Human tear fluid lipidome: from composition to function

We have explored human aqueous tear fluid lipidome with an emphasis to identify the major lipids. We also address the physiological significance of the lipidome. The tears were analysed using thin layer chromatographic, enzymatic and mass spectrometric techniques. To emphasize the physiological aspect of the lipidome, we modelled the spreading of the non-polar tear fluid lipids at air-water interface in macroscopic scale with olive oil and egg yolk phosphatidylcholine. Based on enzymatic analysis the respective concentrations of choline-containing lipids, triglycerides, and cholesteryl esters were 48±14, 10±0, and 21±18 μM. Ultra performance liquid chromatography quadrupole time of flight mass spectrometry analysis showed that phosphatidylcholine and phosphatidylethanolamine were the two most common polar lipids comprising 88±6% of all identified lipids. Triglycerides were the only non-polar lipids detected in mass spectrometric analysis i.e. no cholesteryl or wax esters were identified. The spreading experiments show that the presence of polar lipids is an absolute necessity for a proper spreading of non-polar tear fluid lipids. We provide evidence that polar lipids are the most common lipid species. Furthermore, we provide a physiological rationale for the observed lipid composition. The results open insights into the functional role of lipids in the tear fluid and also aids in providing new means to understand and treat diseases of the ocular surface.     

Differential role of PI3K/Akt pathway in the infarct size limitation and antiarrhythmic protection in the rat heart

Endogenous cardiac protection against prolonged ischemic insult can be achieved by repeated brief episodes of ischemia (hypoxia) or by cardiac adaptation to various stresses such as chronic hypoxia. Activation of phosphatidylinositol 3-kinase (PI3K)/Akt is involved in antiapoptotic effects, however, it is not clear whether it is required for overall heart salvage including protection against myocardial infarction and arrhythmias. We focussed on the potential common role of PI3K/Akt in anti-infarct protection, in the experimental settings of long-term adaptation to chronic intermittent hypobaric hypoxia (IHH; 8 h/day, 25–30 exposures, in vivo rats) and acute ischemic preconditioning (IP; Langendorff-perfused hearts). In addition, we explored the role of PI3K/Akt in susceptibility to ischemic ventricular arrhythmias. In normoxic open-chest rats, PI3K/Akt inhibitor LY294002 (LY; 0.3 mg/kg) given 5 min before test occlusion/reperfusion (I/R) did not affect infarct size (IS) normalized to the size of area at risk (AR). In hypoxic rats, LY partially attenuated IS-limiting effect of IHH (IS/AR 59.7 ± 4.1% vs. 51.8 ± 4.4% in the non-treated rats; p > 0.05) and increased IS/AR to its value in normoxic rats (64.9 ± 5.1%). In the isolated hearts, LY (5 μM) applied 15 min prior to I/R completely abolished anti-infarct protection by IP (IS/AR 55.0 ± 4.9% vs. 15.2 ± 1.2% in the non-treated hearts and 42.0 ± 5.5% in the non-preconditioned controls; p < 0.05). In the non-preconditioned hearts, PI3K/Akt inhibition did not modify IS/AR, on the other hand, it markedly suppressed arrhythmias. In the LY-treated isolated hearts, the total number of ventricular premature beats and the incidence of ventricular tachycardia (VT) was reduced from 518 ± 71 and 100% in the controls to 155 ± 15 and 12.5%, respectively (p < 0.05). Moreover, bracketing of IP with LY did not reverse antiarrhythmic effect of IP. These results suggest that activation of PI3K/Akt cascade plays a role in the IS-limiting mechanism in the rat heart, however, it is not involved in the mechanisms of antiarrhythmic protection.     

Rapid growth and large body size in annual fish populations are compromised by density-dependent regulation

We tested the effect of population density on maximum body size in three sympatric species of annual killifishes Nothobranchius spp. from African ephemeral pools. We found a clear negative effect of population density on body size, limiting their capacity for extremely fast development and rapid growth. This suggests that density-dependent population regulation and the ephemeral character of their habitat impose contrasting selective pressures on the life history of annual killifishes.     

On the origin of eukaryotic cells and their endomembranes.

A novel hypothesis for the origin of eukaryotic cells is presented. It is assumed that the universal ancestor was bounded by two membranes of heterochiral lipid composition. We propose that the prokaryotic cells (the hypothetical host entity for alpha proteic-bacteria), though sharing a common ancestor with Archaea, was bounded by two membranes. The hypothesis suggests that an alpha proteic-bacterial symbiont was enclosed in the prokaryotic cells intermembrane space. In this view, the eukaryotic nuclear membrane and endomembrane system arose from the prokaryotic cells inner membrane while the eukaryotic plasma membrane arose from the prokaryotic cells outer membrane. The outlined scenario agrees with the view that engulfment of an alpha-proteic-bacterial cell by a host entity and its transformation to a mitochondrion was the driving force leading to the appearance of the first eukaryotic cell. The hypothesis seems to be consistent with the pre-cell theory, theory of membrane heredity, and the phagocytosis-late scenario.     

ENZO: a web tool for derivation and evaluation of kinetic models of enzyme catalyzed reactions

We describe a web tool ENZO (Enzyme Kinetics), a graphical interface for building kinetic models of enzyme catalyzed reactions. ENZO automatically generates the corresponding differential equations from a stipulated enzyme reaction scheme. These differential equations are processed by a numerical solver and a regression algorithm which fits the coefficients of differential equations to experimentally observed time course curves. ENZO allows rapid evaluation of rival reaction schemes and can be used for routine tests in enzyme kinetics. It is freely available as a web tool, at http://enzo.cmm.ki.si.     

Skiing efficiency versus performance in double-poling ergometry

This study is on how leg utilisation may affect skiing efficiency and performance in double-poling ergometry. Three experiments were conducted, each with a different style of the double-poling technique: traditional with small knee range-of-motion and fixed heels (TRAD); modern with large knee range-of-motion and fixed heels (MOD1) and modern with large knee range-of-motion and free heels (MOD2). For each style, motion data were extracted with automatic marker recognition of reflective markers and applied to a 3D full-body musculoskeletal simulation model. Skiing efficiency (skiing work divided by metabolic muscle work) and performance (forward impulse) were computed from the simulation output. Skiing efficiency was 4.5%, 4.1% and 4.1% for TRAD, MOD1 and MOD2, respectively. Performance was 111, 143 and 149 Ns for TRAD, MOD1 and MOD2, respectively. Thus, higher lower body utilisation increased the performance but decreased the skiing efficiency. These results demonstrate the potential of musculoskeletal simulations for skiing efficiency estimations.