Simple, reliable and fast spectrofluorometric method for determination of plasma Verteporfin (Visudyne) levels during photodynamic therapy for choroidal neovascularization.

Photodynamic therapy with Verteporfin, a potent photosensitizer dye, is a very effective treatment for age related macular degeneration due to choroidal neovascularization. Photodynamic therapy offers the potential for selective tissue injury in part attributable to preferential localization of Verteporfin, administrated by intravenous infusion, to the choroidal neovascularization complex and irradiation of the complex with non-laser thermal light at 690 nm resulting in at least temporary thrombosis and vessel closure. Verteporfin is a benzoporphyrin derivative monoacid ring A formulated as a unilamellar liposome. In the blood Verteporfin is associated with lipoprotein fractions and is rapidly cleared via a receptor-mediated uptake mechanism due the high expression of LDL receptors in neovascular tissues. Verteporfin was undetectable in plasma 24 hr after infusion of the recommended dose: 6 mg/m2 of body surface area. The main side effect is photosensitivity of skin which is usually short-lived (24-48 hr) with a low incidence (2.3%). As skin photosensitivity depends on circulating rather than tissue drug levels, we investigate the possibility of developing a simple, fast and reliable spectrofluorometric method to measure plasma Verteporfin levels. Fluorescence emission spectrum (550-750 nm) of 1:10 saline diluted plasma with lambda exc=430 nm showed a characteristic emission peak at 692 nm, the height being proportional to the Verteporfin levels. The sensitivity is around 100 ng/ml and the pharmacokinetics of Verteporfin has been studied from 0 to 5 hr after infusion in six patients older than 65 years with age-related macular degeneration.     

Aminophylline enhances contractility of frog skeletal muscle: an effect dependent on extracellular calcium

The effects of aminophylline (10-500 microM) on isometric twitch and tetanic forces were studied in vitro on frog semitendinosus muscle. Two hypotheses were tested: 1) that micromolar concentrations of aminophylline enhanced contractility of isolated skeletal muscle and 2) that the potentiating effect of aminophylline was dependent on the presence of extracellular calcium ions. Muscles were removed, placed in aerated Ringer solution at 20 degrees C, attached to a force transducer, and stimulated directly. Muscles in normal Ringer and aminophylline Ringer were compared throughout the frequency-force relationship from twitches to maximum tetanic force. Aminophylline increased twitch force significantly at concentrations as low as 25 microM. Over a range of stimulation frequencies, but especially at 10 and 20 Hz, aminophylline increased tetanic force. The potentiating effect of aminophylline (100 microM) was reduced or eliminated in calcium-free Ringer containing 10 mM magnesium. We conclude that aminophylline, at therapeutic concentrations, enhances muscle contractility, and the enhancement is dependent on the presence of extracellular calcium. These findings support the concept that aminophylline is effective in improving respiration in humans with airway obstruction by enhancing diaphragmatic contractility.     

Metabolic and functional differences between brain and spinal cord mitochondria underlie different predisposition to pathology

Mitochondrial dysfunctions contribute to neurodegeneration, the locations of which vary among neurodegenerative diseases. To begin to understand what mechanisms may underlie higher vulnerability of the spinal cord motor neurons in amyotrophic lateral sclerosis, compared with brain mitochondria, we studied three major functions of rat brain mitochondria (BM) and spinal cord mitochondria (SCM) mitochondria: oxidative phosphorylation, Ca(2+) sequestration, and production of reactive oxygen species (ROS), using a new metabolic paradigm (Panov et al., J. Biol. Chem. 284: 14448-14456, 2009). We present data that SCM share some unique metabolic properties of the BM. However, SCM also have several distinctions from the BM: 1) With the exception of succinate, SCM show significantly lower rates of respiration with all substrates studied; 2) immunoblotting analysis showed that this may be due to 30-40% lower contents of respiratory enzymes and porin; 3) compared with BM, SCM sequestered 40-50% less Ca(2+), and the total tissue calcium content was 8 times higher in the spinal cord; 4) normalization for mitochondria from 1 g of tissue showed that BM can sequester several times more Ca(2+) than was available in the brain tissue, whereas SCM had the capacity to sequester only 10-20% of the total tissue Ca(2+); and 5) with succinate and succinate-containing substrate mixtures, SCM showed significantly higher state 4 respiration than BM and generated more ROS associated with the reverse electron transport. We conclude that SCM have an intrinsically higher risk of oxidative damage and overload with calcium than BM, and thus spinal cord may be more vulnerable under some pathologic conditions. (250).     

THE ACCLIMATED RESPONSE OF GROWTH,PHOTOSYNTHESIS, COMPOSITION,AND CARBON BALANCE TO TEMPERATURE IN THE PSYCHROPHILIC ICE DIATOM NITZSCHIA SERIATA1

Nitzschia seriata Cleve, a common member of marine bottom ice communities in the Arctic, was grown in unialgal batch cultures to test for compensatory mechanisms for the low temperatures (?1.8° C) typical of its natural habitat. The upper lethal limit for growth was between 12° and 15°C, and the optimum was between 6° and 12° C. The Arrhenius function adequately (R²= 73%) fitted the relationship between growth rate and temperature from – 1.6° up to 10° C, with an average Q₁₀ of 1.9 over the entire range. Light-saturated and light-limited rates of photosynthesis (normalized to chlorophyll a or cell carbon) showed complete compensation from 12° to 4° C. Photosynthetic rates, especially at light saturation, declined rapidly at temperatures below 4° C. Susceptibility to photoinhibition was greatest at the lowest growth temperatures. Cellular composition (chlorophyll a, protein, polysaccharide, and lipid contents) was not systematically related to temperature in any simple way, although cell size (carbon per cell) was maximal at the lowest growth temperature. Dark respiration was unmeasurably low (<0.015 day^?1) at all growth temperatures. The strategy of adaptation in N. seriata may be characterized as optimizing efficiency and compensation, rather than maximization, of growth rate.     

Adrenomedullin inhibits angiotensin AT1A receptor expression and function in cardiac fibroblasts

Adrenomedullin (AM) is a multifunctional peptide hormone with wide-ranging actions related to cardiovascular homeostasis. AM receptors are highly expressed in the heart and AM has antihypertrophic and antiproliferative effects on cardiac myocytes and fibroblasts, respectively. We have investigated the interaction between AM and angiotensin II (Ang II) signalling in neonatal cardiac fibroblast cultures to determine whether the antagonistic effects of AM are mediated via the modulation of Ang II receptors. Cardiac fibroblasts exclusively expressed the Ang II type 1 receptor (AT(1)R) and binding to this site was downregulated by 35% following an 18-h incubation with 100 nM AM. Levels of AT(1A)R mRNA were dose-dependently lowered by AM, with a maximal 40-50% inhibition by 6 h. The decreases in both AT(1)R binding and AT(1A)R mRNA levels were mimicked by 8-Br-cAMP or forskolin, suggesting that the effects of AM were mediated via an elevation of cAMP. In cardiac fibroblasts pretreated with AM, the Ang II induction of collagen biosynthesis was attenuated, although basal collagen synthesis was unaffected. These data suggest that AM mediates the heterologous downregulation of AT(1)R expression via a relatively rapid decrease in AT(1A)R mRNA pools. This interaction may represent a relevant pathophysiological mechanism for modulating Ang II responsiveness in the diseased heart.     

Effects of river stage height and water temperature on diet composition of year‐0 sturgeon (Scaphirhynchus spp.): a multi‐year study

The diet composition was quantified of year‐0 Scaphirhynchus sturgeons during sampling years (2004–2008) from the Middle Mississippi River to evaluate trends in diet. River stage height varied among years and seasons. Water temperature differed among seasons but did not differ among years. The presence of empty stomachs was evident across years, seasons, and size classes of sturgeon. The overall Multivariate Analysis of Variance for frequency of occurrence was significant, suggesting that macroinvertebrate taxa (e.g. Ephemeroptera, Diptera pupae, Chironomidae, ‘Other’) differed among size classes of sturgeon, but not seasons or years. Furthermore, Tukey's pairwise comparisons indicated Ephemeroptera frequency of occurrence differed between medium and large size classes; Diptera pupae differed between small and medium size classes and small and large size classes; Chironomidae differed between small and large size classes and medium and large size classes; the macroinvertebrate taxa category ‘Other’ differed between small and large size classes. Ultimately, because of the link between diet and growth of year‐0 Scaphirhynchus sturgeon, the early life history may affect eventual recruitment to adult stages.