Synthesis and antimicrobial studies of hydrazone derivatives of 4-[3-(2,4-difluorophenyl)-4-formyl-1H-pyrazol-1-yl]benzoic acid and 4-[3-(3,4-difluorophenyl)-4-formyl-1H-pyrazol-1-yl]benzoic acid.

Microbial resistance to antibiotics is an unresolved global concern, which needs urgent and coordinated action. One of the guidelines of the Centers for Disease Control and Preventions (CDC) to combat antibiotic resistance is the development of new antibiotics to treat drug-resistant bacteria. In our effort to find new antibiotics, we report the synthesis and antimicrobial studies of 30 new pyrazole derivatives. These novel molecules have been synthesized by using readily available starting materials and benign reaction conditions. Some of these molecules have shown activity with MIC values as low as 0.78 µg/mL against four bacterial strains; Staphylococcus aureus, methicillin-resistant S. aureus, Bacillus subtilis, and Acinetobacter baumannii. Furthermore, active molecules are non-toxic to mammalian cell line.

Creating a Common Data Model for Comparative Effectiveness with the Observational Medical Outcomes Partnership.

BACKGROUND: Adoption of a common data model across health systems is a key infrastructure requirement to allow large scale distributed comparative effectiveness analyses. There are a growing number of common data models (CDM), such as Mini-Sentinel, and the Observational Medical Outcomes Partnership (OMOP) CDMs. OBJECTIVES: In this case study, we describe the challenges and opportunities of a study specific use of the OMOP CDM by two health systems and describe three comparative effectiveness use cases developed from the CDM. METHODS: The project transformed two health system databases (using crosswalks provided) into the OMOP CDM. Cohorts were developed from the transformed CDMs for three comparative effectiveness use case examples. Administrative/billing, demographic, order history, medication, and laboratory were included in the CDM transformation and cohort development rules. RESULTS: Record counts per person month are presented for the eligible cohorts, highlighting differences between the civilian and federal datasets, e.g. the federal data set had more outpatient visits per person month (6.44 vs. 2.05 per person month). The count of medications per person month reflected the fact that one system's medications were extracted from orders while the other system had pharmacy fills and medication administration records. The federal system also had a higher prevalence of the conditions in all three use cases. Both systems required manual coding of some types of data to convert to the CDM. CONCLUSIONS: The data transformation to the CDM was time consuming and resources required were substantial, beyond requirements for collecting native source data. The need to manually code subsets of data limited the conversion. However, once the native data was converted to the CDM, both systems were then able to use the same queries to identify cohorts. Thus, the CDM minimized the effort to develop cohorts and analyze the results across the sites.

COMPANION ANIMALS SYMPOSIUM: Rendered ingredients significantly influence sustainability, quality, and safety of pet food.

The rendering industry collects and safely processes approximately 25 million t of animal byproducts each year in the United States. Rendering plants process a variety of raw materials from food animal production, principally offal from slaughterhouses, but include whole animals that die on farms or in transit and other materials such as bone, feathers, and blood. By recycling these byproducts into various protein, fat, and mineral products, including meat and bone meal, hydrolyzed feather meal, blood meal, and various types of animal fats and greases, the sustainability of food animal production is greatly enhanced. The rendering industry is conscious of its role in the prevention of disease and microbiological control and providing safe feed ingredients for livestock, poultry, aquaculture, and pets. The processing of otherwise low-value OM from the livestock production and meat processing industries through rendering drastically reduces the amount of waste. If not rendered, biological materials would be deposited in landfills, burned, buried, or inappropriately dumped with large amounts of carbon dioxide, ammonia, and other compounds polluting air and water. The majority of rendered protein products are used as animal feed. Rendered products are especially valuable to the livestock and pet food industries because of their high protein content, digestible AA levels (especially lysine), mineral availability (especially calcium and phosphorous), and relatively low cost in relation to their nutrient value. The use of these reclaimed and recycled materials in pet food is a much more sustainable model than using human food for pets.

NVP-LDE-225 (Erismodegib) inhibits epithelial-mesenchymal transition and human prostate cancer stem cell growth in NOD/SCID IL2Rγ null mice by regulating Bmi-1 and microRNA-128.

Prostate cancer stem cells (CSCs) are defined by their extensive self-renewal, differentiation and tumor initiation properties. It is now clear that CSCs are involved in tumor growth and recurrence, and resistance to conventional treatments. The sonic hedgehog (Shh) pathway has a crucial role in stemness and tumorigenesis. Thus, the strategy that suppresses stemness and consequently tumorigenic potential of CSCs could be considered for the management of prostate cancer. The objectives of this study were to examine the molecular mechanisms, by which NVP-LDE-225/Erismodegib (smoothened inhibitor) regulates stem cell characteristics and tumor growth in prostate cancer. The effects of NVP-LDE-225 on CSC's viability, sphere formation, apoptosis, epithelial-mesenchymal transition (EMT) and tumor growth in NOD/SCID IL2Rγ null mice were examined. NVP-LDE-225 inhibited cell viability and spheroid formation, and induced apoptosis by activation of caspase-3 and cleavage of poly-ADP ribose polymerase (PARP). NVP-LDE-225 induced expression of Bax and Bak, and inhibited the expression of Bcl-2, Bcl-XL, XIAP, cIAP1, cIAP2 and survivin. NVP-LDE-225 inhibited Gli transcriptional activity, Gli-DNA interaction and the expression of Gli1, Gli2, Patched1 and Patched-2 in prostate CSCs. Interestingly, NVP-LDE-225 induced PDCD4 and apoptosis and inhibited cell viability by suppressing miR-21. Furthermore, NVP-LDE-225 inhibited pluripotency-maintaining factors Nanog, Oct-4, c-Myc and Sox-2. The inhibition of Bmi-1 by NVP-LDE-225 was regulated by upregulation of miR-128. NVP-LDE-225 suppressed EMT by upregulating E-cadherin and inhibiting N-cadherin, Snail, Slug and Zeb1 by regulating the miR-200 family. Finally, NVP-LDE-225 inhibited CSC tumor growth, which was associated with the suppression of Gli1, Gli2, Patched-1, Patched-2, Cyclin D1, Bmi-1 and PCNA and cleavage of caspase-3 and PARP in tumor tissues derived from NOD/SCID IL2Rγ null mice. Overall, our findings suggest that inhibition of the Shh signaling pathway could therefore be a novel therapeutic option in treating prostate cancer.

Lead (Pb) hohlraum: target for inertial fusion energy.

Recent progress towards demonstrating inertial confinement fusion (ICF) ignition at the National Ignition Facility (NIF) has sparked wide interest in Laser Inertial Fusion Energy (LIFE) for carbon-free large-scale power generation. A LIFE-based fleet of power plants promises clean energy generation with no greenhouse gas emissions and a virtually limitless, widely available thermonuclear fuel source. For the LIFE concept to be viable, target costs must be minimized while the target material efficiency or x-ray albedo is optimized. Current ICF targets on the NIF utilize a gold or depleted uranium cylindrical radiation cavity (hohlraum) with a plastic capsule at the center that contains the deuterium and tritium fuel. Here we show a direct comparison of gold and lead hohlraums in efficiently ablating deuterium-filled plastic capsules with soft x rays. We report on lead hohlraum performance that is indistinguishable from gold, yet costing only a small fraction.

NIF-scale re-emission sphere measurements of early-time Tr = 100 eV hohlraum symmetry (invited).

The indirect-drive National Ignition Campaign (NIC) proposes to set the first 2 ns of hohlraum radiation symmetry by observing the instantaneous soft x-ray re-emission pattern from a high-Z sphere in place of the ignition capsule. To assess this technique under NIC conditions, we used the Omega Laser Facility to image the re-emission of Bi coated spheres with 200 ps temporal, 50-100 microm spatial, and 30% spectral resolution. The sphere is driven by 70% NIC-scale vacuum Au hohlraums heated to Tr=100 eV using two cones per side laser beam illumination. The experiments have demonstrated the required accuracies of <3%P(2)/P(0) and <3%P(4)/P(0) Legendre mode flux asymmetry at both 900 and 1200 eV re-emission photon energies. The re-emission patterns at 900 and 1200 eV are also consistent with each other and their relative dependence on radiation temperature. We measured the P(2)/P(0) and P(4)/P(0) dependence to laser cone power ratio. View factor calculations are in agreement with the experimentally measured radiation flux and re-emit images when assuming 55% inner beam and 100 % outer beam coupling into x rays at the hohlraum wall.

Recording advances for neural prosthetics.

An important challenge for neural prosthetics research is to record from populations of neurons over long periods of time, ideally for the lifetime of the patient. Two new advances toward this goal are described, the use of local field potentials (LFPs) and autonomously positioned recording electrodes. LFPs are the composite extracellular potential field from several hundreds of neurons around the electrode tip. LFP recordings can be maintained for longer periods of time than single cell recordings. We find that similar information can be decoded from LFP and spike recordings, with better performance for state decodes with LFPs and, depending on the area, equivalent or slightly less than equivalent performance for signaling the direction of planned movements. Movable electrodes in microdrives can be adjusted in the tissue to optimize recordings, but their movements must be automated to be a practical benefit to patients. We have developed automation algorithms and a meso-scale autonomous electrode testbed, and demonstrated that this system can autonomously isolate and maintain the recorded signal quality of single cells in the cortex of awake, behaving monkeys. These two advances show promise for developing very long term recording for neural prosthetic applications.

Analysis of Borrelia burgdorferi vlsE gene expression and recombination in the tick vector.

Expression and recombination of the antigenic variation vlsE gene of the Lyme disease spirochete Borrelia burgdorferi were analyzed in the tick vector. To assess vlsE expression, Ixodes scapularis nymphs infected with the B. burgdorferi strain B31 were fed on mice for 48 or 96 h or to repletion and then crushed and acetone fixed either immediately thereafter (ticks collected at the two earlier time points) or 4 days after repletion. Unfed nymphs also were examined. At all of the time points investigated, spirochetes were able to bind a rabbit antibody raised against the conserved invariable region 6 of VlsE, as assessed by indirect immunofluorescence, but not preimmune serum from the same rabbit. This same antibody also bound to B31 spirochetes cultivated in vitro. Intensity of fluorescence appeared highest in cultured spirochetes, followed by spirochetes present in unfed ticks. Only a dim fluorescent signal was observed on spirochetes at the 48 and 96 h time points and at day 4 postrepletion. Expression of vlsE in vitro was affected by a rise in pH from 7.0 to 8.0 at 34 degrees C. Hence, vlsE expression appears to be sensitive to environmental cues of the type found in the B. burgdorferi natural history. To assess vlsE recombination, nymphs were capillary fed the B. burgdorferi B31 clonal isolate 5A3. Ticks thus infected were either left to rest for 4 weeks (Group I) or fed to repletion on a mouse (Group II). The contents of each tick from both groups were cultured and 10 B. burgdorferi clones from the spirochetal isolate of each tick were obtained. The vlsE cassettes from several of these clones were amplified by PCR and sequenced. Regardless of whether the isolate was derived from Group I or Group II ticks, no changes were observed in the vlsE sequence. In contrast, vlsE cassettes amplified from B. burgdorferi clones derived from a mouse that was infected with B31-5A3 capillary-fed nymphs showed considerable recombination. It follows that vlsE recombination does not occur in the tick vector.

Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium.

A phase I clinical trial was conducted in which recombinant adenovirus containing the cystic fibrosis trans-membrane regulator (CFTR) (Ad2/CFTR) was administered by bronchoscopic instillation or aerosolization to the lungs of cystic fibrosis (CF) patients. In this paper, we evaluate the efficiency of Ad2/CFTR-mediated transduction of bronchial airway cells. The ability of an Ad2/CFTR vector to transduce airway cells was first evaluated in patients to whom the vector was administered by bronchoscopic instillation. Cells at the administration site were collected 2 days after treatment by bronchoscopic brushing. Ad2-specific CFTR DNA was detected in four of five individuals by PCR, and Ad2-specific CFTR RNA was detected in three of five individuals by RT-PCR. Ad2/CFTR-mediated transduction of airway epithelial cells was then determined in CF individuals receiving this vector by aerosol inhalation. Ad2-specific CFTR DNA was detected in 13 of 13 individuals 2 days after aerosolization, and in 3 of 5 individuals 7 days after aerosolization. Ad2-specific RNA was detected in 4 of 13 individuals on day 2, but was not detected in the 5 individuals tested on day 7. The percentage of airway epithelial cells containing nuclear-localized vector DNA was < or =2.4% as determined by fluorescence in situ hybridization (FISH). However, in some cases, a high percentage of nonepithelial mononuclear cells or squamous metaplastic epithelial cells was infected with the adenoviral vector. In conclusion, aerosol administration is a feasible means to distribute adenoviral vectors throughout the conducting airways, but improvements in adenovirus-mediated transduction of airway epithelial cells are necessary before gene therapy for CF will be effective.

Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. I. Methods, safety, and clinical implications.

Cystic fibrosis (CF), an autosomal recessive disorder resulting from mutations in the cystic fibrosis trans-membrane conductance regulator (CFTR) gene, is the most common lethal genetic illness in the Caucasian population. Gene transfer to airway epithelium, using adenoviruses containing normal CFTR cDNA, leads to transient production of CFTR mRNA and, in some studies, to correction of the airway epithelial ion transport defect caused by dysfunctional CFTR. Inflammatory responses to the adenoviral vector have been reported, particularly at high viral titers. We evaluated the effects of adenovirus-mediated CFTR gene transfer to airway epithelium in 36 subjects with CF (34 individuals, 2 of whom received two separate doses of vector), 20 by lobar instillation and 16 by aerosol administration. Doses ranged from 8 x 10(6) to 2.5 x 10(10) infective units (IU), in 0.5-log increments. After lobar administration of low doses there were occasional reports of cough, low-grade temperature, and myalgias. At the highest lobar dose (2.5 x 10(9) IU) two of three patients had transient myalgias, fever, and increased sputum production with obvious infiltrates on CT scan. After aerosol administration there were no significant systemic symptoms until the 2.5 x 10(10) IU dose, when both patients experienced myalgias and fever that resolved within 24 hr. There were no infiltrates seen on chest CT scans in any of the patients in the aerosol administration group. There were no consistent changes in pulmonary function tests or any significant rise in serum IgG or neutralizing antibodies in patients from either group. Serum, sputum, and nasal cytokines, measured before and after vector administration, showed no correlation with adenoviral dose. Gene transfer to lung cells was inefficient and expression was transient. Cells infected with the vector included mononuclear inflammatory cells as well as cuboidal and columnar epithelial cells. In summary, we found no consistent immune response, no evidence of viral shedding, and no consistent change in pulmonary function in response to adenovirus-mediated CFTR gene transfer. At higher doses there was a mild, nonspecific inflammatory response, as evidenced by fevers and myalgias. Overall, vector administration was tolerated but transfer of CFTR cDNA was inefficient and transgene expression was transient for the doses and method of administration used here.

A clinical inflammatory syndrome attributable to aerosolized lipid-DNA administration in cystic fibrosis.

Immunologic reactivity to lipid-DNA conjugates has traditionally been viewed as less of an issue than with viral vectors. We performed a dose escalation safety trial of aerosolized cystic fibrosis transmembrane conductance regulator (CFTR) cDNA to the lower airways of eight adult cystic fibrosis patients, and monitored expression by RT-PCR. The cDNA was complexed to a cationic lipid amphiphile (GL-67) consisting of a cholesterol anchor linked to a spermine head group. CFTR transgene was detected in three patients at 2-7 days after gene administration. Four of the eight patients developed a pronounced clinical syndrome of fever (maximum of 103.3EF), myalgias, and arthralgia beginning within 6 hr of gene administration. Serum IL-6 but not levels of IL-8, IL-1, TNF-alpha, or IFN-gamma became elevated within 1-3 hr of gene administration. No antibodies to the cationic liposome or plasmid DNA were detected. We found that plasmid DNA by itself elicited minimal proliferation of peripheral blood mononuclear cells taken from study patients, but led to brisk immune cell proliferation when complexed to a cationic lipid. Lipid and DNA were synergistic in causing this response. Cellular proliferation was also seen with eukaryotic DNA, suggesting that at least part of the immunologic response to lipid-DNA conjugates is independent of unmethylated (E. coli-derived) CpG sequences that have previously been associated with innate inflammatory changes in the lung.

Borrelia burgdorferi proteins whose expression is similarly affected by culture temperature and pH.

Previously, we had demonstrated the upregulation in the expression of several proteins, including the lipoproteins OspC and P35, of Borrelia burgdorferi in the stationary growth phase. Since the expression of OspC is also known to be affected by culture temperature and pH, we examined the effects of both variables on the expression of the remaining stationary-phase-upregulated proteins. Our study revealed that the expression of each of the remaining stationary-phase-upregulated proteins, P35 included, was also influenced by culture temperature; these proteins were selectively expressed at 34 degrees C but not at 24 degrees C. Significantly, the expression of a majority of these proteins was also affected by culture pH, since they were abundantly expressed at pH 7.0 (resembling the tick midgut pH of 6.8 during feeding) but only sparsely at pH 8.0 (a condition closer to that of the unfed tick midgut pH of 7.4). We propose that this group of B. burgdorferi proteins, which in culture is selectively expressed under conditions of 34 degrees C and pH 7.0, may be induced in the tick midgut during the feeding event. Furthermore, the differential and coordinate expression of these proteins under different environmental conditions suggests that the encoding genes may be coregulated.

Impact of the hydrodyne process on tenderness, microbial load, and sensory characteristics of pork longissimus muscle.

Paired, boneless pork loin muscles were obtained from 76 market hogs to evaluate tenderness, meat quality characteristics, sensory attributes, and microbial characterization of pork muscle exposed to the Hydrodyne Process (H) compared with untreated control (C) loin. A subset of 16 paired loins was randomly selected for use in sensory evaluation and microbial characterization. Loins were vacuum packaged and immersed in a heat shrink tank prior to the H treatment. The Hydrodyne treatment exposed the loin to the pressure equivalent of a 150-g explosive, generating a pressure distribution of approximately 703 kg/cm2 at the surface of the samples. Meat quality assessments taken following treatment included subjective color, firmness/wetness, marbling scores (1 to 5 scale), Minolta reflectance and color readings, drip loss, and lipid content. The P-value for statistical significance for main effects and interactions was set at <.05 in all analyses. Administration of H resulted in a 17% improvement in Warner-Bratzler shear force (2.69 vs. 3.24 kg), with the shear force similar at two end-point cooking times (11 and 16 min) corresponding to approximately 75 and 83 degrees C, respectively. No differences between H and C were observed for color score, firmness score, Minolta L, Minolta Y, or drip loss on uncooked samples. The H loins had lower marbling scores (P<.05) and intramuscular lipid (P<.05) content than the paired C loin. Sensory evaluation on the randomly selected (n = 16) paired loins samples showed no improvement in Warner-Bratzler shear force. Sensory panelists were also unable to detect a difference between H and C loins for both initial and sustained tenderness scores. No differences between H and C loins were found for pork flavor, off-flavor, cohesiveness, or number of chews before swallowing, but H loins had a significantly lower juiciness score and more cooking loss than C loins. Microbial analysis results showed no differences in coliform bacteria counts, aerobic plate counts, and no detectable levels of Escherichia coli bacteria in any loins. The findings support the ability of the Hydrodyne procedure to improve tenderness without impacting other muscle quality attributes of pork.

Cationic lipid-mediated CFTR gene transfer to the lungs and nose of patients with cystic fibrosis: a double-blind placebo-controlled trial.

BACKGROUND: We and others have previously reported significant changes in chloride transport after cationic-lipid-mediated transfer of the cystic fibrosis transmembrane conductance regulator (CFTR) gene to the nasal epithelium of patients with cystic fibrosis. We studied the safety and efficacy of this gene transfer to the lungs and nose of patients with cystic fibrosis in a double-blind placebo-controlled trial. METHODS: Eight patients with cystic fibrosis were randomly assigned DNA-lipid complex (active) by nebulisation into the lungs followed 1 week later by administration to the nose. Eight control patients followed the same protocol but with the lipid alone (placebo). Safety was assessed clinically, by radiography, by pulmonary function, by induced sputum, and by histological analysis. Efficacy was assessed by analysis of vector-specific CFTR DNA and mRNA, in-vivo potential difference, epifluorescence assay of chloride efflux, and bacterial adherence. FINDINGS: Seven of the eight patients receiving the active complex reported mild influenza-like symptoms that resolved within 36 h. Six of eight patients in both the active and placebo groups reported mild airway symptoms over a period of 12 h following pulmonary administration. No specific treatment was required for either event. Pulmonary administration resulted in a significant (p<0.05) degree of correction of the chloride abnormality in the patients receiving active treatment but not in those on placebo when assessed by in-vivo potential difference and chloride efflux. Bacterial adherence was also reduced. We detected no alterations in the sodium transport abnormality. A similar pattern occurred following nasal administration. INTERPRETATION: Cationic-lipid-mediated CFTR gene transfer can significantly influence the underlying chloride defect in the lungs of patients with cystic fibrosis.

What are the livestock industries doing, and what do they need from us?

Livestock industries are facing global competition and revolutionary changes. While facing this global competition, the similarities of many animal meat products require that they compete on a cost-of-production basis. Additional issues include the environmental impact of animal agriculture, the role of animal products in human nutrition, food safety and quality, biotechnology, animal welfare, and market access. Progressive producers are becoming more aware of the needs of their customers and are striving to improve product quality. Checkoff funds are used to finance promotion, research, and consumer information programs and are increasingly used to finance producer education. Industrialization trends in the livestock industries are changing the needs of constituencies, delivery mechanisms, and relationships with the people involved. Characteristics of closed operations include high production cost, outdated technology, smaller size, older operators, and lack of management focus. Successful operations tend to be growing in capacity, are system-oriented, maintain high throughput, keep accurate records, use outside consultants, and control production costs. Modern livestock production has lowered the cost of production by integrating new production and management technologies. In order for producers to be successful in the future, access to technology, capital, and timely information will be critical. Animal scientists have many common objectives with livestock industries. Their work in research, teaching, and extension is critical for continued progress. However, people in the industries sometimes have the perception that academic arrogance, discipline myopia, uncoordinated research, slow technology transfer, increasing research costs, and counter-productive tenure systems prevent animal scientists from being as relevant and responsive as they could be. Support from the industries is essential as animal scientists and academic departments seek political and funding support. This support can be attained by including integrated systems research, improving communication skills, achieving more efficient research budgets, rapidly publishing results, reducing the cost of information distribution, developing flexible research agendas, retraining scientists, acquiring modern methods, and emphasizing critical thinking, communication, and teamwork when teaching.

Depletion of dopamine in the nucleus accumbens prevents the generation of locomotion by metabotropic glutamate receptor activation.

The contribution of dopamine (DA) to the locomotion elicited by activation of nucleus accumbens (NAcc) metabotropic glutamate receptors (mGluRs) was investigated in the rat. Different groups of rats were pretreated with bilateral microinjections of either 6-hydroxydopamine (6-OHDA) or its vehicle into the NAcc and, on separate tests starting 10 days later, were tested for locomotion following microinjections (into the same site) of saline, the mGluR agonist, 1-aminocyclopentane-trans-1,3-dicarboxylic acid [(1S, 3R)-ACPD, 0.5 nmol/side] and amphetamine (AMPH, 6.8 nmol/side). DA levels at the microinjection sites were significantly depleted in 6-OHDA-treated rats (42-99% depletions compared to control values obtained in vehicle-treated rats). In contrast to the increased locomotion observed in non-lesioned animals, rats pretreated with 6-OHDA showed no increase in locomotor activity in response to (1S, 3R)-ACPD or AMPH when these were microinjected into the NAcc. The two groups of rats were indistinguishable when tested following NAcc saline. These findings suggest that, as with AMPH, enhanced locomotion produced by NAcc mGluR activation is dependent on intact DA neurotransmission in this site.

Effect of noninvasive positive-pressure ventilation on survival in amyotrophic lateral sclerosis.

BACKGROUND: Noninvasive positive-pressure ventilation may benefit patients with amyotrophic lateral sclerosis and respiratory insufficiency. OBJECTIVE: To determine 1) whether patients tolerant of noninvasive positive-pressure ventilation have better survival than intolerant patients and 2) whether bulbar symptoms account for intolerance of noninvasive positive-pressure ventilation. DESIGN: Observational cohort study. SETTING: Tertiary care referral center. PATIENTS: 39 patients with amyotrophic lateral sclerosis who were treated with noninvasive positive-pressure ventilation. INTERVENTION: Noninvasive positive-pressure ventilation was started for patients with new orthopnea, new hypercapnia, or both. Patients were divided into two groups: those tolerant of and those intolerant of noninvasive positive-pressure ventilation. RESULTS: The risk for death from onset of respiratory insufficiency was higher for intolerant patients than for tolerant patients (relative risk, 3.1 [95% CI, 1.8 to 9.6]). Moderate or severe bulbar symptoms were more prevalent among intolerant patients than among tolerant patients (67% compared with 33%; P = 0.04). CONCLUSIONS: Among patients with amyotrophic lateral sclerosis, those who are tolerant of noninvasive positive-pressure ventilation have better survival than do those who are intolerant. Bulbar symptoms partially account for intolerance of noninvasive positive-pressure ventilation.

Comparison of DNA-lipid complexes and DNA alone for gene transfer to cystic fibrosis airway epithelia in vivo.

Cationic lipids show promise as vectors for transfer of CFTR cDNA to airway epithelia of patients with cystic fibrosis (CF). However, previous studies have not compared the effect of DNA-lipid to DNA alone. Recently, we developed a formulation of plasmid encoding CFTR (pCF1-CFTR) and cationic lipid (GL-67:DOPE) that generated greater gene transfer in mouse lung than previously described DNA-lipid vectors. Therefore, we tested the hypothesis that DNA-lipid complexes were more effective than DNA alone at transferring CFTR cDNA to airway epithelia in vivo. We administered complexes of DNA-lipid to one nostril and DNA alone to the other nostril in a randomized, double-blind study. Electrophysiologic measurements showed that DNA-lipid complexes partially corrected the Cl- transport defect. Importantly, the pCF1-CFTR plasmid alone was at least as effective as complexes of DNA with lipid. Measurements of vector-specific CFTR transcripts also showed gene transfer with both DNA-lipid and DNA alone. These results indicate that nonviral vectors can transfer CFTR cDNA to airway epithelia and at least partially restore the Cl- transport defect characteristic of CF. However, improvements in the overall efficacy of gene transfer are required to develop a treatment for CF.

Safety of a single aerosol administration of escalating doses of the cationic lipid GL-67/DOPE/DMPE-PEG5000 formulation to the lungs of normal volunteers.

Several groups are assessing the use of cationic lipids for respiratory gene therapy. To date no human data are available regarding the safety of intra-pulmonary cationic lipid delivery. In preparation for a trial of pulmonary delivery of the CFTR gene, we have assessed the safety of nebulised lipid GL-67/DOPE/DMPE-PEG5000 (GL-67A), the cationic lipid formulation to be used in this study. Fifteen healthy volunteers were given incremental doses of GL-67A via a Pari LC Jet nebuliser; three volunteers in each of five dosing cohorts with a week interval between cohorts. Markers of safety included clinical assessment, measurement of lung function, chest CT scan, serological testing and analysis of induced sputum. Measurements were taken before administration and at intervals up to 21 days thereafter. No adverse clinical events were seen or any statistically significant changes in spirometry or gas transfer. There were no clinically significant changes in any of the blood parameters and no CT changes were seen. Comparisons of the cellular subpopulations (neutrophils, eosinophils, lymphocytes and macrophages) in induced sputum showed no significant alterations following administration of the GL-67A. This study suggests that a single application of aerosol formulation of GL-67A does not result in clinically detectable changes when given by nebulisation into the lungs of normal volunteers and provides an indication of a lipid dose tolerated in man.

Analysis of the safety of bronchoscopy after recent acute myocardial infarction.

Acute myocardial infarction (AMI) is generally considered to increase the risk of flexible fiberoptic bronchoscopy (FFB). Currently, to our knowledge, no data in the literature support or challenge this concept. We conducted a retrospective chart review for the years 1986 to 1994 of 20 patients (14 men) who underwent 21 FFBs while hospitalized for an AMI. The mean age was 63.8 years (range, 38 to 83 years). Ten patients underwent revascularization procedures (eight coronary artery bypass grafting and two percutaneous transluminal coronary angioplasty) before FFB. The mean period between the AMI and FFB was 11.7 days (range, 1 to 30 days). Indications for FFB were pulmonary infiltrate (n = 10), hemoptysis (n = 6), atelectasis (n = 4), and to localize a suspected bronchopleural fistula (n = 1). Procedures performed included airway examination (21), BAL (12), transbronchial biopsy (2), endobronchial biopsy (3), and endobronchial brushing (4). No procedure was interrupted as a result of an adverse event, and five patients died during the same hospitalization. Four of the deaths occurred 6 to 15 days postprocedure; 1 patient (who had active ischemia at the time of FFB) died 4 h postprocedure. We conclude that FFB is safe in the immediate post-AMI period as long as the patient does not have active ischemia at the time of the procedure.