Lung volume and interdependence in the pig.

The effect of lung volume on the mechanical interdependence between an obstructed sublobar region of lung and its surrounding tissues was investigated in intact and isolated pig lungs. Interdependence is arbitrarily defined as the effectiveness with which the linkage between the region and surrounding tissue mediates a tendency for inflation to become even whenever it is uneven. We found that when the volume of the surrounding lung (Vl) was high relative to the volume of the obstructed region (Vr), or when they were relatively equal at higher inflation states, interdependence decreased. When Vr was high relative changes in regional shape observed during even and uneven inflation states, we suggest that regional distortion and its effects on regional elastic recoil are important determinants of pulmonary interdependence.     

Electroporation of the vasculature and the lung

Electroporation has proven to be a highly effective technique for the in vivo delivery of genes to a number of solid tissues. In most of the reported methods, DNA is injected into the target tissue and electrodes are placed directly on or in the tissue for application of the electric field. While this works well for solid tissues, there are many tissues and organs that are not amenable to such an approach. In this review I will focus on the development of electroporation protocols for two such tissues: the vasculature and the lung. Several methods for in vivo electroporation of the vasculature have been developed in recent years that deliver DNA to vessel segments from either the inside or outside of the vessel. The advantages and disadvantages of each are discussed, as are the applications for which they have been used. In more recent work, our laboratory has developed a novel method to deliver genes to the rodent lung that results in high level, uniform, gene expression throughout all cell types of the lung. Most importantly, this technique is safe, and causes no inflammatory response or alterations in normal physiology of the organs. Taken together, these studies demonstrate the utility of electroporation for gene transfer to non injectible tissues.     

Regional interdependence in calf lungs

We measured K, an index of interdependence in sublobar segments of both intact and excised calf lungs. In excised and intact lungs, segment volume was held constant while the lung was inflated. K in excised lungs was of small magnitude but was greater in caudal than in cranial and middle lobes. When isolated segments in a variety of regions and with a variety of shapes were studied, K was positively correlated with the interface area between the segment and the remainder of the lung, was negatively correlated with the pleural surface area of the segment, but was not correlated with segment volume. In a given region of lung, however, small segments had a greater interdependence than large segments. In intact lungs of anesthetized calves, K was an order of magnitude greater than in excised lungs, confirming that the chest wall is the major determinant of interdependence. There were no lobar differences in K in intact lungs.     

Interstitial fibrosis and collateral ventilation

Interstitial fibrosis may increase resistance to collateral flow (Rcoll) because of decreased lung volume and destruction of collateral channels or it may decrease Rcoll because of emphysematous changes around fibrotic regions. In addition, if interstitial fibrosis involves a small region of lung periphery, interdependence from surrounding unaffected lung should produce relatively large changes in volume of the fibrotic region during lung inflation. We studied the effects of interstitial fibrosis on collateral airflow by measuring Rcoll at functional residual capacity (FRC) in nine mongrel dogs before and 28 days after the local instillation of bleomycin into selected lung segments. In six of these dogs Rcoll was also measured at a higher lung volume (transpulmonary pressure = 12 cmH2O above FRC pressure). Rcoll increased in fibrotic lung segments following local treatment with bleomycin. With lung inflation (high transpulmonary pressure) Rcoll fell a similar proportion in fibrotic and nonfibrotic lung regions. These observations suggest that collateral resistance increases in fibrotic segments because lung volume decreases or because collateral pathways are involved directly in the fibrotic process. Compensatory increases in collateral communications do not occur. In addition, pulmonary interdependence does not cause disproportionate increases in volume and decreases in Rcoll of the fibrotic region during lung inflation.     

Determination of volatile organic compounds as biomarkers of lung cancer by SPME-GC-TOF/MS and chemometrics

A method for qualitative and quantitative the determination of concentrations volatile organic compounds (VOCs) in human breath samples using solid phase microextraction (SPME) and gas chromatography-time of flight-mass spectrometry (GC-TOF/MS) has been carried out. They are employed for the preconcentration, separation and analysis of biological samples. The technique to rapid determination compounds present in human air, at the level of parts per billion (ppb) is applied. This method was optimized and evaluated. It showed linear correlations ranging from 0.83 to 234.05 ppb, limit of detection in the range of 0.31 to 0.75 ppb and precision, expressed as the RSD, was less then 10.00%. The unique combination of statistical methods allowed reduce the number of compounds to significant ones only and indicate the potential way to find the biomarkers of the lung cancer. Presented an analytical and statistical methods for detection composition of exhaled air could be applied as a potential non-intrusive tool for screening of lung cancer.     

Determination of moxifloxacin (BAY 12-8039) in plasma and lung tissue by high-performance liquid chromatography with ultraviolet detection using a fully automated extraction method with a new polymeric cartridge

The aim of this study was to develop a high-performance liquid chromatographic (HPLC) assay for the determination of moxifloxacin in human plasma and lung tissue. The assay was based on HPLC with a Supelcosil ABZ+ column and ultraviolet detection set at a wavelength of 296 nm. The extraction procedure was characterized by a fully automated liquid–solid extraction using an OASIS column for the solid phase. The assay has been found to be linear and validated over the concentration range 3.2 to 0.025 μg/ml for moxifloxacin in plasma and from 16 to 0.25 μg/g for moxifloxacin in lung tissue. In future, the assay will support the pharmacokinetic study of the penetration of moxifloxacin in human lung tissue.     

Endothelial and epithelial signaling in the lung

In recent years, pulmonary research has focused increasingly on the understanding of proinflammatory endothelial and epithelial signaling pathways underlying lung injury that is well known to be associated with high morbidity and mortality. Although traditionally considered separately, current research shows considerable convergence in epithelial and endothelial signaling mechanisms. This was well emphasized in the featured topic session held during the Experimental Biology 2007 Meeting in Washington, D.C., that addressed the complex interdependence between signaling pathways in the two cellular phenotypes. Several perspectives on endothelial and epithelial signaling, as well as their contributions to pulmonary inflammation and damage, as presented and discussed in the session are summarized here.     

A method of intravital study of pulmonary microcirculation in closed-chest cats

The methods of intravital study of the cat lungs have been updated. A device has been designed for the investigation of microvessels in extended lung areas of spontaneously breathing closed-chest cats. The principles for quantitative analysis of microcirculatory lung parameters have been elaborated. These new methods allow the study of important natural phenomena of the lung microvascular functions.     

How much is there really? Why stereology is essential in lung morphometry.

Quantitative data on lung structure are essential to set up structure-function models for assessing the functional performance of the lung or to make statistically valid comparisons in experimental morphology, physiology, or pathology. The methods of choice for microscopy-based lung morphometry are those of stereology, the science of quantitative characterization of irregular three-dimensional objects on the basis of measurements made on two-dimensional sections. From a practical perspective, stereology is an assumption-free set of methods of unbiased sampling with geometric probes, based on a solid mathematical foundation. Here, we discuss the pitfalls of lung morphometry and present solutions, from specimen preparation to the sampling scheme in multiple stages, for obtaining unbiased estimates of morphometric parameters such as volumes, surfaces, lengths, and numbers. This is demonstrated on various examples. Stereological methods are accurate, efficient, simple, and transparent; the precision of the estimates depends on the size and distribution of the sample. For obtaining quantitative data on lung structure at all microscopic levels, state-of-the-art stereology is the gold standard.     

Mathematical simulation of forced expiration

Flow limitation during forced expiration is simulated by a mathematical model. This model draws on the pressure-area law obtained in the accompanying paper, and the methods of analysis for one-dimensional flow in collapsible tubes developed by Shapiro (Trans. ASME J. Biomech. Eng. 99: 126-147, 1977). These methods represent an improvement over previous models in that 1) the effects of changing lung volume and of parenchymal-bronchial interdependence are simulated; 2) a more realistic representation of collapsed airways is employed; 3) a solution is obtained mouthward of the flow-limiting site by allowing for a smooth transition from sub- to supercritical flow speeds, then matching mouth pressure by imposing an elastic jump (an abrupt transition from super- to subcritical flow speeds) at the appropriate location; and 4) the effects of levels of effort (or vacuum pressure) in excess of those required to produce incipient flow limitation are examined, including the effects of potential physiological limitation.     

Targeted proteomics of solid cancers: from quantification of known biomarkers towards reading the digital proteome maps

The concept of personalized medicine includes novel protein biomarkers that are expected to improve the early detection, diagnosis and therapy monitoring of malignant diseases. Tissues, biofluids, cell lines and xenograft models are the common sources of biomarker candidates that require verification of clinical value in independent patient cohorts. Targeted proteomics – based on selected reaction monitoring, or data extraction from data-independent acquisition based digital maps – now represents a promising mass spectrometry alternative to immunochemical methods. To date, it has been successfully used in a high number of studies answering clinical questions on solid malignancies: breast, colorectal, prostate, ovarian, endometrial, pancreatic, hepatocellular, lung, bladder and others. It plays an important role in functional proteomic experiments that include studying the role of post-translational modifications in cancer progression. This review summarizes verified biomarker candidates successfully quantified by targeted proteomics in this field and directs the readers who plan to design their own hypothesis-driven experiments to appropriate sources of methods and knowledge.     

生态系统服务价值评估研究进展——国外学者观点

生态系统服务的价值评估是目前生态学研究热点和存在较多争议的领域之一。争论的焦点主要集中在生态系统服务货币化评估的必要性和有效性。通过对大量文献的检索,较为系统地总结了国外学者的观点,特别是引起诸多争论的主要原因：一是由于生态系统本身的复杂性,各项服务之间存在着相互依赖的关系,使生态系统服务的分类本身就缺乏严格的标准,同时存在着时间和空间上复杂的尺度转换问题,从而使生态系统服务的价值评估容易出现重复计算;二是由于经济学方法本身存在着一定的局限性,对于不同的评估对象和评估目标往往需要选取不同的评估方法,每一种生态系统服务通常可以有几种不同的评估方法,评估结果在很大的程度上依赖于选择的不同方法．从而使得到的结果间缺乏可比性;三是由于生态系统不同于经济系统。有时用经济学方法对生态系统进行评估时往往很难反映出自然系统的价值,特别是当人类对生态系统服务的偏好随着时间和新信息的出现而发生变化时,可能会出现更大的差异。今后的研究应当着力于上述困难的解决,以便使生态系统的评估结果更加客观可信,为生态补偿机制建立和生态系统管理等提供有力支撑。     

Culture of primary lung tumors using medium conditioned by a lung carcinoma cell line

Medium conditioned by the established lung tumor cell line A549 was used as a supplement to culture cells from primary solid lung tumors. Of 36 cases placed into culture, primary cells were obtained in 33 (91.7%). Of 29 cases in which subcultures were attempted, 18 (62.1%) were successful. Nine cell lines have been established by this technique to date. In growth assays, conditioned medium (CM) was found to stimulate both monolayer colony formation and growth in semi-solid medium of cells cultured from primary solid tumors. CM has been found to contain factors with the properties of both transforming growth factor alpha (TGF alpha) and insulin-like growth factor-I (IGF-I). The addition of a combination of these factors as purified peptides to basal medium at levels found in CM (0.1-0.5 ng/ml) stimulated colony formation of lung tumor cells by up to fourfold. These results indicate that secretion of growth factors may be important in tumor growth in vivo, and that use of CM may be a valuable tool for obtaining cultures from primary solid tumors.     

Nonlinear Dynamical Analysis of the Interdependence Between Central and Autonomic Nervous Systems in Neonates During Sleep

We present in this paper the results of a study of the interdependence between signal characteristic of the central nervous system (electroencephalography) and the autonomic nervous system (heart rate and respiration) in human neonates during sleep. By using methods from nonlinear dynamical systems theory, we show that there exist significant differences in this interdependence with the sleep stage and the electrodes considered. This paves the way for the application of this methodology in clinical practice to study pathologies where this interdependence is altered, such as the sudden infant death syndrome.     

Inhomogeneity during deflation of excised canine lungs. I. Alveolar pressures

Factors both intrinsic and extrinsic to the lung may cause inhomogeneity of alveolar pressures during deflation. Wilson et al. (J. Appl. Physiol. 59: 1924-1928, 1985) predicted that any such inhomogeneity would be limited by interdependence of regional expiratory flows. To test this hypothesis and to explore how the pleural pressure gradient might affect inhomogeneity of alveolar pressures, we deflated at submaximal flows excised canine lobes that first were suspended in air and then were immersed in foams that simulated the vertical gradient of pleural pressure. Interregional inhomogeneity of regional transpulmonary pressures was measured with use of an alveolar capsule technique. Flow-dependent inhomogeneity of alveolar pressures was present, with differences in alveolar pressure quickly relaxing to a constant limiting value at each flow. Foam immersion increased inhomogeneity at a given flow. We conclude that factors intrinsic to the lung cause significant inhomogeneity of alveolar pressures at submaximal expiratory flows and that this inhomogeneity is enhanced by the extrinsic gradient of pleural pressure. These observations are consistent with the interdependence of flow proposed by Wilson et al.     

Automated Measurement of Microcirculatory Blood Flow Velocity in Pulmonary Metastases of Rats

Because the lung is a major target organ of metastatic disease, animal models to study the physiology of pulmonary metastases are of great importance. However, very few methods exist to date to investigate lung metastases in a dynamic fashion at the microcirculatory level, due to the difficulty to access the lung with a microscope. Here, an intravital microscopy method is presented to functionally image and quantify the microcirculation of superficial pulmonary metastases in rats, using a closed-chest pulmonary window and automated analysis of blood flow velocity and direction. The utility of this method is demonstrated to measure increases in blood flow velocity in response to pharmacological intervention, and to image the well-known tortuous vasculature of solid tumors. This is the first demonstration of intravital microscopy on pulmonary metastases in a closed-chest model. Because of its minimized invasiveness, as well as due to its relative ease and practicality, this technology has the potential to experience widespread use in laboratories that specialize on pulmonary tumor research.     

Polyacrylates of noble metals as potential antitumor drugs

The antitumor activity of polyacrylates of the noble metals containing argentum (argacryl), aurum (auracryl) and platinum (platacryl) has been studied using experimental murine solid tumor models (Lewis lung carcinoma and Acatol adenocarcinoma). It has been found that polyacrylates of the noble metals are capable of inhibiting tumor development by 50–90% compared to control. Auracryl that inhibits the growth of Lewis lung carcinoma and Acatol adenocarcinoma by 80 and 90%, respectively, compared to control is the most efficient among the tested compounds and can be recommended for the further profound preclinical studies.     

Airway stability and heterogeneity in the constricted lung.

The effect of bronchoconstriction on airway resistance is known to be spatially heterogeneous and dependent on tidal volume. We present a model of a single terminal airway that explains these features. The model describes a feedback between flow and airway resistance mediated by parenchymal interdependence and the mechanics of activated smooth muscle. The pressure-tidal volume relationship for a constricted terminal airway is computed and shown to be sigmoidal. Constricted terminal airways are predicted to have two stable states: one effectively open and one nearly closed. We argue that the heterogeneity of whole lung constriction is a consequence of this behavior. Airways are partitioned between the two states to accommodate total flow, and changes in tidal volume and end-expiratory pressure affect the number of airways in each state. Quantitative predictions for whole lung resistance and elastance agree with data from previously published studies on lung impedance.     

Heterogeneity of maximal lobar emptying rates in dogs with compensatory lung growth

Five dogs underwent left pneumonectomy at 10 wk of age, whereas four littermates underwent a sham operation. At 26 wk of age the postpneumonectomy dogs had total lung vital capacity (VC) and lung weight similar to controls, but maximum expiratory flow was reduced. Pressure capsules were glued to right lower (RLL) and right cardiac (RCL) lobes, and alveolar pressures (PA) were measured during forced expiration. In postpneumonectomy dogs RLL and RCL both emptied more slowly than in control dogs, and emptying was especially delayed in RCL, which underwent the most growth. When both lobes deflated together, PA in RCL and RLL were similar in control dogs, but in postpneumonectomy dogs PA in RCL exceeded that in RLL by approximately 3 cmH2O from 80 to 20% VC. Because the higher driving pressure in RCL compensated for the relatively high resistance of RCL, the pattern of lobar emptying was relatively uniform over these lung volumes. This result was compatible with interdependence of lobar maximum expiratory flows. In addition, at PA of 6-10 cmH2O in postpneumonectomy dogs, maximum emptying rates of RCL were less when RCL deflated alone than when RCL and RLL emptied together, again demonstrating interdependence of lobar maximum expiratory flow.     

Models of dyadic social interaction

We discuss the logic of research designs for dyadic interaction and present statistical models with parameters that are tied to psychologically relevant constructs. Building on Karl Pearson's classic nineteenth-century statistical analysis of within-organism similarity, we describe several approaches to indexing dyadic interdependence and provide graphical methods for visualizing dyadic data. We also describe several statistical and conceptual solutions to the 'levels of analytic' problem in analysing dyadic data. These analytic strategies allow the researcher to examine and measure psychological questions of interdependence and social influence. We provide illustrative data from casually interacting and romantic dyads.