The lungs were then kept in 100% ethanol for 24 h at 4 °C (Nagase et al., 1996). After fixation, tissue blocks were embedded in paraffin and 4-μm thick slices were cut and mounted. Slides were stained with hematoxylin–eosin. Morphometric analysis was done with an integrating eyepiece with a coherent system made of a 100-point grid consisting of 50 lines,

coupled to a conventional light microscope (Axioplan, Zeiss, Oberkochen, Germany). The volume fraction of collapsed and normal pulmonary areas and the fraction of the lung occupied by large-volume gas-exchanging air spaces (wider than 120 μm) were determined by the point-counting technique (Gundersen et al., 1988 and Weibel, 1990) at a magnification of 200× across 10 selleckchem random, non-coincident microscopic fields. Points falling on collapsed, normal or hyperinflated alveoli were counted and divided by the total number of points hitting alveoli in each microscopic field. Polymorpho- (PMN) and mononuclear (MN) cells were counted at 1000× magnification, and divided by the total number of points falling on tissue area in each microscopic field. Thus, data are reported as the fractional area of pulmonary tissue. Lung parenchyma strips (3 mm × 3 mm × 10 mm) were longitudinally cut from right lungs. Pleural tissue was removed, and the strips were stored in liquid nitrogen for analysis of type-III procollagen (PCIII)

mRNA expression. Total RNA was isolated from the

frozen lung tissue (Chomczynsky and Sacchi, 1987). The relative expression of type-III procollagen mRNA (PCIII mRNA) was small molecule library screening obtained by semi-quantitative reverse-transcription and polymerase chain reaction (RT-PCR). In the PCIII mRNA detection by RT-PCR, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) was used as internal positive control. The semi-quantitative method Terminal deoxynucleotidyl transferase of RT-PCR, used to quantify the PCIII mRNA expression in the experimental rat lung, was validated in preliminary experiments (Garcia et al., 2004 and Farias et al., 2005). All reactions included a negative control RT (-). The identity of the amplification was confirmed by determination of the molecular size on agarose gel electrophoresis with 100 bp DNA molecular markers (Gibco BRL, Grand Island, NY, USA). SigmaPlot 11 software package (SYSTAT, Chicago, IL, USA) was used. To evaluate the consequences of mechanical ventilation, ventilated groups were compared to Non-Vent. In order to analyze the effects of PEEP during OLV with low VT, comparisons between V5P2 and V5P5 were done, while the effects of high VT during OLV with physiological PEEP were assessed by comparisons between V5P2 and V10P2. The normality of the data (Kolmogorov–Smirnov test with Lilliefors’ correction) and the homogeneity of variances (Levene median test) were tested. When both conditions were satisfied one-way ANOVA test followed by Dunnett’s test and Student t-test were used.

Paleoindians relied very heavily on species of the palms Astrocaryum, Attalea, selleckchem Oenocarpus, Maximiliana, and occasionally, in Colombia, on the long-lived palm M. flexuosa (all Arecaceae). The palms whose seeds are hyper-abundant in Paleoindian sites are among those whose distribution is thought to be greatly influenced by people ( Henderson, 1995:17–20, 88–251). They are important foods sources for rural Amazonians today ( Goulding and Smith, 2007, Peters et al., 1989 and Smith

et al., 2007:38–91). Indigenous wetland foragers in the Orinoco used the abundant starch and sap from Moriche’s stout trunk as staples, supplemented with fish and fruits ( Heinen, 1988). Its fallen, rotting trunk becomes a source of plump, storable fatty beetle grubs. Also very common in the Brazilian Paleoindian food remains are the seeds of the tree legume, Hymenaea (Fabaceae), whose pod has an edible sweet, pungent aril. Brazilian Paleoindians also favored the fruits of Sacoglottis guianensis

(Humiriaceae), Talisia esculenta (Sapindaceae), Mouriri apiranga (Melastomataceae), Coccoloba pixuna (Polygonaceae), and forest Muruci (Byrsonima crispa, Malpighiaceae), which are collected and sometimes planted by indigenous and peasant communities in Amazonia ( Cavalcante, 1991 and Smith et al., 2007). More rare were Brazil nut kernels (Bertholletia excelsa [Lecythidaceae]), found only in the Brazilian sites. In one Colombian RG 7204 Tacrolimus (FK506) late Paleoindian site, paleobotanists also identified phytoliths of arrowroot (Maranta arundinacea, Marantaceae) and bottle gourd (Lagenaria siceraria, Cucurbitaceae), but these were in layers intersected by a late prehistoric intrusive pit ( Mora, 2003:126–127). Excavators also recovered seeds of the delectable

piquia fruit (Caryocar, Caryocaraceae), avocado pits (Persea, Lauraceae), and seeds of Podocarpus (Podocarpaceae), a now-rare conifer valued both for fruit and timber nowadays. That Paleoindians worked wood is shown by the heavy cutting tools they cached at some sites ( Gnecco and Mora, 1997:685, Fig. 2; Roosevelt et al., 1996:377–378, Fig. 6I). Paleoindians used forest plants that are sources of drugs or tools. A plant genus used for hallucinogens, Virola (Myristicaceae), was found in Colombian sites, and another, Vitex (Verbenaceae), used for fish bait, was identified at the early Brazilian site. The carbonized plant remains are well-dated evidence that the Paleoindians began a close relationship with numerous tree species that continue to dominate anthropic forests in Amazonia today. And their strong reliance on small fish for the bulk of their faunal diet in Brazil is a pattern that would continue through the entire indigenous human sequence in Amazonia. As a prelude to systematic agriculture, early Amazonian foragers eventually settled down at places favorable for intensive fishing and shell-fishing, especially at high land near rivers and wetlands.