In vitro evaluation of antioxidant and radioprotective properties of a novel extremophile from mud volcano: implications for management of radiation emergencies

A thermophilic bacterium, designated as RH 127, was isolated from mud volcano (Baratang Islands) of Andaman region, India (12°07'N 92°47'E/12.117°N 92.783°E) for the first time. Biochemical tests and 16S rRNA gene sequencing indicate that it belongs to the genus Geobacillus. The strain showed 98% confirmed 16S rRNA gene sequence homology with Geobacillus toebii. The bacteria was extracted in various solvent systems and three different fractions prepared. In the present study, antioxidant and radioprotective activity of extracts (INM-7860, INM-7861, and INM-7862) of bacterium G. toebii (strain RH 127) were evaluated. The fractions were evaluated for their introspective comparison of the relative antioxidant efficiency. The antioxidative activities, DPPH radical scavenging effects, hydroxyl radical scavenging effects, membrane protection, antihemolytic activity, and linoleic acid degradation efficacies were assayed. INM-7861 and INM-7862 activated NF-κB expression, as evidenced by reporter assay studies, and thereby contributed to overall radioprotective effect. INM-7862 exhibited best results. This study explicitly shows that the extracts of G. toebii have immense potential as a radiation countermeasure agent.     

Prioritizing Remnant Forests for the Conservation of Mysore Slender Lorises (<Emphasis Type="Italic">Loris lyddekerianus lyddekerianus</Emphasis>) in Karnataka,India Through Estimation of Population Density

Information on a species’ distribution, abundance, and habitat requirements is important for formalizing a comprehensive management and conservation strategy. This becomes an even higher priority when the distribution of a species lies largely outside demarcated protected areas for wildlife. We used line transect distance sampling to estimate the density of an arboreal, nocturnal, and threatened primate, the slender loris (Loris lyddekerianus lyddekerianus) across diverse habitat types including both reserve forests and production landscapes in the southern part of the state of Karnataka, India. Abundance estimates varied from 0.16 individuals/ha in Devrayandurga State Forest to 2.57 individulas/ha in Ippadi State Forest, with the mean density estimate being highest in the Forest Division of Tumkur, at 165 individuals/km². Based on density estimates for the survey regions, their current habitat status, and perceived threats, we identify Ippadi, Ujjani, Devrayandurga, and Savanadurga state forests as priority areas for conservation of the slender loris. We propose that these areas be declared protected. The required management intervention is preventing further degradation of the habitat and enhancing canopy contiguity to facilitate loris movement.     

In vitro and ex vitro evaluation of long-term micropropagated turmeric as analyzed through cytophotometry,phytoconstituents, biochemical and molecular markers

Turmeric (Curcuma longa L.), a high valued medicinal plant, was micropropagated through induction of multiple shoots using latent axillary buds of rhizome. Cytophotometric and random amplified polymorphic DNA (RAPD) as well as inter simple sequence repeats (ISSR) analysis were used to periodically monitor the genetic stability of micropropagated clones of Curcuma longa conserved in vitro up to 7 years at every 6 months interval. A total of eighteen RAPD and eight ISSR primers gave 45,537 distinct and reproducible bands, monomorphic across all 353 plants analyzed. Micropropagated turmeric after being conserved for 7 years in vitro was transplanted into soil in field. Drug yielding potential of tissue culture derived plants was evaluated in field through estimation of phytoconstituents like curcumin and essential oil contents. The result of 2 years of field trial showed that micropropagated turmeric retained stability in all the characteristics examined when compared with the field performance of conventionally propagated plants. Thus long term conservation of an elite genotype of turmeric with epigenetic and genetic stability is significant for stable supply of drug i.e., curcumin and essential oil to the market.     

Effect of different supplements on bioprocessing of wheat straw by Phlebia brevispora: changes in its chemical composition, in vitro digestibility and nutritional properties

Bioprocessing of wheat straw was carried out by Phlebia brevispora under solid state conditions. Effect of different supplements on lignocellulolytic enzymes production, degradation of straw cell wall fibers and its resultant effect on nutritional quality of wheat straw were studied. Ammonium chloride and malt extract were more effective in terms of ligninolysis and enhanced in vitro digestibility. The concentration of the selected supplements and the moisture content was worked out using response surface methodology in order to minimize the loss in total organic matter so as to selectively degrade lignin. The experiment was scaled up to batches of 200 g under optimized conditions and the degraded substrate was analyzed for its biochemical properties. P. brevispora degraded 290 g/kg of lignin and enhanced the in vitro digestibility from 150 to 268 g/kg (78%). Crude protein, amino acids, total phenolic contents and antioxidant properties were significantly higher in degraded straw.     

Dipyridamole reverses peripheral ischemia and induces angiogenesis in the Db/Db diabetic mouse hind-limb model by decreasing oxidative stress

Dipyridamole anti-platelet therapy has previously been suggested to ameliorate chronic tissue ischemia in healthy animals. However, it is not known if dipyridamole therapy represents a viable approach to alleviating chronic peripheral tissue ischemia associated with type 2 diabetes. Here we examine the hypothesis that dipyridamole treatment restores reperfusion of chronic hind-limb ischemia in the murine B6.BKS-Lepr(db/db) diabetic model. Dipyridamole therapy quickly rectified ischemic hind-limb blood flow to near preligation levels within 3 days of the start of therapy. Restoration of ischemic tissue blood flow was associated with increased vascular density and endothelial cell proliferation observed only in ischemic limbs. Dipyridamole significantly increased total nitric oxide metabolite levels in tissue, which were not associated with changes in endothelial NO synthase expression or phosphorylation. Interestingly, dipyridamole therapy significantly decreased ischemic tissue superoxide and protein carbonyl levels, identifying a dominant antioxidant mechanistic response. Dipyridamole therapy also moderately reduced diabetic hyperglycemia and attenuated development of dyslipidemia over time. Together, these data reveal that dipyridamole therapy is an effective modality for the treatment of chronic tissue ischemia during diabetes and highlights the importance of dipyridamole antioxidant activity in restoring tissue NO bioavailability during diabetes.     

Elevated levels of NO are localized to distal airways in asthma

The contribution of nitric oxide (NO) to the pathophysiology of asthma remains incompletely defined despite its established pro- and anti-inflammatory effects. Induction of the inducible nitric oxide synthase (iNOS), arginase, and superoxide pathways is correlated with increased airway hyperresponsiveness in asthmatic subjects. To determine the contributions of these pathways in proximal and distal airways, we compared bronchial wash (BW) to traditional bronchoalveolar lavage (BAL) for measurements of reactive nitrogen/oxygen species, arginase activation, and cytokine/chemokine levels in asthmatic and normal subjects. Levels of NO were preferentially elevated in the BAL, demonstrating higher level NOS activation in the distal airway compartment of asthmatic subjects. In contrast, DHE(+) cells, which have the potential to generate reactive oxygen species, were increased in both proximal and distal airway compartments of asthmatics compared to controls. Different patterns of cytokines and chemokines were observed, with a predominance of epithelial cell-associated mediators in the BW compared to macrophage/monocyte-derived mediators in the BAL of asthmatic subjects. Our study demonstrates differential production of reactive species and soluble mediators within the distal airways compared to the proximal airways in asthma. These results indicate that cellular mechanisms are activated in the distal airways of asthmatics and must be considered in the development of therapeutic strategies for this chronic inflammatory disorder.     

Current perspectives and challenges in understanding the role of nitrite as an integral player in nitric oxide biology and therapy

Beyond an inert oxidation product of nitric oxide (NO) metabolism, current thinking posits a key role for nitrite as a mediator of NO signaling, especially during hypoxia. This concept has been discussed in the context of nitrite serving a role as an endogenous modulator of NO homeostasis, but also from a novel clinical perspective whereby nitrite therapy may replenish NO signaling and prevent ischemic tissue injury. Indeed, the relatively rapid translation of studies delineating mechanisms of action to ongoing and planned clinical trials has been critical in fuelling interest in nitrite biology, and several excellent reviews have been written on this topic. In this article we limit our discussions to current concepts and what we feel are questions that remain unanswered within the paradigm of nitrite being a mediator of NO biology.     

Click chemistry inspired one-pot synthesis of 1,4-disubstituted 1,2,3-triazoles and their Src kinase inhibitory activity

Two classes of 1,4-disubstituted 1,2,3-triazoles were synthesized using one-pot reaction of α-tosyloxy ketones/α-halo ketones, sodium azide, and terminal alkynes in the presence of aq PEG (1:1, v/v) using the click chemistry approach and evaluated for Src kinase inhibitory activity. Structure-activity relationship analysis demonstrated that insertion of C₆H₅- and 4-CH₃C₆H₄- at position 4 for both classes and less bulkier aromatic group at position 1 in class 1 contribute critically to the modest Src inhibition activity (IC₅₀ = 32-43 μM) of 1,4-disubstituted 1,2,3-triazoles.     

Synthesis of 3-phenylpyrazolopyrimidine-1,2,3-triazole conjugates and evaluation of their Src kinase inhibitory and anticancer activities

A series of two classes of 3-phenylpyrazolopyrimidine-1,2,3-triazole conjugates were synthesized using click chemistry approach. All compounds were evaluated for inhibition of Src kinase and human ovarian adenocarcinoma (SK-Ov-3), breast carcinoma (MDA-MB-361), and colon adenocarcinoma (HT-29). Hexyl triazolyl-substituted 3-phenylpyrazolopyrimidine exhibited inhibition of Src kinase with an IC₅₀ value of 5.6 μM. 4-Methoxyphenyl triazolyl-substituted 3-phenylpyrazolopyrimidine inhibited the cell proliferation of HT-29 and SK-Ov-3 by 73% and 58%, respectively, at a concentration of 50 μM.     

Mitigation of chlorine gas lung injury in rats by postexposure administration of sodium nitrite

Nitrite (NO(2)(-)) has been shown to limit injury to the heart, liver, and kidneys in various models of ischemia-reperfusion injury. Potential protective effects of systemic NO(2)(-) in limiting lung injury or enhancing repair have not been documented. We assessed the efficacy and mechanisms by which postexposure intraperitoneal injections of NO(2)(-) mitigate chlorine (Cl(2))-induced lung injury in rats. Rats were exposed to Cl(2) (400 ppm) for 30 min and returned to room air. NO(2)(-) (1 mg/kg) or saline was administered intraperitoneally at 10 min and 2, 4, and 6 h after exposure. Rats were killed at 6 or 24 h. Injury to airway and alveolar epithelia was assessed by quantitative morphology, protein concentrations, number of cells in bronchoalveolar lavage (BAL), and wet-to-dry lung weight ratio. Lipid peroxidation was assessed by measurement of lung F(2)-isoprostanes. Rats developed severe, but transient, hypoxemia. A significant increase of protein concentration, neutrophil numbers, airway epithelia in the BAL, and lung wet-to-dry weight ratio was evident at 6 h after Cl(2) exposure. Quantitative morphology revealed extensive lung injury in the upper airways. Airway epithelial cells stained positive for terminal deoxynucleotidyl-mediated dUTP nick end labeling (TUNEL), but not caspase-3. Administration of NO(2)(-) resulted in lower BAL protein levels, significant reduction in the intensity of the TUNEL-positive cells, and normal lung wet-to-dry weight ratios. F(2)-isoprostane levels increased at 6 and 24 h after Cl(2) exposure in NO(2)(-)- and saline-injected rats. This is the first demonstration that systemic NO(2)(-) administration mitigates airway and epithelial injury.