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1.

Background

Stress urinary incontinence (SUI) is a relatively common disorder that significantly affects the quality of life. Many conservative and surgical treatment methods have been recommended for SUI, but they have major limitations.

Aims

To assess the use of the CO2 fractional laser in the treatment of SUI.

Methods

This clinical trial included 55 patients with confirmed SUI. Patients underwent fractional CO2 laser treatment 3 times at 30-day intervals. Data on age, smoking history, sexual activity, menopause, and history of hormone replacement therapy (HRT) were collected. Response to treatment was assessed by SUI severity and the level of sexual satisfaction was assessed using the visual analog scale (VAS). Patients were evaluated at 3 different time points: before treatment, and 45 days and 6 months after the last laser treatment.

Results

The mean patient age was 44.4±11.4 years (range: 28 to 68 years). Smoking history was positive in 6 patients (9.1%); 19 (54.3%) were menopausal on HRT. The SUI severity score at baseline (before treatment) was 8.56±0.62 and decreased to 2.28 6 months after treatment (p<0.0001). The sexual satisfaction score was 3±0.94 at baseline and increased to 7.87±0.93 6 months after treatment (day 180) (p<0.0001, slope = + 2.2)

Conclusion

Our findings are in line with a previous study that showed the value of transvaginal CO2 fractional laser treatment for alleviation of SUI symptoms and its potential as an alternative treatment. We also observed improved sexual satisfaction in SUI patients.
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2.

Objectives

To investigate the feasibility of coupling carbonyl cyanide m-chlorophenylhydrazone-regulated photohydrogen production by Tetraselmis subcordiformis in a photobioreactor to an alkaline fuel cell (AFC).

Results

H2 evolution kinetics in the AFC integrated process was characterized. The duration of H2 evolution was prolonged and its yield was improved about 1.5-fold (to 78 ± 5 ml l?1) compared with that of the process without AFC. Improved H2 yield was possibly caused by removal of H2 feedback inhibition by H2 consumption in situ. Decreases in the H2 production rate correlated with the gradual deactivation of PSII and hydrogenase activities. The H2 yield was closely associated with catabolism of starch and protein.

Conclusion

A marine green algal CO2-supplemented culture integrated with in situ H2-consumption by an AFC system was developed as a viable protocol for the H2 production.
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3.

Objectives

To evaluate the influence of hydraulic retention time (HRT) and cheese whey (CW) substrate concentration (15 and 25 g lactose l?1) on the performance of EGSB reactors (R15 and R25, respectively) for H2 production.

Results

A decrease in the HRT from 8 to 4 h favored the H2 yield and H2 production rate (HPR) in R15, with maximum values of 0.86 ± 0.11 mmol H2 g COD?1 and 0.23 ± 0.024 l H2 h?1 l?1, respectively. H2 production in R25 was also favored at a HRT of 4 h, with maximum yield and HPR values of 0.64 ± 0.023 mmol H2 g COD?1 and 0.31 ± 0.032 l H2 h?1 l?1, respectively. The main metabolites produced were butyric, acetic and lactic acids.

Conclusions

The EGSB reactor was evaluated as a viable acidogenic step in the two-stage anaerobic treatment of CW for the increase of COD removal efficiency and biomethane production.
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4.

Background and aims

Saline and alkali soils severely impact plant growth. Endophyte and plant associations are known to significantly modify plant metabolism. This study reports the effects of a type of endophyte on organic acid (OA) accumulation and ionic balance in rice under Na2CO3 stress.

Methods

Rice seedlings with (E+) and without (E-) endophytic infection were subjected to different levels of Na2CO3 stress (0, 5, 10, 15, and 20 mM) for two weeks. Organic acids and mineral elements in the leaves and roots were determined.

Results

Seedlings with endophytic infection accumulated mainly citrate and fumarate, with some malate and succinate in the leaves. In the roots, accumulation of malate and fumarate was enhanced significantly by endophytic infection, while less citrate and succinate was accumulated under Na2CO3 stress, which suggested that leaves and roots use different mechanisms to control OA metabolism. Endophytes reduced the total Na and Na:K ratios, but increased ST values, the percent changes of other measured nutrients, Chl content, and dry weight per plant under Na2CO3 stress.

Conclusions

Endophytic infection plays a key role in maintaining plant growth by improving nutrient uptake and adjusting OA accumulation under Na2CO3 stress. The application of endophytes can enhance the resistance of rice to salinity.
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5.

Introduction

The aphid Rhopalosiphum padi L. is a vector of Barley yellow dwarf virus (BYDV) in wheat and other economically important cereal crops. Increased atmospheric CO2 has been shown to alter plant growth and metabolism, enhancing BYDV disease in wheat. However, the biochemical influences on aphid metabolism are not known.

Objectives

This work aims to determine whether altered host-plant quality, influenced by virus infection and elevated CO2, impacts aphid weight and metabolism.

Methods

Untargeted 1H NMR metabolomics coupled with multivariate statistics were employed to profile the metabolism of R. padi reared on virus-infected and non-infected (sham-inoculated) wheat grown under ambient CO2 (aCO2, 400 µmol mol?1) and future, predicted elevated CO2 (eCO2, 650 µmol mol?1) concentrations. Un-colonised wheat was also profiled to observe changes to host-plant quality (i.e., amino acids and sugars).

Results

The direct impacts of virus or eCO2 were compared. Virus presence increased aphid weight under aCO2 but decreased weight under eCO2; whilst eCO2 increased non-viruliferous (sham) aphid weight but decreased viruliferous aphid weight. Discriminatory metabolites due to eCO2 were succinate and sucrose (in sham wheat), glucose, choline and betaine (in infected wheat), and threonine, lactate, alanine, GABA, glutamine, glutamate and asparagine (in aphids), irrespective of virus presence. Discriminatory metabolites due to virus presence were alanine, GABA, succinate and betaine (in wheat) and threonine and lactate (in aphids), irrespective of CO2 treatment.

Conclusion

This study confirms that virus and eCO2 alter host-plant quality, and these differences are reflected by aphid weight and metabolism.
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6.

Background

Airway epithelium is an active and important component of the immunological response in the pathophysiology of obstructive lung diseases. Recent studies suggest an important role for vitamin D3 in asthma severity and treatment response.

Objective

Our study evaluated the influence of an active form of vitamin D3 on the expression of selected mediators of allergic inflammation in the respiratory epithelium.

Material and Methods

Primary nasal and bronchial epithelial cells were exposed to1,25D3 for 1 hour and were then stimulated or not with IL-4, TNF-α, LPS, and poly I:C. After 24 hours TSLP, IL-33, and IL-25 protein levels were measured in culture supernatants usingELISAandmRNAlevels in cells by real time PCR.

Results

1,25D3 increased TSLP concentration in unstimulated nasal epithelial cells, but did not influence IL-33 and IL-25 expression. In IL-4-stimulated epithelial cell cultures 1,25D3 mostly inhibited TSLP and IL-33 expression. In LPS-treated cultures 1,25D3 decreased IL-33 expression. Simultaneously 1,25D3 augmented IL-25 production in the same model of stimulation.

Conclusion

Our study revealed the dual nature of vitamin D3 manifested in both pro- and anti-inflammatory properties observed in airway epithelial cells.
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7.

Objective

To explore the glycerol utilization pathway in Corynebacterium glutamicum for succinate production under O2 deprivation.

Result

Overexpression of a glycerol facilitator, glycerol dehydrogenase and dihydroxyacetone kinase from Escherichia coli K-12 in C. glutamicum led to recombinant strains NC-3G diverting glycerol utilization towards succinate production under O2 deprivation. Under these conditions, strain NC-3G efficiently consumed glycerol and produced succinate without growth. The recombinant C. glutamicum utilizing glycerol as the sole carbon source showed higher intracellular NADH/NAD+ ratio compare with utilizing glucose. The mass conversion of succinate increased from 0.64 to 0.95. Using an anaerobic fed-batch fermentation process, the final strain produced 38.4 g succinate/l with an average yield of 1.02 g/g.

Conclusions

The metabolically-engineered strains showed an efficient succinate production using glycerol as sole carbon source under O2 deprivation.
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8.

Objective

To investigate the effect of H2O2 on the migration and antioxidant defense of mesenchymal stem cells (MSCs) and the neurotrophic effects of H2O2-treated MSCs on spinal cord injury (SCI).

Results

Sublethal concentrations of H2O2 decreased cell migration and expression of CXCR4 and CCR2 as well as Nrf2 expression in MSCs. In the second phase, transplantation of treated and untreated MSCs to SCI caused minor changes in locomotor dysfunction. There was a significantly difference between cell-treated and spinal cord injury groups in expression of BDNF (brain-derived neurotrophic factor). Transplantation of H2O2-treated cells caused an increase in BDNF expression compared to non-treated cells.

Conclusion

Transplantation of H2O2-treated stem cells may have protective effects against SCI through by increasing neurotrophic factors.
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9.

Background and Aims

Rocket salad (Eruca sativa Mills) is one of the major leafy vegetables produced worldwide and has been characterized as a rich source of chemoprotective glucosinolates (GSL). The relationship between N fertilization and the resulting plant biomass and N status with GSL quantity and quality in rocket leaves was examined.

Methods

A pot experiment was conducted, applying ten different N-rates and destructive sampling was carried out 15, 30 and 45 days after transplanting (DAT). The Mitscherlich equation was used to establish NO3-N critical levels at each growth stage and as an indicator of N demand for relative maximum dry matter accumulation and glucosinolate content and composition was determined.

Results

Glucosinolate content was significantly influenced by N rate, growth stage and their interaction. Different GSL types showed dissimilar responses to N fertilization: aliphatic GSLs were significantly reduced under increased N rates whereas indole GSL showed the reverse. Under excess N fertilization (>1.04 g/plant), dry matter accumulation remained constant, NO3-N was significantly increased and total GSL content was significantly reduced, factors that could lead to an anticipated product quality decline.

Conclusions

The application of the critical NO3-N level approach used to identify optimal N fertilization rates for plant growth could serve as means to obtain optimized GSL content in the edible plant parts.
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10.

Background

Mitochondria exhibit a dynamic morphology in cells and their biogenesis and function are integrated with the nuclear cell cycle. In mitotic cells, the filamentous network structure of mitochondria takes on a fragmented form. To date, however, whether mitochondrial fusion activity is regulated in mitosis has yet to be elucidated.

Findings

Here, we report that mitochondria were found to be fragmented in G2 phase prior to mitotic entry. Mitofusin 1 (Mfn1), a mitochondrial fusion protein, interacted with cyclin B1, and their interactions became stronger in G2/M phase. In addition, MARCH5, a mitochondrial E3 ubiquitin ligase, reduced Mfn1 levels and the MARCH5-mediated Mfn1 ubiquitylation were enhanced in G2/M phase.

Conclusions

Mfn1 is degraded through the MARCH5-mediated ubiquitylation in G2/M phase and the cell cycle-dependent degradation of Mfn1 could be facilitated by interaction with cyclin B1/Cdk1 complexes.
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11.

Background

Near infrared spectroscopy (NIRS) is used to assess muscle oxygenation (MO) within skeletal muscle at rest and during aerobic exercise. Previous investigations have used a single probe placement to measure MO during various forms of exercise. However, regional MO differences have been shown to exist within the same muscle which suggests that different areas of the same muscle may have divergent MO. Thus, the aim of this study was to examine whether regional differences in MO exist within the same muscle during different types of incremental (rest, 25, 50, 75, 100 % of maximum) exercise (1 leg knee extension (KE), 2 leg KE, or cycling).

Methods

Nineteen healthy active males (Mean ± SD: Age 27 ± 4 yrs; VO2max: 55 ± 11 mL/kg/min) performed incremental exercise to fatigue using each mode of exercise. NIRS probes were placed on the distal and proximal portion of right leg vastus lateralis (VL). Results were analyzed with a 3-way mixed model ANOVA (probe × intensity × mode).

Results

Differences in MO exist within the VL for each mode of exercise, however these differences were not consistent for each level of intensity. Comparison of MO revealed that the distal region of VL was significantly lower throughout KE exercise (1 leg KE proximal MO – distal MO = 9.9 %; 2 leg KE proximal MO – distal MO = 13 %). In contrast, the difference in MO between proximal and distal regions of VL was smaller in cycling and was not significantly different at heavy workloads (75 and 100 % of maximum).

Conclusion

MO is different within the same muscle and the pattern of the difference will change depending on the mode and intensity of exercise. Future investigations should limit conclusions on MO to the area under assessment as well as the type and intensity of exercise employed.
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12.

Background and aims

Elemental uptake in serpentine floras in eastern North America is largely unknown. The objective of this study was to determine major and trace element concentrations in soil and leaves of three native pseudo-metallophyte C4 grasses in situ at five sites with three very different soil types, including three serpentine sites, in eastern USA.

Methods

Pseudo-total and extractible concentrations of 15 elements were measured and correlated from the soils and leaves of three species at the five sites.

Results

Element concentrations in soils of pseudo-metallophytes varied up to five orders of magnitude. Soils from metalliferous sites exhibited higher concentrations of their characteristic elements than non-metalliferous. In metallicolous populations, elemental concentrations depended on the element. Concentrations of major elements (Ca, Mg, K) in leaves were lower than typical toxicity thresholds, whereas concentrations of Zn were higher.

Conclusions

In grasses, species can maintain relatively low metal concentrations in their leaves even when soil concentrations are richer. However, in highly Zn-contaminated soil, we found evidence of a threshold concentration above which Zn uptake increases drastically. Finally, absence of main characteristics of serpentine soil at one site indicated the importance of soil survey and restoration to maintain serpentinophytes communities and avoid soil encroachment.
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13.

Aims

Despite extensive studies on effects of elevated CO2 concentration ([CO2]e) on plant growth, few studies have investigated the responses of native grassland plant species to [CO2]e in terms of nutrient acquisition.

Methods

The effects of [CO2]e (769 ± 23 ppm) on Artemisia frigida and Stipa krylovii, two dominant species in Inner Mongolia steppe were investigated by growing them for 7 weeks in Open-Top Chambers (OTC).

Results

Exposure to [CO2]e enhanced shoot and root growth of A. frigida and S. krylovii. Elevated [CO2] increased photosynthetic rates (Pn) by 34 % in A. frigida but decreased Pn by 52 % in S. krylovii. Moreover, root-secreted acid phosphatase activity in A. frigida was stimulated by [CO2]e, while exudation of malate from roots of S. krylovii was suppressed by [CO2]e. Exposure to [CO2]e led to a decrease in P concentration in shoots and roots of A. frigida and S. krylovii, but total amount of P accumulated in shoots and roots of both species was increased by [CO2]e.

Conclusions

The two dominant species in temperate steppes differed in their responses to [CO2]e, such that A. frigida was more adapted to [CO2]e than S. krylovii under low availability of soil P.
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14.

Background

The ESX-1 type VII secretion system is an important determinant of virulence in pathogenic mycobacteria, including Mycobacterium tuberculosis. This complicated molecular machine secretes folded proteins through the mycobacterial cell envelope to subvert the host immune response. Despite its important role in disease very little is known about the molecular architecture of the ESX-1 secretion system.

Results

This study characterizes the structures of the soluble domains of two conserved core ESX-1 components – EccB1 and EccD1. The periplasmic domain of EccB1 consists of 4 repeat domains and a central domain, which together form a quasi 2-fold symmetrical structure. The repeat domains of EccB1 are structurally similar to a known peptidoglycan binding protein suggesting a role in anchoring the ESX-1 system within the periplasmic space. The cytoplasmic domain of EccD1has a ubiquitin-like fold and forms a dimer with a negatively charged groove.

Conclusions

These structures represent a major step towards resolving the molecular architecture of the entire ESX-1 assembly and may contribute to ESX-1 targeted tuberculosis intervention strategies.
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15.

Purpose

The increasing use of engineered nanomaterials (ENMs) in industrial applications and consumer products is leading to an inevitable release of these materials into the environment. This makes it necessary to assess the potential risks that these new materials pose to human health and the environment. Life cycle assessment (LCA) methodology has been recognized as a key tool for assessing the environmental performance of nanoproducts. Until now, the impacts of ENMs could not be included in LCA studies due to a lack of characterization factors (CFs). This paper provides a methodological framework for identifying human health CFs for ENMs.

Methods

The USEtox? model was used to identify CFs for assessing the potential carcinogenic and non-carcinogenic effects on human health caused by ENM emissions in both indoor (occupational settings) and outdoor environments. Nano-titanium dioxide (nano-TiO2) was selected for defining the CFs in this study, as it is one of the most commonly used ENMs. For the carcinogenic effect assessment, a conservative approach was adopted; indeed, a critical dose estimate for pulmonary inflammation was assumed.

Results and discussion

We propose CFs for nano-TiO2 from 5.5E?09 to 1.43E?02 cases/kgemitted for both indoor and outdoor environments and for carcinogenic and non-carcinogenic effects.

Conclusions

These human health CFs for nano-TiO2 are an important step toward the comprehensive application of LCA methodology in the field of nanomaterial technology.
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16.

Purpose

To evaluate the efficiency of corneal collagen cross-linking (CXL) in addition to topical voriconazole in cases with mycotic keratitis.

Design

Retrospective case series in a tertiary university hospital.

Participants

CXL was performed on 13 patients with mycotic keratitis who presented poor or no response to topical voriconazole treatment.

Methods

The clinical features, symptoms, treatment results and complications were recorded retrospectively. The corneal infection was graded according to the depth of infection into the stroma (from grade 1 to grade 3). The visual analogue scale was used to calculate the pain score before and 2 days after surgery.

Main Outcome Measures

Grade of the corneal infection.

Results

Mean age of 13 patients (6 female and 7 male) was 42.4 ± 17.7 years (20–74 years). Fungus was demonstrated in culture (eight patients) or cytological examination (five patients). Seven of the 13 patients (54%) were healed with topical voriconazole and CXL adjuvant treatment in 26 ± 10 days (15–40 days). The remaining six patients did not respond to CXL treatment; they initially presented with higher grade ulcers. Pre- and post-operative pain score values were 8 ± 0.8 and 3.5 ± 1, respectively (p < 0.05).

Conclusions

The current study suggests that adjunctive CXL treatment is effective in patients with small and superficial mycotic ulcers. These observations require further research by large randomized clinical trials.
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17.

Background

Lung protective mechanical ventilation (MV) is the corner stone of therapy for ARDS. However, its use may be limited by respiratory acidosis.This study explored feasibility of, effectiveness and safety of low flow extracorporeal CO2 removal (ECCO2R).

Methods

This was a prospective pilot study, using the Abylcap® (Bellco) ECCO2R, with crossover off-on-off design (2-h blocks) under stable MV settings, and follow up till end of ECCO2R. Primary endpoint for effectiveness was a 20% reduction of PaCO2 after the first 2-h. Adverse events (AE) were recorded prospectively.We included 10 ARDS patients on MV, with PaO2/FiO2?<?150 mmHg, tidal volume?≤?8 mL/kg with positive end-expiratory pressure ≥?5 cmH2O, FiO2 titrated to SaO2 88–95%, plateau pressure ≥?28 cmH2O, and respiratory acidosis (pH <7.25).

Results

After 2-h of ECCO2R, 6 patients had a ≥?20% decrease in PaCO2 (60%); PaCO2 decreased 28.4% (from 58.4 to 48.7 mmHg, p?=?0.005), and pH increased (1.59%, p?=?0.005). ECCO2R was hemodynamically well tolerated. During the whole period of ECCO2R, 6 patients had an AE (60%); bleeding occurred in 5 patients (50%) and circuit thrombosis in 3 patients (30%), these were judged not to be life threatening.

Conclusions

In ARDS patients, low flow ECCO2R significantly reduced PaCO2 after 2 h, Follow up during the entire ECCO2R period revealed a high incidence of bleeding and circuit thrombosis.

Trial registration

https://clinicaltrials.gov identifier: NCT01911533, registered 23 July 2013.
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18.

Objectives

To search for a novel glutamate decarboxylase (GAD) with an optimum pH towards near-neutrality in order to improve production of gamma-aminobutyric acid (GABA) in recombinant hosts.

Results

A novel glutamate decarboxylase, BmGAD, from Bacillus megaterium was overexpressed and purified. BmGAD was approximately 53 kDa by SDS-PAGE analysis. Its optimum activity was at pH 5 and 50 °C. BmGAD had a specific activity of 59 ± 5.2 U mg?1 at pH 6, which is the highest value reported so far. The apparent Km and Vmax values of BmGAD were 8 ± 0.5 mM and 150 ± 4.7 U mg?1, respectively. Through site-directed mutagenesis, two BmGAD mutants (E294R and H467A) showed higher Vmax values than that of wild-type, with the values of 210 ± 6.9 and 180 ± 4.1 U mg?1 at pH 5 and 50 °C, respectively.

Conclusions

The unusual high activity of BmGAD at pH 6 makes it an attractive GABA-producing candidate in industrial application.
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19.

Objective

To investigate the inhibiting effect of formic acid on acetone/butanol/ethanol (ABE) fermentation and explain the mechanism of the alleviation in the inhibiting effect under CaCO3 supplementation condition.

Results

From the medium containing 50 g sugars l?1 and 0.5 g formic acid l?1, only 0.75 g ABE l?1 was produced when pH was adjusted by KOH and fermentation ended prematurely before the transformation from acidogenesis to solventogenesis. In contrast, 11.4 g ABE l?1 was produced when pH was adjusted by 4 g CaCO3 l?1. The beneficial effect can be ascribed to the buffering capacity of CaCO3. Comparative analysis results showed that the undissociated formic acid concentration and acid production coupled with ATP and NADH was affected by the pH buffering capacity of CaCO3. Four millimole undissociated formic acid was the threshold at which the transformation to solventogenesis occurred.

Conclusion

The inhibiting effect of formic acid on ABE fermentation can be alleviated by CaCO3 supplementation due to its buffering capacity.
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20.

Background

In patients with COPD, both laboratory exercise tests and field walking tests are used to assess physical performance. In laboratory tests, peak exercise capacity in watts (W peak) and/or peak oxygen uptake (VO2 peak) are assessed, whereas the performance on walking tests usually is expressed as distance walked. The aim of the study was to investigate the relationship between an incremental shuttle walking test (ISWT) and two laboratory cycle tests in order to assess whether W peak could be estimated from an ISWT.

Methods

Ninety-three patients with moderate or severe COPD performed an ISWT, an incremental cycle test (ICT) to measure W peak and a semi-steady-state cycle test with breath-by-breath gas exchange analysis (CPET) to measure VO2 peak. Routine equations for conversion between cycle tests were used to estimate W peak from measured VO2 peak (CPET). Conversion equation for estimation of W peak from ISWT was found by univariate regression.

Results

There was a significant correlation between W peak and distance walked on ISWT × body weight (r = 0.88, p < 0.0001). The agreement between W peak measured by ICT and estimated from ISWT was similar to the agreement between measured W peak (ICT) and W peak estimated from measured VO2 peak by CPET.

Conclusion

Peak exercise capacity measured by an incremental cycle test could be estimated from an ISWT with similar accuracy as when estimated from peak oxygen uptake in patients with COPD.
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