Studies of Epithelial Electrolyte Transport by Marker Ions

The paper reviews several recent studies in which marker ions, such as Rb and Br, were used to identify ion transport pathways and membrane properties in epithelia. In the frog skin epithelium, using Rb as a substitute for K, Cl transport mechanisms across the basolateral membranes of principal cells were studied. The data suggest that intracellular Cl is maintained above electrochemical equilibrium by an Na-K-2Cl cotransport system which, under non-stimulated conditions, is normally quiescent. In toad and frog skins, the route of transepithelial Cl movement was investigated. A subpopulation of mitochondria-rich cells demonstrated a ready exchange of Br with the apical and basal bathing media, consistent with the view that these cells constitute a transcellular anion shunt. Moreover, voltage-activation resulted in an increased Br uptake from the apical bath. Nevertheless, because of the very small number of these cells, it may be questioned whether the mitochondria-rich cell constitutes the only shuntpathway for Cl. In other studies, Rb uptake was employed to measure the Na/K.-pump activity. In principal cells of the frog skin epithelium, amiloride inhibited Rb uptake and lowered Na concentration, supporting the view that this cell type is engaged in amiloride-sensitive Na transport. In contrast, no significant changes in the Rb, Na, and Cl concentration of mitochondria-rich cells were detectable. Studies with Rb as marker ion in the rabbit urinary bladder revealed that the epithelium behaves like a functional syncytium with regard to transepithelial ion transport

Identification of Ion Transport Compartments in Turtle Urinary Bladder

To identify the turtle urinary bladder cells involved in Na and Cl absorption and Hand HCO3 secretion cellular electrolyte concentrations and uptake of Br and Solutrast were determined using electron microprobe analysis. Whereas inhibition of transepithelial Na transport by ouabain (reversion of short circuit current) led to a pronounced K-Na exchange in granular, and most of the basal cells, surface CA-cells and some basal cells were ouabain insensitive. Surface CA-cells could be divided into a large Cl-rich and a small Cl-poor population. Since the ouabain-induced K-Na exchange could be completely prevented by blocking passive luminal Na entry by amiloride, granular and most of the basal cells seem to form a syncytial Na transport compartment. Luminal uptake of Br only occurred in Cl-poor surface CA-cells, indicating the sole responsibility of these cells for electrogenic and electroneutral Cl absorption and HCO3 secretion. Serosal Br was taken up into all cell types. Whereas H secretion and serosal Br uptake into all cell types could be inhibited by 4-isothiocyano-4\u27-acetamido-2,2\u27-disulfonic stilbene (SITS), blockade of H secretion by lowering luminal pH to 4.5 diminished Br uptake only in Cl-rich surface CA-cells. Theses results indicate: a) Only Cl-rich surface CA-cells have a serosal anion exchanger involved in H secretion and b) granular and basal cells also possess a serosal anion exchanger, possibly responsible for cellular pH regulation. Luminal endocytosis of the I-containing Solutrast was observed in apical regions of Cl-rich surface CA-cells after inhibition of H secretion, but not under steady-state conditions, indicating a transport related but not a constitutive endo-exocytosis

Electron Microprobe Analysis of Electrolytes in Whole Cultured Epithelial Cells

Microprobe analysis was used to determine electrolyte contents in whole epithelial sheets of A6 cells and to investigate the most critical points of this method. Analysis of dextran standard sections of different thickness revealed that low accelerating voltages of about 10 kV are best suited for whole freeze-dried cells on thick supports, since 5 μm thick sections are not penetrated by 10 kV electrons. Washing of A6 cells for 10 sec with distilled water led to cell swelling of about 40%, but the molar concentration ratios and the concentrations per dry weight (dw) were not altered. Washing for 60 sec with distilled water caused a further increase in cell volume (120%) and loss of cellular K and Cl (90 mmol/kg dw). Washing with isotonic NH4-acetate led to a loss of cell Cl already after 10 sec. To characterize the Na transport compartment, A6 cells cultured on permeable supports were washed for 5 sec with distilled water, freeze-dried, and analyzed. Inhibition of transepithelial Na transport by ouabain increased Na/P from 0.15±0.07 to 0.75±0.03 and Cl/P from 0.21±0.001 to 0.38±0.003 while KIP decreased from 0.83±0.08 to 0.32±0.03. The changes in cell Na and K contents can be explained by K/Na exchange; the increase in Cl content indicates some cell swelling. Since the ouabain-induced changes could be prevented by apical amiloride, the apical membrane provides the most important pathway for Na entry in A6 cells

Effect of soil conditions and landscape factors on macro-snail communities in newly created grasslands of restored landfill sites in the UK

Though restored landfill sites provide habitat for a number of taxa, their potential for land snail remains unexplored. In this study, large-sized land snails (> 5 mm) were surveyed using transect sampling on nine restored landfill sites and nine corresponding nature sites in the East Midlands region of the UK during 2008. The effect of restoration was investigated by examining land snail species composition, richness, and diversity (Shannon Weiner index) in relation to habitat and landscape structure. Thirteen macro-snail species were found in total and rarefied species richness and diversity on restored landfill sites was not found to be statistically different to that of reference sites. One third of the snail species, comprising 30% of total abundance, found in the restored landfill sites were non-native species introduced to the UK. Soil electrical conductivity was the strongest predictor for richness and diversity of land snails. Road density was found to have a positive influence on snail species diversity. Given the high percentage of introduced species detected further research is needed in terms of the management implications of restored landfill sites and the dynamics of native versus non-native species

From 2D leg kinematics to 3D full-body biomechanics-the past, present and future of scientific analysis of maximal instep kick in soccer

Biomechanics investigation on soccer kicking has a relatively long history, yet the body of knowledge is still small. This paper reviews articles published from 1960s to 2011, summarizing relevant findings, research trends and method development. It also discusses challenges faced by the field. The main aim of the paper is to promote soccer kicking studies through discussions on problem solving in the past, method development in the present, and possible research directions for the future

Extravascular perivenous fibrin support leads to aneurysmal degeneration and intimal hyperplasia in arterialized vein grafts in the rat

Abstract Background and aims External support of vein grafts by fibrin glue possibly prevents overdistension, vascular remodeling, and neointimal hyperplasia. Previous animal models of neointimal hyperplasia showed conflicting results. Here, long-term effects of external fibrin glue support were studied in a new rat model of jugular vein to abdominal aorta transposition. Materials and methods and methods In male Wistar rats (250-300 g) right jugular vein (1.0-1.5 cm) was transposed to the infrarenal aorta. Fibrin glue (0.25 ml) covered the vein before releasing the vascular clamps (n=6). Control vein grafts were exposed directly to blood pressure. After 16 weeks vein grafts were pressure-fixed for histology

A novel application of motion analysis for detecting stress responses in embryos at different stages of development.

Motion analysis is one of the tools available to biologists to extract biologically relevant information from image datasets and has been applied to a diverse range of organisms. The application of motion analysis during early development presents a challenge, as embryos often exhibit complex, subtle and diverse movement patterns. A method of motion analysis able to holistically quantify complex embryonic movements could be a powerful tool for fields such as toxicology and developmental biology to investigate whole organism stress responses. Here we assessed whether motion analysis could be used to distinguish the effects of stressors on three early developmental stages of each of three species: (i) the zebrafish Danio rerio (stages 19 h, 21.5 h and 33 h exposed to 1.5% ethanol and a salinity of 5); (ii) the African clawed toad Xenopus laevis (stages 24, 32 and 34 exposed to a salinity of 20); and iii) the pond snail Radix balthica (stages E3, E4, E6, E9 and E11 exposed to salinities of 5, 10 and 15). Image sequences were analysed using Sparse Optic Flow and the resultant frame-to-frame motion parameters were analysed using Discrete Fourier Transform to quantify the distribution of energy at different frequencies. This spectral frequency dataset was then used to construct a Bray-Curtis similarity matrix and differences in movement patterns between embryos in this matrix were tested for using ANOSIM

Effects of rising temperature on pelagic biogeochemistry in mesocosm systems: a comparative analysis of the AQUASHIFT Kiel experiments

A comparative analysis of data, obtained during four indoor-mesocosm experiments with natural spring plankton communities from the Baltic Sea, was conducted to investigate whether biogeochemical cycling is affected by an increase in water temperature of up to 6 °C above present-day conditions. In all experiments, warming stimulated in particular heterotrophic bacterial processes and had an accelerating effect on the temporal development of phytoplankton blooms. This was also mirrored in the build-up and partitioning of organic matter between particulate and dissolved phases. Thus, warming increased both the magnitude and rate of dissolved organic carbon (DOC) build-up, whereas the accumulation of particulate organic carbon (POC) and phosphorus (POP) decreased with rising temperature. In concert, the observed temperature-mediated changes in biogeochemical components suggest strong shifts in the functioning of marine pelagic food webs and the ocean’s biological carbon pump, hence providing potential feedback mechanisms to Earth’s climate system

Influence of the postoperative inflammatory response on cognitive decline in elderly patients undergoing on-pump cardiac surgery: a controlled, prospective observational study

BACKGROUND: The role of non-infective inflammatory response (IR) in the aetiology of postoperative cognitive dysfunction (POCD) is still controversial. The aim of this controlled, prospective observational study was to assess the possible relationship between the grade of IR, defined by procalcitonin (PCT) changes, and development of POCD related to cardiac surgery. METHODS: Forty-two patients, who were >/= 60 years of age and scheduled for elective cardiac surgery, were separated into the low inflammatory (LIR) and high inflammatory (HIR) response groups based on their PCT levels measured on the first postoperative day. A matched normative control group of 32 subjects was recruited from primary care practice. The PCT and C-reactive protein (CRP) levels were monitored daily during the first five postoperative days. The cognitive function and mood state were preoperatively tested with a set of five neurocognitive tests and two mood inventories and at the seventh postoperative day. The Reliable Change Index modified for practice (RCIp) using data from normative controls was applied to determine the significant decline in test performance. RESULTS: The LIR (n = 20) and HIR (n = 22) groups differed significantly in the PCT (p 0.05). Additionally, there was no difference in the mood states, anxiety levels and perioperative parameters known to influence the development of POCD. CONCLUSIONS: In this study, the magnitude of the non-infective inflammatory response generated by on-pump cardiac surgery did not influence the development of POCD in the early postoperative period in elderly patients

Practical guidelines for rigor and reproducibility in preclinical and clinical studies on cardioprotection

The potential for ischemic preconditioning to reduce infarct size was first recognized more than 30 years ago. Despite extension of the concept to ischemic postconditioning and remote ischemic conditioning and literally thousands of experimental studies in various species and models which identified a multitude of signaling steps, so far there is only a single and very recent study, which has unequivocally translated cardioprotection to improved clinical outcome as the primary endpoint in patients. Many potential reasons for this disappointing lack of clinical translation of cardioprotection have been proposed, including lack of rigor and reproducibility in preclinical studies, and poor design and conduct of clinical trials. There is, however, universal agreement that robust preclinical data are a mandatory prerequisite to initiate a meaningful clinical trial. In this context, it is disconcerting that the CAESAR consortium (Consortium for preclinicAl assESsment of cARdioprotective therapies) in a highly standardized multi-center approach of preclinical studies identified only ischemic preconditioning, but not nitrite or sildenafil, when given as adjunct to reperfusion, to reduce infarct size. However, ischemic preconditioning—due to its very nature—can only be used in elective interventions, and not in acute myocardial infarction. Therefore, better strategies to identify robust and reproducible strategies of cardioprotection, which can subsequently be tested in clinical trials must be developed. We refer to the recent guidelines for experimental models of myocardial ischemia and infarction, and aim to provide now practical guidelines to ensure rigor and reproducibility in preclinical and clinical studies on cardioprotection. In line with the above guideline, we define rigor as standardized state-of-the-art design, conduct and reporting of a study, which is then a prerequisite for reproducibility, i.e. replication of results by another laboratory when performing exactly the same experiment