolysaccharide production and rate of decline of lung function. In vitro incubation of BCC with ceftazidime and ciprofloxacin but not meropenem caused conversion of BCC from mucoid to nonmucoid., Conclusions: our data suggest an inverse correlation between the quantity of mucoid exopolysaccharide production by BCC bacteria and rate of decline in CF lung function. Certain antibiotics may induce a change in bacterial morphology that enhances their virulence. A simple in vitro test of bacterial mucoidy may be useful in predicting the rate of decline of respiratory function in CF.

The pontomedullary reticular formation (PMRF) of the monkey produces motor outputs to both upper limbs. EMG effects evoked from stimulus-triggered averaging (StimulusTA) were compared with effects from stimulus trains to determine whether both stimulation methods produced comparable results. Flexor and extensor muscles of scapulothoracic, shoulder, elbow, and wrist joints were studied bilaterally in two male M. fascicularis monkeys trained to perform a bilateral reaching task. The frequency of facilitation versus suppression responses evoked in the muscles was compared between methods. Stimulus trains were more efficient (94% of PMRF sites) in producing responses than StimulusTA (55%), and stimulus trains evoked responses from more muscles per site than from StimulusTA. Facilitation (72%) was more common from stimulus trains than StimulusTA (39%). In the overall results, a bilateral reciprocal activation pattern of ipsilateral flexor and contralateral extensor facilitation was evident for StimulusTA and stimulus trains. When the comparison was restricted to cases where both methods produced a response in a given muscle from the same site, agreement was very high, at 80%. For the remaining 20%, discrepancies were accounted for mainly by facilitation from stimulus trains when StimulusTA produced suppression, which was in agreement with the under-representation of suppression in the stimulus train data as a whole. To the extent that the stimulus train method may favor transmission through polysynaptic pathways, these results suggest that polysynaptic pathways from the PMRF more often produce facilitation in muscles that would typically demonstrate suppression with StimulusTA.

Recent neurophysiological studies indicate a role for reticulospinal neurons of the pontomedullary reticular formation (PMRF) in motor preparation and goal-directed reaching in the monkey. Although the macaque monkey is an important model for such investigations, little is known regarding the organization of the PMRF in the monkey. In the present study, we investigated the distribution of reticulospinal neurons in the macaque. Bilateral injections of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) were made into the cervical spinal cord. A wide band of retrogradely labeled cells was found in the gigantocellular reticular nucleus (Gi) and labeled cells continued rostrally into the caudal pontine reticular nucleus (PnC) and into the oral pontine reticular nucleus (PnO). Additional retrograde tracing studies following unilateral cervical spinal cord injections of cholera toxin subunit B revealed that there were more ipsilateral (60%) than contralateral (40%) projecting cells in Gi, while an approximately 50:50 ratio contralateral to ipsilateral split was found in PnC and more contralateral projections arose from PnO. Reticulospinal neurons in PMRF ranged widely in size from over 50 microm to under 25 microm across the major somatic axis. Labeled giant cells (soma diameters greater than 50 microm) comprised a small percentage of the neurons and were found in Gi, PnC and PnO. The present results define the origins of the reticulospinal system in the monkey and provide an important foundation for future investigations of the anatomy and physiology of this system in primates.

In metabolomic studies using liquid chromatography mass spectrometry of urine from children with cystic fibrosis (CF), high levels of metabolites of low molecular weight phthalates were found. Phthalate metabolite excretion was explained by therapy with enteric-coated pancreatic enzyme replacements. Phthalate metabolite identity was confirmed by tandem mass spectrometry. Pancreatic insufficient CF children taking Cotazym ECS(®), which is formulated with diethyl phthalate (DEP), had urinary metabolites of DEP. Children taking Creon(®), which has dibutyl phthalate (DBP), excreted DBP metabolites. The estimated concentrations of free MEP were 2-3 orders of magnitude higher than reported from environmental phthalate exposure. Enteric-coated pancreatic enzymes can expose individuals with CF to incessant, high oral intakes of phthalates. Although adverse effects have neither been shown to be present nor absent, we raise the need to consider that individuals requiring life-long therapy with some current pancreatic enzyme replacements chronically ingest high amounts of phthalates., (Copyright © 2009 Elsevier B.V. All rights reserved.)

Background: Since 2003, it has been routine practice at Children's and Women's Health Centre of British Columbia to monitor serum levels of prolactin in pediatric patients with cystic fibrosis who are receiving domperidone. Although a pharmacologic relationship between domperidone and prolactin has been documented in the literature, there is no information about routine monitoring of prolactin, and guidance on interpretation of prolactin values is lacking., Objectives: To characterize how prolactin levels were being used in monitoring patients with cystic fibrosis who were receiving domperidone therapy at this institution, to evaluate the need for this practice, and to formulate recommendations accordingly., Methods: A chart review was conducted for pediatric patients with cystic fibrosis who had been receiving domperidone therapy and whose serum prolactin levels had been monitored between June 1, 2001, and October 1, 2005., Results: A total of 219 samples had been drawn, from 49 patients, for determination of prolactin level. Of these, 100 (45.7%) were above the normal range. Of the values above the normal range, 86 (86%) led to no dosage adjustment of domperidone and 14 (14%) led to either a decrease in dose or discontinuation of therapy. None of the elevated prolactin levels were associated with supratherapeutic doses of domperidone., Conclusion: The role of routine monitoring of prolactin in this patient population requires further study. In particular, more information is needed about prolactin levels in pediatric patients and the relationship of prolactin level to domperidone dose.

Background: Medium Chain Acyl-CoA Dehydrogenase (MCAD) Deficiency is an autosomal recessive disorder of fatty acid oxidation, with potential fatal outcome. MCAD deficiency is diagnosed by acylcarnitine analysis on newborn screening blood spot cards by tandem mass spectrometry. Early diagnosis of MCAD and presymptomatic treatment can potentially reduce morbidity and mortality., Objectives: To evaluate incidence, clinical outcome, biochemical and molecular phenotype of MCAD cases detected in the first three years of newborn screening in British Columbia (BC)., Methods and Results: Medium chain length acylcarnitines, octanoylcarnitine (C8) and decanoylcarnitine (C10), were measured on newborn screening blood spot cards. Out of 121,000 live births, 17 newborns had C8 values above the screening cut-off of 0.38 umol/L. Ten newborns had elevated C8 on repeat cards and were investigated further. Both C8 and C8/C10 ratios remained abnormal in all confirmed MCAD cases. Positive predictive value of screening was 58% with no false negative results. Seven patients were homozygous for the common c.985A > G MCAD mutation and three others were compound heterozygous for the c.985A > G and a second mutation. Two novel mutations were identified (c.260T > C and c.382T > A). The estimated incidence of MCAD was approximately 1:12,000 live births. Upon frequent feeding and carnitine supplementation, none of the patients had metabolic crises or adverse outcomes., Conclusion: Frequency of MCAD in BC is comparable to reports from other newborn screening programs. Persistence of elevated C8 levels and C8/C10 ratios in confirmed MCAD cases suggest that these are sensitive markers for newborn screening. Early detection and treatment have successfully prevented adverse health outcomes in patients with MCAD.

Novel therapies to target lung inflammation are predicted to improve the lives of people with cystic fibrosis (CF) but specific antiinflammatory targets have not been identified. The goal of this study was to establish whether TLR5 signaling is the key molecular pathway mediating lung inflammation in CF, and to determine whether strategies to inhibit TLR5 can reduce the damaging inflammatory response. The innate immune responses were analyzed in both airway epithelial cells and primary PBMCs from CF patients and matched controls. Additionally, 151 clinical isolates of Pseudomonas aeruginosa from CF patients were assessed for motility and capacity to activate TLR5. Blood and airway cells from CF patients produced significantly more proinflammatory cytokine than did control cells following exposure to the CF pathogens P. aeruginosa and Burkholderia cepacia complex (p < 0.001). Stimulation with pure TLR ligands demonstrated that TLR signaling appears to mediate the excessive cytokine production occurring in CF. Using complementary approaches involving both neutralizing Ab targeting TLR5 and flagellin-deficient bacteria, we established that inhibition of TLR5 abolished the damaging inflammatory response generated by CF airway cells following exposure to P. aeruginosa (p < 0.01). The potential therapeutic value of TLR5 inhibition was further supported by our demonstration that 75% of clinical isolates of P. aeruginosa retained TLR5 activating capacity during chronic CF lung infection. These studies identify the innate immune receptor TLR5 as a novel antiinflammatory target for reducing damaging lung inflammation in CF.

Objective: To describe the clinical presentation and delays in diagnosis of patients with cystic fibrosis (CF) with the goal of raising physicians' awareness of CF and establishing baseline data for comparison with outcomes of patients who undergo newborn screening for CF., Design: Retrospective review of hospital medical records and CF clinic charts of newly diagnosed CF patients younger than 18 years who had attended the CF clinic at the BC Children's Hospital in Vancouver between January 1, 1993, and January 1, 2005. Age at diagnosis of CF was ascertained for 24 adult patients diagnosed during the same period from the CF clinic at St Paul's Hospital in Vancouver, BC., Setting: Cystic fibrosis clinic at the BC Children's Hospital., Participants: All newly diagnosed CF patients from mainland BC and northern Vancouver Island (N = 122)., Main Outcome Measures: Mean age at diagnosis; mean delay in diagnosis; weight and height or length at diagnosis; vitamin E status; mean head circumference; types of symptoms before diagnosis; Pseudomonas aeruginosa status; and number of days spent in tertiary care hospitals before diagnosis., Results: Excluding the adult patients and patients with meconium ileus, mean age at diagnosis of CF was 3.6 years, and mean delay in diagnosis after first symptoms was 2.1 years. Weight at diagnosis was < or = 5th percentile in 37% of cases, and height or length was < or = 5th percentile in 26% of cases. Excluding those with meconium ileus and those taking vitamin E supplementation, 70% of the children were vitamin E deficient at diagnosis. These children had a mean head circumference substantially smaller than that of children who had adequate levels of vitamin E. About 95% of children had gastrointestinal (GI) or malnutrition symptoms before diagnosis; 15% had GI symptoms only. About 81% of patients had respiratory symptoms, but only 4% had respiratory symptoms as the only evidence of CF before diagnosis. Around 9% were colonized with P aeruginosa at diagnosis. Before being diagnosed, 79% of patients had required tertiary care hospitalization for a group total of 320 hospital days., Conclusion: Considerable delays in diagnosis of children with CF occur when the disease is identified solely on clinical presentation. Morbidity is often severe enough to require hospital admission before CF is diagnosed. Symptoms that occurred before diagnosis were often GI or malnutritional in nature rather than respiratory, but all such symptoms were associated with diagnostic delays.

The characteristic elevation of plasma glycine concentrations observed in propionic acidaemia (PA) and other 'ketotic hyperglycinaemias' has been attributed to secondary inhibition of the hepatic glycine cleavage system (GCS) by accumulating CoA derivatives of branched-chain amino acid metabolites. In nonketotic hyperglycinaemia (NKH), cerebrospinal fluid (CSF) and plasma glycine levels and their ratio are increased due to primary deficiency of central nervous system (CNS) as well as hepatic GCS. Whether the GCS in the CNS is also inhibited in PA is unclear, as there are scant data available on CSF glycine levels in this disorder. We studied the relation of CSF and plasma glycine levels in 6 paired samples from 4 PA patients, including one PA patient with bacterial meningitis who underwent ventriculoperitoneal shunting and multiple CSF analyses (n = 26). In contrast to the CSF glycine levels which were generally elevated in all four PA patients, the CSF/plasma glycine concentration ratios in paired samples were normal (0.016-0.029), with the exception of a single sample (0.132) with extremely high CSF protein concentration (2010 mg/L) during the course of meningitis indicating a disturbed blood-brain barrier. This finding of normal CSF/plasma glycine ratio in PA suggests that the observed elevations of CSF glycine levels are a reflection of the concurrent hyperglycinaemia resulting from secondary inhibition of hepatic GCS, but that brain GCS is not affected, in contrast to the situation in NKH. The neurological sequelae in PA are therefore unlikely to be related to disturbed glycine metabolism.

Motor cortex output is capable of considerable reorganization, which involves modulation of excitability within the cortex. Does such reorganization also involve changes beyond the cortex, at the level of throughput from single motor cortex neurons to muscle activity? We examined such throughput during a paradigm that provided incentive for enhancing functional connectivity from motor cortex neurons to muscles. Short-latency throughput from a recorded neuron to muscle activity not present during some behavioral epochs often appeared during others. Such changes in throughput could not always be attributed to a higher neuron firing rate, to more ongoing muscle activity, or to neuronal synchronization, indicating that reorganization of motor cortex output may involve rapid changes in functional connectivity from single motor cortex neurons to alpha-motoneuron pools.

Primary Sertoli cell cultures have been established from several animals including the sheep and rhesus monkey; however, not for the domestic dog, Canis familiaris. Sertoli cells are the only readily accessible cell type in the body which expresses all six type IV collagens. These collagens play key roles in tissue structure, basement membrane formation, and filtration. The study of these genes is necessary to determine their exact roles and regulation in the aforementioned functions and to investigate diseases associated with mutations in these genes. For such studies, a cell culture system is a requisite tool. Therefore, Sertoli cells were targeted, and a culture was established from cells isolated from canine testes. Cultures maintained consistent morphology and steady growth for up to seven passages. Cultured cells were identified as Sertoli cells through positive Western blot results for SOX9 and Clusterin B proteins and transcript sequence verification of SOX9 as well as the presence of type IV collagen transcripts. Primary cultures of canine Sertoli cells will provide a useful tool for study of the function and regulation of collagen genes and will permit new research pertaining to canine health while also serving as a model for the study of human diseases.

Effects in spike-triggered averages (SpikeTAs) of rectified electromyographic activity (EMG) compiled for the same neuron-muscle pair during various behaviors often appear different. Do these differences represent significant changes in the effect of the neuron on the muscle activity? Quantitative comparison of such differences has been limited by two methodological problems, which we address here. First, although the linear baseline trend of many SpikeTAs can be adjusted with ramp subtraction, the curvilinear baseline trend of other SpikeTAs can not. To address this problem, we estimated baseline trends using a form of moving average. Artificial triggers were created in 1 ms increments from 40 ms before to 40 ms after each spike used to compile the SpikeTA. These 81 triggers were used to compile another average of rectified EMG, which we call a single-spike increment-shifted average (single-spike ISA). Single-spike ISAs were averaged to produce an overall ISA, which captured slow trends in the baseline EMG while distributing any spike-locked features evenly throughout the 80 ms analysis window. The overall ISA then was subtracted from the initial SpikeTA, removing any slow baseline trends for more accurate measurement of SpikeTA effects. Second, the measured amplitude and temporal characteristics of SpikeTA effects produced by the same neuron-muscle pair may vary during different behaviors. But whether or not such variation is significant has been difficult to ascertain. We therefore applied a multiple fragment approach to permit statistical comparison of the measured features of SpikeTA effects for the same neuron-muscle pair during different behavioral epochs. Spike trains recorded in each task were divided into non-overlapping fragments of 100 spikes each, and a separate, ISA-corrected, SpikeTA was compiled for each fragment. Measurements made on these fragment SpikeTAs then were used as test statistics for comparison of peak percent increase, mean percent increase, peak width at half maximum, onset latency, and offset latency. The average of each test statistic measured from the fragment SpikeTAs was well correlated with the single measurement made on the overall SpikeTA. The multiple fragment approach provides a sensitive means of identifying significant changes in SpikeTA effects.

Pontomedullary reticular formation (PMRF) neurons (309) were recorded simultaneously with electromyographic activity from arm and shoulder muscles in four monkeys performing arm-reaching tasks. Spike-triggered averages (SpikeTAs) were compiled for 292 neurons (3836 neuron-muscle pairs). Fourteen PMRF neurons located in a region ventral to the abducens nucleus produced 42 significant SpikeTA effects in arm and shoulder muscles. Of these 14 PMRF neurons, nine produced SpikeTA effects bilaterally. Overall, PMRF neurons facilitated ipsilateral flexors and contralateral extensors, while suppressing ipsilateral extensors and contralateral flexors. Spike- and stimulus-triggered averaging effects obtained from the same recording site were similar. These findings indicate that single PMRF neurons can directly influence movements of both upper limbs.

Background: Autosomal recessive hereditary nephropathy (ARHN) in the English Cocker Spaniel is caused by a type IV collagen defect, but the underlying mutation is unknown., Animals: One hundred thirty-four English Cocker Spaniels (12 with ARHN, 8 obligate carriers, and 114 others), 3 mixed breed dogs with X-linked hereditary nephropathy (XLHN), and 7 other dogs without hereditary nephropathy were included., Methods: Diagnosis of ARHN was based on transmission electron microscopy and immunostaining of kidney. Quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR) was used to compare COL4A3, COL4A4, and COL4A5 mRNA concentrations in the renal cortex from ARHN-affected English Cocker Spaniels, XLHN-affected dogs, and dogs without hereditary nephropathy. The entire coding region of COL4A4 was sequenced in 2 ARHN-affected dogs, 2 obligate carriers, 2 English Cocker Spaniels of unknown status, and 2 healthy mixed breed dogs. The exon containing the mutation was sequenced for all 134 English Cocker Spaniels., Results: Quantitative real time RT-PCR implicated COL4A4 as the gene harboring the mutation, and sequencing identified a single nucleotide substitution at base 115 as the cause of ARHN in English Cocker Spaniels. This mutation, which causes a premature stop codon in exon 3 of COL4A4, was segregated with clinical status in all affected dogs and obligate carriers. The mutation also was identified in 39 of 114 other English Cocker Spaniels with previously unknown status., Conclusions and Clinical Importance: The cause of this disease has been identified, and use of a test for the mutation will permit eradication of ARHN in the English Cocker Spaniel.

Background: Liver triacylglycerol accumulation and oxidative stress are common in cystic fibrosis (CF) and also occur in choline deficiency. Previously, we showed an association between elevated plasma homocysteine, reduced ratios of S-adenosylmethionine to S-adenosylhomocysteine (SAM:SAH) and of phosphatidylcholine to phosphatidylethanolamine, and phospholipid malabsorption in children with CF., Objective: The objective was to address a possible relation between altered methionine-homocysteine metabolism and choline metabolism in children with CF., Design: Children with CF were assigned without bias to supplementation with 2 g lecithin/d (n = 13), 2 g choline/d (n = 12), or 3 g betaine/d (n = 10) for 14 d. Plasma concentrations of methionine, adenosine, cysteine, cysteinyl-glycine, glutathione, glutathione disulfide (GSSG), and fatty acids; SAM:SAH; and red blood cell phospholipids were measured within each group of children with CF before and after supplementation. Plasma from healthy children without CF (n = 15) was analyzed to obtain reference data., Results: Children with CF had higher plasma homocysteine, SAH, and adenosine and lower methionine, SAM:SAH, and glutathione:GSSG than did children without CF. Supplementation with lecithin, choline, or betaine resulted in a significant increase in plasma methionine, SAM, SAM:SAH, and glutathione:GSSG and a decrease in SAH (n = 35). Supplementation with choline or betaine was associated with a significant decrease in plasma SAH and an increase in SAM:SAH, methionine, and glutathione:GSSG. Supplementation with lecithin or choline also increased plasma methionine and SAM., Conclusion: We showed that dietary supplementation with choline-related compounds improves the low SAM:SAH and glutathione redox balance in children with CF.

The motor output of the pontomedullary reticular formation (PMRF) was investigated to determine the reticulospinal system's capacity for bilateral control of the upper limbs. Stimulus triggered electromyographic averages (StimulusTA) were constructed from muscles of both upper limbs while two awake monkeys (Macaca fascicularis) performed a reaching task using either arm. Extensor and flexor muscles were studied at the wrist, elbow, and shoulder; muscles acting on the scapula were also studied. Post-stimulus effects (PStEs) resulted from 435 (81%) of 535 sites tested. Of 1611 PStEs analyzed, 58% were post-stimulus suppression (PStS), and 42% were post-stimulus facilitation (PStF). Onset latency was earlier for PStF than PStS, duration was longer for PStS, and amplitude was larger for PStF. Ipsilateral and contralateral PStEs were equally prevalent; bilateral responses were typical. In the ipsilateral forelimb and shoulder, the prevalent pattern was flexor PStF and extensor PStS; the opposite pattern was prevalent contralaterally. Sites producing strong ipsilateral upper trapezius PStF were concentrated in a region caudal and ventral to abducens. The majority of muscles studied had no clear somatotopic organization. Overall, the results indicate the monkey PMRF has the capacity to support bilateral coordination of limb movements using reciprocal actions within a limb and between sides.

Background: Hepatic steatosis and fat malabsorption are common in cystic fibrosis (CF). Choline deficiency results in decreased phosphatidylcholine synthesis through the cytidine diphosphocholine-choline pathway and hepatic steatosis and in increased synthesis of phosphatidylcholine from phosphatidylethanolamine using methyl groups from S-adenosylmethionine. The intestinal absorption of phosphatidylcholine in CF is unknown., Objectives: The objective was to determine whether excretion of choline phosphoglyceride (phosphatidylcholine and lysophosphatidylcholine) is increased in CF and whether loss of fecal choline phosphoglyceride is associated with altered plasma methionine cycle metabolites., Design: A cross-sectional study involved 53 children with CF and 18 control children without CF. Blood was collected from all participants. A subset of 18 children with CF and 8 control children provided 72-h fecal samples and 5-d food records., Results: Fat absorption was significantly lower (x+/- SEM: 86.2 +/- 1.6% and 94.1 +/- 1.2%) and excretion of fecal fat (12.9 +/- 1.7 and 3.9 +/- 0.7 g/d), phospholipid (median: 130 and 47.7 mg/d), phosphatidylcholine (19.6 and 2.1 mg/d), and lysophosphatidylcholine (60.3 and 16.9 mg/d) was significantly higher in children with CF than in control children, respectively (P < 0.05). Choline phosphoglyceride excretion was positively correlated with plasma homocysteine and S-adenosylhomocysteine and inversely related with plasma methionine (P < 0.05)., Conclusions: Choline phosphoglyceride excretion is increased in children with CF and is associated with decreased plasma methionine and increased homocysteine and S-adenosylhomocysteine. These findings suggest choline depletion and an increased choline synthesis by S-adenosylmethionine-dependent methylation in CF, as well as a metabolic link between phosphatidylcholine metabolism and the methionine-homocysteine cycle in humans.

Three monkeys ( M. fascicularis) performed a center-out, two-dimensional reaching task that included an instructed delay interval based on a color-coded visuospatial cue. Neural activity in the medial pontomedullary reticular formation (mPMRF) was recorded along with hand movement. Of 176 neurons with movement-related activity, 109 (62%) had movement-related but not preparatory activity (M cells), and 67 (38%) had both movement-related and preparatory activity (MP cells). EOG analyses indicated that the preparatory activity was not consistent with control of eye movements. There were slight changes in electromyograms (EMG) late in the instructed delay period before the Go cue, but these were small compared with the movement-related EMG activity. Preparatory activity, like the EMG activity, was also confined to the end of the instructed delay period for 14 MP cells, but the remaining 53 MP cells (30%) had preparatory activity that was not reflected in the EMG. Peri-movement neural activity varied with movement direction for 70% of the cells, but this variation rarely fit circular statistics commonly used for studies of directional tuning; directional tuning was even less common in the preparatory activity. These data show that neurons in the mPMRF are strongly modulated during small reaching movements, but this modulation was rarely correlated with the trajectory of the hand. In accord with findings in the literature from other regions of the CNS, evidence of activity related to motor preparation in these cells indicates that this function is distributed in the nervous system and is not a feature limited to the cerebral cortex.

The motor output of the medial pontomedullary reticular formation (mPMRF) was investigated using stimulus-triggered averaging (StimulusTA) of EMG responses from proximal arm and shoulder muscles in awake, behaving monkeys (M. fascicularis). Muscles studied on the side ipsilateral (i) to stimulation were biceps (iBic), triceps (iTri), anterior deltoid (iADlt), posterior deltoid (iPDlt), and latissimus dorsi (iLat). The upper and middle trapezius were studied on the ipsilateral and contralateral (c) side (iUTr, cUTr, iMTr, cMTr). Of 133 sites tested, 97 (73%) produced a poststimulus effect (PStE) in one or more muscles; on average, 38% of the sampled muscles responded per effective site. For responses that were observed in the arm and shoulder, poststimulus facilitation (PStF) was prevalent for the flexors, iBic (8 responses, 100% PStF) and iADlt (13 responses, 77% PStF), and poststimulus suppression (PStS) was prevalent for the extensors, iTri (22 responses, 96% PStS) and iLat (16 responses, 81% PStS). For trapezius muscles, PStS of upper trapezius (iUTr, 49 responses, 73% PStS) and PStF of middle trapezius (iMTr, 22 responses, 64% PStF) were prevalent ipsilaterally, and PStS of middle trapezius (cMTr, 6 responses, 67% PStS) and PStF of upper trapezius (cUTr, 46 responses, 83% PStS) were prevalent contralaterally. Onset latencies were significantly earlier for PStF (7.0 +/- 2.2 ms) than for PStS (8.6 +/- 2.0 ms). At several sites, extremely strong PStF was evoked in iUTr, even though PStS was most common for this muscle. The anatomical antagonists iBic/iTri were affected reciprocally when both responded. The bilateral muscle pair iUTr/cUTr demonstrated various combinations of effects, but cUTr PStF with iUTr PStS was prevalent. Overall, the results are consistent with data from the cat and show that outputs from the mPMRF can facilitate or suppress activity in muscles involved in reaching; responses that would contribute to flexion of the ipsilateral arm were prevalent.

Background: Exercise has been shown to maintain or increase bone mineral density (BMD) in non-CF populations., Objectives: The purpose of our study was to elucidate the relationship between exercise, body composition and dietary intake with BMD in an adult CF population with heterogeneous disease severity., Design: We measured spinal (L1-4) and femoral (femoral neck) BMD by dual energy X-ray absorptiometry (DEXA) in 68 CF adults (24 female, 44 male) with a mean age 30.8(1.7) and 27.4(1.3) (range 18-55) years. We used the average BMD Z score for spine and femoral neck for analyses. Differences in disease severity, exercise capacity, physical activity level, dietary intake, body composition, body mass index (BMI), glucocorticoid use were correlated with BMD scores. Exercise capacity was defined as the maximal amount of oxygen consumed by muscles during maximal exercise (Vo2max). Vertebral and non-vertebral fracture rate were also recorded., Results: Fifty-seven patients were identified with low BMD (Z score < -1). Multiple linear regression identified exercise capacity and BMI as significant predictors of BMD. Later diagnosis of CF was also associated with low adult BMD., Conclusions: Low BMD is common in adult CF patients. Exercise capacity and BMI are predictors of low BMD., (Copyright 2003 European Cystic Fibrosis Society)

Cystic fibrosis (CF) transmembrane conductance regulator (CFTR) genotype does not explain the heterogeneity observed in CF pulmonary disease severity. Modifier genes are implicated for this heterogeneity. alpha1-antitrypsin (alpha1-AT) is one of the few antiproteases capable of inactivating neutrophil elastase. We investigated whether alpha1-AT alleles (Z, S deficiency alleles and the 3' G1237-->A mutation) were associated with increased disease severity and the alpha1-AT acute phase response during pulmonary exacerbations. This was a multicenter Canadian study. Seven hundred sixteen patients with CF (age range, 5.0-63.6 yr) were genotyped for the Z, S, and G1237-->A polymorphisms of the alpha1-AT gene. Stable and acute levels of alpha1-AT were measured on 31 adult patients with CF and were correlated to clinical parameters. There were 69, 13, and 18 patients with CF who were MS, SS, and MZ, respectively. There were 95 and 7 patients with CF heterozygous or homozygous for the A1237 allele, respectively. alpha1-AT genotype did not predict pulmonary disease severity, and was not associated with more severe clinical outcome (death or lung transplantation) or age of onset of Pseudomonas aeruginosa infection. Body mass index was a significant predictor of alpha1-AT levels during exacerbations. alpha1-AT genotype is not a major contributor to the variability of pulmonary disease severity in CF.

Objective: We used a novel approach based on the intersection of phospholipid and methionine metabolism at the S-adenosylmethionine (SAM)-dependent methylation of phosphatidylethanolamine (PE) to study potential alterations in phospholipid metabolism in children with cystic fibrosis (CF). Methyl groups from methionine via SAM are used for sequential methylation of PE to form phosphatidylcholine (PC) with the generation of S-adenosylhomocysteine (SAH) and homocysteine., Study Design: Plasma phospholipids and methionine metabolites and plasma and red blood cell phospholipid fatty acids were determined in 53 children with CF and 18 control children., Results: Plasma methionine and the PC/PE ratio was lower and homocysteine, SAH, and PE were higher in children with CF than in control children (P<.001). Plasma methionine was inversely (P<.05) and SAH and homocysteine were positively (P<.001) correlated with the plasma PE. Docosahexaenoic acid (22:6n-3) was significantly lower in plasma phospholipids and triglycerides and in red blood cell PC and PE of children with CF than in control children (P<.05)., Conclusions: These studies demonstrate that methionine metabolism is altered and associated with alteration of the plasma PC/PE ratio in CF. Altered phospholipid and methionine metabolism may contribute to the clinical complications associated with CF.

Objectives: To use findings from a time-flow study at an academic pediatric practice to restructure practice patterns and to repeat the time-flow study to evaluate consequent changes in patient waiting times, total visit times, and room and nursing time usage rates., Design: A before-and-after time-flow evaluation of patient waiting times, total visit times, and room and nursing time usage rates., Setting: An inner-city academic pediatric practice located at a community health center affiliated with a major urban academic teaching hospital., Patients: All patients visiting the pediatric practice during the weeks of April 26 through 30, 1999, and March 24 through 28, 2000., Interventions: Initial time-flow findings generated in telephone messaging, provider scheduling, nursing location, and provider and preceptor documentation., Main Outcome Measures: Patient waiting times, total visit times, room usage rates, and nursing time usage rates., Results: After the implementing of practice changes, mean total visit time declined from 91.9 to 78.3 minutes. Mean waiting time to be called by the nurse decreased 3.2 minutes, mean time spent with the provider decreased 4.6 minutes, and mean time to wait for a preceptor decreased by 8.8 minutes. Multivariate analysis controlling for visit type, provider type, and the type of postvisit interventions found that mean time of visit decreased by 13.6 minutes. Room and nursing time usage rates, resident satisfaction, and quality-of-care indicators remained largely unchanged., Conclusions: Time-flow studies can be useful instruments for academic ambulatory practices to identify and ameliorate practice inefficiencies without sacrificing quality of teaching or patient care.

Pseudomonas aeruginosa is the most common respiratory pathogen in patients with cystic fibrosis (CF), but the predominant mechanism by which it is acquired is controversial. To determine the frequency of patient-to-patient spread, we evaluated P. aeruginosa isolates from 174 patients treated at the CF clinics in Vancouver, BC, Canada, since 1981. Multiple isolates were obtained from each patient and genetically typed by random amplified polymorphic DNA and pulsed field gel electrophoresis analyses. A total of 157 genetic types of P. aeruginosa was identified, 123 of which were unique to individual patients. A total of 34 types was shared by more than one patient; epidemiologic evidence linked these individuals only in the cases of 10 sibships and 1 pair of unrelated patients. We conclude that there is an extremely low risk in Vancouver for patients with CF to acquire P. aeruginosa from other patients. It appears that prolonged close contact, such as occurs between siblings, is necessary for patient-to-patient spread. The major source of acquisition of P. aeruginosa in CF appears to be from the environment. Considering these observations, we do not recommend segregation of patients with CF on the basis of their colonization status with P. aeruginosa.

Infection with Burkholderia cepacia complex in patients with cystic fibrosis (CF) results in highly variable clinical outcomes. The purpose of this study was to determine if there are genomovar-specific disparities in transmission and disease severity. B. cepacia complex was recovered from 62 patients with CF on > or =1 occasions (genomovar III, 46 patients; genomovar II [B. multivorans], 19 patients; genomovar IV [B. stabilis], 1 patient; genomovar V [B. vietnamiensis], 1 patient; and an unclassified B. cepacia complex strain, 1 patient). Patient-to-patient spread was observed with B. cepacia genomovar III, but not with B. multivorans. Genomovar III strains replaced B. multivorans in 6 patients. Genomovar III strains were also associated with a poor clinical course and high mortality. Infection control practices should be designed with knowledge about B. cepacia complex genomovar status; patients infected with transmissible genomovar III strains should not be cohorted with patients infected with B. multivorans and other B. cepacia genomovars.

Objective: The objective was to evaluate the long-term effects of physiotherapy with an oscillating positive pressure device ("flutter") compared with physiotherapy with the use of a positive expiratory pressure (PEP) mask in patients with cystic fibrosis (CF)., Study Design: Forty children with CF were randomly assigned to performing physiotherapy with the PEP mask or the flutter device for 1 year. Clinical status, pulmonary function, and compliance were measured at regular intervals throughout the study., Results: The flutter group demonstrated a greater mean annual rate of decline in forced vital capacity compared with the PEP group (-8.62 +/- 15.5 vs 0.06 +/- 7.9; P =.05) with a similar trend in forced expiratory volume in 1 second (-10.95 +/- 19.96 vs -1.24 +/- 9.9; P =.08). There was a significant decline in Huang scores (P =.05), increased hospitalizations (18 vs 5; P =.03), and antibiotic use in the flutter group., Conclusion: Flutter was not as effective in maintaining pulmonary function in this group of patients with CF compared with PEP and was more costly because of the increased number of hospitalizations and antibiotic use.

Photoonycholysis in association with a generalized phototoxic reaction or as an isolated event is a well-recognized complication of the tetracycline group of antibiotics. We describe a 14-year-old white girl with cystic fibrosis who developed photoonycholysis of all 20 nails while receiving treatment with doxycycline. Pediatricians who prescribe tetracyclines should be aware of this potential complication.

Variable clinical course has been reported with the acquisition of Burkholderia cepacia in patients who have cystic fibrosis (CF). We hypothesized that the perceived worsening with B. cepacia may reflect the underlying severity of pulmonary disease at the time of acquisition. To test this hypothesis, we matched CF patients colonized with B. cepacia with CF patients not colonized with the organism. Two-year pre- and postacquisition data and long-term data were compared. Patients were matched for gender, age (+/- 1 yr), height (+/- 5 cm), weight (+/- 8 kg), percent predicted forced expiratory volume in one second (% pred FEV(1)) (+/- 10%), and pancreatic sufficiency status. Differences in rates of change pre- and postacquisition for FEV(1), FVC, weight, and frequency of intravenous courses were compared within pairs with the Wilcoxon signed rank test. Two-year and long-term survival was compared within pairs with the McNemar test. No significant differences were observed in mean annual rates of change in weight (0.33 and -0.28 kg/yr), % pred FEV(1) (-0.36 and -1.74%/yr), and percent predicted forced vital capacity (% pred FVC) (-3.80 and -2.32%/yr) between B. cepacia and control pairs in 2-yr and long-term postacquisition interval, respectively. Similar rates of change were noted for pre- to postacquisition intervals within pairs for weight (0.17 kg/yr), % pred FEV(1) (-0.16%/yr), % pred FVC (5.02 %/yr). There was a significantly higher rate of intravenous antibiotic courses in B. cepacia cases in the 2-yr and long-term postacquisition interval. Higher mortality was observed in the B. cepacia cases in the long term (p < 0.05). We conclude that colonization with B. cepacia does not necessarily adversely affect pulmonary status, but is associated with reduced long term survival. Whereas previous associations may be attributed to a propensity to colonize those who had more advanced disease, specific strain types of B. cepacia may have enhanced pathogenicity.

The mechanism of the G0/G1 arrest and inhibition of proliferation by quinidine, a potassium channel blocker, was investigated in a tissue culture cell line, MCF-7, derived from a human breast carcinoma. The earliest measurable effect of quinidine on the cell cycle was a decrease in the fraction of cells in S phase at 12 hr, followed by the accumulation of cells in G1/G0 phases at 30 hr. Arrest and release of the cell cycle established quinidine as a cell synchronization agent, with a site of arrest in early G1 preceding the lovastatin G1 arrest site by 5-6 hr. There was a close correspondence among the concentration-dependent arrest by quinidine in G1, depolarization of the membrane potential, and the inhibition of ATP-sensitive potassium currents, supporting a model in which hyperpolarization of the membrane potential and progression through G1 are functionally linked. Furthermore, the G1 arrest by quinidine was overcome by valinomycin, a potassium ionophore that hyperpolarized the membrane potential in the presence of quinidine. With sustained exposure of MCF-7 cells to quinidine, expression of the Ki67 antigen, a marker for cells in cycle, decreased, and apoptotic and necrotic cell death ensued. We conclude that MCF-7 cells that fail to progress through the quinidine-arrest site in G1 die.

Objective: We report the results of a long-term comparative trial of physiotherapy by the positive expiratory pressure (PEP) technique with a PEP mask (Astra Meditec) versus conventional postural drainage and percussion (PD&P). Forty patients, ages 6 to 17 years, with Shwachman scores between 52 and 93, attending the cystic fibrosis clinic were enrolled in the study and randomly assigned to one of two groups. Group A (control) continued to perform physiotherapy by using PD&P for a 1-year period, whereas patients assigned to group B performed physiotherapy with the PEP technique for the same period. Compliance with physiotherapy was closely monitored for both groups throughout the study. Clinical status and pulmonary function (forced vital capacity [FVC], FEV1, and FEF25-75) were measured at 3-month intervals. Group B (PEP) demonstrated improved pulmonary function in all parameters as measured by change in percent predicted value for age, gender, and height. The changes in pulmonary function over the study period were: FVC, +6.57; FEV1, +5.98; and FEF25-75, +3.32. This improvement was significantly different from that of group A (PD&P) whose pulmonary function declined in all parameters (FVC, -2.17; FEV1, -2.28; FEF25-75, -0.24). The differences between treatment groups were statistically significant for the changes in FVC (p = 0.02) and FEV(1) (p = 0.04). Our results indicate that for our patients with cystic fibrosis, pulmonary physiotherapy with the PEP technique was superior to conventional physiotherapy with the PD&P technique.

Airway clearance is a major component of the management of cystic fibrosis. In North America, the airway clearance technique of choice has been physiotherapy using postural drainage combined with chest percussion, deep breathing and vibration. The disadvantages of this technique, the introduction of newer airway clearance techniques, and a greater emphasis on exercise has sparked a growing trend to prescribing individualized programs combining exercise with strategies adapted to the needs of each patient.

The course of a 12-year-old girl with cystic fibrosis (CF) and with recurrent hemoptysis since age 8 years is described. Conservative measures failed to control her bleeding. Hemoptysis was only partially controlled by repeated bronchial arterial embolizations. However, the addition of tranexamic acid (TXA) resulted in complete cessation of bleeding. Attempts to withdraw TXA therapy resulted in recurrence of hemoptysis; this patient has, therefore, been continuously maintained on this therapy for the past 4 years. No side effects of long-term TXA treatment have been noted.

One hundred and six specimens from 90 patients with cystic fibrosis were evaluated for the presence of Burkholderia cepacia using a current routine diagnostic protocol as well as a research protocol involving polymyxin B-MacConkey agar without crystal violet, PC agar, OFPVL agar, and a selective brain-heart infusion broth. Ten specimens from eight patients (8.9%) were positive by any method. The selective enrichment broth was the only medium that yielded B cepacia from all 10 positive samples, although the routine protocol was successful for eight of these. Transient carriage was identified in one patient. Epidemiological studies may be better served by the use of selective enrichment rather than selective solid media alone. Carrier status for B cepacia requires more strict definition if positive carrier status is to be accepted as having medical importance.

The effective dose and schedule of enzyme replacement therapy for Gaucher's disease have not been definitely established. We report a case of mixed chimeric state in an allogeneic BMT patient and followed her clinical and laboratory progress. The result shows that a low but sustained glucocerebrosidase level may provide symptomatic relief for this lysosomal disorder.

Background: Altered retinoblastoma (RB [also known as RB1]) gene expression was initially found in a small cohort study to occur in five (22%) of 23 patients with primary stage I and II non-small-cell lung carcinomas (NSCLCs). Putative mutation of the p53 gene (also known as TP53) has also been found to occur frequently in stage I and II NSCLCs and to be associated with more aggressive disease and a poorer prognosis., Purpose: Our purpose was to determine the Rb protein status in the same cohort that had been previously studied for their p53 protein status and to document whether loss of Rb protein expression was also an important factor in overall survival., Methods: One hundred one stage I or II NSCLC specimens were analyzed by immunohistochemical staining. These paraffin-embedded tumor sections were obtained from individual paraffin blocks prepared for each patient in the previous study. Patient survival status was obtained from hospital and tumor registry records., Results: Altered Rb protein expression was found in 24 of 101 stage I and II NSCLCs. The median survival was 32 months for patients with Rb-positive (Rb+) tumors and 18 months for individuals in whom expression of Rb protein was absent or altered (Rb-) in tumor cells. Log-rank analysis of the differences in overall survival was statistically significant (P = .007). When these results were combined with the p53 status in the same tumor, the median survival was 12 months for those individuals who had theoretically the worst pattern (Rb-/p53+) and 46 months for those patients with theoretically the best pattern (Rb+/p53-) (P < .001). The Rb+ and Rb- groups in this cohort were well balanced with respect to the distribution of age, disease stage, histologic types, p53 status, and sex. Using a multivariate proportional hazards regression model, both altered Rb and p53 status were found to be significantly associated with poor prognosis (P = .005 and .012, respectively) in the overall cohort., Conclusion: Altered Rb protein expression is an independent prognostic marker for overall decreased survival in early-stage NSCLC as detected by absence of nuclear Rb protein staining. There appears to be a poorer prognosis when loss of Rb protein function and mutated p53 protein occur in the same tumor., Implications: If these findings can be confirmed in larger prospective studies, the results would suggest that both the Rb and p53 status should be utilized as independent prognostic factors in early-stage NSCLC.