Tag Archives: GSK503

The main drawback of NSC based anti

The main drawback of NSC-based anti-tumor therapies is that, despite the effective tumor tropism exhibited by NSCs, only small portions of transplanted cells can migrate towards the tumor. Several recent publications have revealed that 70–80% tumor volume reduction can be achieved in various orthotopic GBM xenograft models (Aboody et al., 2006; Lee et al., 2009) even when only 20–30% of implanted NSCs are able to effectively migrate from their implantation site to the tumor area (Ahmed et al., 2011a; Lee et al., 2009; Zhao et al., 2008; Jeon et al., 2008; Kim et al., 2010). Enhancing such homing capacity will likely be one of the critical goals for the fulfillment of the preclinical promise of NSC-based anti-cancer therapeutic strategies. The mechanisms that guide selective tumor-tropic NSC migration are yet to be completely understood. Recent data from our lab and others have suggested that chemokines and pro-angiogenic factors produced by the tumor microenvironment may serve as chemoattractants (Ahmed et al., 2011a; Imitola et al., 2004). It has been shown that NSCs preferentially distribute within hypoxic areas in intracranial glioma xenografts. In addition, hypoxia inducible factor 1 alpha (HIF-1α) knockdown in glioma cells prevented the hypoxia-induced recruitment of NSCs (Zhao et al., 2008). This impaired migration was due to decreased vascular endothelial growth factor (VEGF), stromal cell-derived factor-1 (SDF-1), and urokinase-type plasminogen activator (uPA) expression in tumor cells (Zhao et al., 2008). It was also demonstrated that intratumoral upregulation of VEGF induced a long-range attraction of transplanted human NSCs from distant sites in the adult GSK503 (Schmidt et al., 2005). In addition to VEGF, activation of MMPs has also been pointed as a mechanism of hypoxia-mediated NSC tropism to malignant areas (Zhao et al., 2008; Xu et al., 2007; Lamszus et al., 2000). Taken together, these data indicated that tumor-derive hypoxia-inducible chemo-attractant factors play a critical role in promoting tumor-tropic migration of NSCs. However, the downstream signaling pathways involved in this targeted migration remain to be elucidated. Therefore, the goal of this study is to understand why only certain implanted NSCs possess migratory capacity. We also aim to establish a mechanism that can be leveraged as a possible target to enhance NSC migration and payload delivery capacity to the diffuse tumor burdens through selective overexpression of such molecule. Here we examined both the migratory and non-migratory fraction of NSC populations with functional assays, both in vitro and in vivo. Based on an array based analysis data presented here suggest MMP14 as a novel functional downstream target of VEGFA and VEGFR2 signaling, driving NSC migration and homing to targeted tumor areas.

Materials and methods

Results

Discussion
The role of NSCs as therapeutic carriers against malignant brain tumors has been widely studied and well established (Aboody et al., 2000; Ahmed et al., 2013; Auffinger et al., 2013; Thaci et al., 2012; Ahmed et al., 2011b). Due to their safety profile, inherent tumor-tropism, and immunosuppressive properties, HB1.F3.CD NSCs are considered one of the optimal carriers for anti-cancer therapeutics and have recently entered a phase I clinical trial for patients with recurrent malignant gliomas (NCT01172964). However, the major obstacle for the clinical application of NSC-based anti-cancer therapies is only a small fraction of carrier cells that possess tumor-tropic migratory capacities in preclinical animal models (Ahmed et al., 2011a; Lee et al., 2009; Zhao et al., 2008; Jeon et al., 2008; Kim et al., 2010). Here, our main goal is to understand why some NSCs can effectively migrate and deliver their therapeutic payload to targeted tumor areas while others are not. To better comprehend this process, we examined both migratory and non-migratory NSC populations in vitro and in vivo. We closely assessed their mechanism and route of migration, and through gene expression analysis we identified two essential genes were driving this process: VEGF-A and MMP14.

Recent evidence suggests OPG deficiency may promote breast cancer development

Recent evidence suggests OPG deficiency may promote breast cancer development. Serum OPG analysis in >6000 healthy subjects (Vik et al., 2015) showed an inverse relation in younger women (<60years) between serum OPG and risk of incident cancer at all sites, but primarily in the breast and reproductive tract (Vik et al., 2015). High OPG levels protected against breast cancer and mortality (Vik et al., 2015). An independent analysis of ~4500 primary breast cancers showed no prognostic value for RANK/RANKL expression, but a substantially better prognosis in oestrogen receptor-positive breast cancer cases showing high OPG expression (Sanger et al., 2014). A “suppressed” RANK pathway was likewise associated with better prognosis in another large breast cancer set (Santini et al., 2011). Denosumab, an antibody against RANKL that mimics the physiological role of OPG, is currently in clinical use for osteoporosis and for the treatment of skeletal related events in patients with bone metastasis resulting from solid tumours. In patients with breast cancer, denosumab has recently been shown to reduce the risk of clinical fractures in postmenopausal women with breast cancer receiving aromatase inhibitors and is currently being studied for any effect on prolonging bone metastasis free survival in women with early-stage breast cancer who are at high risk for disease recurrence (Gnant et al., 2015). While our previous data highlighted the potential for antiprogestogens for breast cancer prevention, the current data additionally support a clinical breast cancer prevention trial using denosumab to compensate for the low OPG levels throughout the menstrual GSK503 in BRCA-mutation carriers.

Contributors

Declaration of Interests
The following are the supplementary data related to this article.

Acknowledgements
This work was funded by Amgen Inc. and the Eve Appeal (http://www.eveappeal.org.uk/) and undertaken at UCLH/UCL, which received a proportion of its funding from the Department of Health NIHR Biomedical Research Centres (BRC) funding scheme. The UKFOCSS was core funded by Cancer Research UK (grants C315/A2621 and C1005/A6383), the UK Department of Health and the Eve Appeal with additional support from grants MRC8804/A/A7058, CA152990 & CA086381 from the NCI Early Detection Research Network. This work was aided by grant number R01 CA133117 from the US National Institutes of Health, by Organon Pharmaceuticals, and the Martin and Sharleen Cohen Foundation for Biomedical Research. Editing and formatting support was provided by Albert Y. Rhee (Amgen Inc).

Introduction
The immune system recognizes and eliminates tumor cells (Dunn et al., 2002), which defend themselves by different mechanisms (Villalba et al., 2013). The lymphocyte lineage natural killer (NK) cell belongs to the innate immune system (Lanier, 2008; Vivier et al., 2008) and show strong anti-leukemia activity when engrafted in allogeneic settings in hematological cancer patients (Velardi, 2008; Ruggeri et al., 2007; Anel et al., 2012). However, the presence of an anti-leukemia NK cell population in patients with hematological malignancies has not been proven.
NK cells are not a homogenous population and different subsets have different physiological activities. Moreover, different stimuli (e.g., cytokines vs targets cells) give rise to different immunophenotypes (Fujisaki et al., 2009; Sanchez-Martinez et al., 2014). In peripheral blood, human NK cells are mostly CD3−CD56dim cells with high cytotoxic activity, while CD3−CD56brigth cells excel in cytokine production (Bryceson et al., 2011). In vitro evidence indicates that CD56bright NK cells are precursors of CD56dim NK cells and this might also be the case in vivo (Domaica et al., 2012). In addition, combined analysis of CD56 and CD16 expression during NK cell development indicates that their profiles change as follows: CD56brigthCD16−→CD56brigthCD16dim→CD56dimCD16dim→CD56dimCD16+. Additional markers can be used to identify specific subsets within these NK cell populations (Moretta, 2010; Freud et al., 2014).

br Data The data represent the

Data
The data represent the result of metagenomic shotgun-sequencing of human gut microbiota at 3 time points: before the H. pylori eradication therapy, immediately after 2 weeks of therapy and several weeks after the treatment. The dataset contains 15 metagenomic samples in raw reads format with 30.4 ± 10.7 mln of reads per sample (mean ± SD).
These data together with those described in [2] were involved in the study devoted to the gut microbiome changes caused by antimicrobial therapy [1].
Detailed description of samples is given in Table 1.

Experimental design, materials and methods

Acknowledgements
This work was financially supported by the Ministry of Education and Science of Russian Federation (Agreement no. 14.575.21.0076, ID RFMEFI575I4×0076).

Specifications Table

Value of the data

Data
Along with the data set, we provide information about the experimental details on the hypoxic cell culture setup that determine the actual pericellular pO2, both in a normoxic and hypoxic incubator. Culture medium volume as well as vessel surface area and geometry all influence culture medium depth above the settled GSK503 in an unstirred culture. Because of poor solubility in the liquid phase and continuous cellular consumption, the pO2 in the culture medium decreases with depth toward the cells at the bottom of a standard culture dish where its actual value is not known [5]. In addition, we provide a step by step protocol for using the mirVana buffer system in combination with Pure Link micro kit filter columns for extraction of total RNA from cells. We tested the quality of the deposited microarray data for each of the 20 samples (Fig. 1, Supplementary material). Further, we represent the global comparison between the four experimental conditions (Figs. 2 and 3). Fig. 1 depicts the box plots for all microarray experiments after normalization. We illustrate differential gene expression through experimental factors (IL-15 priming and hypoxia) using Venn diagram (Fig. 2) and heatmap representations (Figs. 3 and 4). The differential gene expression data is available as Supplementary material 1.

Experimental design, materials and methods

Acknowledgements
This work was supported by intramural funds and by the foundation Klaus Tschira Stiftung, Germany​, grant number 00.277.2015.

Data
The dataset of this article describes delayed IFNγ production in adult mice infected with RSV as well as the immune response and viral clearing effects of IFNγ when delivered intranasally compared to PBS alone. Fig. 1 shows RSV-mediated weight changes and viral clearance in adult mice treated with intranasal IFNγ or PBS. Figs. 2–5 show changes in innate and adaptive immunity in RSV-infected adult mice treated with IFNγ or PBS.

Experimental design, materials and methods
Balb/cJ mice aged 6–8 weeks, were ordered from The Jackson Laboratory, Bar Harbor, ME and were maintained in pathogen-free facilities in the Division of Laboratory Animal Resources at the University of Pittsburgh (Pittsburgh, PA). Experiments and animal handling were performed according to protocols approved by The University of Pittsburgh Institutional Animal Care and Use Committee. Where indicated, mice were infected intranasally (i.n.) with RSV Line 19 (RSV L19, Martin Moore, Emory University, Atlanta, GA) (5 × 105pfu/g, ~1.5 × 106 pfu in 100μl) under isoflurane anesthesia. On one day post infection (dpi) 50μl of recombinant murine IFNγ (16ng/g) (Peprotech, Rocky Hill, NJ) or vehicle only (PBS) were delivered intranasally to RSV-infected mice under light isoflurane anesthesia GSK503 on 1, 3, and 5dpi. Mice were weighed daily; percent change from baseline weight was reported. At the indicated times post-infection, at least 5 mice per group were culled for tissue collection. Lungs were lavaged with HBSS-EDTA, then right lungs were harvested and processed for flow cytometry and left lungs were snap frozen for viral plaque assays as previously described [2].

According to our results AE PE AQ and PQ

According to our results AE, PE, AQ and PQ are not correlated with CH and CRF. Corneal elevation indices are important for early diagnosis of keratoconus, to differentiate keratoconus-suspect eyes from normal and to diagnose early corneal ectasia after refractive surgery. However, our results showed that viscoelastic characteristics of cornea measured by ORA are not related to these topographic features of the cornea.

Conflict of interest

Acknowledgment

Introduction
Visual impairment in children has a broad negative impact on their quality of life and of course will affect their social and educational development.
Keratoconus is a degenerative disorder characterized by ectasia and thinning of the axial or para-axial region of the cornea resulting in irregular astigmatism, myopia and scarring with mild to marked impairment in the quality of vision. Pediatric keratoconus displays a higher ratio of keratoconus eyes being about 88%. It is also diagnosed in more advanced stage (stage IV) comparing to adult patients (27.8% versus 7.8%). Hence, keratoconus in children progresses aggressively with a higher rate of acute hydrops as compared to the adult group.
Early treatment to stop the progression and to avoid future keratoplasty is of greater benefit in long run in those patients. For this reason, corneal cross-linking (CXL) has been widely utilized and evaluated in children after its success in adult keratoconus patients.
The only treatment that is believed to be able to stop or decrease the keratoconus progression is collagen cross-linking. The overall treatment time of CXL is still a drawback of this process, so a GSK503 in the operation time and shorter UVA exposure time (accelerated UVA exposure) to a few minutes are currently being investigated in the pediatric group for better cooperation and comfort.

Subjects and methods

Results
Thirty-three eyes of 25 children (8 bilateral and 17 unilateral) with confirmed keratoconus were included in this prospective study. The mean age was 12±2.02years (range: 8–15years). Eighty percent of the study population was boys, and 20% was girls. Of these, 29 eyes showed progression, as defined by an increase in anterior surface (K max) readings of at least 1.00 diopter (D) in serial corneal topographies over a maximum of one year.
Visual acuity was measured using the Landolt C or Tumbling E metric charts and transformed into logarithm of the Minimum Angle of Resolution (Log MAR) for further statistical analysis as recommended by Holladay. . Table 1 summarizes the UAVA and AVA data, expressed in Log MAR and covering 12-month follow-up period. There was a statistically significant improvement from the preoperative values (P<0.001). The preoperative mean UAVA was 0.54±0.2 which changed to 0.46±0.2 at 3months, 0.35±0.23 at 6months and 0.34±0.22 at one year. Eight eyes (24.2%) maintained the preoperative UAVA; twelve eyes (36.4%) gained one line, eight eyes (24.2%) gained two lines and five (15.2%) eyes gained three to four lines. None of the eyes lost lines of the preoperative UAVA. ( The preoperative AVA, at 3 and 6months, and one-year postoperative examinations is shown in Table 1. There was a statistically significant (P<0.001) improvement in AVA between the preoperative and 1-year evaluations. Out of the 33 eyes were evaluated at one year, 17 eyes (51.5%) experienced gained one line of AVA and 10 eyes (30.3%) gained 2–4 lines. Five eyes (15.2%) experienced no change in AVA. There was no statistically significant change from the 6-months examination values and one-year values (P>0.05). The preoperative mean AVA was 0.36±0.2 and changed to 0.31±0.17 at 3months, 0.19±0.13 at 6months and 0.17±0.15 at one year.
Corneal astigmatism values at the end of one-year showed a significant change. The values at 1-year examination (mean 2.01±0.8 D) were statistically significantly less than the preoperative values (mean 2.4±1.01 D) (P<0.001). The changes in refractive errors at 1-year compared with the preoperative baseline are presented in Table 2.

br Materials and methods br Results and discussion br

Materials and methods

Results and discussion

Conclusion
This study demonstrated the worth of Taguchi orthogonal array design combined with a response surface methodological approach for the development of a robust stability indicating HPLC method for the GLP. This approach exposed the two-way interactions between factors, which were impossible to detect with the Taguchi design alone. The critical factors that are significantly (p<0.05) affecting the studied responses were identified and controlled to achieve the predetermined method goal. The best method having improved system suitability parameters was employed in the routine assay of GLP from an S-SNEDDS formulation. Lastly, the presented data in the future will help analysts to diagnose any problems along with corrective action, which may come across during the life cycle of the stated stability indicating method, if appliedfor other purposes involving quantitative estimation of glimepiride provided specificity is justified for the intended purpose.
Acknowledgements

Introduction
Inflammation is a normal, natural protective and defense mechanism of the organism to tissue injury caused by various factors which include physical trauma, injurious stimuli, chemical action or microbial infections (Ashley et al., 2012). It is a very common symptom of many chronic diseases such as arthritis, osteoarthritis, inflammatory bowel disease, and chronic GSK503 which put enormous burden on the economy of the countries. The prevalence of inflammatory diseases is on rise across the world, mostly affecting elderly population (Gautam and Jachak, 2009). Few epidemiological studies conducted elsewhere have also linked inflammation to pathogenesis of stroke, cardiovascular diseases, various types of cancer and to some extent neurodegenerative diseases. Mantovani and Pierotti in 2008 reported that inflammatory reactions and underlying infections are involved in 15–20% of all cancer deaths (Mantovani and Pierotti, 2008).
Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most commonly prescribed medicines for the management and treatment of various inflammatory conditions. These drugs interfere with the production of lipid autacoids known as prostaglandins (PGs), which play an important role in eliciting inflammatory reactions and its sign and symptoms (Ricciotti and Fitzgerald, 2011). NSAIDs block the biosynthesis of PGs primarily by inhibiting the arachidonic acid metabolism via inhibition of several enzymes involved in their synthesis including cyclooxygenase enzyme (COX-1 and COX-2) (Tan et al., 1992). Commonly used NSAIDs exhibit higher selectivity toward COX-1, an enzyme that is involved in the cytoprotection of the gastrointestinal tract (GIT), than COX-2 which is principally responsible to cause inflammation. A majority of the commonly used NSAIDs are non selective inhibitors of both the isozyme forms of cyclooxygenase and thus are associated with undesirable GI effects such as gastric irritation, ulceration, bleeding and renal disorders (Allison et al., 1992; Agnihotri et al., 2010). In order to overcome GI problems, highly selective COX-2 inhibitors (celecoxib, rofecoxib, etc.) were developed and marketed as gastro-protective NSAIDs (Lanza, 1998). However, long term use of some selective COX-2 inhibitors has shown potential limitations including cardiovascular complications, aggravation of ulcers among high-risk patients, delay in healing process of gastro-duodenal ulcers, prostacyclin deficiency leading to thrombosis and kidney toxicity (Verrico et al., 2003; Buttgereit et al., 2001). Hence, selective COX-2 inhibitors because of their high cost and undesirable side effects are not the ideal candidates for the treatment/management of various chronic inflammatory disorders and therefore, efforts should be made for the development of new orally active, potent, improved and safer NSAIDs with low or no GI side effects.
Imidazolidines (saturated imidazoles), also known as tetrahydroimidazoles are biologically active nitrogen containing heterocyclic moiety which have been reported to shown wide array of significant bioactivities such as anti-inflammatory, analgesic, α-adrenergic receptor agonist, antimicrobial, antiparasitic, oral hypoglycemic and anticonvulsant activities (Marki et al., 1984; Sharma and Khan, 2001; Caterina et al., 2008; Saczewski et al., 2009; Neves et al., 2010; Robert et al., 2010). They have also been considered as important scaffolds and intermediates for designing and synthesis of medicinal compounds with potential cyclooxygenase-2 (COX-2) inhibition activity (Patel et al., 2004). Several substituted-imidazolidine derivatives have been shown to be potential anti-edema agents in animal models of inflammation. Khan and Chawla, reported them to be promising group of NSAIDs with potential anti-inflammatory activities (Khan and Chawla, 2002).

Several ABC variants have been developed to improve

Several ABC variants have been developed to improve its optimization performance. One significant improvement is the introduction of PSO-based search equation (Zhu and Kwong, 2010), which allows a powerful global search in the early stage by incorporating the information of the gbest solution into ABC. Similarly, Banharnsakun et al. (2011) presented a modified search equation for the onlooker bees. In their method, the new candidate solutions are more likely to be close to the current best solution. Gao et al. (2013a,b) proposed an efficient and robust ABC variant based on modified search equation and orthogonal learning strategies, which demonstrated its high effectiveness and efficiency. Another interesting approach by (Gao et al., 2013a,b) is using the Powell’s method as a local search tool to enhance the exploitation of the algorithm. In this method, ABC good at exploration ensures the search is less likely to be trapped in local optima while it enjoys the merits of fine local search by Powell’s method. Hybridization of ABC with other operators has also been studied widely. For example, Kang et al. (2011) used the Rosenbrock’s rotational direction method to implement the exploitation phase and proposed the Rosenbrock ABC algorithm. Coelho and Alotto (2011) developed a novel alternative search equation in which a parameter is responsible for the balance between the Gaussian and the uniform distribution.
Inspired by previous works, this paper presents a novel optimization algorithm called comprehensive learning artificial bee colony optimizer (CLABC), which synergizes the idea of extended life-cycle evolving model with a pool of local searching strategies (Liu, 2013). The main motive of CLABC is to enrich artificial bee foraging behaviors in ABC model by combining GSK503 initialization based on orthogonal Latin squares approach, Powell’s pattern search method, life-cycle, and crossover-based social learning strategy, which contributes in the following aspects:
This work adopted the moving peaks benchmark (MPB) to illustrative the inherent adaptive mechanism in the proposed algorithm of surviving in a changing environment. The proposed CLABC has been compared with its classical counterpart, the classical ABC algorithm (Karaboga, 2005) over dynamic benchmarks with respect to the statistical performance measures of solution quality and convergence speed.
The rest of the paper is organized as follows. In Section 2, the proposed comprehensive learning artificial bee colony (CLABC) algorithm is given. Section 3 presents the experimental studies of the proposed CLABC and the other algorithms with descriptions of the involved benchmark functions, experimental settings, and experimental results. Finally, Section 4 outlines the conclusion.

Comprehensive learning artificial bee colony algorithm

Benchmark test

Conclusions

Introduction
In many fields of science and engineering, there are always multiple conflicting objectives, which are formulated as multi-objective (MO) optimization problems in order to minimize or maximize Anticoding strand conflicting objective functions simultaneously. In MO optimization domain, the set of Pareto GSK503 optimal solutions, namely several optimal solutions with different trade-offs in the objective space, is called the Pareto optimal front (Fonseca and Fleming, 1998; Cruz et al., 2014). Optimal power flow (OPF) is one of the most important MO problems in power system. The main goal of OPF is to find the optimal adjustments of the control variables to minimize the selected objective function while satisfying various physical and operational constraints imposed by equipment and network limitations (Kumari and Maheswarapu, 2010). Since the real power generation levels and voltage magnitudes are continuous variables whereas the transformer winding ratios and shunt capacitors are discrete variables, the OPF problem is considered as a non-linear multi-modal optimization problem with a combination of the discrete and continuous variables (Abou El Ela et al., 2010).

Based on MALDI TOF MS identification of colonies displaying distinct

Based on MALDI-TOF MS identification of colonies displaying distinct colony appearance on Slanetz–Bartley agar, E. faecium (50%) was detected as the most prevalent enterococcal species, followed by E. faecalis (27%), E. hirae (16%) and E. avium (9%). AMP resistance was detected in E. faecium and E. faecalis isolated from 12% and 3% of the dogs, respectively. The faecal concentrations of AMPR enterococci varied from 8×101 to 2.1×103cfu/g for E. faecium (<1–100% of total E. faecium) and from 103 to 6×103cfu/g for E. faecalis (<1–1.7% of total E. faecalis). Each laboratory replicate alone had a sensitivity of 100% for identification of E. faecalis, AMPRE. faecalis and E. avium/raffinosus carriers. The same sensitivity was observed using one of the replicates for identification of E. coli, CTXRE. coli and E. hirae, whereas the other replicate would have missed between one and two carriers for each of these bacterial targets (Se=98%, 88% and 94%, respectively). Use of a single replicate for detection of the remaining bacterial targets (AMPRE. coli and total and AMPRE. faecium) would have missed between one and four carriers (Se=92–96%). Data analysis showed that total enterococci were less frequent amongst dogs with gastrointestinal problems (p=0.05, OR=0.07, 95% CI 0.003-0.7). A borderline significant association was found between carriage of AMPR enterococci and AMPRE. coli (p=0.07, OR=3.2, 95% CI=0.9–1.3).
Discussion
The resistance phenotypes investigated in this GSK503 study are of high clinical relevance in small animal veterinary practice in view of the importance of β-lactams for treatment of common infections in dogs such as urinary tract infections, which are often associated to E. coli and to a lesser extent to enterococci. Various studies have shown that recent treatment with β-lactams is a risk factor for carriage of resistant bacteria in dogs (Gibson et al., 2011; Lawrence et al., 2013). The results of this study indicate that dogs without a recent history of antimicrobial treatment shed resistant bacteria such as AMPRE. coli (40%), AMPRE. faecium (12%), CTXRE. coli (8%) and AMPRE. faecalis (3%) at different frequencies and concentrations. The prevalence of AMPRE. coli carriers (40%) in Danish dogs was two to four times higher than those reported by previous studies in other countries (Murphy et al., 2009; Wedley et al., 2011), whereas the prevalence of CTXRE. coli carriers (8%) fell within the range of expected carriage frequencies (1–18.5%) (Haenni et al., 2014; Murphy et al., 2009). However, comparison between different studies is difficult due to biases associated to geographical, temporal and methodological factors. In addition, dogs included from a University veterinary facility may bias the results toward a higher proportion of carriers. However, our study population represented both primary and secondary/tertiary cases with 57% of dogs being primary cases and 1/3 of the dogs being healthy individuals.
Quantitative microbiological risk assessment is advised to assess the role of animals as a source of antimicrobial resistance in humans (Snary, 2008). The median faecal load of AMPRE. coli (3.2×104cfu/g) in dog carriers was slightly higher to that of CTXRE. coli (8.6×103cfu/g). However, CTXRE. coli are resistant bacteria of higher impact on public health due to the importance of third generation cephalosporins in the therapy of severe E. coli infections in humans. Faecal shedding of these bacteria was extremely variable amongst dogs (8×101 to 2×105cfu/g) and even higher variability (1×102 to 6×1010cfu/g) was reported by a recent study in The Netherlands (Baede et al., 2015). The average faecal concentration of CTXRE. coli in Danish dogs was close to those found in weaners and finishers (105 and 103, respectively) but lower than the average in piglets (107) in Danish pig farms positive for ESBL-producing E. coli (Hansen et al., 2013).

GSK503 br Discussion br Our study determined the

Discussion

Our study determined the average excursion of the diaphragms during tidal breathing in a standing position in a health screening center cohort using dynamic chest radiography (“dynamic X-ray phrenicography”). These findings are important because they provide reference values of diaphragmatic motion during tidal breathing useful for the diagnosis of diseases related to respiratory kinetics. Our study also suggests that dynamic X-ray phrenicography is a useful method for the quantitative evaluation of diaphragmatic motion with a radiation dose comparable to conventional posteroanterior chest radiography (22).

Our study demonstrated that the average excursions of the bilateral GSK503 during tidal breathing (right: 11.0 mm, 95% CI 10.4 to 11.6 mm; left: 14.9 mm, 95% CI 14.2 to 15.5 mm) were numerically less than those during forced breathing in previous studies using other modalities 2; 7 ;  8. Using fluoroscopy, Alexander reported that the average right excursion was 27.5 mm and the average left excursion was 31.5 mm during forced breathing in the standing position in 127 patients (2). Using ultrasound, Harris et al. reported that the average right diaphragm excursion was 48 mm during forced breathing in the supine position in 53 healthy adults (7). Using MR fluoroscopy, Gierada et al. reported that the average right excursion was 44 mm and the average left excursion was 42 mm during forced breathing in the supine position in 10 healthy volunteers (8). The difference in diaphragmatic excursion during tidal breathing versus forced breathing is unsurprising.

Our study showed that the excursion and peak motion speed of the left diaphragm are significantly greater and faster than those of the right. With regard to the excursion, the results of our study are consistent with those of previous reports using fluoroscopy in a standing position 2 ;  3. However, in the previous studies evaluating diaphragmatic motion in the supine position, the asymmetric diaphragmatic motion was not mentioned 7 ;  8. The asymmetric excursion of the bilateral diaphragm may be more apparent in the standing position, but may not be detectable or may disappear in the supine position. Although we cannot explain the reason for the asymmetry in diaphragmatic motion, we speculate that the presence of the liver may limit the excursion of the right diaphragm. Regarding the motion speed, to the best of our knowledge this study is the first to evaluate it. The faster motion speed of the left diaphragm compared to that of the right diaphragm would be related to the greater excursion of the left diaphragm.

We found that higher BMI and higher tidal volume were independently associated with the increased excursions of the bilateral diaphragm by both univariate and multivariate analyses, although the strength of these associations was weak. We cannot explain the exact reason for the correlation between BMI and the excursion of the diaphragm. However, a previous study showed that BMI is associated with peak oxygen consumption (23), and the increased oxygen consumption in an obese participant may affect diaphragmatic movement. Another possible reason is that lower thoracic compliance due to higher BMI may cause increased movement of the diaphragm for compensation. Regarding the correlation between tidal volume and excursion of the diaphragm, given that diaphragmatic muscle serves as the most important respiratory muscle, the result is to be expected. Considering our results, the excursion evaluated by dynamic X-ray phrenicography could potentially predict tidal volume.

Our study has several limitations. First, we included only 172 volunteers, and additional studies on larger participant populations are required to confirm these preliminary findings. Second, we evaluated only the motion of the highest point of the diaphragms for the sake of simplicity, and three-dimensional motion of the diaphragm could not be completely reflected in our results. However, we believe that this simple method would be practical and more easily applicable in a clinical setting.

br Our study has several limitations First

Our study has several limitations. First, we included only 172 volunteers, and additional studies on larger participant populations are required to confirm these preliminary findings. Second, we evaluated only the motion of the highest point of the diaphragms for the sake of simplicity, and three-dimensional motion of the GSK503 could not be completely reflected in our results. However, we believe that this simple method would be practical and more easily applicable in a clinical setting.

Conclusions

The time-resolved quantitative analysis of the diaphragms with dynamic X-ray phrenicography is feasible. The average excursions of the diaphragms are 11.0 mm (right) and 14.9 mm (left) during tidal breathing in a standing position in our health screening center cohort. The diaphragmatic motion of the left is significantly larger and faster than that of the right. Higher tidal volume and BMI are associated with increased excursions of the bilateral diaphragm.

AcknowledgmentsThe authors acknowledge the valuable assistance of Hideo Ogata, MD, PhD, Norihisa Motohashi, MD, PhD, Misako Aoki, MD, Yuka Sasaki, MD, PhD, and Hajime Goto, MD, PhD, from the Department of Respiratory Medicine; Yuji Shiraishi, MD, PhD, from the Department of Respiratory Surgery; and Masamitsu Ito, MD, PhD, Atsuko Kurosaki, MD, Yoichi Akiyama, RT, Kenta Amamiya, RT, and Kozo Hanai, RT, PhD, from the Department of Radiology, Fukujuji Hospital, for their important suggestions. The authors also acknowledge the valuable assistance of Alba Cid, MS, for editorial work on the manuscript. Yoshitake Yamada, MD, PhD, is a recipient of a research fellowship from the Uehara Memorial Foundation.

Appendix. Supplementary DataThe following is the supplementary data to this article:
To view the video inline, enable JavaScript on your browser. However, you can download and view the video by clicking on the icon belowVideo S1.
 A representative video of sequential chest radiographs obtained by chest dynamic radiography for the motion of the diaphragms (“dynamic X-ray phrenicography”). A board-certified radiologist placed a point of interest (red point) on the highest point of each diaphragm on the radiograph at the resting end-expiratory position. These points were automatically traced by the template-matching technique throughout the respiratory phase. Based on locations of the points on sequential radiographs, the vertical excursions and the peak motion speeds of the bilateral diaphragm were calculated (Fig 2c).Help with MP4 filesOptionsDownload video (1042 K)
Data S1.
 Multivariate analysis of associations between the excursions and participant demographics using age, gender, BMI, tidal volume, VC, FEV1, and smoking history as factors (Model 2).Help with DOCX filesOptionsDownload file (23 K)

The bilateral diaphragm is the most important respiratory muscle. Diaphragmatic dysfunction is an underappreciated cause of respiratory difficulties and may be due to a wide variety of issues, including surgery, trauma, tumor, and infection (1). Several previous studies have evaluated diaphragmatic motion using fluoroscopy 2; 3; 4 ;  5, ultrasound 6 ;  7, magnetic resonance (MR) fluoroscopy (dynamic MR imaging [MRI]) 8; 9; 10; 11 ;  12, and computed tomography (CT) 13; 14; 15 ;  16. However, the data of the previous studies using ultrasound, MR fluoroscopy, or CT were obtained in a supine position 6; 7; 8; 9; 10; 11; 12; 13; 14; 15 ;  16, not in a standing position. Also, while the data of the previous studies using fluoroscopy were obtained in a standing position, the data were assessed under forced breathing 2 ;  3, not under tidal or resting breathing. Thus, diaphragmatic motion in a standing position during tidal breathing remains unclear, even though it is essential for understanding respiratory physiology in our daily life. Furthermore, the evaluation of diaphragmatic motion using fluoroscopy, ultrasound, dynamic MRI, or CT has not been used as a routine examination because of limitations, including high radiation dose, small field of view, low temporal resolution, and/or high cost.

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Discussion

Our study determined the average excursion of the diaphragms during tidal breathing in a standing position in a health screening center cohort using dynamic chest radiography (“dynamic X-ray phrenicography”). These findings are important because they provide reference values of diaphragmatic motion during tidal breathing useful for the diagnosis of diseases related to respiratory kinetics. Our study also suggests that dynamic X-ray phrenicography is a useful method for the quantitative evaluation of diaphragmatic motion with a radiation dose comparable to conventional posteroanterior chest radiography (22).

Our study demonstrated that the average excursions of the bilateral GSK503 during tidal breathing (right: 11.0 mm, 95% CI 10.4 to 11.6 mm; left: 14.9 mm, 95% CI 14.2 to 15.5 mm) were numerically less than those during forced breathing in previous studies using other modalities 2; 7 ;  8. Using fluoroscopy, Alexander reported that the average right excursion was 27.5 mm and the average left excursion was 31.5 mm during forced breathing in the standing position in 127 patients (2). Using ultrasound, Harris et al. reported that the average right diaphragm excursion was 48 mm during forced breathing in the supine position in 53 healthy adults (7). Using MR fluoroscopy, Gierada et al. reported that the average right excursion was 44 mm and the average left excursion was 42 mm during forced breathing in the supine position in 10 healthy volunteers (8). The difference in diaphragmatic excursion during tidal breathing versus forced breathing is unsurprising.

Our study showed that the excursion and peak motion speed of the left diaphragm are significantly greater and faster than those of the right. With regard to the excursion, the results of our study are consistent with those of previous reports using fluoroscopy in a standing position 2 ;  3. However, in the previous studies evaluating diaphragmatic motion in the supine position, the asymmetric diaphragmatic motion was not mentioned 7 ;  8. The asymmetric excursion of the bilateral diaphragm may be more apparent in the standing position, but may not be detectable or may disappear in the supine position. Although we cannot explain the reason for the asymmetry in diaphragmatic motion, we speculate that the presence of the liver may limit the excursion of the right diaphragm. Regarding the motion speed, to the best of our knowledge this study is the first to evaluate it. The faster motion speed of the left diaphragm compared to that of the right diaphragm would be related to the greater excursion of the left diaphragm.

We found that higher GSK503 BMI and higher tidal volume were independently associated with the increased excursions of the bilateral diaphragm by both univariate and multivariate analyses, although the strength of these associations was weak. We cannot explain the exact reason for the correlation between BMI and the excursion of the diaphragm. However, a previous study showed that BMI is associated with peak oxygen consumption (23), and the increased oxygen consumption in an obese participant may affect diaphragmatic movement. Another possible reason is that lower thoracic compliance due to higher BMI may cause increased movement of the diaphragm for compensation. Regarding the correlation between tidal volume and excursion of the diaphragm, given that diaphragmatic muscle serves as the most important respiratory muscle, the result is to be expected. Considering our results, the excursion evaluated by dynamic X-ray phrenicography could potentially predict tidal volume.

Our study has several limitations. First, we included only 172 volunteers, and additional studies on larger participant populations are required to confirm these preliminary findings. Second, we evaluated only the motion of the highest point of the diaphragms for the sake of simplicity, and three-dimensional motion of the diaphragm could not be completely reflected in our results. However, we believe that this simple method would be practical and more easily applicable in a clinical setting.