Another approach is to study the organisms living at natural CO2 seeps which can be considered as a natural analogue for CO2 leakage. This volume presents data from three such sites; a deep water site in the northern Gulf of California, Mexico ( Pettit et al., 2013), a shallow water site near Vulcano Island in Italy ( Calosi et al., 2013 and Boatta et al., 2013) and a tropical site in Papua New Guinea ( Russell et al., 2013). To support the safe implementation of CCS, impact data gathered from laboratory and field experiments and from studies at analogue sites will need to be

used within a framework for environmental risk assessment. De Vries et al. (2013) explore a method to quantify the ecological risk associated with elevated CO2 levels using a Species Sensitivity Distribution (SSD); an established approach for assessing risks from toxicants. The final key element in understanding consequence is to understand http://www.selleckchem.com/Bcl-2.html the water volume or sea

floor area impacted by harmful pH changes for given leakage scenarios. If deleterious impacts are spatially restricted then environmental concerns diminish and vice versa. Whilst defining leakage scenarios is problematic, due to lack of previous events’ it is possible Roxadustat to model hypothetical scenarios. Dewar et al. (2013) show how bubble plumes of CO2 could be expected to disperse and impact the surrounding water column. While this special issue does not seek to deliver the ‘last word’ on the subject of the biological consequences of CCS leakage, the papers it contains do constitute state-of-the-art understanding, combining as they do laboratory and field investigations. It is our hope that they will act as a springboard for further work into this pressing issue, but also provide

enough of a background to inform political decision makers, and public understanding, in terms of predicting, and managing the effects of future leaks, if such leaks do occur. As a word of caution, we remind readers that when contemplating the likely environmental risks associated with leakage MG-132 it is all too easy to focus solely on the severity of any biological impacts observed. However, a comprehensive appreciation of risk must also consider the likelihood that leakage will happen, the spatial and temporal extent over which any such leak would occur and the potential recovery of organisms and ecosystems once the leak has ceased. Whilst none of these issues are considered within the current issue, this should not detract from their importance. Finally, when weighing up the environmental risks associated with CO2 leakage from CCS we must not forget that if this CO2 had not been placed into geological reservoirs it would have most likely have been released into the atmosphere, contributing to climate change, from where it will have been absorbed by the oceans thus also exacerbating ocean acidification.

Each of the 102 samples was run on the same plate in triplicate. All mRNA levels are presented relative to the geometric mean of the three control genes. PHLDA2 expression levels were quantified by Real-time PCR (QPCR) against three reference genes: tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide (YWHAZ), ubiquitin C (UBC) and topoisomerase

(TOP1) [28]. Summary data are presented as mean (SD) or median (inter-quartile range) depending on whether or not the data were normally distributed. Variables not normally distributed were transformed logarithmically. To investigate associations between PHLDA2 expression and parental body composition, fetal growth rates and infants body composition, Pearson’s and Spearman’s selleck screening library www.selleckchem.com/products/pci-32765.html correlation coefficients were calculated where appropriate. Differences in PHLDA2 expression levels between different categories of maternal lifestyle were tested by t-test or one-way

analysis of variance. Neonatal anthropometric measurements were adjusted for sex and gestational age and neonatal DXA measurements were adjusted for sex, gestational age and age at DXA. As there was a question regarding sex differences in mRNA levels between male and female placentas all mRNA data were adjusted for the sex of the baby [29]. Within group Z-scores were generated for femur length and abdominal circumference at 19 and 34 weeks. Royston models were fitted to fetal measurements to create z-scores for size and conditional growth rates [30]. To investigate whether there were sex differences in the relationship between PHLDA2 expression and the variables sex was included in regression analyses as appropriate and where an interaction was found data were analyzed separately by sex. Data were analyzed using Stata

version 11.0 (Statacorp, Texas, USA). In this study, PHLDA2 gene expression was examined in the placentas from 102 infants collected as part of the Southampton Women’s Survey. All were singleton, term deliveries (37 weeks gestation or greater). 53 of the infants were male and 49 were female. Descriptive statistics are given in Table 1. Within this cohort of 102 infants, no association was Parvulin found between the placental expression level of PHLDA2 and birth weight, placental weight or other neonatal anthropometric or body composition measurements at birth ( Table 2). Longitudinal fetal ultrasound data was available at both 19 and 34 weeks for 58 fetuses within the cohort of 102 infants. There were no differences in the birth parameters between this subset of 58 pregnancies and the 43 pregnancies without full fetal scan data (data not shown). A lower 19–34 week femur length z-score change (linear growth velocity) was significantly associated with higher term placental PHLDA2 mRNA levels ( Table 3, Fig. 1).

7 O uso do método bidimensional já foi sistematizado. As imagens são captadas por meio de um transdutor transvaginal, em tempo real Ulixertinib e em duas dimensões. A CFA com ultrassom 2 D é iniciada com a identificação do primeiro ovário, seguida por uma varredura da gônada em uma única direção de seus principais eixos em busca de imagens hipoecogênicas com diâmetro de 2 a 10 mm. Essas imagens hipoecogênicas

são contadas como folículos antrais nos dois ovários e, ao ser identificadas, são medidas em suas maiores dimensões.8 O Sono AVC (Sono Automatic Volume Calculation or Count: GE Medical Systems, Zipf, Áustria) é um novo software que identifica e quantifica regiões hipoecoicas de um ovário dentro de um conjunto de dados em três dimensões. O programa fornece estimativas automáticas das dimensões absolutas, como diâmetro e volume das imagens hipoecoicas. Na tela do ultrassom percebe‐se que a cada imagem hipoecogênica é atribuída uma cor específica e suas dimensões são

medidas automaticamente: volume (de acordo com see more o volume real de uma esfera) e os três diâmetros (x, y e z). Os volumes são exibidos em ordem decrescente. Um número ilimitado de folículos é rastreado e quantificado.9,10 Um folículo é uma estrutura tridimensional (3 D) e seu volume é a medida mais precisa para medir seu tamanho. Com o uso do diâmetro como um substituto para o volume, os folículos assumem estruturas

de esferas. Além disso, não há um padrão universal para medir o diâmetro folicular.6 Um trabalho publicado recentemente sugere que o Sono AVC fornece medições automáticas de diâmetro e volume folicular Osimertinib mais confiáveis e precisas do que as estimativas feitas com a ultrassonografia bidimensional (2 D). Esse estudo levantou a hipótese de que a medição automatizada com o uso do Sono AVC seria mais confiável e mais rápida do que medições com o método convencional 2 D.11 O presente trabalho tem o objetivo de fazer uma revisão da literatura sobre a confiabilidade da contagem de folículos antrais ovarianos com o uso da ultrassonografia bidimensional e tridimensional. Foi feita uma revisão sistemática da literatura dos trabalhos publicados de janeiro de 2000 a fevereiro de 2013 nas bases de dados eletrônicas Medical Literature Analysis and Retrieval System Online (Medline), Scientific Eletronic Library Online (Scielo) e Literatura Latino‐Americana e do Caribe (Lilacs). Como descritores foram usados: contagem de folículos antrais, reserva ovariana, cálculo automatizado de volume, ultrassom 3 D e Sono AVC. Após a leitura dos resumos foram selecionados artigos relevantes em relação à confiabilidade da contagem de folículos antrais com o uso de ultrassom bidimensional e tridimensional.

The refinery residue was obtained from ASA Indústria e Comércio LTDA (Recife-PE, Brazil). The composition of the refinery residue was previously

described [22]. The inoculums were prepared in an Erlenmeyer flask with a capacity of 250 ml containing 50 ml of YMB and were inoculated using a microbial loop, incubated in an orbital shaker at 150 rpm and 28 °C for 24 h. The pH of the culture medium was adjusted to 5.7 by addition 1 M NaOH solution or 1 M HCl solution. All fermentations were conducted in 250 ml Erlenmeyer flasks containing 50 ml of the production medium. Immediately after inoculation of 5% of 108 cells/ml, Atezolizumab price the flasks were incubated for 72 h at 28 °C in an orbital shaker at 150 rpm. The ABT-737 datasheet 72 h culture was filtered through Whatman No. 1 filter paper and centrifuged at 5000 × g for 20 min. The cell-free broth was concentrated (500 ml) by freeze-drying and extracted two times with chloroform (1:1, by vol.) in a separator funnel at 28 ̊C [23]. Surface tension was determined on cell-free broth obtained by centrifuging the cultures at 5000 × g for 20 min with a Tensiometer model Sigma 70 (KSV Instruments LTD, Finland) using the Du Nouy ring method at room temperature. The surface tension

was measured by the ring method using a DuNouy Tensiometer model Sigma 70 (KSV Instruments LTD, Finland) at room temperature. The concentration at which micelles began to form was represented as the CMC. At the CMC, sudden changes in surface tension, electrical conductivity and detergency were observed. The CMC was automatically determined by measuring the surface tensions of the purified biosurfactant in distilled water up to a constant value of surface tension. The antimicrobial activity of the crude biosurfactant produced by C. lipolytica UCP 0988 against several microbial strains was determined by the microdilution method in 96-well flat-bottom plastic tissue culture plates (Greiner Bio-One GmbH, Frickenhausen, Germany) [20]. For each strain, appropriate medium and temperature were used (as previously described); briefly, 125 μl of sterile, double-strength culture medium

were placed into the first column of the 96-well microplate and 125 μl of sterile, single-strength culture medium in the remaining Tenoxicam wells. Subsequently, 125 μl of biosurfactant solution in phosphate buffer saline at a 100 mg/ml concentration (PBS: 10 mM KH2PO4/K2HPO4 and 150 mM NaCl with pH adjusted to 7.0) (100 mg/ml) were added to the first column of the microplate and mixed with the medium; this results in a biosurfactant concentration of 50 mg/ml; serially, 125 μl were transferred to the subsequent wells, discarding 125 μl of the mixture in the tenth column, so that the final volume for each well was 125 μl. This process results in twofold serial dilutions of the biosurfactant in the first 10 columns (50–0.09 mg/ml). Columns 11 and 12 did not contain biosurfactant and served as negative and growth controls, respectively.

A long dsRNA molecule (e.g., pre-mature miRNA) is processed into a shorter dsRNA (e.g., miRNA) and then one strand is retained – the guide strand – to direct protein complexes to target mRNA molecules and prevent their translation (cytoplasmic pathways), or to target and chemically modify DNA sequences by addition of methyl groups and cause modification of DNA-associated histone proteins (the nuclear pathway).

The nuclear pathway is known to inhibit transcription and to seed heterochromatin formation (Ahlenstiel et al., 2012, Grewal and Elgin, 2007, Reyes-Turcu and Grewal, 2012 and Zhang and Zhu, 2012). Once a silencing OSI-906 order effect is initiated, the effect may be inherited. The biochemistry of this process varies depending on the organism and remains an area of active research with many unknown aspects. Nevertheless, it is known for example that human cells can maintain the modifications necessary for TGS, creating actual or potential http://www.selleckchem.com/products/U0126.html epigenetic inheritance within tissues and organisms (Hawkins et al., 2009). In some cases the dsRNA pathways induce RNA-dependent DNA methylation and chromatin changes (TGS) that persist through reproduction or cell division, and in other cases the cytoplasmic pathways remain active in descendents (Cogoni and Macino, 2000). Unintended

gene silencing is a common outcome of the genetic engineering process. Indeed, most cells initially engineered using in vitro nucleic acid techniques ultimately “silence” the gene inserted because of the engineering-associated production of dsRNA ( Carthew and Sontheimer, 2009, Hannon, 2002 and Weld et al., 2001). The new RNA sequence may be created when the DNA strand that

is not normally used as a co-factor (or “template”) for transcription is used as such. The resulting single-stranded RNA may bind to the target mRNA to create regions of linear dsRNA that can be processed into siRNA ( Fig. 1). Another possibility is that the insert contributes to the formation of a stem-loop, from which the “stem” may be processed into an miRNA-like molecule ( Fig. 1). dsRNAs are remarkably stable in the environment; a property perhaps overlooked based on the relative instability of single stranded species of RNA (Parrott et al., 2010). Insects and worms that feed on plants that make dsRNA can Pyruvate dehydrogenase lipoamide kinase isozyme 1 take in the dsRNA through their digestive system, where it remains intact (Gordon and Waterhouse, 2007 and Mao et al., 2007). RNAi has been induced through oral exposure in several insect pests (Chen et al., 2010 and Whyard et al., 2009) and oral exposure to dsRNA has been shown to reduce the lethal effects of the Israeli Acute Paralysis Virus on honey bees (Maori et al., 2009). Worms can absorb dsRNA through their skin when dsRNA is suspended in liquid (Cogoni and Macino, 2000 and Tabara et al., 1998). Once taken up, the dsRNA can circulate throughout the body and alter gene expression in the animal (Mello and Conte, 2004).

, 2013). While non-natives are usually not prevalent in mixed conifer forests, non-native plants generally have increased

in western North America ( Keeley, 2006 and Abella and Fornwalt, 2014). This increases chance that some will become established in mixed conifer forest, combined with expanding wildland-urban interfaces likely increasing opportunities for seed transport. Moreover, with reintroducing open stand structures and fire, sustainability of the current low invasion status of mixed conifer forests could be uncertain ( Keeley, 2006). It should be noted, however, that untreated forest that burns in stand-replacing wildfire can become heavily invaded over time ( McGlone and Egan, 2009). These observations suggest that: (1) monitoring non-native plant dynamics is warranted, (2) consideration could be given PD0325901 ic50 to proactively treating incipient infestations of priority species as a precautionary approach, and (3) non-native abundance after severe wildfire is likely an appropriate benchmark against which to compare non-native abundance after tree cutting and prescribed fire treatments ( Abella, 2014). Few studies of post-wildfire

dynamics have been conducted in mixed conifer forests, and few of these met our inclusion criteria. The main unmet criterion was including either pre-fire data (difficult for unplanned events such as wildfires) or comparisons to unburned areas. Some studies not meeting inclusion criteria compared fire severities SB203580 clinical trial within a burned area, but this does not provide insight into actual effect of burning (relative to no burning), which was the focus of our analysis. We suggest that wherever possible, studies of wildfires include unburned areas for comparison that also can be monitored through time. On large wildfires exceeding tens of thousands of hectares, unburned areas may not exist nearby, yet measuring unburned areas as close as possible Cyclin-dependent kinase 3 would represent unburned forest now extant on the landscape. Some preliminary expectations for wildfire effects developed from extant research of wildfire influences

on mixed conifer understories include reductions in shrub soil seed banks (Stark et al., 2006 and Knapp et al., 2012), variable responses of shrub cover which might hinge on the pre-fire shrub community (Donato et al., 2009, Knapp et al., 2012, Crotteau et al., 2013 and Walker et al., 2013), increased total species richness and forb abundance (Donato et al., 2009 and Walker et al., 2013), and contingency of effects upon fire severity likely partly mediated through overstory tree mortality (Stark et al., 2006 and Crotteau et al., 2013). Research also suggests probable increases in understory native plant cover and richness after severe burning where tree overstories are mostly or completely removed (Newland and DeLuca, 2000, Laughlin and Fulé, 2008 and Fornwalt and Kaufmann, 2014).

Different simulation models for poplar SRWC assume a mortality

of all fine roots (Fr) after the coppice of the aboveground biomass (Garten et al., 2011 and Werner et al., 2012). This confers a huge input of C into the soil after coppice, and it presents an important control on soil C sequestration (Garten et al., 2011). This assumption has, however, never been validated empirically. A recent study on oaks showed that forest interventions often result in an increase of Fr mortality and in a reduction of Fr biomass (Ma et al., 2013). Only a few studies have addressed the effect of the total aboveground removal on the vertical and the temporal SCH 900776 cell line distribution of fine roots, in particular on the annual production and turnover rate (Dickmann et al., 1996 and Dipesh and Schuler, 2013). In case all Fr would die after the harvest, this would result in a tremendous C input into the soil and it should be reflected in larger C stocks in the soil. Recent empirical research, however, has indicated that poplar SRWC did not increase the C stock in the soil (Walter et al., 2014). A SRWC potentially not only sequesters C into the soil, but also in the belowground biomass (Pacaldo et al., 2014). The belowground organs such as the stump, coarse roots (Cr) and Fr

remain in the soil after coppice, and also contribute to the C sequestration. Moreover, the allocation of Y27632 C belowground and its partitioning over different root compartments (Cr and Fr) and soil depths are important controls of the soil C sequestration (Jandl et al., 2007 and Franklin et al., 2012). This C sequestration potential could also be influenced by the initial soil C and nutrient contents of the former land use. Within the framework of SRWC we were particularly interested in the effects of the removal of aboveground Amoxicillin biomass through coppice on: (i) the seasonal and the vertical dynamics of Fr biomass and necromass, (ii) the C allocation patterns over Fr and Cr, and (iii) the C sequestration potential of the belowground organs of two contrasting Populus genotypes. Within this context our hypotheses

were: (i) harvesting of aboveground biomass decreases Fr productivity and increases Fr mortality in trees; (ii) the root:shoot ratio changes when trees are coppiced and change from a single-stem to a multi-shoot culture; (iii) the former land use (cropland or pasture) influences the belowground traits. The answers to these two hypotheses are analyzed within the context of a higher soil resource use efficiency and of the potential of SRWC for C sequestration. The experimental field site of this study is located in Lochristi, Belgium (51°06′N, 03°51′E), at an altitude of 6.25 m above sea level with a flat topography, and consists of a high-density SRWC plantation with poplar (Populus). The long-term average annual temperature at the site is 9.5 °C and the average annual precipitation is 726 mm (Royal Meteorological Institute of Belgium).

Considering its disease-inducing nature and capacity, F. cf. incarnatum may have potentials to become an important causal agent of ginseng root rot. Bacillus species are usually found in diverse natural environments of soil, water, and air and have antifungal

effects against several kinds of plant fungal pathogens [21], [23] and [40]. They also show controlling capacities for root rots and Phytophthora blight of ginseng caused by Cylindrocarpon destructans and Phytophthora cactorum, respectively [22] and [33]. In our study, a bacterial isolate identified as SP600125 B. amyloliquefaciens B2-5 had a strong antagonistic activity against the causal pathogen of ginseng root rot, F. cf. incarnatum, showing strong inhibitory activity against mycelial growth and conidial germination that

play important roles in the infection cycle of the pathogen [17]. These attributes may make the bacterium useful for controlling the ginseng root rot caused by this fungal pathogen. The bacterial isolate B2-5 had the highest control efficiency of ginseng root rot caused by F. cf. incarnatum when it was applied 2 d prior to pathogen inoculation (by pretreatment); significantly lowered PARP inhibitor control efficacies were observed in the simultaneous treatment and post-treatment. This suggests that the proper application time of the bacterial isolate may be any time prior to the disease occurrence as Bacillus spp. are durable in harsh environments due to endospore formation [41], which may be an advantage for easy formulation of the bacterial isolate for the commercialization of microbial fungicidal products. The mycelial growth of F. cf. incarnatum increased selleck inhibitor with temperature increase; however, the antagonistic activity of the bacterial isolate to the pathogen was enhanced much more than the fungal growth increase with a temperature increase up to

25°C, at which temperature the growth of the pathogen treated with antagonistic bacterium was reduced the most. This suggests that the antagonistic bacterium may exert its full disease-control capacity at a range of optimum temperatures in controlling the growths of the fungal pathogen and the half-heliophobus ginseng plant, and accordingly may lead to improved efficacy for the control of the root rot caused by F. cf. incarnatum. The inhibition of the conidial germination by the bacterial culture filtrate and the hyphal damages with no noticeable parasitism following the bacterial treatment as viewed by microscopy, suggest that bacterial antibiotics and other toxic compounds present in bacterial metabolites or a direct interaction might be responsible for the inhibition of the pathogen growth, for which antibiosis is the major action mode that exhibits instant disease control effects [42].

Various validated systems for testing the components of HPV E1 helicase and viral DNA replication using transient transfection of E1 and E2 expression plasmids or using purified enzymes in vitro have been reported ( Liu et al., 1995, Kuo et al., 1994 and Fradet-Turcotte et al., 2010). Further research is also needed in understanding the effects of CDV on the productive replicative cycle of low-risks HPVs and the organotypic epithelial raft cultures appear to be the ideal system to perform these investigations as they reproduce epithelial differentiation in an ex vivo system. A fully productive 3-dimensional tissue culture system

for production of high yields of infectious HPV-18 virions this website was first described in 2009, with multiple published applications since then (Wang et al., 2009). This system appears to be also more appropriate to analyse drug-metabolism because nucleoside metabolism in cell monolayer cultures (especially with immortalized and transformed cells)

are considerably abnormal compared to 3-dimensional tissues, where most cells are quiescent. Moreover, uptake of small molecules is substantially altered in rapidly dividing monolayer cells that do not have cell–cell junctions. Nucleotide synthetic pathways have exquisitely coordinated balancing of de novo production of the ribonucleoside and the deoxyribonucleoside RAD001 purchase triphosphates, and these regulatory responses are also heavily influenced by salvage of nucleosides from broken down RNA and DNA or from the general circulation. Exogenous agents such as inhibitors of these synthetic or salvage pathways (eg. hydroxyurea, methotrexate) or from nucleoside analogues (eg. 5-FU) can substantially alter this balancing network. Whether CDV or other ANP’s

have an impact on the normal distribution of ribo- and deoxyribo-nucleosides and their phosphorylated derivatives should be investigated. How CDV and other ANPs impact ribonucleoside diphosphate reductase, the main source PIK3C2G of deoxynucleotide synthesis in virally infected cells should be considered, as well as the consequences of cell growth in the presence of CDV with respect to ribosomal RNA transcription and processing. One of the major findings regarding CDV-antitumor activities points to the potential use of the drug in the therapy of non-viral induced tumors such as glioblastomas. Also, further research will be necessary to elucidate the effects of CDV in several signalling pathways compared to PME derivatives and other chemotherapeutics in order to highlight (dis)similarities and understand their mechanisms of action. We are grateful to the Geconcerteerde Onderzoeksacties (GOA), Krediet no. 10/014 and to the Program Financing (PF-10/08) of the KU Leuven for funding. “
“Integrase inhibitors (INIs) are an important addition to the HIV infection treatment armamentarium. Licensed in 2007, raltegravir (RAL, Merck Laboratories) is the first INI approved for clinical use (FDA, 2007).

Thereafter, a constant flow ventilator provided artificial ventilation (Samay VR15, Universidad de la Republica, Montevideo, Uruguay) with an inspired oxygen fraction of 0.21. The physiological PEEP level

was determined as follows: before the pleural space was opened, the airways were occluded at end expiration. After pleural incision, the increase Ivacaftor nmr in airway pressure corresponds to the elastic recoil pressure of the lung at relaxation volume. Thereafter, the same pressure was applied to the lung, 2 cm H2O on the average (Saldiva et al., 1992), except in V5P5 group that received 5 cm H2O of PEEP. The anterior chest wall was then surgically removed. An arterial cannula was inserted into the femoral artery for the determination of arterial partial pressure of oxygen (PaO2PaO2) (AVL Biomedical Instruments, MK-1775 cell line Roswell, GA, USA). PaO2PaO2 was measured at the beginning of the experiment and at the end of 1-h OLV (Fig. 1). The experimental protocol is depicted in Fig. 1. Two-lung volume-controlled ventilation was first established. After stabilization of the mechanical parameters under two-lung ventilation, the tracheal cannula was further introduced into the right main stem bronchus in order to exclude the left lung from ventilation. As seen in Fig. 1, pulmonary mechanics were measured in three occasions: immediately after stabilization of two-lung ventilation (TLV), immediately after

stabilization of one-lung ventilation (OLV PRE) and 1 h after the second measurement (OLV POST). Pulmonary mechanics were measured by the end-inflation occlusion method (Bates et al., 1985). In an open-chest preparation tracheal pressure reflects transpulmonary pressure. Driving pressure [difference between plateau pressure (Pplat) and PEEP], viscoelastic/inhomogeneous pressure (ΔP2) and static compliance (Cst) were measured. Cst was corrected by end-expiratory lung volume (EELV) in order to obtain specific compliance (Csp), enabling the comparison between one- and two-lung ventilation.

Pulmonary mechanics were measured 10 times in each animal in each occasion. All data were analyzed using ANADAT data analysis software (RHT InfoData, Montreal, QC, Canada). A laparotomy was performed immediately after the determination of lung mechanics, and heparin (1000 IU) was intravenously injected (abdominal vena cava). The trachea (Non-Vent group) or the right main stem bronchus (V5P2, V5P5, and Acesulfame Potassium V10P2 groups) was clamped at end-expiration, and the abdominal aorta and vena cava were sectioned, yielding a massive hemorrhage that quickly killed the animals. The lungs (Non-Vent) or the right lung (V5P2, V5P5, and V10P2 groups) were removed and weighed. End-expiratory lung volume (EELV) was determined by volume displacement (Scherle, 1970). To perform the morphometrical study, the middle lobe of the right lung was isolated at EELV, quick-frozen by immersion in liquid nitrogen, and fixed with Carnoy’s solution (ethanol:chloroform:acetic acid, 70:20:10) at −70 °C.