Efficient in vitro propagation of <Emphasis Type="Italic">Artemisia nilagirica</Emphasis> var. <Emphasis Type="Italic">nilagirica</Emphasis> (Indian wormwood) and assessment of genetic fidelity of micropropagated plants

A reliable protocol has been established for in vitro propagation of Artemisia nilagirica var. nilagirica (Indian wormwood), a valuable medicinal plant from India. A highly proliferating organogenic callus was obtained on Murashige and Skoog (MS) medium supplemented with 2.5 µM IAA when nodal explants were cultured on MS medium supplemented with various growth regulators. Further, highest regeneration frequency (83.3 %) of adventitious shoots was observed, when the callus was sub-cultured on MS medium supplemented with 6-benzylaminopurine (BAP; 2.5 µM) along with 7.5 µM 2-isopentenyl adenine (2-iP). An optimal of 10.16 ± 2.24 shoots were regenerated on medium supplemented with 2.5 µM BAP + 7.5 µM 2-iP. Quarter strength MS medium supplemented with 10 µM IBA was effective for rooting of the shoots. Ex-vitro plants were normal and were established successfully. Cytological and molecular marker studies showed that regenerated plants showed genetic stability in micro-propagated plants.     

Heavy metal (Pb) accumulation in metallophytes as influenced by the variations in rhizospheric and non-rhizospheric soils physico-chemical characteristics

Activities at root-soil interface determine the solubility and uptake of metals by plants. Metal accumulation in plant species (Imperata cylindrical, Cynodon dactylon, Eleucine indica, Gomphrena celosoides, Sporobolus pyramidalis, Chromolaena odorata and Rhynchospora corymbosa) growing on Pb contaminated site as influenced by variations in physico-chemical characteristics, dissolved organic matter (DOM), Pb fractionation and different functional groups (using Fourier Transmittance Infra-red) of rhizospheric and non-rhizospheric soils was assessed. The electrical conductivity (2660–5520 µs) and Pb concentrations (51390.0–64080.0 mg/kg) were more in non-rhizospheric than rhizospheric soils having 276 µs to 3160 µs EC and 3289.0 to 44850.0 mg/kg Pb. More nutrients, DOM and carbohydrates functional groups (C-O; 1100 -1000 and O-H; 3700–3600) were found in rhizospheric compared to non-rhizospheric soils. The pH was slightly acidic (5.0–5.54) and E. indica with the lowest pH (5.0) accumulated highest Pb concentrations in shoot (8030 mg/kg) and root (16380 mg/kg) while C. odorata with highest values of pH, P, Ca and Mg in rhizospheric soil accumulated the least (root; 331.6 and shoot: 209.0 mg/kg). Pb was more in organic and residual fractions of rhizospheric and non-rhizospheric soils respectively. Reduction in pH, EC coupled with nutrients and DOM availability increased Pb uptake by plants.     

High Throughput Discovery of Solar Fuels Photoanodes in the CuO–V2O5 System

Solar photoelectrochemical generation of fuel is a promising energy technology yet the lack of an efficient, robust photoanode remains a primary materials challenge in the development and deployment of solar fuels generators. Metal oxides comprise the most promising class of photoanode materials, but no known material meets the demanding requirements of low band gap energy, photoelectrocatalysis of the oxygen evolution reaction (OER), and stability under highly oxidizing conditions. Here, the identification of new photoelectroactive materials is reported through a strategic combination of combinatorial materials synthesis, high‐throughput photoelectrochemistry, optical spectroscopy, and detailed electronic structure calculations. Four photoelectrocatalyst phases, α ‐Cu₂V₂O₇, β ‐Cu₂V₂O_7, γ ‐Cu₃V₂O₈, and Cu₁₁V₆O₂₆, are reported with band gap energy at or below 2 eV. The photoelectrochemical properties and 30 min stability of these copper vanadate phases are demonstrated in three different aqueous electrolytes (pH 7, pH 9, and pH 13), with select combinations of phase and electrolyte exhibiting unprecedented photoelectrocatalytic stability for metal oxides with sub‐2 eV band gap. Through integration of experimental and theoretical techniques, new structure‐property relationships are determined and establish CuO–V₂O₅ as the most prominent composition system for OER photoelectrocatalysts, providing crucial information for materials genomes initiatives and paving the way for continued development of solar fuels photoanodes.     

Identification of a pH sensor in the furin propeptide that regulates enzyme activation

The folding and activation of furin occur through two pH- and compartment-specific autoproteolytic steps. In the endoplasmic reticulum (ER), profurin folds under the guidance of its prodomain and undergoes an autoproteolytic excision at the consensus furin site Arg-Thr-Lys-Arg107/ generating an enzymatically masked furin-propeptide complex competent for transport to late secretory compartments. In the mildly acidic environment of the trans-Golgi network/endosomal system, the bound propeptide is cleaved at the internal site 69HRGVTKR75/, unmasking active furin capable of cleaving substrates in trans. Here, by using cellular, biochemical, and modeling studies, we demonstrate that the conserved His69 is a pH sensor that regulates the compartment-specific cleavages of the propeptide. In the ER, unprotonated His69 stabilizes a solvent-accessible hydrophobic pocket necessary for autoproteolytic excision at Arg107. Profurin molecules unable to form the hydrophobic pocket, and hence, the furin-propeptide complex, are restricted to the ER by a PACS-2- and COPI-dependent mechanism. Once exposed to the acidic pH of the late secretory pathway, protonated His69 disrupts the hydrophobic pocket, resulting in exposure and cleavage of the internal cleavage site at Arg75 to unmask the enzyme. Together, our data explain the pH-regulated activation of furin and how this His-dependent regulatory mechanism is a model for other proteins.     

Reconstitution of allogeneic hemopoietic stem cells: the essential role of FcR gamma and the TCR beta-chain-FCp33 complex

Transplantation of purified allogeneic hemopoietic stem cells (SC) alone is characterized by a decreased risk of graft-vs-host disease but increased incidence of engraftment failure. It has been established that the facilitating cell (FC) promotes allogeneic SC reconstitution and results in donor-specific transplantation tolerance across MHC disparities, without graft-vs-host disease. Although the requirements for this facilitating function are not well-characterized, it is known that facilitation is dependent on FC expression of a unique heterodimer consisting of the TCR beta-chain (TCRbeta) and a 33-kDa protein, FCp33. The current study confirms that CD3epsilon and TCRbeta expression are present on the FC at the time of transplantation and demonstrates that the majority of cells in the FC population express the TCR signaling molecule, FcRgamma, rather than the more conventional CD3zeta receptor. Of particular significance, we have now demonstrated that FC-mediated allogeneic SC reconstitution is critically dependent on FcRgamma expression and that FcRgamma coprecipitates with the TCRbeta-FCp33 heterodimer. The mandatory requirement of TCRbeta and FcRgamma for FC function provides the first evidence of a previously undescribed role for FcRgamma in the facilitation of allogeneic SC reconstitution and establishes that FcRgamma is part of the TCRbeta-FCp33 complex uniquely expressed on FC.     

Synthesis of antimicrobial 2,9,10-trisubstituted-6-oxo-7,12-dihydro-chromeno[3,4-b]quinoxalines

A new series of 2,9,10-trisubstituted-6-oxo-7,12-dihydro-chromeno[3,4-b]quinoxalines was synthesized and submitted to antibacterial and antifungal activities. Result of the antimicrobial screening showed the compound 4j being the most effective among the various treatments in antimicrobial screening. Compounds 4c, 4d, 4k, and 4l showed moderate activity against the microorganisms tested.     

A role for SSRP1 in recombination‐mediated DNA damage response

A possible role for structure‐specific recognition protein 1 (SSRP1) in replication‐associated repair processes has previously been suggested based on its interaction with several DNA repair factors and the replication defects observed in SSRP1 mutants. In this study, we investigated the potential role of SSRP1 in association with DNA repair mediated by homologous recombination (HR), one of the pathways involved in repairing replication‐associated DNA damage, in mammalian cells. Surprisingly, over‐expression of SSRP1 reduced the number of hprt⁺ recombinants generated via HR both spontaneously and upon hydroxyurea (HU) treatment, whereas knockdown of SSRP1 resulted in an increase of HR events in response to DNA double‐strand break formation. In correlation, we found that the depletion of SSRP1 in HU‐treated human cells elevated the number of Rad51 and H2AX foci, while over‐expression of the wild‐type SSRP1 markedly reduced HU‐induced Rad51 foci formation. We also found that SSRP1 physically interacts with a key HR repair protein, Rad54 both in vitro and in vivo. Further, branch migration studies demonstrated that SSRP1 inhibits Rad54‐promoted branch migration of Holliday junctions in vitro. Taken together, our data suggest a functional role for SSRP1 in spontaneous and replication‐associated DNA damage response by suppressing avoidable HR repair events. J. Cell. Biochem. 108: 508–518, 2009. © 2009 Wiley‐Liss, Inc.     

Optimized production of isoflavones in cell cultures of Psoralea corylifolia L. Using elicitation and precursor feeding

The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in cell suspension cultures of corylifolia L. Untreated cell suspension (control) accumulated 1.66% dry wt of daidzein and 0.165% dry wt of genistein. In precursor feeding experiment, phenylalanine at 0.5 mM concentration led to 1.3 fold higher production of daidzein (1.99% dry wt) and genistein (0.22% dry wt). In biotic elicitors, yeast extract (100 mg/L) was found to be the most efficient elicitor to induce higher production levels of daidzein (2.21% dry wt) and genistein (0.293% dry wt) in suspension cultures. Salicylic acid (signaling molecule) at 1 mM concentration stimulated the maximum accumulation of daidzein (3.4% dry wt) and genistein (0.41% dry wt) 2 days after elicitation. In case of polyamines, spermidine (100 mM) resulted in highest accumulation of daidzein (3.2% dry wt) and genistein (0.475% dry wt) after 7 days of addition, which was 2.4 fold of that in control. This is the first report on kinetics of isoflavone production in response to elicitation in cell suspension of P. corylifolia.     

Enhanced production of phytoestrogenic isoflavones from hairy root cultures of Psoralea corylifolia L. Using elicitation and precursor feeding

The effect of biotic elicitors (yeast extract, chitosan), signaling molecule (salicylic acid), and polyamines (putrescine and spermidine) was studied with respect to isoflavones accumulation in hairy root cultures of Psoralea corylifolia L. Untreated hairy roots (control) accumulated 1.55% dry wt of daidzein and 0.19% dry wt of genistein. In precursor feeding experiment, phenylalanine at 2 mM concentration led to 1.3 fold higher production of daidzein (1.91% dry wt) and genistein (0.27% dry wt). In biotic elicitors, chitosan (2 mg/L) was found to be the most efficient elicitor to induce daidzein (2.78% dry wt) and genistein (0.279% dry wt) levels in hairy roots. Salicylic acid at 1 mM concentration stimulated the maximum accumulation of daidzein (2.2% dry wt) and genistein (0.228% dry wt) 2 days after elicitation. In case of polyamines, putrescine (50 mM) resulted in highest accumulation of daidzein (3.01% dry wt) and genistein (0.227% dry wt) after 5 days of addition. Present results indicated the effectiveness of elicitation and precursor feeding on isoflavones accumulation in hairy roots of P. corylifolia. This is the first report of elicitation on isoflavones production by hairy roots of P. corylifolia.     

Synthesis and anti-inflammatory activity of some 3-(4,6-disubtituted-2-thioxo-1,2,3,4-tetrahydropyrimidin-5-yl) propanoic acid derivatives

A series of 3-(4,6-disubtituted-2-thioxo-1,2,3,4-tetrahydropyrimidin-5-yl) propanoic acid derivatives has been synthesized by condensation of thiourea, 5-(4-subtituted phenyl)-5-oxopentanoic acid and substituted aldehyde. The synthesized compounds were screened for their anti-inflammatory activity using rat paw edema method. Most of the compounds from the series showed significant (p <0.05) anti-inflammatory activity.