Predictors of Quality of Life Decline in Patients with Oligometastases treated with Stereotactic Ablative Radiotherapy: Analysis of the Population-Based SABR-5 Phase II Trial.

AIMS: Most patients experience stable quality of life (QoL) after stereotactic ablative radiotherapy (SABR) treatment for oligometastases. However, a subset of patients experience clinically relevant declines in QoL on post-treatment follow-up. This study aimed to identify risk factors for QoL decline. MATERIALS AND METHODS: The SABR-5 trial was a population-based single-arm phase II study of SABR to up to five sites of oligometastases. Prospective QoL was measured using treatment site-specific tools at pre-treatment baseline and 3, 6, 9, 12, 15, 18, 21, 24, 30 and 36 months after treatment. The time to persistent QoL decline was calculated as the time from SABR to the first decline in QoL score meeting minimum clinically important difference with no improvement to baseline score on subsequent assessments. Univariable and multivariable logistic regression analyses were carried out to determine factors associated with QoL decline. RESULTS: One hundred and thirty-three patients were included with a median follow-up of 32 months (interquartile range 25-43). Thirty-five patients (26%) experienced a persistent decline in QoL. The median time until persistent QoL decline was not reached. The cumulative incidence of QoL decline at 2 and 3 years were 22% (95% confidence interval 14.0-29.6) and 40% (95% confidence interval 28.0-51.2), respectively. In multivariable analysis, disease progression (odds ratio 5.23, 95% confidence interval 1.59-17.47, P = 0.007) and adrenal metastases (odds ratio 9.70, 95% confidence interval 1.41-66.93, P = 0.021) were associated with a higher risk of QoL decline. Grade 3 or higher (odds ratio 3.88, 95% confidence interval 0.92-16.31, P = 0.064) and grade 2 or higher SABR-associated toxicity (odds ratio 2.24, 95% confidence interval 0.85-5.91, P = 0.10) were associated with an increased risk of QoL decline but did not reach statistical significance. CONCLUSIONS: Disease progression and adrenal lesion site were associated with persistent QoL decline following SABR. The development of grade 3 or higher toxicities was also associated with an increased risk, albeit not statistically significant. Further studies are needed, focusing on the QoL impact of metastasis-directed therapies.

Prospective Longitudinal Assessment of Quality of Life After Stereotactic Ablative Radiotherapy for Oligometastases: Analysis of the Population-based SABR-5 Phase II Trial.

AIMS: To evaluate longitudinal patient-reported quality of life (QoL) in patients treated with stereotactic ablative radiotherapy (SABR) for oligometastases. MATERIALS AND METHODS: The SABR-5 trial was a population-based single-arm phase II study of SABR to up to five sites of oligometastases, conducted in six regional cancer centres in British Columbia, Canada from 2016 to 2020. Prospective QoL was measured using treatment site-specific QoL questionnaires at pre-treatment baseline and at 3, 6, 9, 12, 15, 18, 21, 24, 30 and 36 months after treatment. Patients with bone metastases were assessed with the Brief Pain Inventory (BPI). Patients with liver, adrenal and abdominopelvic lymph node metastases were assessed with the Functional Assessment of Chronic Illness Therapy-Abdominal Discomfort (FACIT-AD). Patients with lung and intrathoracic lymph node metastases were assessed with the Prospective Outcomes and Support Initiative (POSI) lung questionnaire. The two one-sided test procedure was used to assess equivalence between the worst QoL score and the baseline score of individual patients. The mean QoL at all time points was used to determine the trajectory of QoL response after SABR. The proportion of patients with 'stable', 'improved' or 'worsened' QoL was determined for all time points based on standard minimal clinically important differences (MCID; BPI worst pain = 2, BPI functional interference score [FIS] = 0.5, FACIT-AD Trial Outcome Index [TOI] = 8, POSI = 3). RESULTS: All enrolled patients with baseline QoL assessment and at least one follow-up assessment were analysed (n = 133). On equivalence testing, the patients' worst QoL scores were clinically different from baseline scores and met MCID (BPI worst pain mean difference: 1.8, 90% confidence interval 1.19 to 2.42]; BPI FIS mean difference: 1.68, 90% confidence interval 1.15 to 2.21; FACIT-AD TOI mean difference: -8.76, 90% confidence interval -11.29 to -6.24; POSI mean difference: -4.61, 90% confidence interval -6.09 to -3.14). However, the mean FIS transiently worsened at 9, 18 and 21 months but eventually returned to stable levels. The mean FACIT and POSI scores also worsened at 36 months, albeit with a limited number of responses (n = 4 and 8, respectively). Most patients reported stable QoL at all time points (range: BPI worst pain 71-82%, BPI FIS 45-78%, FACIT-AD TOI 50-100%, POSI 25-73%). Clinically significant stability, worsening and improvement were seen in 70%/13%/18% of patients at 3 months, 53%/28%/19% at 18 months and 63%/25%/13% at 36 months. CONCLUSIONS: Transient decreases in QoL that met MCID were seen between patients' worst QoL scores and baseline scores. However, most patients experienced stable QoL relative to pre-treatment levels on long-term follow-up. Further studies are needed to characterise patients at greatest risk for decreased QoL.

Approaching the motional ground state of a 10-kg object.

The motion of a mechanical object, even a human-sized object, should be governed by the rules of quantum mechanics. Coaxing them into a quantum state is, however, difficult because the thermal environment masks any quantum signature of the object's motion. The thermal environment also masks the effects of proposed modifications of quantum mechanics at large mass scales. We prepared the center-of-mass motion of a 10-kilogram mechanical oscillator in a state with an average phonon occupation of 10.8. The reduction in temperature, from room temperature to 77 nanokelvin, is commensurate with an 11 orders-of-magnitude suppression of quantum back-action by feedback and a 13 orders-of-magnitude increase in the mass of an object prepared close to its motional ground state. Our approach will enable the possibility of probing gravity on massive quantum systems.

Approximate numerical abilities and mathematics: Insight from correlational and experimental training studies.

Humans have the ability to nonverbally represent the approximate numerosity of sets of objects. The cognitive system that supports this ability, often referred to as the approximate number system (ANS), is present in early infancy and continues to develop in precision over the life span. It has been proposed that the ANS forms a foundation for uniquely human symbolic number and mathematics learning. Recent work has brought two types of evidence to bear on the relationship between the ANS and human mathematics: correlational studies showing individual differences in approximate numerical abilities correlate with individual differences in mathematics achievement and experimental studies showing enhancing effects of nonsymbolic approximate numerical training on exact, symbolic mathematical abilities. From this work, at least two accounts can be derived from these empirical data. It may be the case that the ANS and mathematics are related because the cognitive and brain processes responsible for representing numerical quantity in each format overlap, the Representational Overlap Hypothesis, or because of commonalities in the cognitive operations involved in mentally manipulating the representations of each format, the Operational Overlap hypothesis. The two hypotheses make distinct predictions for future work to test.

COMBINED DELAY AND GRAPH EMBEDDING OF EPILEPTIC DISCHARGES IN EEG REVEALS COMPLEX AND RECURRENT NONLINEAR DYNAMICS.

The dynamical structure of the brain's electrical signals contains valuable information about its physiology. Here we combine techniques for nonlinear dynamical analysis and manifold identification to reveal complex and recurrent dynamics in interictal epileptiform discharges (IEDs). Our results suggest that recurrent IEDs exhibit some consistent dynamics, which may only last briefly, and so individual IED dynamics may need to be considered in order to understand their genesis. This could potentially serve to constrain the dynamics of the inverse source localization problem.

Adaptive considerations of temperature dependence of neuromuscular function in two species of summer- and winter-caught Crab (Carcinus maenas and Cancer pagurus).

The aim of this study was to determine seasonal differences in the temperature dependence of neuromuscular parameters of the dactylopodite walking leg closer muscle in two species of freshly caught summer and winter decapod crabs. The relatively stenothermal Cancer pagurus (Cp) and eurythermal Carcinus maenas (Cm) muscle resting potential (RP) hyperpolarised significantly with increasing experimental temperature. The muscle RP in Cm was seasonally dependent at acute temperatures above 20 °C whereas in Cp no seasonal effect was observed. The latent period of the muscle excitatory junction potential (EJP) following tonic motor nerve stimulation was significantly longer in winter-caught crabs in both species, although the effect was significantly more marked in Cp than Cm. Summer-caught Cp had larger excitatory junction potentials (EJPs) than did winter-caught crabs, a seasonal effect not seen in Cm. In contrast, marked seasonal differences were found in the EJP decay time constant in Cm having significantly longer time constants in winter-caught crabs, where no seasonal difference was found in Cp. These results suggest that different seasonal effects of neuromuscular parameters between Cm and Cp may reflect different strategies of response to their different seasonal temperature environments.

Early life exposure to allergen and ozone results in altered development in adolescent rhesus macaque lungs.

In rhesus macaques, previous studies have shown that episodic exposure to allergen alone or combined with ozone inhalation during the first 6 months of life results in a condition with many of the hallmarks of asthma. This exposure regimen results in altered development of the distal airways and parenchyma (Avdalovic et al., 2012). We hypothesized that the observed alterations in the lung parenchyma would be permanent following a long-term recovery in filtered air (FA) housing. Forty-eight infant rhesus macaques (30 days old) sensitized to house dust mite (HDM) were treated with two week cycles of FA, house dust mite allergen (HDMA), ozone (O3) or HDMA/ozone (HDMA+O3) for five months. At the end of the five months, six animals from each group were necropsied. The other six animals in each group were allowed to recover in FA for 30 more months at which time they were necropsied. Design-based stereology was used to estimate volumes of lung components, number of alveoli, size of alveoli, distribution of alveolar volumes, interalveolar capillary density. After 30 months of recovery, monkeys exposed to HDMA, in either group, had significantly more alveoli than filtered air. These alveoli also had higher capillary densities as compared with FA controls. These results indicate that early life exposure to HDMA alone or HDMA+O3 alters the development process in the lung alveoli.

Infection rates following hip and knee joint arthroplasty: large referral centre versus a small elective-only hospital.

PURPOSE: The aim of this study was to investigate deep infection rates following hip and knee arthroplasty at a large referral hospital and to compare these rates with a smaller hospital where only elective surgery was performed. Both hospitals were administered by the same public institution. METHODS: A search of the medical records was performed for all deep infections following elective primary hip and knee arthroplasty; revision procedures were excluded as were total hip replacement and hemiarthroplasty following trauma. To be considered, a deep infection cases must have had bacterial growth confirmed on deep tissue surgical specimens or on aspiration of the joint within 1 year of the index procedure. RESULTS: There were 14 infections confirmed following 1,160 arthroplasties at the larger hospital and 1 infection for the elective-only hospital following 466 arthroplasties. Statistical analysis showed there was a 7.06 greater chance of having an infection at the larger campus compared with the smaller campus CI (1.3, 130.7). Although there was a trend towards a greater number of infections at the larger hospital, the result was not statistically significant (P = 0.06). We acknowledge there were some differences between the two study populations. CONCLUSION: We found a trend towards, but not a statistically significant difference, between infection rates at the elective-only hospital compared to the larger institution. Given the low overall rate of infection, studies with improved statistical power are needed to determine whether there is a difference in infection rates at smaller elective-only hospitals versus larger hospitals providing elective and non-elective services. The reasons for the difference are likely to be multifactorial. We hypothesise that infection rates are increased in the larger hospital where there is more procedures, both clean and contaminated being performed in the operating theatres, as well as a greater number of inpatient beds and where the hospital admits non-elective cases via its emergency department. LEVEL OF EVIDENCE: Level-two cohort study.

Lung effects of inhaled corticosteroids in a rhesus monkey model of childhood asthma.

BACKGROUND: The risks for infants and young children receiving inhaled corticosteroid (ICS) therapy are largely unknown. Recent clinical studies indicate that ICS therapy in pre-school children with symptoms of asthma result in decreased symptoms without influencing the clinical disease course, but potentially affect postnatal growth and development. The current study employs a primate experimental model to identify the risks posed by ICS therapy. OBJECTIVE: To (1) establish whether ICS therapy in developing primate lungs reverses pulmonary pathobiology associated with allergic airway disease (AAD) and (2) define the impact of ICS on postnatal lung growth and development in primates. METHODS: Infant rhesus monkeys were exposed, from 1 through 6 months, to filtered air (FA) with house dust mite allergen and ozone using a protocol that produces AAD (AAD monkeys), or to FA alone (Control monkeys). From three through 6 months, the monkeys were treated daily with ICS (budesonide) or saline. RESULTS: Several AAD manifestations (airflow restrictions, lavage eosinophilia, basement membrane zone thickening, epithelial mucin composition) were reduced with ICS treatment, without adverse effects on body growth or adrenal function; however, airway branching abnormalities and intraepithelial innervation were not reduced. In addition, several indicators of postnatal lung growth and differentiation: vital capacity, inspiratory capacity, compliance, non-parenchymal lung volume and alveolarization, were increased in both AAD and Control monkeys that received ICS treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Incomplete prevention of pathobiological changes in the airways and disruption of postnatal growth and differentiation of airways and lung parenchyma in response to ICS pose risks for developing primate lungs. These responses also represent two mechanisms that could compromise ICS therapy's ability to alter clinical disease course in young children.

Sci-Thur AM: Planning - 05: Lung SBRT: Dosimetric accuracy of the Analytical Anisotropic Algorithm (AAA) for 6MV FFF RapidArc planning.

PURPOSE: Stereotactic Body Radiation Therapy (SBRT) requires the delivery of a high biologically effective dose in only a few fractions. These large doses per fraction can necessitate long treatment times. The Varian Truebeam is capable of RapidArc delivery and also has the optional Flattening Filter Free (FFF) modes which greatly increase the dose rate. We have commissioned the 6MV FFF beam (1400 MU/min) for RapidArc lung SBRT, and verified heterogeneous dose calculations with Monte Carlo (MC). METHODS: The standard commissioning data was acquired for Varian's Analytical Anisotropic Algorithm (AAA) beam model. Measurements were acquired with the IBA Blue Phantom, using the CC13 and CC01 ion chambers and PTW diode. MLC-defined fields were also acquired for model verification. The Dosimetric Leaf Gap (DLG) was measured and then optimized using RapidArc lung SBRT plans, matching Eclipse with ion chamber measurements. Heterogeneous dose calculations were independently verified using MC. RESULTS: There were some discrepancies regarding leaf transmission and penumbra, but the AAA model was generally well within 2% and 2 mm. A nominal DLG value of 1.6 mm was chosen. A representative lung SBRT case utilizing FFF RapidArc was calculated with MC. For the high dose region, 99% matched Eclipse within 3% and 3 mm. The mean dose difference of the PTV was 0.7%. CONCLUSIONS: Although we have observed some minor infield discrepancies between the AAA and Monte Carlo calculations in heterogeneous media, the Eclipse AAA is reasonably accurate for complex FFF, RapidArc, SBRT lung planning.

Sci-Fri PM: Delivery - 03: Spine SBRT: Treating multiple vertebrae using cone-beam CT image-guidance and the hexapod robotic couch.

PURPOSE: The intent of spinal SBRT is to maximize the biological effective dose and improve local control, while sparing the adjacent spinal cord. We report on the spinal SBRT positional accuracy for multiple, consecutive vertebrae in a single course, using the Elekta Synergy-S and BodyFIX immobilization systems. METHODS: After initial patient adjustment, verification cone-beam CT (CBCT) images were acquired before, during and after treatment. These images were used to assess immobilization and correct any misalignment exceeding 1 mm or 1°, in all six degrees-of-freedom using the HexaPOD robotic couch. This analysis is based on 415 verification images from 67 consecutive courses of treatment. These treatment courses comprised 25 single thoracic vertebrae, 16 multiple thoracic vertebrae, 20 single lumbar vertebrae and 6 multiple lumbar vertebrae. RESULTS: The absolute intra-fraction motion averaged over all directions (±std dev.) for the T-single, T-multiple, L-Single and L-Multiple was 0.54 (±0.73) mm, 0.54 (±0.88) mm, 0.36 (±0.57) mm, and 0.47 (±0.63) mm respectively. The percentage that exceeded the 1.5 mm planning margin was 3.8%, 4.0%, 1.0% and 0.85% respectively. T-spine treatments were out-of-tolerance more frequently than the L-spine. There was a statistically significant difference between single and multiple lumbar treatments (unpaired t-test, p<0.01), but this was not clinically significant as 99% were within our 1.5 mm margin. CONCLUSIONS: Near-rigid immobilization with the acquisition of intra-fraction CBCT images and the correction of misalignments in all six degrees-of-freedom provides the necessary precision to safely perform SBRT of consecutive spinal metastases within one course of treatment.

Case report. Fractionated Helical Tomotherapy as an alternative to radiosurgery in patients unwilling to undergo additional radiosurgery for recurrent brain metastases.

Our clinic routinely treats brain metastases with stereotactic radiosurgery using a 6 megavoltage (MV) linear accelerator, cones, and a surgically attached head frame. Four patients declined repeat radiosurgery for new lesions due to their previous discomfort and a fifth patient could not complete radiosurgery because of uncontrolled nausea. Instead patients were treated with Helical Tomotherapy (HT). This report discusses the spatial dose distribution of HT as measured in a head phantom and the clinical course of these five patients. The planning target volume (PTV) was a 3 mm geometric expansion of the gross tumour volume (GTV). The prescribed dose to the PTV was 27 Gy in five daily fractions with the distribution optimised to deliver 30 Gy to the GTV. Patients were immobilised with a mask and the lesions were targeted by MV computerised tomography, an inherent feature of the system. One patient died six weeks later from systemic disease; the remaining patients survived eight to 16 months. No patient experienced an exacerbation of neurological symptoms following Helical Tomotherapy. These results suggest that fractionated Helical Tomotherapy for brain metastases may be a viable alternative to radiosurgery in patients unable or unwilling to undergo that procedure.

Airway generation-specific differences in the spatial distribution of immune cells and cytokines in allergen-challenged rhesus monkeys.

BACKGROUND: Accumulation of immune cell populations and their cytokine products within tracheobronchial airways contributes to the pathogenesis of allergic asthma. It has been postulated that peripheral regions of the lung play a more significant role than proximal airways with regard to inflammatory events and airflow obstruction. OBJECTIVE: To determine whether immune cell populations and associated cytokines are uniformly distributed throughout the conducting airway tree in a non-human primate model of allergic asthma. METHODS: We used a stereologic approach with a stratified sampling scheme to measure the volume density of immune cells within the epithelium and interstitium of trachea and 4-5 intrapulmonary airway generations from house dust mite (HDM) (Dermatophagoides farinae)-challenged adult monkeys. In conjunction with immune cell distribution profiles, mRNA levels for 21 cytokines/chemokines and three chemokine receptors were evaluated at four different airway generations from microdissected lungs. RESULTS: In HDM-challenged monkeys, the volume of CD1a+ dendritic cells, CD4+ T helper lymphocytes, CD25+ cells, IgE+ cells, eosinophils, and proliferating cells were significantly increased within airways. All five immune cell types accumulated within airways in unique patterns of distribution, suggesting compartmentalized responses with regard to trafficking. Although cytokine mRNA levels were elevated throughout the conducting airway tree of HDM-challenged animals, the distal airways (terminal and respiratory bronchioles) exhibited the most pronounced up-regulation. CONCLUSION: These findings demonstrate that key effector immune cell populations and cytokines associated with asthma differentially accumulate within distinct regions and compartments of tracheobronchial airways from allergen-challenged primates.

Zebrafish pitx3 is necessary for normal lens and retinal development.

The human PITX3 gene encodes a bicoid-like homeodomain transcription factor associated with a variety of congenital ocular conditions, including anterior segment dysgenesis, Peter's anomaly, and cataracts. We identified a zebrafish pitx3 gene encoding a protein (Pitx3) that possesses 63% amino acid identity with human PITX3. The zebrafish pitx3 gene encompasses approximately 16.5kb on chromosome 13 and consists of four exons, which is similar to the genomic organization of other pitx genes. Expression of the zebrafish pitx3 gene was studied by in situ mRNA hybridization and RT-PCR. The pitx3 transcripts were detected throughout development with the greatest level of expression occurring in the developing lens and brain at 24hpf. In adults, the highest expression was detected in the eye. Morpholinos were used to knockdown expression of the Pitx3 protein and a control morpholino that contains five mismatched bases was used to confirm the specificity of the phenotypes. The morphants had small eyes, misshapen heads and reduced jaws and fins relative to controls. The morphants exhibited abnormalities in lens development and their retinas contained pyknotic nuclei accompanied by a reduction in the number of cells in different neuronal classes. This suggests the lens is required for retinal development or Pitx3 has an unexpected role in retinal cell differentiation or survival. These results demonstrate zebrafish pitx3 represents a true ortholog of the human PITX3 gene and the general function of the Pitx3 protein in lens development is conserved between mammals and the teleost fish.

Smooth muscle hypertrophy in distal airways of sensitized infant rhesus monkeys exposed to house dust mite allergen.

BACKGROUND: Airway smooth muscle hypertrophy is closely associated with the pathophysiology of hyper-reactive airways in allergic asthma. OBJECTIVE: To determine whether repeated exposure to allergens during postnatal lung development promotes remodelling of airway smooth muscle. METHODS: Infant, male rhesus monkeys (30-day-old) were sensitized to house dust mite allergen (HDMA) and then exposed to HDMA aerosol periodically over 5 months. Smooth muscle mass and bundle size and abundance in conducting airways were measured and compared with age-matched control (filtered air-exposed) monkeys. RESULTS: Total smooth muscle mass and average bundle size were significantly greater in the conducting airways of monkeys exposed to HDMA. Smooth muscle bundle abundance was not affected by exposure to HDMA. CONCLUSION: Repeated cycles of allergen exposure alter postnatal morphogenesis of smooth muscle, affecting both total mass and bundle size, in conducting airways of infant monkeys.

Effects of immersion water temperature on whole-body fluid distribution in humans.

AIM: In this study, we quantified acute changes in the intracellular and extracellular fluid compartments during upright neutral- and cold-water immersion. We hypothesized that, during short-term cold immersion, fluid shifts would be wholly restricted to the extracellular space. METHODS: Seven males were immersed 30 days apart: control (33.3 degrees SD 0.6 degrees C); and cold (18.1 degrees SD 0.3 degrees C). Posture was controlled for 4 h prior to a 60-min seated immersion. RESULTS: Significant reductions in terminal oesophageal (36.9 degrees +/- 0.1 degrees -36.3 degrees +/- 0.1 degrees C) and mean skin temperatures (30.3 degrees +/- 0.3 degrees -23.0 degrees +/- 0.3 degrees C) were observed during the cold, but not the control immersion. Both immersions elicited a reduction in intracellular fluid [20.17 +/- 6.02 mL kg(-1) (control) vs. 22.72 +/- 9.90 mL kg(-1)], while total body water (TBW) remained stable. However, significant plasma volume (PV) divergence was apparent between the trials at 60 min [12.5 +/- 1.0% (control) vs. 6.1 +/- 3.1%; P < 0.05], along with a significant haemodilution in the control state (P < 0.05). Plasma atrial natriuretic peptide concentration increased from 18.0 +/- 1.6 to 58.7 +/- 15.1 ng L(-1) (P < 0.05) during cold immersion, consistent with its role in PV regulation. We observed that, regardless of the direction of the PV change, both upright immersions elicited reductions in intracellular fluid. CONCLUSION: These observations have two implications. First, one cannot assume that PV changes reflect those of the entire extracellular compartment. Second, since immersion also increases interstitial fluid pressure, fluid leaving the interstitium must have been rapidly replaced by intracellular water.

The remodelled tracheal basement membrane zone of infant rhesus monkeys after 6 months of recovery.

BACKGROUND: In previous studies, we showed that repeated exposure to (1) house dust mite allergen (HDMA) (Dermatophagoides farinae) caused thickening of the basement membrane zone (BMZ) and (2) HDMA+ozone (O3) caused depletion of BMZ perlecan and atypical development of BMZ collagen (irregular thin areas<2.0 microm in width). OBJECTIVE: The purpose of this study was to determine if these remodelling changes were reversible after 6 months of recovery. METHODS: Rhesus monkeys were exposed to a regimen of HDMA and or O3 or filtered air (FA) for 6 months. After the exposure protocol was completed FA and O3 groups were allowed to recover in FA for 6 months. The HDMA and HDMA+O3 exposure groups recovered in a modified environment. They were re-exposed to HDMA aerosol for 2 h at monthly intervals during recovery in order to maintain sensitization for pulmonary function testing. To detect structural changes in the BMZ, collagen I and perlecan immunoreactivity were measured and compared to data from the previous papers. RESULTS: The remodelled HDMA group had a significantly thicker BMZ and after 6 months of recovery the width had not regressed. In the remodelled BMZ of the HDMA+O3 group, perlecan had returned to the BMZ after 6 months of the recovery protocol, and the thin, irregular, collagen BMZ had been resolved. CONCLUSION: In summary, this study has shown that: (1) The width of the remodelled HDMA BMZ did not regress during a recovery protocol that included a sensitizing dose of HDMA. (2) The atypical collagen BMZ in the HDMA+O3 BMZ was resolved in the absence of O3. (3) Depletion of perlecan from the BMZ by O3 was reversed by recovery in the absence of O3.

Cold-water acclimation does not modify whole-body fluid regulation during subsequent cold-water immersion.

We investigated the impact of cold-water acclimation on whole-body fluid regulation using tracer-dilution methods to differentiate between the intracellular and extracellular fluid compartments. Seven euhydrated males [age 24.7 (8.7) years, mass 74.4 (6.4) kg, height 176.8 (7.8) cm, sum of eight skinfolds 107.4 (20.4) mm; mean (SD)] participated in a 14-day cold-water acclimation protocol, with 60-min resting cold-water stress tests [CWST; 18.1 (0.1) degrees C] on days 1, 8 and 15, and 90-min resting cold-water immersions [18.4 (0.4) degrees C] on intervening days. Subjects were immersed to the 4th intercostal space. Intracellular and extracellular fluid compartments, and plasma protein, electrolyte and hormone concentrations were investigated. During the first CWST, the intracellular fluid (5.5%) and plasma volumes were reduced (6.1%), while the interstitial fluid volume was simultaneously expanded (5.4%). This pattern was replicated on days 8 and 15, but did not differ significantly among test days. Acclimation did not produce significant changes in the pre-immersion distribution of total body water, or changes in plasma osmolality, total protein, electrolyte, atrial natriuretic peptide or aldosterone concentrations. Furthermore, a 14-day cold-water acclimation regimen did not elicit significant changes in body-fluid distribution, urine production, or the concentrations of plasma protein, electrolytes or the fluid-regulatory hormones. While acclimation trends were not evident, we have confirmed that fluid from extravascular cells is displaced into the interstitium during acute cold-water immersion, both before and after cold acclimation.

Total number and mean size of alveoli in mammalian lung estimated using fractionator sampling and unbiased estimates of the Euler characteristic of alveolar openings.

Estimation of alveolar number in the lung has traditionally been done by assuming a geometric shape and counting alveolar profiles in single, independent sections. In this study, we used the unbiased disector principle to estimate the Euler characteristic (and thereby the number) of alveolar openings in rat lungs and rhesus monkey lung lobes and to obtain robust estimates of average alveolar volume. The estimator of total alveolar number was based on systematic, uniformly random sampling using the fractionator sampling design. The number of alveoli in the rat lung ranged from 17.3 x 10(6) to 24.6 x 10(6), with a mean of 20.1 x 10(6). The average number of alveoli in the two left lung lobes in the monkey ranged from 48.8 x 10(6) to 67.1 x 10(6) with a mean of 57.7 x 10(6). The coefficient of error due to stereological sampling was of the order of 0.06 in both rats and monkeys and the biological variation (coefficient of variance between individuals) was 0.15 in rat and 0.13 in monkey (left lobe, only). Between subdivisions (left/right in rat and cranial/caudal in monkey) there was an increase in variation, most markedly in the rat. With age (2-13 years) the alveolar volume increased 3-fold (as did parenchymal volume) in monkeys, but the alveolar number was unchanged. This study illustrates that use of the Euler characteristic and fractionator sampling is a robust and efficient, unbiased principle for the estimation of total alveolar number in the lung or in well-defined parts of it.