Screening of antioxidant potential of Arctic lichens

Antioxidants are compounds that scavenge the free radicals produced in living organisms. The antioxidant potential of eight Arctic lichen species was evaluated in vitro using free radical scavenging activity (FRS), inhibition of lipid peroxidation (ILP), and Trolox equivalent antioxidant capacity assay (TEAC). FRS activities of lichen species in various organic solvents such as methanol, ethanol, acetone, and dimethyl sulphoxide (DMSO) were in the range 9.6–51.77%, while ILP activities in these solvents ranged from 32.5 to 82.43%. Pseudophebe pubescens showed the highest ILP (82.43%) and FRS (51.77%) activities as compared to other lichen species and the standard antioxidants butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). The TEAC value was also found to be higher in all species compared to the standard water soluble vitamin E analog Trolox (3.9 mM). The order of antioxidative activities in lichen species was Pseudophebe pubescens > Cladonia amaurocraea > Cladonia mediterranea > Physcia caesia > Flavocetraria nivalis > Cetraria fastigata > Xanthoria elegans > Umbilicaria hyperborea. This is the first report of the measurement of antioxidant potential in Arctic lichens.     

Comparative effects of alpha-receptor stimulation and nitrergic inhibition on bronchovascular tone.

Adrenergic agonists are known to influence bronchial blood flow and bronchovascular resistance. Recently, the nitrergic system has also been implicated in the control of bronchovascular tone. In this study, we compared the effects of the nitric oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) and the alpha(1)-receptor agonist phenylephrine on bronchovascular resistance in anesthetized sheep (n = 9). Bronchial blood flow, cardiac output, and systemic and pulmonary arterial pressures were continuously monitored. Phenylephrine (1.2-3.4 microg. kg(-1). min(-1)) was infused intravenously to increase mean systemic arterial pressure above 95 Torr for 10 min and then was discontinued. When hemodynamic parameters returned to baseline, nebulized phenylephrine (10 mg) was given over 10 min. When parameters again normalized, L-NAME (30 mg/kg) was infused intravenously over 1 min. Intravenous phenylephrine increased systemic vascular resistance by 40% at 10 min with no concurrent increase in bronchovascular resistance, but inhaled phenylephrine increased bronchovascular resistance by 66% at 10 min. By comparison, intravenous L-NAME produced a rapid and sustained fivefold increase in bronchovascular resistance at 10 min. We conclude that, although alpha-agonist stimulation has some influence on bronchovascular resistance in sheep, the nitrergic system has predominant control of bronchovascular tone.     

Novel screening protocol for precise phenotyping of salt-tolerance at reproductive stage in rice

The present study reports an unequivocal and improved protocol for efficient screening of salt tolerance at flowering stage in rice, which can aid phenotyping of population for subsequent identification of QTLs associated with salinity stress, particularly at reproductive stage. To validate the new method, the selection criteria, level and time of imposition of stress; plant growth medium were standardized using three rice genotypes. The setup was established with a piezometer placed in a perforated pot for continuous monitoring of soil EC and pH throughout the period of study. Further, fertilizer enriched soil was partially substituted by gravels for stabilization and maintaining the uniformity of soil EC in pots without hindering its buffering capacity. The protocol including modified medium (Soil:Stone, 4:1) at 8 dS m^?1 salinity level was validated using seven different genotypes possessing differential salt sensitivity. Based on the important selection traits such as high stability index for plant yield, harvest index and number of grains/panicle and also high K⁺ concentration and low Na⁺– K⁺ ratio in flag leaf at grain filling stage were validated and employed in the evaluation of a mapping population in the modified screening medium. The method was found significantly efficient for easy maintenance of desired level of soil salinity and identification of genotypes tolerant to salinity at reproductive stage.     

The effects of N(omega)-propyl-L-arginine on reperfusion injury of skeletal muscle.

N(omega)-Propyl-L-arginine (NPA) is reported to be a highly selective inhibitor of neuronal nitric oxide synthase (nNOS). This in vivo study observed its role in ischemia/reperfusion (I/R) injury in rat skeletal muscle. Our results showed that NPA infusion significantly increased vessel diameters and blood flow in reperfused cremaster muscle, and slightly increased contractile function in reperfused extensor digitorum longus (EDL) muscle. In addition, NPA treatment slightly increased I/R-mediated downregulation of nNOS and eNOS mRNA and protein levels. Although NPA showed a beneficial role in I/R injury, our in vivo data do not support NPA as a selective nNOS inhibitor. Also, our data do not provide any insight into the mechanism of NPA. Thus, the in vivo mechanism of action of NPA needs to be further identified, and the role of nNOS in skeletal muscle I/R still remains to be determined.     

Cellular origin of chlorinated diketopiperazines in the dictyoceratid sponge Dysidea herbacea (Keller)

The tropical marine sponge Dysidea herbacea (Keller) contains the filamentous unicellular cyanobacterium Oscillatoria spongeliae (Schulze) Hauck as an endosymbiont, plus numerous bacteria, both intracellular and extracellular. Archaeocytes and choanocytes are the major sponge cell types present. Density gradient centrifugation of glutaraldehyde-fixed cells with Percoll as the support medium has been used to separate the cyanobacterial symbiont from the sponge cells on the basis of their differing densities. The protocol also has the advantage of separating broken from intact cells of O. spongeliae. The lighter cell preparations contain archaeocytes and choanocytes together with damaged cyanobacterial cells, whereas heavier cell preparations contain intact cyanobacterial cells, with less than 1% contamination by sponge cells. Gas chromatography/mass spectrometry analysis has revealed that the terpene spirodysin is concentrated in preparations containing archaeocytes and choanocytes, whereas nuclear magnetic resonance analysis of the symbiont cell preparations has shown that they usually contain the chlorinated diketopiperazines, dihydrodysamide C and didechlorodihydrodysamide C, which are the characteristic metabolites of the sponge/symbiont association. However, one symbiont preparation, partitioned by a second Percoll gradient, has been found to be devoid of chlorinated diketopiperazines. The capability to synthesize secondary metabolites may depend on the physiological state of the symbiont; alternatively, there may be two closely related cyanobacterial strains within the sponge tissue.     

Antibody Detection and Molecular Characterization of Toxoplasma gondii from Bobcats (Lynx rufus), Domestic Cats (Felis catus), and Wildlife from Minnesota,USA

Little is known of the epidemiology of toxoplasmosis in Minnesota. Here, we evaluated Toxoplasma gondii infection in 50 wild bobcats (Lynx rufus) and 75 other animals on/near 10 cattle farms. Antibodies to T. gondii were assayed in serum samples or tissue fluids by the modified agglutination test (MAT, cut‐off 1:25). Twenty nine of 50 bobcats and 15 of 41 wildlife trapped on the vicinity of 10 farms and nine of 16 adult domestic cats (Felis catus) and six of 14 domestic dogs resident on farms were seropositive. Toxoplasma gondii oocysts were not found in feces of any felid. Tissues of all seropositive wild animals trapped on the farm were bioassayed in mice and viable T. gondii was isolated from two badgers (Taxidea taxus), two raccoons (Procyon lotor), one coyote (Canis latrans), and one opossum (Didelphis virginiana). All six T. gondii isolates were further propagated in cell culture. Multi‐locus PCR‐RFLP genotyping using 10 markers (SAG1, SAG2 (5′‐3′SAG2, and alt.SAG2), SAG3, BTUB, GRA6, c22‐8, c29‐2, L358, PK1, and Apico), and DNA from cell culture derived tachyzoites revealed three genotypes; #5 ToxoDataBase (1 coyote, 1 raccoon), #1 (1 badger, 1 raccoon, 1 opossum), and #2 (1 badger). This is the first report of T. gondii prevalence in domestic cats and in bobcats from Minnesota, and the first isolation of viable T. gondii from badger.