Assessment of concordance between diffusion of carbon monoxide through the lung using the 10 s breath-hold method, and the simultaneous NO/CO technique, in healthy participants.

INTRODUCTION: There is limited, large sample size, healthy control data comparing measurement of diffusing capacity of the lungs for carbon monoxide (DLCO) via the 10 s single-breath carbon monoxide uptake method (DLCO10) and using a DLCO-DLNO double diffusion test performed with a 5 s time of apnoea (DLCO5). OBJECTIVES: The primary objective was to compare DLCO5 and DLCO10 in healthy participants. The secondary objective was to evaluate the reproducibility of DLCO5. MATERIAL AND METHODS: We included medical students at Caen University Hospital, from 2008 to 2011. We performed a standard single-breath carbon monoxide uptake and combined DLCO and DLNO measurement for each participant. The combined test was repeated one week later. RESULTS: Among the 153 study participants, there was no statistically significant difference between the mean values of DLCO10 (10.2 ±â€¯2.2 mmol.min-1 kPa-1) and DLCO5 (10.3 ±â€¯2.2 mmol.min-1 kPa-1; paired t-test p = 0.19). Corrected for the same FiO2, DLCO5 was calculated at 10.5 ±â€¯2.3 mmol.min-1 kPa-1 and was significantly different from DLCO10 (paired t-test p < 0.001). DLCO5 deviates from 1,6 mmol.min-1 kPa-1 (4,6 mL.min-1. mmHg-1) or 15 % of DLCO10 (17 % above and 13% below, for 95 % of the subjects). Forty-seven participants were included in the DLCO5 reproducibility test. The 2 test sessions were carried out at 6 ±â€¯2 day intervals. Reproducibilities for DLCO, DLNO, DmCO and Vc was respectively 1.2 (11 %), 6.8 (13%), 16.5 (32 %), 12.5 (17 %) mmol.min-1 kPa-1. CONCLUSION: In healthy participants, discrepancies between DLCO measured during the double diffusion and DLCO measured on an apnoea of 10 s are quite large. It may be an indication that the Roughton and Forster interpretation to describe this type of measurements is inadequate.

Nitric Oxide Releasing Nanoparticles as a Strategy to Improve Current Onychomycosis Treatments.

Topical antimicrobials are the ideal mode of onychomycosis treatment for efficient drug delivery and avoidance of sytemic effects associated with oral medications. However, high treatment costs, tissue penetration limitations, and low cure rates have continued to pose major challenges. To capitalize on the progress made by topical efinaconazole solution, efinaconazole was combined with inexpensive, previously-characterized nitric oxide releasing nanoparticles (NO-np), which have been shown to offer sustained nitric oxide release over time and enhanced barrier penetration, while exerting broad spectrum antimicrobial and immunomodulating properties. NO-np were combined with efinaconazole in varying concentrations and applied against reference strains of Trichophyton rubrum using a checkerboard method. Results demonstrated synergism of NO-np+efinaconazole against T. rubrum, which is noteworthy given the barriers present in the topical treatment of onychomycosis, and the multiple potential benefits offered by NO-np. Overall, this study illustrates the untapped potential of nanotechnology in the treatment of disorders of the skin, hair, and nails where drug delivery remains a challenge. J Drugs Dermatol. 2018;17(7):717-720.

DNA fragmentation dynamics allows the assessment of cryptic sperm damage in human: evaluation of exposure to ionizing radiation, hyperthermia, acidic pH and nitric oxide.

Sperm DNA fragmentation (SDF) is not a static seminal parameter, since the longevity of sperm DNA decreases progressively with time following ejaculation or thawing. While the dynamics of SDF is a species-specific characteristic, in the case of humans, there is still significant variation within patients. To evaluate the suitability of the dynamic SDF assay to assess the adverse effects of agents that cause genetic damage, fresh semen samples from different donors were exposed in vitro to (1) increasing acute doses of ionizing radiation, (2) elevated temperature (41 °C and 45 °C), (3) acidic pH (pH 4) and (4) the nitric oxide (NO) donor sodium nitroprusside (SNP). Sperm DNA fragmentation was analyzed after an incubation period of chronic (24h), or acute (1h) exposure to each treatment followed by incubation at 37 °C over a period of 24h. SDF was assessed using the sperm chromatin dispersion (SCD) test. Dynamic SDF for each treatment was analyzed using Kaplan-Meier survival curves. All agents, except for ionizing radiation, accelerated SDF kinetics following chronic exposure over a 24h period. Transient exposure to NO and heat but not acidic pH increased the basal (T0) level of SDF. Despite the removal of the three toxicants, the remaining sperm following acute exposure showed a decrease in their expected DNA longevity. It is concluded that the assessment of sperm DNA fragmentation dynamics is an effective methodological approach for revealing latent damage associated with toxicants that is not initially expressed following a single initial observation of SDF.

Current and future therapeutic options for persistent pulmonary hypertension in the newborn.

Persistent pulmonary hypertension of the newborn (PPHN) is a potentially life-threatening condition that is characterized by supra-systemic pulmonary vascular resistance causing right-to-left shunting through the ductus arteriosus and/or foramen ovale, leading to a vicious cycle of hypoxemia, acidosis and further pulmonary vasoconstriction. Advances in neonatology including surfactant instillation, high-frequency ventilation, extracorporeal membrane oxygenation and, most importantly, inhaled nitric oxide (INO), have revolutionized the management of PPHN. However, given that INO does not improve oxygenation in a significant proportion (30-40%) of cases, there is an urgent need to consider other therapeutic options for PPHN. The issue is more important for developing nations with a higher PPHN-related health burden and limited resources. This article discusses the evidence about INO in term and preterm neonates in brief, and focuses mainly on the potential alternative drugs in the management of PPHN.

Is lowering blood pressure hazardous in patients with significant ipsilateral carotid stenosis and acute ischaemic stroke? Interim assessment in the 'Efficacy of Nitric Oxide in Stroke' trial.

BACKGROUND: High blood pressure (BP) in acute stroke patients is both common and associated with a poor outcome, although best management remains unclear. Particular uncertainty exists in patients with carotid stenosis in whom lowering BP might reduce cerebral perfusion and worsen outcome. METHODS: Efficacy of Nitric Oxide in Stroke (ENOS) is an international, randomized controlled trial investigating the effect of lowering BP with glyceryl trinitrate in 5000 patients with acute stroke. This analysis is based on patients with ischaemic stroke for whom information on the carotid status was available. Neurological impairment (Scandinavian Stroke Scale) and rate of recurrent stroke were assessed on day 7, and the functional outcome (modified Rankin score) was determined on day 90. ENOS is ongoing, therefore analyses are blinded to treatment. RESULTS: At the time of analysis, 565 patients with ischaemic stroke had been randomized into ENOS and data on carotid status were available in 394 (70%) of these patients. Ipsilateral stenosis > or =50% was present in 50 patients (13%). Six of 344 (2%, 95% confidence interval: 0.7, 4%) patients with ipsilateral stenosis <50% had a recurrent stroke by 7 days as compared with none of 50 patients (0%, 95% confidence interval: 0, 9%) (P=0.73) with stenosis > or =50%. No significant difference in impairment was present on day 7; mean Scandinavian Stroke Scale with stenosis 38.3 versus no stenosis 43.2 (P=0.48). Adjusted functional outcome after 90 days was worse in those with a baseline carotid stenosis > or =50%; median modified Rankin score 3.0 versus 2.0 (P=0.03). CONCLUSION: Interim data provide reassurance that it is reasonable to continue including patients with carotid stenosis into trials of acute BP lowering (such as ENOS).

Nitric oxide accelerates the degradation of tetrahydrobiopterin but not total neopterin in cerebrospinal fluid; potential implications for the assessment of tetrahydrobiopterin metabolism.

Assessment of total neopterin and tetrahydrobioterin (BH4) concentrations in cerebrospinal fluid (CSF) can be used to identify potential disorders of BH4 biosynthesis. In this study, we demonstrate that exposure of CSF to nitric oxide leads to an accelerated degradation of BH4 but does not affect the total neopterin concentration. These data suggest that in those conditions associated with increased nitric oxide formation, perturbation of the total neopterin to BH4 ratio could occur. Under such circumstances a putative diagnosis of a defect in BH4 biosynthesis may erroneously be proposed. Assessment of central nitric oxide generation may therefore be a useful adjunct to the determination of CSF pterin status.

Myocardial oxygen consumption regulation in isolated mouse heart: assessment by intracoronary administration of exogenous nitric oxide.

In our previous work we have shown that in mouse heart basal level of endothelial produced nitrite, as a marker of nitric oxide (NO) formation, was 9.7 nmol l(-1). Bradykinin (10 microl l(-1)) induced a 5-fold rise in nitrite release, the coronary venous effluent concentration being 58 nmol l(-1), but there was no effect on myocardial oxygen consumption (MVO2). The aim of this study was to assess the levels of authentic nitric oxide solution, exogenously applied, on myocardial oxygen consumption. Isolated mouse hearts (n=36) were paced (500 imp./min) and perfused at constant flow (16.0 +/- 0.3 ml g(-1) min(-1)). When coronary vasculature resistance was carefully controlled by adenosine (1 micromol l(-1)), authentic nitric oxide solution, in a concentration less than 5 micromol l(-1) did not alter myocardial oxygen consumption. Only concentrations of nitric oxide higher than 5 micromol l(-1) induced reduction in myocardial oxygen consumption. Thus in the saline perfused mouse heart, with carefully controlled vasodilatation, modulating myocardial nitric oxide levels using an arterial application of authentic nitric oxide, concentrations higher than 5 micromol l(-1) of nitric oxide were required to induce a decrease in myocardial oxygen consumption.

Magnetic resonance imaging guided catheterisation for assessment of pulmonary vascular resistance: in vivo validation and clinical application in patients with pulmonary hypertension.

OBJECTIVES: To validate in vivo a magnetic resonance imaging (MRI) method for measurement of pulmonary vascular resistance (PVR) and subsequently to apply this technique to patients with pulmonary hypertension (PHT). METHODS AND RESULTS: PVR was assessed from velocity encoded cine MRI derived pulmonary artery (PA) flow volumes and simultaneously determined invasive PA pressures. For pressure measurements flow directed catheters were guided under magnetic resonance fluoroscopy at 1.5 T into the PA. In preliminary validation studies (eight swine) PVR was determined with the thermodilution technique and compared with PVR obtained by MRI (0.9 (0.5) v 1.1 (0.3) Wood units.m2, p = 0.7). Bland-Altman test showed agreement between both methods. Inter-examination variability was high for thermodilution (6.2 (2.2)%) but low for MRI measurements (2.1 (0.3)%). After validation, the MRI method was applied in 10 patients with PHT and five controls. In patients with PHT PVR was measured at baseline and during inhalation of nitric oxide. Compared with the control group, PVR was significantly increased in the PHT group (1.2 (0.8) v 13.1 (5.6) Wood units.m2, p < 0.001) but decreased significantly to 10.3 (4.6) Wood units.m2 during inhalation of nitric oxide (p < 0.05). Inter-examination variability of MRI derived PVR measurements was 2.6 (0.6)%. In all experiments (in vivo and clinical) flow directed catheters were guided successfully into the PA under MRI control. CONCLUSIONS: Guidance of flow directed catheters into the PA is feasible under MRI control. PVR can be determined with high measurement precision with the proposed MRI technique, which is a promising tool to assess PVR in the clinical setting.

A novel method for the assessment of cellular composition and beta-cell viability in human islet preparations.

Current methodologies to evaluate islet cell viability are largely based on tests that assess the exclusion of DNA-binding dyes. While these tests identify cells that have lost selective membrane permeability, they do not allow us to recognize apoptotic cells, which do not yet stain with DNA-binding dyes. Furthermore, current methods of analysis do not discriminate between cell subsets in the preparation and, in particular, they do not allow for selectively defining beta-cell viability. For these reasons we have developed novel methods for the specific assessment of beta-cell content and viability in human islets based on cellular composition analysis through laser scanning cytometry (LSC) coupled with identification of beta-cell-specific apoptosis at the mitochondrial level. Our novel analytical methods hold promise to prospectively analyze clinical islet transplantation preparations and predict functional performance, as suggested by the observed correlation with in vivo analysis of islet potency in immunodeficient rodents.

Assessment of a dual regulatory role for NO in liver regeneration after partial hepatectomy: protection against apoptosis and retardation of hepatocyte proliferation.

The role of hepatic nitric oxide (NO) in liver regeneration after partial hepatectomy (PH) was studied in animals carrying a nitric oxide synthase-2 transgene under the control of the phospho(enol)pyruvate carboxykinase promoter. These mice expressed NOS-2 in liver cells under fasting conditions. Liver mass recovery and molecular parameters related to cell proliferation were determined after PH. Preexisting hepatic NO synthesis, as well as NO delivery by NO-donors, impaired early signaling (for example, attenuated NF-kappaB activation and TNF-alpha and IL-6 release). The regenerative process was also impaired as a result of an insufficient proliferative response, but mouse survival after surgery was not compromised. However, NO exerted a protective role against apoptosis in transgenic hepatectomized mice. Local production of NO in liver cells, achieved by hydrodynamic-based transfection with a NOS-2-encoding plasmid, also resulted in delayed liver recovery after PH and also protected against Fas-mediated apoptosis. These data show that sustained presence of NO after PH exerts a dual role: attenuating liver regeneration while efficiently protecting against liver apoptosis.

Nitric oxide-induced capacitation of cryopreserved bull spermatozoa and assessment of participating regulatory pathways.

The effect of nitric oxide (NO*) on the capacitation rates of cryopreserved bull spermatozoa and the participation of protein kinases in the capacitation process were evaluated. A pool of spermatozoa from four bulls were incubated in TALP medium in the presence of heparin (10 IU/ml) or sodium nitroprusside (SNP, 0.05-100 microM), a NO* donor. The participation of NO* was confirmed by the use of scavengers, i.e. methylene blue (50,100 microM) and hemoglobin (20-40 microg/ml). The role of nitric oxide synthase in heparin-induced capacitation was evaluated using enzyme inhibitors Nomega-nitro-L-arginine methyl ester (L-NAME) and Nomega-nitro-L-arginine (L-NA) in concentrations ranging from 1 to 500 microM. The effects of protein kinase A (PKA), protein kinase C (PKC) and protein tyrosine kinase (PTK), on NO*-induced capacitation were evaluated by incubation with specific inhibitors of these enzymes (H-89, 50 microM; bisindolylmaleimide I, 0.1 microM and genistein, 3 microM). The role of hydrogen peroxide or superoxide anion in NO*-induced capacitation was evaluated by incubation with catalase (20-100 microg/ml) or superoxide dismutase (SOD, 0.05-0.5 mg/ml), respectively. Capacitation percentages were determined by the fluorescence technique with chlortetracycline (CTC). SNP concentrations employed had no effect on progressive motility or sperm viability. Capacitation values of the 0.05 microM SNP treatment (31 +/- 5.15%) were similar to those of heparin treated samples (33 +/- 4.27%). Inhibitors of nitric oxide synthase (NOS) diminished capacitation percentages in a dose-dependent manner as did the addition of NO*- scavengers (P <0.05). The presence of PKA, PKC and PTK inhibitors likewise decreased capacitation percentages (6.25 +/- 0.71, 12.75 +/- 1.41, 9.00 +/- 1.41%, respectively). The presence of catalase or SOD in the incubation medium had no effect on capacitation percentages. These results indicate that NO* may be generated by a sperm NOS during heparin-induced capacitation and that exogenous NO* acts as a capacitation inducer and involves the participation of PKA, PKC and PTK as part of the intracellular mechanisms that lead to capacitation in cryopreserved bull spermatozoa.

Avaliação temporal da regulação do tônus vascular e da produção de superóxido induzido por purinas em aorta isolada de ratos Wistar endotoxêmicos / Analysis of purinergic modulation of vascular reactivity and superoxide production in aorta from endotoxemic rats

Pacientes sépticos podem evoluir para choque séptico, destes 40 por cento sobrevivem. Caracterizamos o modelo experimental, avaliamos fatores envolvidos na inflamação e avaliamos a modulação causada por purinas (ATP/ADP) na quantificação de superóxido (O2-) e na reatividade vascular da aorta isolada. Os resultados sugerem que na aorta isolada de animais endotoxêmicos, ATP e ADP aumentam a síntese de óxido nítrico (NO), porém somente o ATP reduz a biodisponibilidade de O2-, provavelmente pelo reacoplamento da NO sintase endotelial / Septic patients can evolve for septic shock and 40 per cent of these survive. We characterize the experimental model we evaluate involved factors in the inflammation and evaluate the modulation caused by purines (ATP/ADP) in the superoxide quantification (O2-) and in the vascular reactivity of isolated aorta. The results suggest that in isolated aorta of endotoxemics rats, ATP and ADP increase the endothelial nitric oxide synthase (NOS) however just ATP reduces the bio availability of O2-, probably for the re-couples of the endothelial NOS synthase...

Physiology and pathophysiology of the interstitial cell of Cajal: from bench to bedside. III. Interaction of interstitial cells of Cajal with neuromediators: an interim assessment.

Interstitial cells of Cajal (ICC) control gastrointestinal motility; some pace slow waves and others act in enteric neurotransmission. This review asks the question, does either class of ICC receive and respond to messages carried by neuromediators from these nerves? Relevant evidence includes the presence of receptors or responses to exogenous neuromediators and responses to endogenous neuromediators. Some pacemaking ICC networks have receptors for or respond to some exogenous neuromediators. None is known to respond to endogenous neuromediators. Intramuscular ICC have receptors for and respond to some neuromediators and are required in mice for responses to the exogenous and endogenous neuromediators nitric oxide and acetylcholine. The mechanisms underlying this requirement remain unclear. ICC pathologies exist, but their origins are unknown.

Metabolic coronary-flow regulation and exogenous nitric oxide in human coronary artery disease: assessment by intravenous administration of nitroglycerin.

We sought to evaluate the effect of intravenous administration of the nitric oxide--donor substance nitroglycerin (NTG) on metabolic coronary-flow regulation in patients with coronary artery disease (CAD). In 12 patients with stable CAD, we measured coronary sinus blood flow and myocardial oxygen supply and consumption (MVO2) at sinus rhythm and during atrial pacing (30 beats/min above sinus rate), both at control and during infusion of NTG, 1 microg/kg/min, and NTG, 2 microg/kg/min. To study metabolic coronary vasodilation, changes in myocardial oxygen supply were related to pacing-induced changes in MVO2, by using standard regression analysis. The myocardial oxygen supply/consumption ratio (i.e., the slope of the regression line at control, characterizing physiological metabolic coronary flow regulation) was compared with the ratios obtained during infusion of NTG. Compared with control measurements, NTG, 1 microg/kg/min, and NTG, 2 microg/kg/min, attenuated pacing-induced increases in MVO2 by 29 and 60%, respectively, whereas coronary blood flow during pacing remained unchanged. At control, normal metabolic coronary-flow regulation resulted in a myocardial oxygen supply/demand ratio of 1.39 (95% CI, 1.29-1.49). This ratio did not change during NTG, 1 microg/kg/min: 1.44 (95% CI, 1.33-1.56). However, during NTG, 2 microg/kg/min, this ratio significantly increased to 1.84 (95% CI, 1.63-2.05; p<0.01). Intravenous administration of high-dose NTG, a donor of exogenous NO, blunts pacing-induced increases in MVO2 and may increase metabolic coronary vasodilation in patients with CAD.

Avaliação da hemostasia e do óxido nítrico (NO) na doença hipertensiva específica da gravidez (DHEG) / The assessment of the hemostasia and nitric oxide (NO) in preeclampsia

A Doença Hipertensiva Específica da Gravidez (DHEG), na forma pura, caracteriza-se pelo aparecimento em grávida normotensa, após a vigésima semana de gestação, da tríade sintomática: hipertensão, proteinúria e edema. A DHEG está associada a vasoconstrição generalizada, a disfunção da célula endotelial e a consideráveis alterações hemostáticas. O óxido nítrico (NO) constitui um importante vasodilatador sintetizado pelo endotélio vascular. A determinação laboratorial de NO é extremamente complexa devido à sua ínfima concentração e meia-vida. Estudos anteriores sugerem que o NO está intimamente envolvido em todas as etapas da gravidez. A hipótese que norteou este estudo admite uma redução do NO em gestantes que desenvolvem a DHEG. Outros objetivos incluem a avaliação do estado hemostático/trombótico na DHEG e a definição de parâmetros hemostáticos que tenham um valor preditivo para o diagnóstico e prognóstico da doença...

Inhaled nitric oxide: clinical applications, indications, and toxicology.

PURPOSE: Although the analogy of nitric oxide (NO) to Endothelium-derived Relaxing Factor remains controversial, medical use of exogenous NO gas by inhalation has grown exponentially. This review presents the mechanisms of action of inhaled NO in pulmonary hypertension, hypoxaemia, inflammation and oedema, as well as its therapeutic and diagnostic indications with emphasis on acute respiratory distress syndrome (ARDS) and toxicology. SOURCE: Two medical databases (Current Contents, Medline) were searched for citations containing the above-mentioned key words to December 1996. Moreover, many presentations in congresses such as 4th International Meeting of Biology of Nitric Oxide, 52nd and 53rd Annual Meeting of Canadian Anaesthetists' Society or 10th Annual Meeting of European Association of Cardiothoracic Anaesthesiologists were used. PRINCIPAL FINDINGS: Inhaled NO is now recognized as an invaluable tool in neonatal and paediatric critical care, and for heart/lung surgery. Other clinical applications in adults, such as chronic obstructive pulmonary disease and ARDS, require a cautious approach. The inhaled NO therapy is fairly inexpensive, but it would seem that it is not indicated for everybody with regards to the paradigm of its efficiency and potential toxicity. The recent discovery of its anti-inflammatory and extrapulmonary effects open new horizons for future applications. CONCLUSION: Clinical use of inhaled NO was mostly reported in case series, properly designed clinical trials must now be performed to establish its real therapeutic role. These trials would permit adequate selection of the cardiopulmonary disorders, and subsequently the patients that would maximally benefit from inhaled NO therapy.

Assessment of ischemia and reperfusion injury.

Direct videomicroscopy of the rat cremaster muscle microcirculation supplemented by animal models of replantation, vascular crushing, and muscle function after injury and recovery were used to investigate the occurrence of reperfusion failure. It is evident that failure of blood reflow may be induced by multiple factors that can be grouped into categories of ischemia, intimal damage, and systemic or local responses, which are referred to as the no reflow triad. The components comprising the 3 sides of the no reflow triad can interact with one another in an intricate manner, and any single factor or combination of factors is capable of triggering the events leading to reperfusion failure. The pronounced regional nature of reperfusion injury and the direct relationship between the severity of the observed vascular alterations and increasing duration of ischemia have been documented. The dynamic changes and histopathology of the microcirculation included constriction of the arteries, swelling of endothelial and leukocytes, and erythrocyte rouleaux formation during ischemia. As ischemia duration was lengthened, the degree of these changes increased correspondingly. The changes on reperfusion were disruption of blood flow patterns, vortex formation, regional stasis, adhesion and migration of leukocytes, focal hemorrhage, edema, vasospasm, and platelet aggregation. The deleterious effects of systemic acidosis, interstitial hemorrhage, denervation, and prolonged venous occlusion were subsequently documented. The application of information gained from this series of laboratory experiments has resulted in continued improvement in the success rate in clinical microvascular surgery.