Lipidomic Analyses of Female Mice Lacking Hepatic Lipase and Endothelial Lipase Indicate Selective Modulation of Plasma Lipid Species

Hepatic lipase (HL) and endothelial lipase (EL) share overlapping and complementary roles in lipoprotein metabolism. The deletion of HL and EL alleles in mice raises plasma total cholesterol and phospholipid concentrations. However, the influence of HL and EL in vivo on individual molecular species from each class of lipid is not known. We hypothesized that the loss of HL, EL, or both in vivo may affect select molecular species from each class of lipids. To test this hypothesis, we performed lipidomic analyses on plasma and livers from fasted female wild-type, HL-knockout, EL-knockout, and HL/EL-double knockout mice. Overall, the loss of HL, EL, or both resulted in minimal changes to hepatic lipids; however, select species of CE were surprisingly reduced in the livers of mice only lacking EL. The loss of HL, EL, or both reduced the plasma concentrations for select molecular species of triacylglycerol, diacylglycerol, and free fatty acid. On the other hand, the loss of HL, EL, or both raised the plasma concentrations for select molecular species of phosphatidylcholine, cholesteryl ester, diacylglycerol, sphingomyelin, ceramide, plasmanylcholine, and plasmenylcholine. The increased plasma concentration of select ether phospholipids was evident in the absence of EL, thus suggesting that EL might exhibit a phospholipase A₂ activity. Using recombinant EL, we showed that it could hydrolyse the artificial phospholipase A₂ substrate 4-nitro-3-(octanoyloxy)benzoic acid. In summary, our study shows for the first time the influence of HL and EL on individual molecular species of several classes of lipids in vivo using lipidomic methods.     

Multifaceted Target Specificity Analysis as a Tool in Antimicrobial Drug Development: Type III Pantothenate Kinases as a Case Study

The research and development of a new antimicrobial drug using a target-based approach raises the question of whether any resulting hits will also show activity against the homologous target in other closely related organisms. While an assessment of the similarities of the predicted interactions between the identified inhibitor and the various targets is an obvious first step in answering this question, no clear and consistent framework has been proposed for how this should be done. Here we developed Multifaceted Target Specificity Analysis (MTSA) and applied it to type III pantothenate kinase (PanK_III) – an essential enzyme required for coenzyme A biosynthesis in a wide range of pathogenic bacteria – as a case study to establish if targeting a specific organism's PanK_III would lead to a narrow- or broad-spectrum agent. We propose that MTSA is a useful tool and aid for directing new target-based antimicrobial drug development initiatives.     

Isotopic disequilibrium during uptake of atmospheric CO2 into mine process waters: implications for CO2 sequestration

Dypingite, a hydrated Mg-carbonate mineral, was precipitated from high-pH, high salinity solutions to investigate controls on carbon fixation and to identify the isotopic characteristics of mineral sequestration in mine tailings. δ(13)C values of dissolved inorganic carbon content and synthetic dypingite are significantly more negative than those predicted for equilibrium exchange of CO(2) gas between the atmosphere and solution. The measured δ(13)C of aqueous carbonate species is consistent with a kinetic fractionation that results from a slow diffusion of atmospheric CO(2) into solution. During dypingite precipitation, dissolved inorganic carbon concentrations decrease and δ(13)C values become more negative, indicating that the rate of CO(2) uptake into solution was outpaced by the rate of carbon fixation within the precipitate. This implies that CO(2) gas uptake is rate-limiting to CO(2) fixation. δ(13)C of carbonate mineral precipitates in mine tailings and of DIC in mine process waters display similar (13)C-depletions that are inconsistent with equilibrium fractionation. Thus, the rate of carbon fixation in mine tailings may also be limited by supply of CO(2). Carbon sequestration could be accelerated by increasing the partial pressure of CO(2) in tailings ponds or by using chemicals that enhance the uptake of gaseous CO(2) into aqueous solution.     

Ultrastructural Variation in Beef M. longissimus dorsi as an Explanation of the Variation in Beef Tenderness

ABSTRACT: The objective of this experiment was to quantify variation in bovine M. longissimus dorsi tenderness and determine the extent such variation is explained by variation in the ultrastructure of muscles after different postmortem treatments. Eight muscles were hot-boned and aged for 2 d at 2 °C (T1) to achieve very contracted actomyosin crossover and tough beef. Eight carcass sides were tenderstretched for 10 h at 10 °C and a further 38 h at 2 °C (T2) to achieve lengthened actomyosin crossover and tender beef. Both T1 and T2 were compared with conventionally hung carcasses, which underwent similar chilling regimes, C1 ( n = 8) and C2 ( n = 8), respectively. Measurements of sarcomere length, pH, Warner Bratzler shear force (WBSF), and sensory tenderness were taken, and transmission electron microscopy images analyzed. Variances of attributes were analyzed on Bartlett's test. Variances of the 4 groups were homogenous for all attributes except for pH after 24 h postmortem (with T1 [0.00] having lower variances than C1 = 0.04, T2 = 0.06, and C2 = 0.05) and WBSF after 2 d aging (with T2 [74.33] having lower variances than T1 = 236.76, C1 = 398.82, and C2 = 856.74). The variation in the tenderness of beef was quantified through ultrastructural variation in bovine muscle, with tenderstretched moderately chilled beef having the least variable tenderness as a result of more uniform overlap between actin and myosin filaments. Variation in the eating quality of beef was not reduced by hot-boning with fast chilling or conventional hanging with fast or moderate chilling. The development of the uniformity within filaments of tenderstretched muscle requires further analysis as residual variation remains.     

Assessing How Much Couples Work at Their Relationship: The Behavioral Self-Regulation for Effective Relationships Scale.

It is widely believed that satisfying couple relationships require work by the partners. The authors equated the concept of work to relationship self-regulation and developed a scale to assess this construct. A factor analysis of the scale in a sample of 187 newlywed couples showed it comprised 2 factors of relationship strategies and effort. The factor structure was replicated in an independent sample of 97 newlywed couples. In both samples the scale had good internal consistency and high convergent validity between self- and partner-report forms. Self-regulation accounted for substantial variance in relationship satisfaction in both newlywed samples and in a 3rd sample of 61 long-married couples. The self-regulation and satisfaction association was independent of mood or self-report common method variance. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Impression cytology of a case of conjunctival metaplasia associated with oral carbamazepine use?

PURPOSE: To report a case of conjunctival squamous metaplasia associated with oral carbamazepine use. METHODS: Following completion of an ocular comfort questionnaire, biomicroscopy and a phenol red thread test, impression cytology from the inter-palpebral zone (nasal) of the bulbar conjunctival surface was undertaken using a Millcell-CM filter after topical anaesthesia with oxybuprocaine 0.4%. The filter was stained with Giemsa and colour images taken at 400x magnification by light microscopy. The images were graded and also a 35 mm was prepared. From the projected image, an overlay method was used to outline the borders such that the cell and nucleus area, and the longest and shortest dimensions could be measured by planimetry. RESULTS: A male subject, added 22 years, presented with slight conjunctival injection but no substantial symptoms and only slight surface staining with fluorescein. The subject reported use of oral carbamazepine (200mg, b.d.s.). Impression cytology showed large sheets of squamous cells (grade 2-3) with few goblet cells. The average cell area was 1509 microm(2), the long:short dimension ration averaged 1.42 and the average nucleus/cytoplasm (N/C) value was just 0.092 (or 1:11.5 as a ratio). CONCLUSIONS: Since the drug has been reported to be excreted in the tear film, and with no other risk factors (such as contact lens wear, smoking or dry eye disease), the squamous metaplasia is attributed to the use of carbamazepine.     

Information processing by simple molecular motifs and susceptibility to noise

Biological organisms rely on their ability to sense and respond appropriately to their environment. The molecular mechanisms that facilitate these essential processes are however subject to a range of random effects and stochastic processes, which jointly affect the reliability of information transmission between receptors and, for example, the physiological downstream response. Information is mathematically defined in terms of the entropy; and the extent of information flowing across an information channel or signalling system is typically measured by the ‘mutual information’, or the reduction in the uncertainty about the output once the input signal is known. Here, we quantify how extrinsic and intrinsic noise affects the transmission of simple signals along simple motifs of molecular interaction networks. Even for very simple systems, the effects of the different sources of variability alone and in combination can give rise to bewildering complexity. In particular, extrinsic variability is apt to generate ‘apparent’ information that can, in extreme cases, mask the actual information that for a single system would flow between the different molecular components making up cellular signalling pathways. We show how this artificial inflation in apparent information arises and how the effects of different types of noise alone and in combination can be understood.