A hypothesis on the mechanism of trauma of lung tissue subjected to impact load

When a compressive impact load is applied on the chest, as in automobile crash or bomb explosion, the lung may be injured and show evidences of edema and hemorrhage. Since soft tissues have good strength in compression, why does a compression wave cause edema? Our hypothesis is that tensile and shear stresses are induced in the alveolar wall on rebound from compression, and that the maximum principal stress (tensile) may exceed critical values for increased permeability of the epithelium to small solutes, or even fracture. Furthermore, small airways may collapse and trap gas in alveoli at a critical strain, causing traumatic atelectasis. The collapsed airways reopen at a higher strain after the wave passes, during which the expansion of the trapped gas will induce additional tension in the alveolar wall. To test this hypothesis, we made three new experiments: (1), measuring the effect of transient overstretch of the alveolar membrane on the rate of lung weight increase; (2) determining the critical pressure for reopening collapsed airways of rabbit lung subjected to cyclic compression and expansion; (3) cyclic compression of lung with trachea closed. We found that in isolated rabbit lung overstretching increases the rate of edema fluid formation, that the critical strain for airway reopening is higher than that for closing, and that these critical strains are strain-rate dependent, but independent of the state of the trachea, whether it is open or closed. Furthermore, a theoretical analysis is presented to show that the maximum principal (tensile) stress is of the same order of magnitude as the maximum initial compressive stress at certain localities of the lung.(ABSTRACT TRUNCATED AT 250 WORDS)     

Functional cues in the development of osseous tooth support in the pig, Sus scrofa

Alveolar bone supports teeth during chewing through a ligamentous interface with tooth roots. Although tooth loads are presumed to direct the development and adaptation of these tissues, strain distribution in the alveolar bone at different stages of tooth eruption and periodontal development is unknown. This study investigates the biomechanical effects of tooth loading on developing alveolar bone as a tooth erupts into occlusion. Mandibular segments from miniature pigs, Sus scrofa, containing M₁ either erupting or in functional occlusion, were loaded in compression. Simultaneous recordings were made from rosette strain gages affixed to the lingual alveolar bone and the M₂ crypt. Overall, specimens with erupting M₁s were more deformable than specimens with occluding M₁s (mean stiffness of 246 vs. 944 MPa, respectively, p=0.004). The major difference in alveolar strain between the two stages was in orientation. The vertically applied compressive loads were more directly reflected in the alveolar bone strains of erupting M₁s, than those of occluding M₁s, presumably because of the mediation of a more mature periodontal ligament (PDL) in the latter. The PDL interface between occluding teeth and alveolar bone is likely to stiffen the system, allowing transmission of occlusal loads. Alveolar strains may provide a stimulus for bone growth in the alveolar process and crest.     

Quantitative measurements of released amines from individual exocytosis events

Chemical analysis of single cells is an area of great interest in the biological sciences. Single-cell systems are being utilized as a model to understand in vivo processes better. One method that is moving to the forefront in cellular analysis is electrochemistry. Owing to their rapid response time and small dimensions, voltammetric microelectrode techniques, such as amperometry and fast-scan voltammetry, have made it possible to monitor minute amounts of biological compounds and transiently occurring chemical events in cellular systems. The application of these methods to the quantitation of individual vesicular release events from single cells is overviewed here. The application of electrochemical monitoring to several types of cultured cells, including bovine adrenal chromaffin cells, rat pheochromocytoma (PC12) cells, beige mouse mast cells, superior cervical ganglion neurons, and human pancreatic β-cells, as well as to the invertebrate systems, the leechHirudo medicinalis, and pond snailPlanorbis corneus has provided a wealth of new information concerning exocytosis. Results obtained from the studies highlight the potential of electrochemical techniques in cellular analysis to contribute to our understanding of molecular and pharmacological effects on exocytosis. This article overviews work done on all the above cell types with an emphasis on PC12 cells.     

The development and in vitro characterisation of an intracellular nanosensor responsive to reactive oxygen species

Advances in sensor technologies have enhanced our understanding of the roles played by reactive oxygen species (ROS) in a number of physiological and pathological processes. However, high inter-reactivity and short life spans has made real-time monitoring of ROS in cellular systems challenging. Fluorescent dyes capable of intracellular ROS measurements have been reported. However, these dyes are known to be intrinsically cytotoxic and thus can potentially significantly alter cellular metabolism and adversely influence in vitro data. Reported here is the development and in vitro application of a novel ROS responsive nanosensor, based on PEBBLE (Probes Encapsulated By Biologically Localised Embedding) technology. The ROS sensitive fluorescent probe dihydrorhodamine 123 (DHR 123) was employed as the sensing element of the PEBBLE through entrapment within a porous, bio-inert polyacrylamide nanostructure enabling passive monitoring of free radical flux within the intracellular environment. Successful delivery of the nanosensors into NR8383 rat alveolar macrophage cells via phagocytosis was achieved. Stimulation of PEBBLE loaded NR8383 cells with phorbol-12-myristate-13-acetate (PMA) enabled real time monitoring of ROS generation within the cell without affecting cellular viability. These data suggest that PEBBLE nanosensors could offer significant advantages over existing technologies used in monitoring the intracellular environment.     

Radiographic evaluation of alveolar ridge heights of dentate and edentulous patients

doi: 10.1111/j.1741‐2358.2010.00391.x Radiographic evaluation of alveolar ridge heights of dentate and edentulous patients Objective: To evaluate the reduction of residual alveolar ridge height on panoramic radiographs and the differences between denture wearers and non‐denture wearers. Materials and methods: The study consisted of 147 individuals (74 men and 73 women) [50 were denture wearers and 50 non‐denture wearers (examination groups) and 47 of them were dentate (control group)]. Individuals having diseases impacting on bone were excluded. Vertical measurements were made at 15 sites (central incisors, first premolars and molars at the left and right of both jaws and the distance between the zygoma/orbit). MANOVA (multi‐variate analysis of variation) was used for the statistical analysis of the results. Results: There were significant differences between the alveolar ridge heights of dentate and edentulous groups (p < 0.001). Between the denture wearer and the non‐denture wearer groups, there was significant difference in the lower jaw (p < 0.001), but no significant difference in the upper jaw (p = 0.635). There were also differences between men and women (p < 0.005) and upper and lower jaws at every measurement sites (p < 0.01). Conclusion: Reduction in residual alveolar ridge height was in close relation with gender, denture usage and edentulousness.     

A novel mouse Cre‐driver line targeting Perilipin 2‐expressing cells in the neonatal lung

Pulmonary diseases such as chronic obstructive pulmonary disease, lung fibrosis, and bronchopulmonary dysplasia are characterized by the destruction or malformation of the alveolar regions of the lung. The underlying pathomechanisms at play are an area of intense interest since these mechanisms may reveal pathways suitable for interventions to drive reparative processes. Lipid‐laden fibroblasts (lipofibroblasts) express the Perilipin 2 (Plin2) gene‐product, PLIN2, commonly called adipose‐differentiation related protein (ADRP). These cells are also thought to play a role in alveolarization and repair after injury to the alveolus. Progress in defining the functional contribution of lipofibroblasts to alveolar generation and repair is hampered by a lack of in vivo tools. The present study reports the generation of an inducible mouse Cre‐driver line to target cells of the ADRP lineage. Robust Cre‐mediated recombination in this mouse line was detected in mesenchymal cells of the postnatal lung, and in additional organs including the heart, liver, and spleen. The generation and validation of this valuable new tool to genetically target, manipulate, and trace cells of the ADRP lineage is critical for assessing the functional contribution of lipofibroblasts to lung development and repair.     

Microbial systems for study of the biotransformations of drugs

The metabolic fate of drugs and other xenobiotics in mammalian organisms represents an area of intense contemporary interest. Traditionally, it is a difficult area of research becausethe biological systems which are used to study biotransformations are capable of yielding only minute quantities of metabolites. Recent developments in comparative biochemistry have made itpossible to link diverse metabolic systems through similarities in the pathways by which they alter foreign organic compounds. The potential thus exists for utilizing microbial metabolic systems to study and possibly predict the metabolic fate of a drug or other foreign compound in mammals. The ease with which microbial systems may be used to obtain large amounts of metabolites is an obvious Advantage. We havhe attemped to review the ways in which mammalian and microbialorganisms metabolize a variety of organic compounds. Attention has been focused on the similarities and differences in the mechanisms by which these living systems metabolize xenobiotics. Particular emphasis has been given to four types of reactions which are important in drug biotransformations: aromatic hydroxylationl; N- and O-dealkylations; and sulfur oxygenations.     

Criteria for the emergence and evolution of life in the solar system

During the past years we have explored most of the bodies of the solar system by means of the Apollo, Venera, Viking, Voyager, and other space missions. We are now in a better position to be able to compare the conditions of other planets and satellites with those of the Earth in order to determine what is unique about our planet which permitted the emergence and evolution of life on it. On the basis of this and other available scientific information we have arrived at the conclusion that there are at least some twentyfive specific conditions or requirements which have to be fulfilled in order for life as we know it to appear and evolve in a planetary system such as ours. Most of these necessary conditions or requirements are mutually interdependent, but in order to discuss their role in depth they have been divided into five major general areas which are discussed in some detail herein. Planetary criteria, which relate to the physical properties of the planet as it is formed and as it becomes a differentiated cosmic body and potential abode of life. The mass, orbital characteristics and energetic relationships with the central star as well as the discrete separation of gas, liquid and solid phases of the planet are of utmost importance. Chemical criteria, which are concerned with the composition, availability of effective energy sources, and chemical constrainst (solvent, pH range, redox potential) of the environment(s) where reactions take place for the prebiological formation of biochemical compounds. Protobiological criteria, which relate to the prebiologically synthesized oligomeric and polymeric biomolecules, how they interact cooperatively to form protobiological structures and functions (replication, catalysis, information transfer, etc.) and self-assemble to give rise to a living system. Evolutionary criteria, which are concerned with the processes responsible for the increase in complexity of organisms by genomic multiplication, symbiotic integration and cellular differentiation, as well as with the negentropic ability of organisms to continuously recycle all the volatile biogenic elements. Altogether these processes made possible the development and evolution of life from the simplest prokaryotic cell ancestor to a cognitive and manipulative multicellular organism (man).In order to extend this inquiry to other systems beyond our solar system a fifth set of requirements based on astronomical observations is also discussed, namely, theStellar criteria, which relate to the elemental composition mass, lifetime, and other features of Main Sequence stars which may be surrounded by planetary systems similar to our own. Finally, a brief review is made on the probability of the existence of extraterrestrial life as well as of civilizations capable of interstellar communication in our Galaxy.Paper presented at the 6th College Park Colloquium, October 1981.     

Colonisation of experimentally immersed wood in south eastern Australia: responses of feeding groups to changes in riparian vegetation

We investigated macroinvertebrate abundance and functional feeding groups colonising experimentally-positioned woody substrates of different species in streams with three different riparian vegetation types. Native Eucalyptus forest formed a dense closed canopy over our streams; introduced (exotic, alien) pine plantation forest did not fully shade the streams, and grassland streams were completely open, although with woody riparian vegetation well upstream of our sites. Macroinvertebrate assemblages varied taxonomically and functionally with both wood species and riparian vegetation composition. Two specialist feeding groups responded clearly to riparian vegetation: wood gougers were most common in forested streams, and algal grazers in more open streams. Gougers colonised native Eucalyptus wood in preference to alien species. Other feeding groups responses showed complex interactions between vegetation and wood type. Our results indicate the importance of sampling appropriate substrates when assessing questions of this type – if seeking shifts in functional organisation, the substrates on which the feeding groups of interest occur must be sampled. The composition of the riparian strip may influence xylophilous communities as much as the structure (i.e. whether closed or open).     

A strategy for determining arterial blood gases on the summit of Mt. Everest

Background  

Climbers on the summit of Mt. Everest are exposed to extreme hypoxia, and the physiological implications are of great interest. Inferences have been made from alveolar gas samples collected on the summit, but arterial blood samples would give critical information. We propose a plan to insert an arterial catheter at an altitude of 8000 m, take blood samples above this using an automatic sampler, store the samples in glass syringes in an ice-water slurry, and analyze them lower on the mountain 4 to 6 hours later.     

Cellular dynamics of conjugation in the ciliate euplotes aediculatus. II. Cellular membranes

The formation and subsequent dissolution of a common bridge of cytoplasm between conjugating ciliated protozoan cells provides an excellent opportunity to follow the dynamics of the cellular membrane systems involved in this process. In particular, separation of conjugant partners offers the chance to observe, at a fixed site on the cell surface, how the ciliate surface complex of plasma and alveolar membranes (collectively termed the “pellicle”) is constructed. Consequently, cortical and cellular membranes of Euplotes aediculatus were studied by light and electron microscopy through the conjugation sequence. A conjugant fusion zone of shared cytoplasm elaborates between the partner cells within their respective oral fields (peristomes) to include microtubules, cytosol, and a concentrated endoplasmic reticulum (heavily stained by osmium impregnation techniques) that may also be continuous with cortical ER of each cell. Cortical membranes displacd by fusion are autolyzed in acid phosphatase-positive lysosomes in the fusion zone. As conjugants separate, expansion of the plasma membrane may occur through the fusion of vesicles with the plasma membrane, presumably at bare membrane, presumably at bare membrane patches near the fusion zone. The underlying cortical alveolar membranes and their plate-like contents are reconstructed beneath the plasma membrane, apparently by multiple fusions of dense-cored alveolar precursor vesicles (APVs). These precursor vesicles themselves appear to condense directly from the smooth ER present in the fusion zone. No Golgi apparatus was visible in the fusion zone cytoplasm, and no step of APV maturation that might involve the Golgi complex was noted.     

In vitro study of gas effects on alveolar macrophages

Summary To evaluate the biological effects of gas pollutants on alveolar macrophages several in vitro systems ave been developed. We described here an original method of cell culture in aerobiosis, which permitted direct contact between the atmosphere and the target cells. We studied the long term (24 h) and short term (30 min) effects of NO₂ on alveolar macrophages. Our results demonstrated that exposure of alveolar macrophages to gas pollutants may be responsible for either cell injury or cell activation associated with the release of various bioactive mediators (superoxide anion, neutrophil chemotactic activity). Cell culture in aerobiosis opens new ways for the research on the biological effects of gas pollutants.Abbreviations AM alveolar macrophages - CL Chemiluminescence     

A Derivative Matching Approach to Moment Closure for the Stochastic Logistic Model

Continuous-time birth-death Markov processes serve as useful models in population biology. When the birth-death rates are nonlinear, the time evolution of the first n order moments of the population is not closed, in the sense that it depends on moments of order higher than n. For analysis purposes, the time evolution of the first n order moments is often made to be closed by approximating these higher order moments as a nonlinear function of moments up to order n, which we refer to as the moment closure function. In this paper, a systematic procedure for constructing moment closure functions of arbitrary order is presented for the stochastic logistic model. We obtain the moment closure function by first assuming a certain separable form for it, and then matching time derivatives of the exact (not closed) moment equations with that of the approximate (closed) equations for some initial time and set of initial conditions. The separable structure ensures that the steady-state solutions for the approximate equations are unique, real and positive, while the derivative matching guarantees a good approximation, at least locally in time. Explicit formulas to construct these moment closure functions for arbitrary order of truncation n are provided with higher values of n leading to better approximations of the actual moment dynamics. A host of other moment closure functions previously proposed in the literature are also investigated. Among these we show that only the ones that achieve derivative matching provide a close approximation to the exact solution. Moreover, we improve the accuracy of several previously proposed moment closure functions by forcing derivative matching.     

Fish communities in temporarily open/closed estuaries from the warm- and cool-temperate regions of South Africa: A review

The majority of estuaries along the coastline of southern Africa are termed temporarily open/closed estuaries (TOCEs) and are closed off from the sea for varying periods by a sandbar which forms at the mouth. It is therefore important to understand the processes occurring within TOCEs and their importance to fishes in order to make sound management recommendations. Estuaries along the coast of South Africa and their associated fish assemblages are biogeographically distinct and occur in either a subtropical, warm-temperate or cool-temperate zone. There are 125 TOCEs found within the cool-temperate and warm-temperate zones. Most fish species found in TOCEs are the juveniles of marine taxa that breed at sea. Permanently open estuaries generally have a higher diversity of species than TOCEs, but TOCEs still provide important nursery areas for many marine species and numerically often have a higher proportion of estuarine resident species. Important taxa in terms of abundance and biomass in warm-temperate TOCEs include the sparids Rhabdosargus holubi and Lithognathus lithognathus, several mugilid species, estuarine residents (particularly Gilchristella aestuaria and Atherina breviceps) and the freshwater cichlid Oreochromis mossambicus. The diversity of fishes in cool-temperate TOCEs is low when compared with warm-temperate systems and Liza richardsonii tends to dominate catches by number and mass in most systems. Several species recorded in TOCEs show clear longitudinal distribution trends. For example Atherina breviceps is generally more abundant in the lower reaches of estuaries. Mouth state, particularly the frequency, timing and duration of mouth opening plays a key role in determining species richness, composition, diversity and abundance in TOCEs. Mouth state is directly linked to freshwater input. Reduced river inflow leads to prolonged mouth closure and shorter open phases, which inhibits immigration and emigration of marine fish species between estuaries and the sea. Understanding of the effects of various processes occurring within these systems, particularly variation in freshwater input, on the biota of these important systems facilitates the development of informed management recommendations.     

Electron microscopic study of the open circulatory system of the shrimp,Caridina japonica. I. Gill capillaries

The ultrastructure of the phyllobranchiate type gill of the shrimp, Caridina japonica, was studied. The most characteristic feature of the open circulatory system of Cardina is the vascular lumen of the gill capillaries which is considered to be the interstitial space. The following observations substantiate this view: (1) a thin fibrous layer forms the innermost structure of the walls of gill capillaries and is in direct contact with the blood stream; (2) filaments in the fibrous layer are assumed to correspond to the reticular fibers in the interstitial space of the alveolar wall of mammals; (3) the absence of the endothelium as well as the endothelial basal lamina which are the essential structural components of the closed circulatory system in vertebrates. The gill epithelium contains intermediate, septate and tight junctions. The first two form a junctional complex near the apical cell border and may function as a permeability barrier by occluding the intercellular space as well as functioning in electrical coupling and cellular adhesion. The tight junction is spot-like and may serve no role in the function of the permeability barrier.     

Quantifying the strength of the linear causal coupling in closed loop interacting cardiovascular variability signals

 The coherence function measures the amount of correlation between two signals x and y as a function of the frequency, independently of their causal relationships. Therefore, the coherence function is not useful in deciding whether an open-loop relationship between x and y is set (x acts on y, but the reverse relationship is prevented) or x and y interact in a closed loop (x affects y, and vice versa). This study proposes a method based on a bivariate autoregressive model to derive the strength of the causal coupling on both arms of a closed loop. The method exploits the definition of causal coherence. After the closed-loop identification of the model coefficients, the causal coherence is calculated by switching off separately the feedback or the feedforward path, thus opening the closed loop and fixing causality. The method was tested in simulations and applied to evaluate the degree of the causal coupling between two variables known to interact in a closed loop mainly at a low frequency (LF, around 0.1 Hz) and at a high frequency (HF, at the respiratory rate): the heart period (RR interval) and systolic arterial pressure (SAP). In dogs at control, the RR interval and the SAP are highly correlated at HF. This coupling occurs in the causal direction from the RR interval to the SAP (the mechanical path), while the coupling on the reverse causal direction (the baroreflex path) is not significant, thus pointing out the importance of the direct effects of respiration on the RR interval. Total baroreceptive denervation, by opening the closed loop at the level of the influences of SAP on RR interval, does not change these results. In elderly healthy men at rest, the RR interval and SAP are highly correlated at the LF and the HF. At the HF, a significant coupling in both causal directions is found, even though closed-loop interactions are detected in few cases. At the LF, the link on the baroreflex pathway is negligible with respect to that on the reverse mechanical one. In heart transplant recipients, in which SAP variations do not cause RR interval changes as a result of the cardiac denervation, the method correctly detects a significant coupling only on the pathway from the RR interval to the SAP. Received: 28 June 2001 / Accepted in revised form: 23 October 2001     

MADS-box out of the black box

The compelling elegance of using genome‐wide scans to detect the signature of selection is difficult to resist, but is countered by the low demonstrated efficacy of pinpointing the actual genes and traits that are the targets of selection in nonmodel species. While the difficulty of going from a suggestive signature to a functional nucleotide polymorphism should not prevent researchers from using genome scans, it does lessen their long‐term utility within and across study systems. In a new study published in this issue of Molecular Ecology ( Mariac et al. 2011 ), researchers have gone a long way towards increasing the relevance of genome‐wide scans for selection via two approaches: (i) they tailored the markers used in the scan to target a family of developmental genes that were good candidates for controlling a trait of interest and (ii) they used an independent mapping population to confirm the association of the gene with polymorphism in the trait of interest. All of this was completed in the nonmodel system of pearl millet (Pennisetum glaucum) and may provide a road map for other researchers hoping to pin down solid candidate genes for selected traits in natural or cultivated systems. Outside of these broad methodological innovations, the paper specifically focuses on a trait (flowering time) that varies across an environmental gradient (rainfall). This environmental gradient potentially serves as a model for environmental change over time, and allele frequencies at the gene can therefore be used to track how populations of pearl millet will adapt to future climate shifts at the genetic level.     

Poisson-Boltzmann model studies of molecular electrostatic properties of the cAMP-dependent protein kinase

Protonation equilibria of residues important in the catalytic mechanism of a protein kinase were analyzed on the basis of the Poisson-Boltzmann electrostatic model along with a cluster-based treatment of the multiple titration state problem. Calculations were based upon crystallographic structures of the mammalian cAMP-dependent protein kinase, one representing the so called closed form of the enzyme and the other representing an open conformation. It was predicted that at pH 7 the preferred form of the phosphate group at the catalytically essential threonine 197 (P-Thr197) in the closed form is dianionic, whereas in the open form a monoanionic ionization state is preferred. This dianionic state of P-Thr197, in the closed form, is stabilized by interactions with ionizable residues His87, Arg165, and Lys189. Our calculations predict that the hydroxyl of the Ser residue in the peptide substrate is very difficult to ionize, both in the closed and open structures of the complex. Also, the supposed catalytic base, Asp166, does not seem to have a pK _a appropriate to remove the hydroxyl group proton of the peptide substrate. However, when Ser of the peptide substrate is forced to remain ionized, the predicted pK _a of Asp166 increases strongly, which suggests that the Asp residue is a likely candidate to attract the proton if the Ser residue becomes deprotonated, possibly during some structural change preceding formation of the transition state. Finally, in accord with suggestions made on the basis of the pH-dependence of kinase kinetics, our calculations predict that Glu230 and His87 are the residues responsible for the molecular pK _a values of 6.2 and 8.5, observed in the experiment. Received: 19 October 1998 / Revised version: 12 April 1999 / Accepted: 15 April 1999     

Correlative Nanoscale 3D Imaging of Structure and Composition in Extended Objects

Structure and composition at the nanoscale determine the behavior of biological systems and engineered materials. The drive to understand and control this behavior has placed strong demands on developing methods for high resolution imaging. In general, the improvement of three-dimensional (3D) resolution is accomplished by tightening constraints: reduced manageable specimen sizes, decreasing analyzable volumes, degrading contrasts, and increasing sample preparation efforts. Aiming to overcome these limitations, we present a non-destructive and multiple-contrast imaging technique, using principles of X-ray laminography, thus generalizing tomography towards laterally extended objects. We retain advantages that are usually restricted to 2D microscopic imaging, such as scanning of large areas and subsequent zooming-in towards a region of interest at the highest possible resolution. Our technique permits correlating the 3D structure and the elemental distribution yielding a high sensitivity to variations of the electron density via coherent imaging and to local trace element quantification through X-ray fluorescence. We demonstrate the method by imaging a lithographic nanostructure and an aluminum alloy. Analyzing a biological system, we visualize in lung tissue the subcellular response to toxic stress after exposure to nanotubes. We show that most of the nanotubes are trapped inside alveolar macrophages, while a small portion of the nanotubes has crossed the barrier to the cellular space of the alveolar wall. In general, our method is non-destructive and can be combined with different sample environmental or loading conditions. We therefore anticipate that correlative X-ray nano-laminography will enable a variety of in situ and in operando 3D studies.