Grass-specific CD4(+) T-cells exhibit varying degrees of cross-reactivity, implications for allergen-specific immunotherapy.

BACKGROUND: Conceptually, allergic responses may involve cross-reactivity by antibodies or T-cells. While IgE cross-reactivity among grass-pollen allergens has been observed, cross-reactivity at the allergen-specific T-cell level has been less documented. Identification of the patterns of cross-reactivity may improve our understanding, allowing optimization of better immunotherapy strategies. OBJECTIVES: We use Phleum pratense as model for the studying of cross-reactivity at the allergen-specific CD4(+) T cell level among DR04:01 restricted Pooideae grass-pollen T-cell epitopes. METHODS: After in vitro culture of blood mono-nucleated cells from grass-pollen-allergic subjects with specific Pooideae antigenic epitopes, dual tetramer staining with APC-labelled DR04:01/Phleum pratense tetramers and PE-labelled DR04:01/Pooideae grass homolog tetramers was assessed to identify cross-reactivity among allergen-specific DR04:01-restricted T-cells in six subjects. Direct ex vivo staining enabled the comparison of frequency and phenotype of different Pooideae grass-pollen reactive T-cells. Intracellular cytokine staining (ICS) assays were also used to examine phenotypes of these T-cells. RESULTS: T-cells with various degrees of cross-reactive profiles could be detected. Poa p 1 97-116 , Lol p 1 221-240 , Lol p 5a 199-218 , and Poa p 5a 199-218 were identified as minimally cross-reactive T-cell epitopes that do not show cross-reactivity to Phl p 1 and Phl p 5a epitopes. Ex vivo tetramer staining assays demonstrated T-cells that recognized these minimally cross-reactive T-cell epitopes are present in Grass-pollen-allergic subjects. CONCLUSIONS: Our results suggest that not all Pooideae grass epitopes with sequence homology are cross-reactive. Non-cross-reactive T-cells with comparable frequency, phenotype and functionality to Phl p-specific T-cells suggest that a multiple allergen system should be considered for immunotherapy instead of a mono-allergen system.

Comparative investigations of anatomy and physiology in mammalian noses (Homo sapiens--Artiodactyla).

BACKGROUND: Knowledge of airflow in animal noses is sparse. Such knowledge could be important for selection of animal models used in environmental studies. From the phylogenetic and ontogenetic point of view, a comparison between the animal and human nose is interesting. METHOD: Nose models of 5 even-toed ungulate species (he-goat, sheep, cow, roebuck, wild boar) and two humans (new born infant and adult) were examined. Anatomical and physiological features of the nasal cavities of all species were compared. All models were rinsed with water and the flow was visualized for observation. Geometric and rhinoresistometric measurements were then performed. RESULTS AND CONCLUSIONS: Even-toed ungulates have two turbinates directly in the main part of the nasal airflow (respiratory turbinates) and a different number of turbinates in a so-called dead space of the nasal airflow above the nasopharyngeal duct (ethmoidal turbinates). The latter correspond with the upper and middle turbinate in analogy to the human nose. Respiratory turbinates of even-toed ungulates insert immediately behind the external nasal ostium. Thus, the whole nasal cavity acts as a functional area with the exception of a small area acting as dead space only detectable in ruminants, possibly indicating a small evolutionary progress from suinae to bovidae. The shape of the animal nasal cavity is stretched and flat. The airflow runs nearly completely turbulent through the nose. The nasal cavity in the adult human is relatively short and high. The area between the external nasal ostium and the head of the inferior turbinate is called inflow area. It distributes the airflow over the whole nasal cross section and generates a turbulent flow. So the airflow is prepared to contact the mucosa in the functional area (turbinate area). The morphology of the inflow area is approximately formed by the shape of the external nose. The nasal cavity of a newborn child is also stretched and flat and more similar to the nasal shape of the investigated animals. The inflow area in the newborn nose is not yet developed and corresponds with the growing external newborn nose. One can hypothesize that the inflow area in human noses is a morphological adaptation in the changed length-height-ratio of the nasal cavity.

Noradrenergic modulation of hypoglossal motoneuron excitability: developmental and putative state-dependent mechanisms.

Hypoglossal (XII) motoneurons (MNs) contribute to diverse behaviors. Their innervation of the genioglossus muscle, a tongue protruder, plays a critical role in maintaining upper airway patency during breathing. Indeed, reduced activity in these motoneurons is implicated in sleep related disorders of breathing such as obstructive sleep apnea (OSA). The excitability of these MNs is modulated by multiple neurotransmitter systems. The focus of this review is on the modulation of XII MN excitability by norepinephrine (NE), which increases MN excitability through a variety of mechanisms. The level of noradrenergic drive, however, is very dynamic, varying on developmental, sleep-wake and even millisecond timescales relevant to transitions between behaviours. Here we review and provide new data on the maturation of the noradrenergic modulatory system, focusing on those elements specifically relevant to XII MN excitability including the: i) ontogeny of the noradrenergic cell group that provides the majority of the noradrenergic innervation to the XII nucleus, the Locus subcoeruleus (LsC); ii) time course over which the XII nucleus is innervated by noradrenergic nerve fibres, and; iii) ontogeny of XII MN sensitivity to NE. In the context of state-dependent changes in noradrenergic cell activity, we review mechanisms of NE action most relevant to its role in the muscle atonia of REM sleep. We conclude with a discussion of the hypothesis that the dynamics of MN modulation by NE extend to the spatial domain and recent data suggesting that noradrenergic modulation of the dendritic tree is not uniform but compartmentalized. Implications for information processing are discussed.

Risk factors for goiter in a previously iodine-deficient region.

OBJECTIVE: Little information exists from formerly iodine-deficient areas regarding gender-specific risk factors for goiter and their synergisms. The aim of the present study was to investigate such gender-specific risk factors and their interactions in a large population-based sample. METHODS AND RESULTS: The Study of Health in Pomerania (SHIP) comprised 4310 randomly selected participants, aged 20 - 79 years. SHIP was performed in a previously iodine-deficient region. Data from 3915 participants with no known thyroid disorders were analyzed. Goiter was determined by thyroid ultrasound. Sociodemographic characteristics, smoking and alcohol drinking habits, marital status, education level, urine thiocyanate concentrations, and specifically in women, parity and previous or current use of oral contraceptives and hormone replacement therapy, were considered as candidate risk factors for multivariable statistical tests. Only two variables, an advanced age and current smoking, were independently associated with an increased risk for goiter in both genders. Analyses further revealed specific risk factor profiles for goiter which were different among men, pre- and postmenopausal women. CONCLUSION: We conclude that besides previous iodine deficiency, other risk factors for goiter exist which differ between gender. Among the avoidable risk factors, current smoking was strongly associated with the risk of goiter in men and women. These findings should influence activities which are intended to prevent thyroid disease.

Investigations for the diagnostic recording of nasal wing collapse.

BACKGROUND: The inspiratory medial movement of the nasal wing at high flow velocities is a protective physiologic mechanism. If this collapse of the nasal wing occurs at lower flow velocities, it may result in nasal obstruction. "Nasal wing collapse" is generally a clinical diagnosis. However, in the pressure-flow relationship of rhinomanometry, the medial movement of the nasal wing can be documented in the inspiratory arm of the graph. The diagnostic impact of this hysteresis was investigated. METHODS: The pressure-flow curves of three box models and three nasal models with a moveable valve (analogous to the nasal wing) in the entrance area as well as three volunteers with unstable nasal wings were investigated. We recorded synchronously the pressure-flow relationship and by endoscopy the movement of the valve in the box models and the nasal wing in the volunteers on video. For evaluation, we used the frame by frame analysis of the tape. RESULTS AND CONCLUSIONS: The medial movement of the nasal wing causes a hysteresis in the inspiratory arm of the pressure-flow curve. At this point, the graph runs on or between two border curves, termed the "no collapse curve" (for the maximally opened valve or a stable nasal wing) and the "collapse curve" (for the subtotally closed valve or a collapsed nasal wing), respectively. Analogous to the nasal wing motion, the descending course of hysteresis runs in two phases, and the ascending course runs in three phases. The medial movement of the nasal wing is expressed by a deviation of the graph from the "no collapse curve." The flow, at which the graph leaves this curve, depends on the elasticity module of the nasal wing. The extent of nasal wing collapse is reflected by the approximation of the pressure-flow curve to the "collapse curve" of the graph. The hysteresis appears because of a late opening of the collapsed nasal wing.

[Fluoroscopy time -- an overestimated factor for patient radiation exposure in invasive cardiology]. / Durchleuchtungszeit -- Ein überschätzter Parameter der Patientendosis in der invasiven Kardiologie.

PURPOSE: To analyze the effects of an optimized fluoroscopy time on patient radiation exposure in the course of coronary angiography (CA) and percutaneous coronary interventions (PTCA), in comparison to those with consistent collimation to the region of interest (ROI). Furthermore, to analyze efforts concerning reduction of radiographic frames as well as concerning adequate instead of best possible image quality. MATERIAL AND METHODS: For 3,115 elective CAs and 1,713 PTCA performed by one interventionist since 1997, we documented the radiographic dose-area products (DAP (R)) and fluoroscopic dose-area products (DAP (F)), the number of radiographic frames and the fluoroscopy times during selected 2-month intervals. Under conditions of constant image intensifier entrance dose, levels of DAP (R)/frame and DAP (F)/s represent valid parameters for consistent collimation. RESULTS: In 1997, the mean baseline values of DAP for elective CA and PTCA amounted to 37.1 and 31.6 Gy x cm (2), respectively. A reduction of mean fluoroscopy times from 264 to 126 seconds for CA and from 630 to 449 seconds for PCI, both resulted in an overall DAP-reduction of merely 20 %. Optimization of mean radiographic frames from 543 to 98 for CA and from 245 to 142 for PTCA enabled reductions of 53 and 13 %, respectively. By restriction to adequate instead of best-possible image quality for coronary angiography in clinical routine, we achieved an optimized radiographic DAP/frame of 30.3 to 13.3 mGy x cm (2), which enabled a 45 % reduction of overall DAP. Most efficient however was a consistent collimation to the ROI, which resulted in a remarkable radiation reduction by 46 % for CA and by 65 % for PTCA. CONCLUSIONS: Radiation-reducing educational efforts in the clinical routine of invasive cardiology should -- against widely held opinion -- focus less exclusively toward a reduction of fluoroscopy time but more efficiently toward consistent collimation to the region of interest, reduction of radiographic frames and restriction to an adequate instead of best-possible image quality.

Dosimetric evaluation of lung tumor immobilization using breath hold at deep inspiration.

PURPOSE: To examine the dosimetric benefit of self-gated radiotherapy at deep-inspiration breath hold (DIBH) in the treatment of patients with non-small-cell lung cancer (NSCLC). The relative contributions of tumor immobilization at breath hold (BH) and increased lung volume at deep inspiration (DI) in sparing high-dose lung irradiation (> or = 20 Gy) were examined. METHODS AND MATERIALS: Ten consecutive patients undergoing radiotherapy for Stage I-IIIB NSCLC who met the screening criteria were entered on this study. Patients were instructed to BH at DI without the use of external monitors or breath-holding devices (self-gating). Computed tomography (CT) scans of the thorax were performed during free breathing (FB) and DIBH. Fluoroscopy screened for reproducible tumor position throughout DIBH, and determined the maximum superior-inferior (SI) tumor motion during both FB and DIBH. Margins used to define the planning target volume (PTV) from the clinical target volume included 1 cm for setup error and organ motion, plus an additional SI margin for tumor motion, as determined from fluoroscopy. Three conformal treatment plans were then generated for each patient, one from the FB scan with FB PTV margins, a second from the DIBH scan with FB PTV margins, and a third from the DIBH scan with DIBH PTV margins. The percent of total lung volume receiving > or = 20 Gy (using a prescription dose of 70.9 Gy to isocenter) was determined for each plan. RESULTS: Self-gating at DIBH was possible for 8 of the 10 patients; 2 patients were excluded, because they were not able to perform a reproducible DIBH. For these 8 patients, the median BH time was 23 (range, 19-52) s. The mean percent of total lung volume receiving > or = 20 Gy under FB conditions (FB scan with FB PTV margins) was 12.8%. With increased lung volume alone (DIBH scan with FB PTV margins), this was reduced to 11.0%, tending toward a significant decrease in lung irradiation over FB (p = 0.086). With both increased lung volume and tumor immobilization (DIBH scan with DIBH PTV margins), the mean percent lung volume receiving > or = 20 Gy was further reduced to 8.8%, a significant decrease in lung irradiation compared to FB (p = 0.011). Furthermore, at DIBH, the additional benefit provided by tumor immobilization (i.e., using DIBH instead of FB PTV margins) was also significant (p = 0.006). The relative contributions of tumor immobilization and increased lung volume toward reducing the percent total lung volume receiving > or = 20 Gy were patient specific; however, all 8 of the patients analyzed showed a dosimetric benefit with this DIBH technique. CONCLUSION: Compared to FB conditions, at DIBH the mean reduction in percent lung volume receiving > or = 20 Gy was 14.3% with the increase in lung volume alone, 22.1% with tumor immobilization alone, and 32.5% with the combined effect. The dosimetric benefit seen at DIBH was patient specific, and due to both the increased lung volume seen at DI and the PTV margin reduction seen with tumor immobilization.

Development of the ventilatory response to hypoxia in Swiss CD-1 mice.

We examined developmental changes in breathing pattern and the ventilatory response to hypoxia (7.4% O(2)) in unanesthetized Swiss CD-1 mice ranging in age from postnatal day 0 to 42 (P(0)-P(42)) using head-out plethysmography. The breathing pattern of P(0) mice was unstable. Apneas were frequent at P(0) (occupying 29 +/- 6% of total time) but rare by P(3) (5 +/- 2% of total time). Tidal volume increased in proportion to body mass ( approximately 10-13 ml/kg), but increases in respiratory frequency (f) (55 +/- 7, 130 +/- 13, and 207 +/- 20 cycles/min for P(0), P(3), and P(42), respectively) were responsible for developmental increases in minute ventilation (690 +/- 90, 1,530 +/- 250, and 2,170 +/- 430 ml. min(-1). kg(-1) for P(0), P(3), and P(42), respectively). Between P(0) and P(3), increases in f were mediated by reductions in apnea and inspiratory and expiratory times; beyond P(3), increases were due to reductions in expiratory time. Mice of all ages showed a biphasic hypoxic ventilatory response, which differed in two respects from the response typical of most mammals. First, the initial hyperpnea, which was greatest in mature animals, decreased developmentally from a maximum, relative to control, of 2.58 +/- 0.29 in P(0) mice to 1. 32 +/- 0.09 in P(42) mice. Second, whereas ventilation typically falls to or below control in most neonatal mammals, ventilation remained elevated relative to control throughout the hypoxic exposure in P(0) (1.73 +/- 0.31), P(3) (1.64 +/- 0.29), and P(9) (1. 34 +/- 0.17) mice but not in P(19) or P(42) mice.

Increased peak oxygen consumption of trained muscle requires increased electron flux capacity.

The importance of the training-induced increase in mitochondrial capacity in realizing the increase in maximal O2 consumption (VO2max) of trained muscle was evaluated using an isolated perfused rat hindlimb preparation at a high blood flow (approximately 80 ml.min-1.100 g-1) during tetanic contractions. Rats trained for 8-12 wk by treadmill running exhibited an approximately 25% increase in muscle VO2max (5.62 +/- 0.31 to 7.06 +/- 0.64 mumol.min-1.g-1), an increase in mitochondrial enzyme activity (approximately 70% for cytochrome oxidase and approximately 55% for NADH cytochrome-c reductase), and an increase in tissue capillarity (14%) that is expected to increase the O2 exchange capacity of the tissue. Muscle VO2max of sedentary (n = 34) and trained (n = 30) animals was determined, and electron transport capacity was acutely managed with myxothiazol, a tight-binding inhibitor of complex III. Inhibition of complex III was similar among 1) the low- and high-oxidative fibers and 2) the superficial and deep mitochondrial populations within muscle. Inhibition of NADH cytochrome-c reductase activity resulted in reductions in muscle VO2max with similar dose responses (mean effective dose of approximately 0.2 microM) of myxothiazol added to the perfusion medium. The extraction of O2 by the contracting muscle decreased as VO2max declined. The increase in muscle VO2max observed in the muscle of trained animals was eliminated when its electron transport capacity was reduced to that observed in normal sedentary rat muscle. Thus, the exercise-induced adaptation of an increased muscle mitochondrial content appears to be essential for trained muscle to exhibit its increased O2 flux capacity. The results of the present experiment illustrate the importance of mitochondrial adaptations in muscle remodeled by exercise training.

Serological evidence for a high incidence of transmission of Rickettsia tsutsugamushi in two Orang Asli settlements in Peninsular Malaysia.

Two communities of Orang Asli (aborigines) in Peninsular Malaysia were observed for evidence of Rickettsia tsutsugamushi infection over periods of 1-8 mo. Sequential sera were examined for antibody by the indirect immunofluorescence test. The incidence of infection in the two self-selected populations in the two communities was calculated to be 3.9% per month and 3.2% per month.

Adaptation of a microimmunofluorescence test to the study of human Rickettsia tsutsugamuskh antibody.

A microimmunofluorescence (micro-IFA) test has been adapted to the study of naturally occurring antibody to Rickettsia tsutsugamushi in humans. The micro-IFA test was comparable to the present immlnofluorescent assay in sensitivity and reproducibility, but offered distinct advantages in the quantity of reagents necessary. Also the micro-IFA could be used to simultaneously titer a serum to as many as nine strains of the organism resulting in a great saving of time.

Identification and antigenic typing of Rickettsia tsutsugamushi in naturally infected chiggers (Acarina: Trombiculidae) by direct immunofluorescence.

Rickettsia tsutsugamushi organisms were detected and typed antigenically by direct immunofluorescence in mites from laboratory-maintained infected colonies of Leptotrombidium (Leptotrombidium) fletcheri and L. (L.) arenicola. Rickettsiae were identified most readily in unengorged larvae, but were also discernable in engorged larvae and all post-larval stages of the vectors.

A method for activating neurons using endogenous synaptic waveforms.

Neuronal input-output functions are traditionally studied using rectangular or ramp waveforms of injected current. These waveforms are easy to produce and responses to them easy to quantify; thus they have been central to our understanding of the roles that membrane properties play in controlling repetitive firing. However, since smooth rectangular step and ramp waveforms lack the dynamic features of endogenous synaptic input, their use has the potential to underemphasize the importance of input patterns in controlling physiological patterns of neuronal output. To activate neurons with current waveforms that replicate natural synaptic input, we developed a method for acquiring, digitally manipulating and reinjecting endogenous synaptic currents. We demonstrate, by applying this technique to phrenic motoneurons (PMNs) in rhythmically-active in vitro preparations from neonatal rats, that stimulation of neurons with endogenous current waveforms produces responses that mimic those produced by spontaneous synaptic inputs. Acquired waveforms can be reinjected repeatedly to produce consistent responses, and can also be amplified or filtered prior to reinjection to yield a range of information including standard descriptors of firing behavior such as frequency/current plots. This technique provides a valuable tool for analysing characteristics of the synaptic waveform important in generating neuronal output and how synaptic factors interact with membrane properties to control repetitive firing.

Optimized tissue compensators.

The fabrication of retracted missing tissue compensators which account only for missing primary attenuation produces compensation which is less than optimum. Experimental results can be accurately reproduced with theoretical calculations based on primary and first-order scattered radiation. This method of analysis may be extended to predict the shape of a compensator which will produce the desired dose distribution at a given depth in phantom. An optimized compensator is constructed based on these theoretical considerations and the analysis of its performance is presented.

Modeling of asymmetric compensator geometries.

Dose distributions arising from the use of near unit density retracted missing tissue compensators in symmetric geometries have been successfully modeled on the basis of primary and first-order scattered radiation. This method of analysis has been extended to both low- and high-density materials in asymmetric geometries. Good agreement is achieved between theory and experiment.

Missing tissue compensators: evaluation and optimization of a commercial system.

A commercial system for producing retracted compensators has been adapted to suit local needs, and is evaluated here. It comprises a magnetic field surface digitizer and computer-driven milling machine. Improvements in dose distributions, resulting in standard deviations of the mean dose between 2% and 3%, have been achieved for treatment fields in wax phantoms simulating the head and neck regions. Optimization of compensator shape to allow for changes in the amount of scattered radiation has resulted in a further improvement in dose uniformity, particularly near the field borders; for these compensators the standard deviation was as low as 1.6%. The system using the basic algorithm has been in clinical use since July.

Scrub typhus: a common cause of illness in indigenous populations.

An explanation was sought for the disparity between the low reported incidence of scrub typhus and the high prevalence of antibody to Rickettsia tsutsugamushi in the rural population of Malaysia. A combination of isolation of the organism, titration of antibody by indirect immunofluorescence, and the Weil-Felix test was used to confirm infections. Scrub typhus was found to be very common, causing 23% of all febrile illnesses at one hospital. The infection was particularly prevalent in oil-palm workers, causing an estimated 400 cases annually in a population of 10,000 people living on one plantation. The clinical syndrome, whether mild or severe, was difficult to distinguish from that due to other infections. Eschars, rashes and adenopathy were uncommon. When used to examine early sera, the Weil-Felix test failed to confirm the diagnosis in most infections.20

P2 receptors modulate respiratory rhythm but do not contribute to central CO2 sensitivity in vitro.

Multiple brainstem sites are proposed to contribute to central respiratory chemosensitivity, however, the underlying molecular mechanisms remain unknown. P2X2 subunit-containing ATP receptors, which mediate pH-sensitive currents, appear to contribute to central chemosensitivity in vivo [J. Physiol. 523 (2000) 441]. However, recent data from P2X2 knockout mice [J. Neurosci. 23 (2003) 11315] indicate that they are not essential. To further explore the role of P2 receptors in central chemosensitivity, we examined the effects of P2 receptor agonists/antagonists on respiratory-related activity and CO2-sensitivity of rhythmically-active in vitro preparations from neonatal rat. Our main findings: (i) that putative chemosensitive regions of the ventrolateral medulla are immunoreactive for the P2X2 subunit; (ii) that ATP potentiates respiratory frequency in a dose-dependent, and PPADS-sensitive (P2 receptor antagonist), manner; and (iii) that the increase in burst frequency produced by increasing CO2 is unaffected by PPADS, indicate that ATP is a potent modulator of respiratory activity, but that P2 receptors do not contribute to central chemosensitivity in vitro.

Training intensity of elite male distance runners.

To quantify training intensity in 13 nationally ranked male distance runners, training heart rates, environmental factors, and motivational factors were recorded throughout a 6-8 wk period of normal training. Variation in motivational and environmental factors such as intended effort, terrain, and running with companions altered training session mean heart rates by up to 4.min-1 (standard deviation). Heart rates and blood lactate concentrations, recorded in a series of steady-state treadmill runs, were used to convert training session mean heart rates to mean training speeds (TS) and to derive a measure of the anaerobic threshold (AT, the treadmill speed at a blood lactate concentration of 4 mmol.l-1). TS (15.6 +/- 1.4 km.h-1, mean +/- SD) was considerably lower than AT (20.2 +/- 1.1 km.h-1) in all subjects (P less than 0.001). Relative TS (TS expressed as a fraction of AT) differed significantly (P less than 0.001) between subjects and correlated significantly with the distance of the event for which the subject was training (r = 0.59, P less than 0.05). Relative TS may therefore be determined by the subject's or coach's perception of the appropriate intensity for the event. If the AT is the optimum training intensity, these subjects have considerable scope for improvement.

Monoenergetic approximately of a polyenergetic beam: a theoretical approach.

There exist numerous occasions in which it is desirable to approximate the polyenergetic beams employed in radiation therapy by a beam of photons of a single energy. In some instances, commonly used rules of thumb for the selection of an appropriate energy may be valid. A more accurate approximate energy, however, may be determined by an analysis which takes into account both the spectral qualities of the beam and the material through which it passes. The theoretical basis of this method of analysis is presented in this paper. Experimental agreement with theory for a range of materials and beam qualities is also presented and demonstrates the validity of the theoretical approach taken.