The cost responsibility category included such contractual elemen

The cost responsibility category included such contractual elements as each party’s responsibilities for liability/indemnity, insurance, security, and restitution/repairs. Elements such

as sanitation, other facility maintenance responsibilities, and state/local law compliance fell Z-VAD-FMK chemical structure under the sustainability category. Finally, elements that defined the range of program services to be provided, specific spaces/facilities to be utilized, and use periods of the school grounds/facilities were grouped under the scope category. Agreements were also analyzed by type of mechanism used and whether the SUA included programmatic and/or open-gate elements. To provide supplemental context to the 18 SUA reviews, we calculated the potential number of residents reached by each agreement intervention, using geographic information systems (GIS) and the 2010 Census data (U.S. Census, 2010). Mapping of the 49 SUA school locations, for example, was carried out using a 1-mile buffer placed around each of the shared-use school sites with the assumption that community members may travel up to

1 mile to use the open space or facilities. When reviewing the literature, we found a lack of consensus on an acceptable distance that people are willing to travel to for recreation, ranging from 1/8th of a mile to 1 mile (Harnik and Simms, 2004). Although we believe people are not likely to walk more than 1/2 mile to a park or recreation space, given the commuter culture of LAC and the lack of recreational facilities MLN8237 solubility dmso in the targeted communities, we believe 1 mile is an acceptable distance for people to travel. Population in the surrounding community was estimated for each of the census tracts

within the 1-mile radius (buffer region), assuming uniform population numbers throughout the census tract. When appropriate, we calculated a ratio of CPPW funds invested to community members reached, based on the total expenditures or investments made by the JUMPP Task Force to construct and implement SUAs across the seven school districts. DPH’s institutional review board reviewed and approved all study protocols, procedures, and materials prior to fieldwork. Eighteen SUAs met the criteria for inclusion (JUMPP-assisted, physical activity-related, focus Suplatast tosilate on children and adults). Of the eight school representatives that completed the school site and community partner survey, approximately half (50%) reported safety, vandalism, and staffing as their top concerns. A little over one-third (37.5%) considered operational/maintenance issues as a challenge. Approximately 62.5% indicated that their school district would be amendable to opening outdoor school facilities for community use outside of regular school hours; about half would work with third parties (e.g., sports leagues, government agencies, and community organizations) to operate programs (e.g.

For children over 12 months of age, there were 4 cases of inpatie

For children over 12 months of age, there were 4 cases of inpatient pneumonia in children who had received the 12 month PPV-23 compared with 7 cases in those that had not during the same follow up period. There were no cases of IPD throughout the study period. This study has shown that 1, 2, or 3 doses of PCV-7 in infancy primed infants sufficiently elicit an excellent booster response to the PPV-23 at 12 months GW786034 of age for all PCV-7 serotypes. Furthermore, there were good antibody responses to the 16 non-PCV-7

serotypes following PPV-23 at 12 months. The antibody concentrations for all 23 serotypes remained significantly higher at 17 months of age in the PPV-23 group compared to the group that had not received PPV-23. In addition, this study has shown that priming with a single PCV-7 dose in infancy produced the greatest booster (memory) response for most serotypes following PPV-23 at 12 months compared with 2 or 3 PCV-7 doses. Responses following the PPV-23 were similar for those children that had received either 2 or 3 PCV-7 doses in infancy and lower than that in children

who received a single PCV-7 dose. The immunological explanation for the single PCV-7 dose having a better booster response is not clear. Post booster antibody concentrations are PI3K activation usually higher in those that have had a stronger primary response [34]. One study found that a stronger primary response was more likely following higher doses of antigen and/or a higher concentration of carrier protein, possibly through the enhanced induction of antibody producing plasma cells [35]. However this would not explain the findings in our study of a better booster response in the single dose group as our previously published data has shown that a single PCV-7 dose (lower antigen dose) administered at 14 weeks of age induced a weaker primary see more response [29]. In that previous study, a significant immunological response was found in the single dose group compared with an unvaccinated control group, but significantly lower

GMC for all PCV-7 serotypes compared to 2 or 3 PCV-7 doses [29]. Another possible explanation for the better booster response in the single PCV-7 dose group may be that a single antigen challenge rather than multiple antigen exposures, may preferentially drive the induction of memory B cells (which are required for a booster response), rather than plasma cells [36]. Having a greater pool of memory B cells would subsequently elicit a greater booster response. A fewer dose (single PCV-7 dose) primary series may preferentially induce B cell differentiation away from plasma cells, towards memory B cells compared to repeated antigen exposure associated with 2 or 3 PCV-7 dose primary series [8] and [11].

2) (35) These results indicate that NOSs in bone marrow cells

2) (35). These results indicate that NOSs in bone marrow cells

exert an inhibitory effect on vascular lesion formation caused by blood flow disruption in mice in vivo, UMI-77 demonstrating a novel vasculoprotective role of NOSs in bone marrow-derived vascular progenitor cells. During 11 months of follow-up, all (100%) of the wild-type mice lived, whereas only 15% of the triple NOSs null mice survived (Fig. 3A) (33). The survival rate was significantly worse in accordance with the number of disrupted NOS genes in the order of single, double, and triple NOSs null mice. Postmortem examination revealed that 55% of the triple NOSs null mice died of myocardial infarction (Fig. 3 and Fig. 4A) (33). This is the first demonstration to show that a deficiency of NOSs leads to the development of spontaneous myocardial infarction. In the coronary arteries of the dead triple NOSs null mice, marked intimal formation, medial thickening, and mast cell infiltration were noted, while intra-coronary thrombus was rarely observed

learn more (Fig. 4A–C) (33). Histamine released from adventitial mast cells is thought to cause coronary vasospasm with resultant myocardial infarction in humans (36). It is thus possible that coronary intimal hyperplasia, medial thickening, and vasospasm are involved in the pathogenesis of myocardial infarction in the triple NOSs null mice. Although human myocardial infarction mainly results from rupture of atherosclerotic plaques and subsequent thrombus formation, the triple NOSs null mice seem to be a model of non-atherosclerotic forms of acute myocardial infarction in humans. In the triple NOSs null mice, there was a complete lack of endothelium-dependent relaxations to acetylcholine, which is a physiological Linifanib (ABT-869) eNOS activator, and contractions to phenylephrine, which is an α1 adrenergic

receptor agonist, were markedly potentiated (33). Thus, vascular dysfunction could also be involved in the pathogenesis of myocardial infarction in the triple NOSs null mice. The renin-angiotensin system was markedly activated in the triple NOSs null mice, and long-term treatment with an angiotensin II type 1 (AT1) receptor blocker olmesartan potently inhibited coronary arteriosclerotic lesion formation, vascular mast cell infiltration, and the occurrence of myocardial infarction in those mice, with a resultant improvement of the prognosis (33). These results suggest that the AT1 receptor pathway is involved in the occurrence of spontaneous myocardial infarction in the triple NOSs null mice.

2 N sodium hydroxide

(NaOH)] Eppendorf tubes were inverte

2 N sodium hydroxide

(NaOH)] Eppendorf tubes were inverted five times gently, and allowed to stand at room temperature for 5 min. Subsequently, incorporated 0.3 ml ice-cold solution 3 (3 M Potassium acetate and 5 M glacial acetic acid) into each tube and inverted five times gently, and allowed to stand on ice for 10 min. After centrifugation (14,000 rpm, 2 min) pellet was dissolved in 0.5 ml of TE (Tris–EDTA, 0.05 M, pH 8.0) and incubated for 5 min at 65 °C, added 0.5 ml of Phenol–Chloroform–Isoamyl alcohol (25:24:1) and shaken thoroughly for 10 min and then solution was centrifuged at 14,000 rpm for 3 min at 4 °C. Supernatant was transferred to another tube Y-27632 mw and added 1 ml of ice-cold 70% ethanol and centrifuged at 4 °C for 7 min at 7500 rpm. The pellet was air dried and suspended in an appropriate volume of Tris–EDTA buffer. DNA purity and concentration were assayed in a spectrophotometer (260/280). The vanA gene was detected using previously reported primers. 18 Primers were obtained from Sigma Aldrich Chemicals Pvt. Ltd., Banglore, India. Primer used for vanA-F-5′-CATGAATAGAATAAAAGTTGCAATA-3′ and vanA-R-5′-CCCCTTTAACGCTAATACGACGATCAA-3′ BMS-354825 cell line that amplify a fragment

of about 1030 bp. PCR assay was performed in a total volume of 20 microliter (μl) containing 200 picogram (pg) of DNA, 0.5 mM of deoxynucleotide triphosphates (dNTPs), 1.25 micromolar (μM) of each primer and 1.5 U of Taq polymerase (Banglore Genei). PCR amplification was carried out on an Eppendorf thermocycler (Germany)

with cycling conditions: initial denaturation at 94 °C for 10 min followed by 30 cycles each of denaturation (94 °C for 30 s), annealing (50 °C for 45 s), extension (72 °C for 30 s) and final extension (72 °C for 10 min), for the amplification of vanA gene. The PCR products were analyzed in 1% (w/v) agarose gel containing 25 μg of ethidium bromide in Tris–EDTA buffer and the gel was photographed under ultraviolet illumination using gel documentation system (Bio-Rad, USA). After electrophoresis, density of during PCR product bands were measured by Image J software. Conjugation study was done by a broth mating method as described elsewhere.13 Briefly, donor (vanA positive VRSA) and recipient (vanA negative S. aureus) cells at a concentration of 106 cfu/ml cells were mixed in one to nine ratio (0.1 ml donor cells and 0.9 ml recipient cells), and was swirled for a few minutes and then incubated at 37 °C for 6 h in M-H broth (without shaking). Transconjugants were selected by plating 0.2 ml on MH agar plate containing 16 μg/ml vancomycin and 2.5 μg/ml ciprofloxacin. Colonies were counted after 48 h of incubation. Donor and recipient cells were also plated separately to check their disability to grow on the vancomycin plus ciprofloxacin plate, because the donor was ciprofloxacin-sensitive and the recipient was susceptible to vancomycin. The transfer of vanA was also confirmed by vanA gene amplification in transconjugants.

, 2007) For IL-6, the PCR primers and sequencing probe were desi

, 2007). For IL-6, the PCR primers and sequencing probe were designed

to target sites within a CpG island located in the promoter region of the gene using the Pyromark Assay Design Software Version 2.0 (Qiagen). The sequences were as follows: TTTTGAGAAAGGAGGTGGGTAG (Forward PCR primer), ACCCCCTTAACCTCAAATCTACAATACTCT (5′ biotinylated Reverse PCR primer), and AAGGAGGTGGGTAGG (Sequencing primer). The coefficients of variation (CV) for the LINE-1 methylation assay range from 0.5 to 2.6% and the CVs for IL-6 promoter methylation assay range 3-MA ic50 between 5.3 and 14.8%. We administered the validated 108-item Block food frequency questionnaire (FFQ), (Block et al., 1990 and Subar et al., 2001) and the Block Adult Energy Expenditure Survey (Block et al., 2009). The nutrient and energy expenditure computations of the de-identified questionnaires

were performed by NutritionQuest, the distributor of the two questionnaires. We first compared the demographics between car drivers and PT users. Linear regression was used to estimate the difference and associated 95% confidence intervals (95%CI). We then compared the median and interquartile range (IQR) of daily intakes of foods and nutrients between the two groups. To construct dietary patterns, we performed factor analysis of 13 food groups using the principal factor method followed by an OSI 744 orthogonal rotation. Based on the scree test results, the proportion of variance accounted and the interpretability criteria, we identified two factors, i.e. two dietary patterns. For each subject, we estimated factor scores for the two dietary patterns by summing the frequency consumption of PD184352 (CI-1040) each food group weighted by their scoring coefficients. Subjects were then categorized into quartiles of factor scores for two dietary patterns, with high scores corresponding to a better adherence to a particular dietary pattern. We also estimated the car-vs-PT mean differences in factor scores for each of the two dietary patterns and associated 95%CIs using the beta coefficients of linear regression models and their standard

errors. Next, we compared the median levels of reported daily physical activities between car drivers and PT users. Using linear regression, we also evaluated whether two groups differed in their adherence to physical activity guidelines by assessing the proportion of subjects meeting the U.S. Department of Agriculture 2005 Dietary Guidelines for Americans (DGA) for physical activity (i.e., engaged in approximately 60 min of moderate- to vigorous-intensity activity on most days of the week), or meeting the Healthy People 2010 Guidelines for physical activity (i.e., engaged in moderate physical activity for at least 30 min on at least 5 days a week, or engaged in vigorous physical activity for 20 min on at least 3 days/week). We used logistic regression to compare differences in distributions across quartiles of durations of the various types of physical activity.

Vaccinomics

provides powerful tools to select antigens fr

Vaccinomics

provides powerful tools to select antigens from pathogens with large genomes, while systems biology offers novel approaches to understanding the complexity of immune responses after infection or vaccination [47], [48] and [49]. However, many click here scientific barriers still need to be overcome. Raising awareness on the fact that STIs are a major global cause of acute illness, infertility, long-term disability and death with serious medical and psychological consequences of millions of men, women and infants is crucial. In this regard, the WHO initiative to convene a Technical Consultation on STI Vaccine Development and Implementation is an important step forward. The disease burden needs to be reviewed and evaluated, not only in terms of numbers of infection or mortality, but also in terms of number of complications and sequelae and of economic and psycho-social impact. This requires improving diagnosis of STIs and case-definition of complications, defining criteria for evaluation of psychological distress and social disruption caused by STIs. Epidemiological studies could help identify geographical variations http://www.selleckchem.com/products/Trichostatin-A.html in incidence, prevalence, and strain circulation, and identify communities

at higher risk of STIs where clinical trials could be carried out. In parallel, building on the experience with HPV vaccine, the public health community could develop programs focusing on advocacy and education on STI prevention, approach public health authorities as well as funding agencies to prepare the introduction of STI vaccines in both developed and developing countries. Identifying what may constitute a protective immune response against STIs in humans is a key issue that requires further research. One approach could be the assessment of cohorts of patients with varying severity of symptoms and outcomes, which calls for improved methods in the diagnosis of subclinical

and clinical STIs. Blood and cell banks from these cohorts could be made available to the scientific community in order to study the mechanisms of pathology and define predictive markers already of outcomes and protection. Comparative studies of the pathogens and host factors, including immune responses from these different groups of patients would contribute to finding correlates of protection in humans. A recently published study [50] clearly identified a cohort of women who acquired immunity post chlamydia infection and further studies along this line could reveal important knowledge. Further research should also be conducted on the mechanisms of immune responses in relation with the specificity of the genital tract, integrating data on the microbiome and hormonal status [for a review, see the article by Brotman et al., in this issue [51].

For this purpose, 50 μL of Acamprosate D12 ((IS) concentration of

For this purpose, 50 μL of Acamprosate D12 ((IS) concentration of 50 ng/mL) 250 μL plasma (respective concentration of plasma sample) was added into riavials then vortexed approximately. Followed by 1000 μl of water was added and vortexed for 2 min. These samples were added into SPE Catridges (Agilent polymer SAX,

3 Ml, 60 mg, 60 μm) which were pre conditioned with 1 ml methanol, followed BIBW2992 price by 1 ml water. After that, the samples which were in SPE, were washed with 1 ml water, followed by 1 ml Methanol. Elute the cartridges with 2 ml of 20% formic acid solution into separate glass cultured tubes and evaporate at 70 °C. Then these samples were reconstituted with 100 μL of 20% formic acid solution PH-3.5 and vortexed. Finally, 900 μL of acetonitrile was added to each sample and vortexed for 2 min. At last, these

samples were centrifuged at 4000 rpm at 20 °C for 5 min. learn more Then transferred the sample into auto sampler vials with caps and 20 μL of sample from each autosampler was allowed to instrument at optimized chromatographic conditions. Six different screened lots of human plasma samples were selected from different donors for selectivity. These screened lots were used for validation experiments to test for interference at the retention time of analyte internal standard. The matrix effect due to the plasma matrix was used to evaluate the ion suppression/enhancement in a signal when comparing the absolute response of QC samples after pretreatment (SPE) with the reconstitution samples extracted blank plasma sample spiking with analyte. Experiments were performed at LQC and HQC levels in triplicate with six different plasma lots with the acceptable precision (%CV) of ≤15%. It was determined by replicate analysis of quality control samples (n = 6) at LLOQ (lower limit of quantification), LQC (low quality control), MQC (medium quality control), HQC (high quality control) and ULOQ (upper limit of quantification) levels. Precision and accuracy should be within 15% for all the standards except LLOQ. For LLOQ it should be within 20%. The recovery

was carried out between extracted area to non extracted area of each concentration. however For Acamprosate recovery was proved at LQC, MQC, HQC level and for Acamprosate D12 recovery was proved at single concentration at respective standards. During real subject sample analysis, some unknown sample concentrations may fall above ULOQ and below MQC Level. To evaluate the actual concentration of those unknown samples, dilution integrity test was performed at 1.5 times of ULOQ concentrations were prepared and performed at six replicates from each level (½, ¼ of ULOQ) and calculated by applying dilution factor 2 and 4 with freshly prepared standards. Stability of the drug was proved in stock solution, and in plasma samples. Stability of internal standard was proved in stock solution.

SSD received fellowship from Department of Biotechnology (DBT), G

SSD received fellowship from Department of Biotechnology (DBT), Government of India. This experimental work in S. album in the author’s laboratory was supported under the project – Prospecting of novel genes and molecules of S. album L. (NGM), sponsored

by DBT, Government of India. “
“Cefpodoxime proxetil (CP) is an orally absorbed, broad spectrum, third generation cephalosporin ester. This prodrug ester is hydrolyzed in vivo into its active metabolite, cefpodoxime. In human, the absolute bioavailability of CP administered as a 130 mg tablet (equivalent 100 mg of cefpodoxime) is about 50%. 1 However, the high solubility, chemical and enzymatic stability, and absorption profile of CP in acidic pH values of stomach, points to the potential of a gastroretentive (GR) dosage form

in altering the absorption profile of CP. 2 Mucoadhesive drug delivery systems for its potential http://www.selleckchem.com/products/ABT-737.html CX-5461 as optimize localized drug delivery, by retaining a dosage form at the site of action or systemic delivery, by retaining a formulation in intimate contact with the absorption site. 3 and 4 Despite the mucoadhesion, the advantage of using microspheres as oral mucoadhesive drug delivery system is that the small size microspheres can be trapped in the reductus of the stomach and stay there longer. Besides, when poorly soluble drugs were loaded in the mucoadhesive microspheres, there were either adsorbed at the surface of the microspheres or highly dispersed in the inner part of the microspheres which may help enhance the solubility of the drugs, results in improved bioavailability. 5 Chitosan (CS), a cationic polymer and an interesting material for microparticulate systems because

of its good mucoadhesive and biodegradable properties. 6 It is well established that traditional experimentation involves a good deal of efforts and time especially when complex formulations secondly are to be developed. In addition to the art of formulation, the technique of factorial design is an efficient method of indicating the relative significance of a number of variables and their interactions. 7 The objective of the present work is to improve the oral bioavailability of CP by formulating gastroretentive mucoadhesive microspheres which will provide protection from intestinal milieu using CS and to characterize for in vitro and in vivo parameters. A 32 full factorial design (two variables in three levels) was employed to evaluate the combined effect of the selected independent variables: CP to CS ratio (A) and amount of glutaraldehyde (GA) (B) on dependent variables such as drug entrapment efficiency, swelling index, percentage mucoadhesion and time for 50% drug dissolution (t50). Cefpodoxime proxetil was received as a gift sample from Orchid Chemicals and Pharmaceuticals Ltd, Chennai. Chitosan (≥75% deacetylated) obtained from Sigma Aldrich (Mumbai, India). Dioctyl sodium sulfo succinate (DOSS), petroleum ether (S.

The plant P oleracea L was proved to show the muscle relaxant ac

The plant P. oleracea L was proved to show the muscle relaxant activity, 3 anti-inflammatory effect, 4 in some Middle East countries, it is considered as beneficial for small tumors and inflammation, urinary disorders, liver obstruction and ulcer of mouth and stomach. Several researchers have shown that P. oleracea L is having anti-hyperglycemic activity, anti-tumor activity and anti-ulcer activity. 5 This plant has also proved for gastric anti-ulcer activity. 6 The plant P. BI 2536 cost oleracea L (Purslane) is commonly known as Porsulane a herbaceous weed. This plant is an annual succulent prostrate herb; stem is about 15.30 cm long, reddish, swollen at the nodes, quite glabrous. Leaves are freshly, sub-sessile, 6.25 mm long

alternate or sub-opposite. Flower few together, in sessile terminal heads. Microscopic analysis of the leaf powder invariably shows spherical mineral crystals, sieve plants, tracheas with spiral, annular and scalariform thickening and vessels with bordered pits. 7 The aim of the present study is to evaluate anti-ovulatory activity, anti-estrogenic activity, effect on uterine

muscle weight and ovary weight and biochemical analysis of ovary and uterus of ethanol extract of P. oleracea L in female albino rats. The healthy aerial part of the plant of P. oleracea L was collected from around Gulbarga university campus during the month of June 2011. The plant material was identified and authenticated at the Department of Botany Gulbarga University Gulbarga Karnataka (India), voucher specimen (No. HGUG-5013) has deposited find more in herbarium of the same department. Methanol, ethanol, ethyl acetate, petroleum ether, diethyl ether, H2SO4, chloroform, HCl, KOH, hexane, silica gel 60–120 mesh, Tween 80 phosphate buffer saline, Folin–Ciocalteu reagent,

all the chemical, solvents and reagents used were analytical grade and obtained from Hi media. The plant material was dried in shade, ground and extracted with 95% ethanol by soxhlet extraction at 90 °C for 12 h until the color of elute should colorless. The extract was taken and solvent was evaporated at room temperature so as to get crud drug and stored at 4 °C for further use. The presence of flavonoids Montelukast Sodium was confirmed by specific tests for flavonoids like shinoda test, lead acetate test, sodium hydroxide test, sulfuric acid test, aqueous test. These are the specific tests, for detection of flavonoids.8 Experiment was performed on virgin female albino rats aged about seven weeks (100 g) obtained from Luqman Pharmacy College, Gulbarga. The animals were acclimatized for 1–2 weeks before being used for the experiment. Fed with Standard palliated diet (Amrut laboratory animal feed diet, Pune, Maharashtra, India) and water was given ad libitum. They were housed under standard condition of temperature (24 °C), humidity (65%) light and dark cycle (14:10 L), respectively. The initial body weight of each animal was recorded.

The exercises check

The exercises VX-770 for SCI are useful, well described, and task-oriented. The section related to infants and children is also full of interesting exercise possibilities that are task-oriented. While most of the exercises related to stroke are potentially useful, some may benefit from review as outlined above. It may be useful to note that our comments on stroke are based on the guidelines for exercise and training after stroke that we have developed over many years, based on current scientific understanding

in the areas of brain plasticity, motor learning, exercise science, and current clinical evidence. “
“Nanoparticles (NPs) play a decisive role in industrial applications on the one hand, and on the other hand, NPs are Androgen Receptor signaling pathway Antagonists gaining in interest for biomedical research (drug and gene delivery) [1]. Regarding an entry of NPs via inhalation, the alveolar region of the lung with a surface area of 100–140 m2 make it an interesting target for drug and gene delivery, but at the same time, the lung represents a significant portal of entry for harmful nanomaterials. Inhaled silica nanoparticles (SNPs), for example, embody a serious health-risk characterised by environmental and occupational lung diseases (silicosis)

[2]. It has been proposed that pulmonary release of cytokines and mediators into the circulation, that are triggered by inhaled NPs, cause extrapulmonary effects [3]. Epidemiological studies revealed that particulate air pollution (PM10: Particulate matter <10 μm) increased the frequency of cardiac diseases [4] and [5]. However, plausible explanations from the biological perspective are still lacking. It is also suggested that the resulting systemic effects are caused by an excess of inhaled PM10 that migrate into the systemic circulation and then translocate to different organs [6] and [7]. Thus, if a lung application is envisaged, toxic effects and the cellular pathways as well as the further disposition of inhaled NPs need to be addressed to gain more insight concerning the above mentioned

hypotheses. Cytotoxicity and cellular uptake/trafficking of nanoparticles in the lower respiratory tract are still poorly understood. One reason for STK38 this is that the alveolar-capillary barrier of the deep lung is difficult to access by in vivo studies. Therefore, we have inspected nanoparticle interactions on an in vitro coculture model of the alveolar-capillary. This in vitro model consists of the epithelial cell line, NCI H441 (with characteristics of type II pneumocytes and Clara cells) and the human microvascular endothelial cell (MEC) line, ISO-HAS-1, which are seeded on opposite sides of a transwell filter membrane. Both cell types in coculture (CC) reach a more differentiated and polarised phenotype than if the cells are kept under conventional monoculture (MC) conditions [8] and [9]. Therefore, it more closely mimics the in vivo situation of the deep lung.