Purification of carbonic anhydrase from dog erythrocytes and investigation of in vitro inhibition by various compounds

The enzyme carbonic anhydrase (E.C. 4.2.1.1) has a stimulatory effect on glaucoma, an eye disease that has a risk to dogs, which are models for the human eye disease, that is similar to that in humans.

In this study, some sulfonamide derivatives, 2-(3-cyclohexene-1-carbamido)-1,3,4-thiadiazole-5-sulfonamide (CCTS), 4-(3-cyclohexene-1-carbamido) methyl-benzenesulfonamide (CCBS), 2-(9-octadecenoylamido)-1,3,4-thiadiazole-5-sulfonamide (ODTS), 2-(4,7,10-trioxa-tetradecanoylamido)-1,3,4-thiadiazole-5-sulfonamide (TDTS), and 2-(8-methoxycoumarine-3-carbamido)-1,3,4-thiadiazole-5-sulfonamide (MCTS), as well as some anionic compounds (perchlorate and chloride) and existing medicines (dorzolamide-HCl, gentamicine sulphate, tropicamide, and procaine-HCl) were assayed for their inhibition of dog carbonic anhydrase (dCA), which was purified from erythrocytes on an affinity gel of L-tyrosine-sulfonamide-Sepharose 4B. ODTS showed the highest potency amongst the synthetic compounds with IC₅₀ value 1.18 × 10^{? 5} M. Amongst the medicines tested, only dorzolamide showed inhibition with IC₅₀ value 5.05 × 10^{? 4} M. Procaine and tropicamide actually showed an activatory effect, whereas gentamicine sulfate had no significant effect. The inhibitory effects of anionic compounds such as perchlorate and chloride were also investigated; whereas perchlorate showed inhibition, chloride did not.     

Differential expression of genes participating in cardiomyocyte electrophysiological remodeling via membrane ionic mechanisms and Ca2+-handling in human heart failure

Molecular and Cellular Biochemistry - Excitation–contraction coupling in normal cardiac function is performed with well balanced and coordinated functioning but with complex dynamic...     

Description of two new species of the genus Gobio (Teleostei: Cyprinidae) from the Black Sea coast of Turkey

Thesystematics of the genus Gobio, especially of the populations on the southern Black Sea coast, seems to be still far from being resolved. Seven species were rec-orded from the northern Black Sea coast while three species were recognized from the southern Black Sea coast. We examined in total 43 specimens from K?z?l?rmak River (southern Black Sea basin), 80 specimens from Çoruh River (the Western Cau-casus of the Black Sea basin), 14 specimens from Rioni River (the Western Caucasus of the Black Sea basin), and 6 specimens from Kherota and Shakhe Rivers (the Western Caucasus of the Black Sea basin). Based on an analysis of 32 metric and 5 meristic data as well as the number of vertebrae, we concluded that the K?z?l?rmak and Coruh populations are two distinct, unnamed species, and we describe them here as Gobio kizilirmakensis from the K?z?l?rmak River and G. artvinicus from the Çoruh River.

http://www.zoobank.org/urn:urn:lsid:zoobank.org:pub:1ED54FB1-588F-4412-B376-4BE4E4516EB1     

Olfactory and Behavioral Mechanisms Underlying Enhanced Mating Competitiveness Following Exposure to Ginger Root Oil and Orange Oil in Males of the Mediterranean Fruit Fly, Ceratitis capitata (Diptera: Tephritidae)

Males of the Mediterranean fruit fly, Ceratitis capitata, are strongly attracted to various plant odors, and previous work has demonstrated that male exposure to certain odors, including the scent of orange oil (OO) and ginger root oil (GRO), increases their mating success relative to non-exposed males. However, the mechanism(s) underlying this mating increase is not known. Here, we describe several experiments that further investigate the association between GRO- and OO-exposure and male signaling activity, pheromone attractiveness, and mating success in male medflies. Exposure to GRO or OO increased time spent pheromone calling but did not accelerate the rate of male sexual maturation. Using a wind tunnel, we compared female attraction to the pheromone of control, non-exposed males versus males previously exposed to OO or GRO. There was no evidence that GRO exposure enhanced the attractiveness of the male pheromone. The data for OO were inconclusive: females tended to spend more time on spheres emanating pheromone from OO-exposed males than on spheres emanating pheromone from non-exposed males, but the number of female landings did not differ between the two types of pheromone sources. Female choice tests confirmed that GRO- and OO-exposure boost male mating success relative to non-exposed males. Application of GRO directly to the abdomen reduced male mating success, whereas similar application of OO boosted male mating success. The potential role and mode of action of plant chemicals in the mating behavior of male medflies are evaluated in light of these findings.     

Does hydatid disease have protective effects against lung cancer?

We hypothesized that solid tumors rarely occur in patients with hydatid disease. We obtained the serum of 14 patients diagnosed with hydatid disease, the serum of 10 patients who did not have a history of hydatid disease, and the hydatid cyst fluid from six patients. These sera and fluid samples were added at different concentrations to NCI-H209/An1 human lung small cell carcinoma cells and L929 mouse fibroblasts as a control group. Sera of patients with hydatid diseases had cytotoxic effects on NCI-H209/An1 cells, but they did not have cytotoxic effects on fibroblast cells. Sera from healthy subjects did not have a cytotoxic effect on the tumor cell line or control fibroblasts. Cyst fluid, also, did not have toxic effects on the NCI-H209/An1 cell line, but was toxic to fibroblasts up to a 1:32 dilution. Sera from patients with hydatid disease had cytotoxic effects on human small cell lung cancer cells in vitro.     

Identifying Fishes through DNA Barcodes and Microarrays

Background

International fish trade reached an import value of 62.8 billion Euro in 2006, of which 44.6% are covered by the European Union. Species identification is a key problem throughout the life cycle of fishes: from eggs and larvae to adults in fisheries research and control, as well as processed fish products in consumer protection.

Methodology/Principal Findings

This study aims to evaluate the applicability of the three mitochondrial genes 16S rRNA (16S), cytochrome b (cyt b), and cytochrome oxidase subunit I (COI) for the identification of 50 European marine fish species by combining techniques of “DNA barcoding” and microarrays. In a DNA barcoding approach, neighbour Joining (NJ) phylogenetic trees of 369 16S, 212 cyt b, and 447 COI sequences indicated that cyt b and COI are suitable for unambiguous identification, whereas 16S failed to discriminate closely related flatfish and gurnard species. In course of probe design for DNA microarray development, each of the markers yielded a high number of potentially species-specific probes in silico, although many of them were rejected based on microarray hybridisation experiments. None of the markers provided probes to discriminate the sibling flatfish and gurnard species. However, since 16S-probes were less negatively influenced by the “position of label” effect and showed the lowest rejection rate and the highest mean signal intensity, 16S is more suitable for DNA microarray probe design than cty b and COI. The large portion of rejected COI-probes after hybridisation experiments (>90%) renders the DNA barcoding marker as rather unsuitable for this high-throughput technology.

Conclusions/Significance

Based on these data, a DNA microarray containing 64 functional oligonucleotide probes for the identification of 30 out of the 50 fish species investigated was developed. It represents the next step towards an automated and easy-to-handle method to identify fish, ichthyoplankton, and fish products.     

Pentoxifylline attenuates cardiac dysfunction and reduces TNF-alpha level in ischemic-reperfused heart

Although pentoxifylline (PTXF), a phosphodiesterase inhibitor, has been reported to exert beneficial effects in cardiac bypass surgery, its effect and mechanisms against ischemia-reperfusion (I/R) injury in heart are poorly understood. Because I/R is known to increase the level of tumor necrosis factor (TNF)-alpha in myocardium and PTXF has been shown to depress the production of TNF-alpha in failing heart, this study examined the hypothesis that PTXF may attenuate cardiac dysfunction and reduce TNF-alpha content in I/R heart. For this purpose, isolated rat hearts were subjected to global ischemia for 30 min followed by reperfusion for 2-30 min. Although cardiac dysfunction due to ischemia was not affected, the recovery of heart function upon reperfusion was markedly improved by PTXF treatment. This cardioprotective effect of PTXF was dose dependent; maximal effect was seen at a concentration of 125 microM. TNF-alpha, nuclear factor-kappaB (NF-kappaB), and phosphorylated NF-kappaB contents were decreased in ischemic heart but were markedly increased within 2 min of starting reperfusion. The ratio of cytosolic-to-homogenate NF-kappaB was decreased, whereas the ratio of particulate-to-homogenate NF-kappaB was increased in I/R hearts. These changes in TNF-alpha and NF-kappaB protein contents as well as in NF-kappaB redistribution due to I/R were significantly attenuated by PTXF treatment. The results of this study indicate that the cardioprotective effects of PTXF against I/R injury may be due to reductions in the activation of NF-kappaB and the production of TNF-alpha content.     

YUPI<Superscript>®</Superscript>, a regional footprint calculator

Purpose

Many tools to quantify the environmental impact of human decisions have been developed, but all of them seem to have a limited application at the regional or local level. A free-of-charge, Argentina-based personal footprint calculator software (YUPI^®) has been developed in order to raise awareness among local citizens about the environmental impacts generated by their daily habits. The extensive use of the tool will generate information suitable for future scientific studies based on local data.

Methods

The software calculates the ecological, carbon, and water footprints of individuals, implementing specific regional data from Argentina developed by the CLIOPE group, complemented with data from the Water Footprint Network and the Global Footprint Network. The calculator was developed focusing on interface attractiveness, ease of use, language simplicity, and a good trade-off between completion time and fullness.

Results and discussion

The YUPI^® software allows its users to understand at a glance their contribution to the environmental impacts of modern society and to quantify the reduction opportunities they have at hand. The program’s language and variables reflect local lifestyle choices, making the filling process accessible for children. The calculator was placed online as an educational tool for teachers and students from all educational levels, and it was also used by visitors in local science and educational fairs. Valuable data was collected for future initiatives on impact mitigation.

Conclusions

Amplified by the mass media, the new tool has helped raise awareness and discussion about the individual environmental footprint, both in the educational and in the domestic terrain. The strategy of creating a simple, easily administered, and widely available quiz helped bridge the gap between the academy and the people, making available to them the continuously updated information generated by the research groups. This is facilitating citizen not only to understand the complexity of the environmental problems but also to take informed actions leading to their mitigation.