Neural stem cells from human cord blood on bioengineered surfaces--novel approach to multiparameter bio-tests.

Stem cell technology combined with emerging surface nano/micro-technologies provides a new tool for better understanding of the mechanisms involved in cell fate decisions and compound-induced adverse reactions. This article provides state-of-the-art on the development of modern multiparameter bio-tests based on interactions between neural stem cells derived from human cord blood and bioengineered surfaces. Cell growth platforms with controlled content, geometry and spatial distribution of bioactive and stem cell attractive areas were fabricated either by micro-contact printing or piezoelectric spotting of polycationic biomolecules or extracellular matrix proteins (ECM) on cell-repellent surfaces. HUCB-NSCs were shown to adhere, differentiate and respond to neurotoxic MeHgCl on functional domains in a manner dependent on protein type and concentration, cell density and serum conditions. While receptor-mediated interactions with ECM proteins under absence of serum promote neuronal differentiation, non-specific adhesion to polycationic molecules maintain cells attached to the surface in non-differentiated stage. Functional domains were further engineered to create "smart" microenvironment by immobilizing to the surface signaling molecules together with ECM proteins. Stimulation of selected intracellular pathways by molecules of Wnt, Shh, CNTF or Notch type resulted in differentiation of HUCB-NSC to either neuronal or astroglial lineage. Sensor techniques applied to HUCB-NSC included measurements of electrical activity using multielectrode array chips. Spontaneous electrical field potentials of HUCB-NSCs were dependent upon developmental stage of tested cells. Bioengineered surfaces, on protein microarrays and micro-electrode array chips provide a novel approach to the multiparameter bio-tests by adding an important information on the sensitivity of certain molecular pathways and functional cellular responses to selected neurotoxins.

Pt(x)Ru(1-x)/Ru(0001) surface alloys-formation and atom distribution.

The formation of PtRu surface alloys by deposition of submonolayer Pt films on a Ru(0001) substrate and subsequent annealing to about 1350 K and the distribution of the Pt atoms in the surface layer were investigated by scanning tunneling microscopy. Quantitative statistical analysis reveals (i) negligible losses of Pt into subsurface regions up to coverages close below 1 monolayer, (ii) a homogeneous distribution of the Pt atoms over the surface, and (iii) the absence of a distinct long-range or short-range order in the surface layer. In addition, the density of specific adsorption ensembles is analyzed as a function of Pt surface content. Possible conclusions on the process for surface alloy formation are discussed. The results are compared with the properties of PtRu bulk alloys and the findings in previous adsorption studies on similar surface alloys (H. Rauscher, T. Hager, T. Diemant, H. Hoster, F. Bautier de Mongeot and R. J. Behm, Surf. Sci., 2007, 601, 4608; T. Diemant, H Rauscher and R. J. Behm, J. Phys. Chem. C, in press).

Poly(N-isopropylacrylamide) grafted on plasma-activated poly(ethylene oxide): thermal response and interaction with proteins.

Thermoresponsive polymer layers offer the possibility of preparing smart surfaces with properties that are switchable through a phase transition, usually close to the lower critical solution temperature of the polymer. In particular, poly( N-isopropylacrylamide) (pNIPAM) has gained a great deal of attention because it has such a phase transition in a physiologically interesting temperature range. We have prepared ultrathin thermoresponsive coatings by grafting pNIPAM on a plasma-CVD-deposited, poly(ethylene oxide)-like polymer substrate that was activated in an Ar plasma discharge to initiate the grafting. The presence and integrity of pNIPAM was verified by XPS and ToF-SIMS, and a dramatic change in the wettability during the phase transition was identified by temperature-dependent contact angle measurements. The transition from the hydrated to the collapsed conformation was analyzed by temperature-dependent QCM measurements and by AFM. An unusual, reversible behavior of the viscoelastic properties was seen directly at the phase transition from the swollen to the collapsed state. The phase transition leads to a switching from protein repulsion to a state that allows the adsorption of proteins.

Formation of viscoelastic protein droplets on a chemically functionalized surface.

Droplet formation during adsorption of the protein lactoferrin from an aqueous solution on a surface functionalized by plasma deposited poly(acrylic acid) is studied using quartz crystal microbalance and atomic force microscopy. The formation of protein droplets is particularly favored at pH values close to the isoelectric point of lactoferrin, where the molecules carry little excess charge and intermolecular attraction exceeds the molecule-surface interaction. By combining topographic data with information on the system dynamics, it is possible to describe the viscoelastic properties of the adsorbate within a quantitative model for nonhomogeneous layers.

Direct identification of critical clusters in chemical vapor deposition.

The kinetics of heteroepitaxial island nucleation on the Si(111) surface during ultrahigh vacuum chemical vapor deposition with the precursor GeH4 was studied by scanning tunneling microscopy. The results can be described within the framework of rate-equation based nucleation theory, modified by an additional energetic barrier for the attachment of adatoms at steps. This barrier results from the passivation of steps by dissociation products from the GeH4 precursor. A critical nucleus size of 9 atoms is derived. Scanning tunneling microscopy images provide direct evidence for the existence of stable clusters consisting of 10 atoms and allow the unequivocal identification of their structure.

Nasal continuous positive airway pressure for nonapneic snoring?

The feasibility of nasal continuous positive airway pressure (CPAP) for heavy snoring associated with daytime sleepiness was studied in 118 consecutive patients with an apnea hypopnea index below 5. Fifty-nine of them reported daytime sleepiness in a questionnaire and were offered treatment with nasal CPAP. Whereas 48 patients refused it, the remaining 11 (19%) accepted nasal CPAP for home therapy. Acceptors and refusers did not differ in sleep structure, but acceptors had slightly more sleep-disordered breathing events per hour of sleep than refusers. The pressure needed to abolish snoring in these 11 patients was 7.3 +/- 1.6 cm H2O. Six months after prescription, the built-in time counters of the patients' devices were read. By dividing the hours of operation by the days since initiation of treatment, we found a mean daily use time of only 2.8 +/- 1.5 h. Nevertheless, eight patients (73%) reported that their sleepiness had improved with therapy. We conclude that only a minority of nonapneic snorers accept treatment with nasal CPAP on a long-term basis and that this subgroup is not predictable from polysomnography.

Chest X-ray in collagen vascular diseases. A comparison of chest X-ray with bronchoalveolar lavage and transbronchial forceps biopsy.

The diagnostic value of chest X-ray following the ILO standards was compared with bronchoalveolar lavage (BAL) and the histology of transbronchial forceps biopsy in 83 patients with collagen vascular diseases. BAL in the middle lobe was considered abnormal in case of increased cell count per milliliter and/or lymphocytosis and/or neutrophil granulocytosis and this was found in 32 out of 42 cases (76.2%) with, and in 33 of out 41 cases (80.5%) without abnormal radiological finding. Pathological changes in the histology were found in 14 out of 20 cases (70.0%) with and in 40 out of 63 cases (63.5%) without abnormal chest X-ray in the upper lobe from which the transbronchial forceps biopsy specimens were obtained. In histological specimens obtained from transbronchial forceps biopsy, only fibrosis correlated with abnormal radiological findings in this region. Other inflammatory processes defied prediction by chest X-ray. This suggests that, regardless of chest X-ray findings, BAL should be performed together with transbronchial forceps biopsy for the histological examination of patients with collagen vascular diseases in which interstitial lung involvement is suspected.

Inspiratory muscle performance relative to the ventilatory threshold in healthy subjects.

Inspiratory muscle performance, ventilation, and gas exchange were studied during exercise in healthy subjects to look for typical changes of pattern of contraction at the ventilatory threshold (VT). The steepening of the slope of carbon dioxide output (VCO2) vs oxygen uptake (VO2) at the VT was accompanied by a nonlinear increase of the mean rate of esophageal pressure development (Pes/TI) vs the esophageal pressure time index (PTIes) reflecting both the relative force (Pbreath/Pesmax) and duration (TI/TTOT) required for inspiration. The esophageal pressure time integral within one breath (Pbreath.dTI) was one of the best single predictors of the ventilatory equivalent for oxygen (VE/VO2) at the VT. Moreover, we presented inspiratory muscle load indices as a mirror image of breathing pattern, with the obvious advantage that the ventilation component can be compared with better established methods of presenting ventilatory output. Inspiratory muscle performance during exercise should link the increased metabolic rate to ventilatory output. We conclude that 1) there exists an inspiratory muscle threshold that is well correlated to commonly used gas exchange thresholds, and 2) the efficiency of ventilation and gas exchange during exercise could be linked to pressure and timing of inspiratory muscle contraction.

Inspiratory muscle performance relative to the anaerobic threshold in patients with COPD.

Rehabilitation programmes in chronic obstructive pulmonary disease (COPD) require exercise training above the anaerobic threshold. However, not all COPD patients develop metabolic acidosis during exercise. The hypothesis of this study was that non-exercise variables, characterizing the mechanical load on the inspiratory muscles during breathing at rest, can be used to reliably predict which patients with COPD are not able to develop metabolic acidosis during exercise. Thirty participants with COPD performed a symptom-limited cycle ergometer test. The oesophageal pressure/time index (PTIoes: the product of pressure magnitude and duration), the mean rate of pressure development during inspiration (Poes/TI), and the mean airway resistance (Raw)/maximal oesophageal pressure (Poesmax) ratio served as indices for the mechanical load on the inspiratory muscles. The oxygen uptake (VO2) at which plasma standard bicarbonate was seen to decrease from its baseline value was taken as the anaerboic threshold (AT). Mean Raw was significantly higher in those patients in whom the AT could not be detected. No other lung function parameters measured at rest allowed the accurate selection of those patients who did or did not develop exercise metabolic acidosis. On the other hand, Raw/Poesmax, PTIoes and Poes/TI were significantly different in the two patient groups. Additionally, whereas in the patient group with identifiable AT exercise hyperpnoea produced a non-linear increase of Poes/TI with respect to PTIoes above the AT, in the patient group without identifiable AT there was a linear relationship between Poes/TI and PTIoes throughout exercise. We conclude that the determination of inspiratory muscle load indices at rest may be useful in pulmonary rehabilitation programmes, for identifying those patients with COPD who do not develop exercise induced metabolic acidosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Self-reported vs measured compliance with nasal CPAP for obstructive sleep apnea.

To estimate reliability of self-reported compliance with nasal continuous positive airway pressure (CPAP) therapy for obstructive sleep apnea (OSA), we studied 63 OSA patients aged 53.7 +/- 1.2 years (mean +/- SEM) with an apnea hypopnea index (AHI) of 50.8 +/- 2.9 and lowest sleep SaO2 of 65.6 +/- 2.3 percent receiving nasal CPAP for 539 +/- 44 days. During a follow-up polysomnography (PSG) on the pressure prescribed for home therapy (10.3 +/- 0.3 cm H2O), the hours of operation shown on the built-in time counter of the patients' devices were read to determine objective compliance by dividing the run time by the days since initiation of therapy. This parameter was compared with subjective compliance reported in a self-administered questionnaire. Mean measured use time was 4.9 +/- 0.3 h per night, whereas reported daily use time calculated from reported nights a week and hours a night was 6.1 +/- 0.3 h per night. As predominantly patients with poor compliance misestimated daily use time, we conclude that self-reports are unable to distinguish between compliant and noncompliant patients.

Nasal CPAP and weight loss in hypertensive patients with obstructive sleep apnoea.

BACKGROUND: The high prevalence of obstructive sleep apnoea (OSA) in patients with systemic hypertension and of hypertension in patients with OSA suggests a causal link between the two disorders. This study was carried out to determine whether nasal continuous positive airway pressure (CPAP) and weight loss affect daytime hypertension in OSA. METHODS: Sixty hypertensive patients with OSA took part in the study; 33 accepted nasal CPAP and used their machine for 5.7 (0.2) hours per night, and the remaining 27 patients refused nasal CPAP and upper airway surgery so the only therapeutic intervention was a recommendation of weight loss. A significant change in hypertension during follow up was defined as either a change in mean blood pressure of at least 10 mm Hg (or more than 8%) without a change in drug treatment, or a reduction in drug dosage with mean blood pressure within these limits. Weight loss was defined as a body mass index of at least 5% below the baseline value. RESULTS: After 512 (41) days, hypertension had become less severe in seven of 12 patients (58%) treated with weight loss only, in eight of 28 patients (29%) with nasal CPAP only, in two of five patients with nasal CPAP and weight loss, and in one of 15 patients without nasal CPAP or weight loss. Multivariate analysis of variance with the outcome of hypertension at follow up as the dependent variable revealed that only the percentage change in body mass index significantly contributed to the course of hypertension. CONCLUSION: The course of hypertension in OSA is more closely linked to weight loss than to elimination of sleep apnoea by nasal CPAP.

Model for investigating snorers with suspected sleep apnoea.

BACKGROUND: Overnight polysomnography is expensive and time consuming. An approach based on a logistic regression model and overnight pulse oximetry has been developed to determine which of the snorers referred to our sleep laboratory need polysomnography. METHODS: The variables entered in the regression model were derived from questionnaires completed by 95 habitual snorers and 89 patients with obstructive sleep apnoea. The resulting regression equation included weight, height, sex, witnessed episodes of apnoea, and reports of falling asleep when reading. This prediction equation was applied to a sample of 116 consecutive patients referred for investigation of heavy snoring. Pulse oximetry data on the 116 test subjects were obtained during polysomnography and analysed separately. Pulse oximetry was judged to indicate obstructive sleep apnoea when it showed cyclic oscillations of oxyhaemoglobin saturation or heart rate, or both, for more than 30 minutes during the study night. RESULTS: A cut off probability of 0.31 gave the prediction model a sensitivity of 94% to predict an apnoea-hypopnoea index above 10, with a specificity of 45%. When this cutoff point was used to predict an apnoea-hypopnoea index of over 20 sensitivity was 95% and specificity 41%. Combined with oximetry our regression model had a sensitivity of 100% for predicting an apnoea-hypopnoea index of more than 10. On the other hand, all patients with negative results from oximetry and a probability value below 0.31 had an apnoea-hypopnoea index lower than 10 according to polysomnography. CONCLUSIONS: It is concluded that snorers with negative results from oximetry classified as not having obstructive sleep apnoea according to this model do not need polysomnography.

How much brushing is enough for the diagnosis of lung tumors?

BACKGROUND: Bronchoscopic investigations of lung tumors require high diagnostic accuracy. Sometimes the combination of brush biopsy with cytologic and histologic examination of forceps-obtained biopsy specimens fails to diagnose tumors. Techniques with a minimum risk and low cost when repeated several times could increase the efficiency of tumor diagnosis and help to avoid rebronchoscopy. METHODS: Repeated brush biopsies were done during one bronchoscopic examination in 270 patients with pulmonary neoplasias using a flexible fiberoptic bronchoscope guided by radiographic video fluoroscopy. The results of up to five brush biopsies were compared for their diagnostic sensitivity. RESULTS: Singly, 68-77% of the specimens showed malignant findings. With repeated brushing, the sensitivity of the diagnostic accuracy increased to 89.6%. In the periphery of the bronchial tree, the sensitivity of brush biopsy was slightly lower in bronchoscopically invisible tumors. In 222 of 242 (91.7%) patients with positive results of brush biopsy, there was agreement in the final typing of tumor morphology. CONCLUSION: For routine bronchoscopy, repeated brush biopsy should be done to obtain the highest diagnostic yield.

Systemic hypertension in snorers with and without sleep apnea.

To investigate the impact of sleep-disordered breathing events on daytime hypertension (HT) in patients with increased upper airway resistance during sleep, we studied 191 male snorers aged 49.9 +/- 0.8 years. In 116 of them, an apnea-hypopnea index (AHI) above 10--defined as the presence of obstructive sleep apnea (OSA)--was found; the other 75 subjects had an AHI lower than 10 and were classified as habitual snorers (HSN). Prevalence of HT was not different between OSA (56 of 116 = 48 percent) and HSN (33 of 75 = 44 percent) and there was also no difference in systolic, diastolic, and mean blood pressures between the two groups. Hypertensive OSA patients had higher body mass index (BMI) than normotensive OSA subjects (31.4 +/- 0.7 vs 29.4 +/- 0.6; p less than 0.05), but there was no difference in age, AHI, and nocturnal oxygenation parameters. The same was true for the HSN group, with hypertensive subjects being more obese than normotensive subjects (BMI: 30 +/- 0.8 vs 27.3 +/- 0.8; p less than 0.05), but no difference in age and polysomnographic features. Discriminant analysis with HT as the classification variable and age, BMI, AHI, mean, and lowest nocturnal oxyhemoglobin saturation as independent variables, revealed an independent influence on HT only for BMI (F-prob = 0.001). Thus, our results stand against the hypothesis of a causal relationship between sleep-disordered breathing events and daytime hypertension. We conclude that the high prevalence of HT in male snorers is more directly linked to obesity than to sleep apnea, but an independent effect of snoring per se cannot be excluded.

Prediction of interstitial lung involvement in rheumatoid arthritis. The value of clinical data, chest roentgenogram, lung function, and serologic parameters.

We initiated the present study to predict interstitial lung involvement in rheumatoid arthritis (RA) by means of logistic regression analysis of clinical data, lung function, chest roentgenogram, and serologic parameters. Fifty-eight nonsmoking patients with RA were randomized from the rheumatologic unit and sent for investigation to the pulmonary department. Bronchoalveolar lavage (BAL) was performed in the middle lobe and the BAL fluid was considered abnormal in case of increased cell count per milliliter and/or lymphocytosis and/or neutrophil granulocytosis; these findings or combinations thereof were found in 42 (72.4 percent) of 58 cases. The patients' data that had an impact on the normality of BAL were the sex (p = 0.001), vital capacity (p = 0.028), peripheral blood T-helper cells (OKT4+) (p = 0.025), DR(+)-lymphocytes (p = 0.002), and antinuclear antibodies (p = 0.025). By means of the logistic regression analysis, it was possible to reach high significance in the prediction of interstitial lung involvement, with a sensitivity of 92.9 percent and a specificity of 75.0 percent (p less than 10(-6)). The efficiency of prediction was 87.9 percent. From these results, we conclude that interstitial lung involvement in RA is predictable from laboratory findings that have been yielded by noninvasive diagnostic techniques. These data should be used in clinical routine monitoring and they may help to facilitate the assessment of whether bronchoscopy is indicated.

Influences of the cellular and humoral immune system in bronchoalveolar lavage on lung function in pulmonary sarcoidosis.

We investigated the changes in the cellular and humoral immune system in bronchoalveolar lavage (BAL) performed in 22 patients with pulmonary sarcoidosis and in 14 normal control subjects and their interactions with lung function parameters. Lymphocytosis, the increase in OKT4+ lymphocytes and OKT4+OKDR+ lymphocytes correlated with the increase in immunoglobulins, especially IgG, IgA and kappa chain assembled immunoglobulins. The transferrin levels obtained in BAL were found to be higher in patients with sarcoidosis, and they correlated with the cellular and, more closely, with other humoral findings. A negative correlation existed between the ventilatory parameters and the cell count and humoral findings. In addition, we found a negative correlation between the diffusing capacity for carbon monoxide and other cellular findings, which was most pronounced with reference to lymphocytes, OKT4+ lymphocytes and the OKT4+/OKT8+ ratio. These results underscore the role of OKT4+ lymphocytes, activated OKT4+OKDR+ lymphocytes and transferrin in the increase in immunoglobulins, mainly kappa chain isotypes. Because of the relationship between these changes and ventilatory parameters, and the diffusing capacity, the above results also reveal the clinical relevance of our findings.

Acceptance of CPAP therapy for sleep apnea.

Although NCPAP is the most efficient nonsurgical treatment for patients with OSA, many patients do not accept sleeping with a nose mask. To determine the factors influencing acceptance, treatment with NCPAP was offered to 95 patients with an AHI greater than 15. After the first night on NCPAP, 47 of 65 patients decided to have NCPAP as a home therapy. Excessive daytime sleepiness was more frequently reported by acceptors than refusers. The frequency of complaints about psychomental symptoms such as poor mental performance and bad memory, was not different between the two groups. There was a close correlation between the rate of acceptance and the AHI as well as the number of positive answers to questions about symptoms of daytime sleepiness in a questionnaire, which correlated with the number and length of apneas. Acceptance of NCPAP was found to be dependent on the subjective feeling of impairment by hypersomnolence due to OSA.

Breathing during sleep in patients treated for obstructive sleep apnea. Nasal CPAP for only part of the night.

To determine whether long-term NCPAP therapy influences severity of sleep disordered breathing during the second part of a night when NCPAP is applied for only the first four hours of sleep, we studied 21 patients with OSA receiving NCPAP therapy for 253 +/- 41.6 days. Results from polysomnography for the period after withdrawal from NCPAP (night B) were compared to the corresponding period of sleep prior to initiation of NCPAP therapy (night A). There was no significant change in RDI from night A (53.9 +/- 8.6) to night B (28.7 +/- 3.3), but maximal apnea length diminished from 55 +/- 2.9 s to 40 +/- 2.9 s (p less than 0.05). Whereas daytime Po2 and the amplitude of desaturations during sleep remained equal, overall oxygenation during sleep improved slightly (mean SaO2 night A = 90.6 +/- 0.9 percent; night B = 92.8 +/- 0.5 percent; p less than 0.05). Differences between nights A and B were more prominent the more severe sleep apnea had been prior to treatment and could not be explained by weight loss. There was strong correlation between improvements in oxygenation measurements and the daily time of NCPAP use. In conclusion, we found a subgroup of OSA patients receiving long-term NCPAP therapy with less disturbed ventilation during sleep following use of NCPAP for only the first part of the night, but in the majority of patients, sleep disordered breathing off NCPAP remained unchanged.

Quantification of sleep disordered breathing by computerized analysis of oximetry, heart rate and snoring.

Intermittent snoring and cyclic oscillations of heart rate and oxyhaemoglobin saturation (Sao2) are characteristic features of the obstructive sleep apnoea syndrome (OSAS). Thus, overnight recordings of laryngeal sounds and heart rate by a portable device (MESAM) and of Sao2 by oximetry are applicable to screen outpatients for the presence of OSAS. Computerized analysis for time intervals of constant heart rate and intervals between snoring sounds is used by MESAM to quantify respiratory disturbances during sleep. Rapid increases in Sao2 during the postapnoeic hyperventilation period together with the number of desaturations are used by a new software for quantitative analysis of oximetry. To elucidate reliability of results from automatically scored MESAM and oximetry recordings, we compared the four computer calculated respiratory disturbance indices from heart rate (RDIH), snoring (RDIS), resaturations (RDIR) and desaturations (RDID) with the apnoea plus hypopnoea index (AHI) from simultaneously performed polysomnography. The study population consisted of 53 snorers with an AHI of 19.0 +/- 2.6 (median +/- SEM; range 0.7-87.8). Whereas both RDI's from MESAM correlated rather weakly with the AHI from polysomnography (RDIH: r = 0.32, p less than 0.05; RDIS: r = 0.33, p less than 0.05), this correlation was much better for the RDI's from oximetry (RDIR: r = 0.951, RDID: r = 0.93; p much less than 0.0001). Accepting a plus/minus 30 percent difference from the AHI, the RDIR classified 77% of patients correctly, the RDID 62%, the RDIS 32% and the RDIH 23%. In conclusion, results from computerized analysis of oximetry for desaturations and rapid resaturations correlate more closely with polysomnography than those from automatic scoring of MESAM recordings.