Influence of semisynthetic modification of the scaffold of a contact domain of HbS on polymerization: role of flexible surface topology in polymerization inhibition

A new variant of HbS, HbS-Einstein with a deletion of segment α_23–26 in the B-helix, has been assembled by semisynthetic approach. B-helix of the α chain of cis αβ-dimer of HbS plays dominant role in the quinary interactions of deoxy HbS dimer. This B-helix is the primary scaffold that provides the orientation for the side chains of contact residues of this intermolecular contact domain. The design of HbS-Einstein has been undertaken to map the influence of perturbation of molecular surface topology and the flexibility of surface residues in the polymerization. The internal deletion exerts a strong inhibitory influence on Val-6 (β)-dependent polymerization, comparable to single contact site mutations and not for complete neutralization of Val-6(β)-dependent polymerization. The scaffold modification in cis-dimer is inhibitory, and is without any effect when present on the trans dimer. The flexibility changes in the surface topology in the region of scaffold modification apparently counteracts the intrinsic polymerization potential of the molecule. The inhibition is close to that of Le Lamentin mutation [His-20 (α) → Gln] wherein a mutation engineered without much change in flexibility of the contact domain. Interestingly, the chimeric HbS with swine–human chimeric α chain with multiple non-conservative mutations completely inhibits the Val-6(β)-dependent polymerization. The deformabilities of surface topology of chimeric HbS are comparable to HbS in spite of the multiple contact site mutations in the α-chain. We conclude that the design of antisickling Hbs for gene therapy of sickle cell disease should involve multiple mutations of intermolecular contact sites.     

Biolistic Transformation of Elite Indica Rice (Oryza sativa L.) Cultivars Through Semi-Solid and Liquid Medium Selection Systems

Following microprojectile mediated delivery of a plasmid construct (pAHC-25) encoding bar (bialophos resistance) gene into five-day-old scutellar calli derived from mature embryos, the effectiveness of selection procedure for bar-gene expressing tissue was compared for two indica rice cultivars (IR-64 and Karnal Local). While IR-64 transformants could be selected through the generally used semi-solid selection medium, the same procedure was not effective in the basmati cultivar Karnal Local. In the latter case, while lower concentrations (2–4 mg 1^?1) of the selective agent phosphinothricin (PPT) yielded only escapes, higher concentrations (6–8 mg l^?1) inhibited proliferation of transformed as well as untransformed sectors. For Karnal Local, a liquid medium based selection system was successfully utilized for recovering transformed sectors and, eventually, regenerants. The study demonstrates the generation of transformants of two elite indica cultivars using the environment-independent system of mature embryos from seeds.     

Spatial Variability of Biofuel Production Potential and Hydrologic Fluxes of Land Use Change from Cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.) to Alamo Switchgrass (<Emphasis Type="Italic">Panicum virgatum</Emphasis> L.) in the Texas High Plains

Bioenergy crop production has the potential to protect marginal crop lands that generate high surface runoff and produce poor crop yields. Long-term evaluation of the impacts of such land use change on hydrologic fluxes and biofuel production potential is necessary before adopting such strategies on a large scale. In this study, the hydrologic impacts of replacing cotton (Gossypium hirsutum L.) on marginal lands in an intensive agricultural watershed in the Texas High Plains with Alamo switchgrass (Panicum virgatum L.) as a bioenergy crop were evaluated using the Agricultural Policy/Environmental eXtender (APEX) model. The surface runoff to cotton yield ratio was used as a criterion to identify marginal cotton subareas (homogenous spatial units delineated by APEX) in the study watershed, and three replacement scenarios (low (9 %), medium (33 %), and high (57 %) extents of cotton acreage replaced by switchgrass) were implemented in the scenario analysis. The average (1994–2009) annual surface runoff decreased by about 84 and 66 %, and the percolation increased by 106 and 57 % in the irrigated and dryland subareas, respectively, when cotton was replaced by switchgrass under the high replacement scenario. Spatial analysis showed that switchgrass was a feasible bioenergy crop for replacing cotton, especially in the western part of the study watershed, due to its higher water use efficiency and better water conservation effects compared to cotton. It is estimated that 193 and 381 million liters of ethanol could be produced from the dryland and irrigated subareas of the study watershed, respectively, under the high replacement scenario.     

Gastroprotective effect of Terminalia arjuna bark on diclofenac sodium induced gastric ulcer

AIM: The present study was aimed to evaluate the effect of methanolic extract of Terminalia arjuna (TA) on diclofenac sodium induced gastric ulcer in experimental rats. METHODS: Animals were induced for gastric ulcer with diclofenac sodium (DIC) (80mg/kg bodyweight in water, orally) and treated orally with TA in various doses ranging from 100mg/kg bodyweight to 500mg/kg bodyweight. The effective dose was 400mg/kg bodyweight, since this dose elicited a maximum reduction in lesion index. The gastroprotective effect of TA was assessed from volume of gastric juice, pH, free and total acidity, pepsin concentration, acid output in gastric juice, the levels of non-protein sulfhydryls (NP-SH), lipid peroxide (LPO), reduced glutathione (GSH), and activities of enzymic antioxidants--super oxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and myeloperoxidase (MPO) in gastric mucosa. The levels of DNA, protein bound carbohydrate complexes--hexose, hexoseamine, sialic acid, fucose in gastric mucosa and gastric juice and the levels of RNA in gastric mucosa were assessed. The stomach tissues were used for adherent mucus content and also for the histological examination. RESULTS: A significant reduction in lesion index was observed in ulcer induced animals treated with TA (DIC+TA) compared to ulcerated rats (DIC). A significant increase was observed in pH, NP-SH, GSH, enzymic antioxidants, protein bound carbohydrate complexes, adherent mucus content, nucleic acids with a significant decrease in volume of gastric juice, free and total acidity, pepsin concentration, acid output, LPO levels and MPO activities in DIC+TA rats compared to DIC rats. Histological studies confirmed the gastroprotective activity of TA. CONCLUSION: From the data presented in this study it could be concluded that T. arjuna acts as an gastroprotective agent probably due to its free radical scavenging activity and cytoprotective nature.     

Assessing the hydrologic and water quality impacts of biofuel‐induced changes in land use and management

The Southern High Plains (SHP) of Texas, where cotton (Gossypium hirsutum L.) is grown in vast acreage, and the Texas Rolling Plains (TRP), which is dominated by an invasive brush, honey mesquite (Prosopis glandulosa) have the potential for biofuel production for meeting the U.S. bioenergy target of 2022. However, a shift in land use from cotton to perennial grasses and a change in land management such as the harvesting of mesquite for biofuel production can significantly affect regional hydrology and water quality. In this study, APEX and SWAT models were integrated to assess the impacts of replacing cotton with Alamo switchgrass (Panicum virgatum L.) and Miscanthus × giganteus in the upstream subwatershed and harvesting mesquite in the downstream subwatershed on water and nitrogen balances in the Double Mountain Fork Brazos watershed in the SHP and TRP regions. Simulated average (1994–2009) annual surface runoff from the baseline cotton areas decreased significantly (P < 0.05) by 88%, and percolation increased by 28% under the perennial grasses scenario compared to the baseline cotton scenario. The soil water content enhanced significantly under the irrigated switchgrass scenario compared to the baseline irrigated cotton scenario from January to April and August to October. However, the soil water content was depleted significantly under the dryland Miscanthus scenario from April to July relative to the baseline dryland cotton scenario. The nitrate‐nitrogen (NO₃‐N) and organic‐N loads in surface runoff and NO₃‐N leaching to groundwater reduced significantly by 86%, 98%, and 100%, respectively, under the perennial grasses scenario. Similarly, surface runoff, and NO₃‐N and organic‐N loads through surface runoff reduced significantly by 98.9%, 99.9%, and 99.5%, respectively, under the post‐mesquite‐harvest scenario. Perennial grasses exhibited superior ethanol production potential compared to mesquite. However, mesquite is an appropriate supplementary bioenergy source in the TRP region because of its standing biomass and rapid regrowth characteristics.     

A Synthetic Uric Acid Analog Accelerates Cutaneous Wound Healing in Mice

Wound healing is a complex process involving intrinsic dermal and epidermal cells, and infiltrating macrophages and leukocytes. Excessive oxidative stress and associated inflammatory processes can impair wound healing, and antioxidants have been reported to improve wound healing in animal models and human subjects. Uric acid (UA) is an efficient free radical scavenger, but has a very low solubility and poor tissue penetrability. We recently developed novel UA analogs with increased solubility and excellent free radical-scavenging properties and demonstrated their ability to protect neural cells against oxidative damage. Here we show that the uric acid analog (6, 8 dithio-UA, but not equimolar concentrations of UA or 1, 7 dimethyl-UA) modified the behaviors of cultured vascular endothelial cells, keratinocytes and fibroblasts in ways consistent with enhancement of the wound healing functions of all three cell types. We further show that 6, 8 dithio-UA significantly accelerates the wound healing process when applied topically (once daily) to full-thickness wounds in mice. Levels of Cu/Zn superoxide dismutase were increased in wound tissue from mice treated with 6, 8 dithio-UA compared to vehicle-treated mice, suggesting that the UA analog enhances endogenous cellular antioxidant defenses. These results support an adverse role for oxidative stress in wound healing and tissue repair, and provide a rationale for the development of UA analogs in the treatment of wounds and for modulation of angiogenesis in other pathological conditions.     

Ligand- and structure-based in silico studies to identify kinesin spindle protein (KSP) inhibitors as potential anticancer agents

Kinesin spindle protein (KSP) belongs to the kinesin superfamily of microtubule-based motor proteins. KSP is responsible for the establishment of the bipolar mitotic spindle which mediates cell division. Inhibition of KSP expedites the blockade of the normal cell cycle during mitosis through the generation of monoastral MT arrays that finally cause apoptotic cell death. As KSP is highly expressed in proliferating/cancer cells, it has gained considerable attention as a potential drug target for cancer chemotherapy. Therefore, this study envisaged to design novel KSP inhibitors by employing computational techniques/tools such as pharmacophore modelling, virtual database screening, molecular docking and molecular dynamics. Initially, the pharmacophore models were generated from the data-set of highly potent KSP inhibitors and the pharmacophore models were validated against in house test set ligands. The validated pharmacophore model was then taken for database screening (Maybridge and ChemBridge) to yield hits, which were further filtered for their drug-likeliness. The potential hits retrieved from virtual database screening were docked using CDOCKER to identify the ligand binding landscape. The top-ranked hits obtained from molecular docking were progressed to molecular dynamics (AMBER) simulations to deduce the ligand binding affinity. This study identified MB-41570 and CB-10358 as potential hits and evaluated these experimentally using in vitro KSP ATPase inhibition assays.     

Involvement of notch signaling in wound healing

The Notch signaling pathway is critically involved in cell fate decisions during development of many tissues and organs. In the present study we employed in vivo and cell culture models to elucidate the role of Notch signaling in wound healing. The healing of full-thickness dermal wounds was significantly delayed in Notch antisense transgenic mice and in normal mice treated with gamma-secretase inhibitors that block proteolytic cleavage and activation of Notch. In contrast, mice treated with a Notch ligand Jagged peptide showed significantly enhanced wound healing compared to controls. Activation or inhibition of Notch signaling altered the behaviors of cultured vascular endothelial cells, keratinocytes and fibroblasts in a scratch wound healing model in ways consistent with roles for Notch signaling in wound healing functions all three cell types. These results suggest that Notch signaling plays important roles in wound healing and tissue repair, and that targeting the Notch pathway might provide a novel strategy for treatment of wounds and for modulation of angiogenesis in other pathological conditions.     

E-, L- and A-values for estimation of plant-available soil phosphorus

Summary E-, L-values growing mungbean and A-values growing maize, mungbean, urdbean and cowpea were assessed in P and farmyard manure enriched soil of permanent manurial trial at Pantnagar, Nainital of India. E- and L-values were found to increase in farmyard manure and P enriched soil while L-values remained constant at different dates of harvesting. A-values varied with fertility status of soil and the kip of crop.Maize gave higher A-values than pulses. In all crops, higher A-values were found with enrichment of farmyard manure and phosphorus. E-, L- and A-values gave significant correlations with yield and P uptake in pot experiments. E-, L- and A-values were good in assessing the availability of soil phosphorus. However, E-value is a rapid technique as it measured in laboratory without involvement of plant.A part of Ph.D. Dissertation (Agronomy) of N. Venkat Reddy.     

Therapeutic efficacy of green tea polyphenols on cellular thiols in 4-Nitroquinoline 1-oxide-induced oral carcinogenesis

In cancer, a high flux of oxidants not only depletes the cellular thiols, but damages the whole cell as well. Epidemiological studies suggest green tea may mitigate cancers in human and animal models for which several mechanisms have been proposed. In the present investigation, the levels of cellular thiols such as reduced glutathione (GSH), oxidised glutathione (GSSG), protein thiols (PSH), total thiols, lipid peroxidation product conjugated dienes and the activity of gamma glutamyl transferase (GGT) were assessed in tongue and oral cavity. In 4-Nitroquinoline 1-oxide- (4-NQO) induced rats, there was a decrease in the levels of GSH, PSH and total thiols and an increase in the levels of GSSG, conjugated dienes and the activity of GGT. On supplementation of green tea polyphenols (GTP) for 30 days (200 mg/kg) for the oral cancer-induced rats, there was a moderate increase in the levels of GSH, PSH and total thiols and a decrease in the levels of GSSG, conjugated dienes and the activity of GGT. Thus, GTP reduces the oxidant production thereby maintains the endogenous low molecular weight cellular thiols in oral cancer-induced rats. From the results, it can be concluded that GTP supplementation enhances the cellular thiol status thereby mitigate oral cancer.