Application of Two-and Three-Parameter Isotherm Models: Biosorption of Acid Red 88 onto Azolla microphylla

Biosorption potential of Azolla microphylla for acid red 88 from aqueous solution was investigated under laboratory conditions as a function of initial pH and temperature. The algal biomass exhibited the highest dye sorption capacity at optimum conditions of pH 3 and temperature 30°C. The experimental isotherms were analyzed using five two-parameter models (Langmuir, Freundlich, Temkin, Dubinin-Radushkevich, and Flory-Huggins) and five three-parameter models (Redlich-Peterson, Sips, Khan, Radke-Prausnitz, and Toth). Three error analysis methods were used to evaluate the experimental data: correlation coefficient, residual root mean square error (RMSE), and chi-square test to find the best fitting isotherm. In particular, Langmuir (two-parameter) and Khan (three-parameter) models described the dye biosorption isotherm data well at all pH and temperature conditions examined.     

Determining the degree of methylesterification of pectin by ATR/FT-IR: Methodology optimisation and comparison with theoretical calculations

The application of FT-IR to the study of the structure and interactions of the major plant-cell wall polysaccharide pectin has been reported for many decades. Nevertheless, here we show that the generally reported methodology for one of its most commonly utilised applications, the measurement of the degree of methylesterification (DM), requires careful interpretation and sample handling; including consideration of the moisture content and ionisation state. We propose instead a different methodology based on the assessment of the magnitude of C–H stretches in the methyl groups relative to those in the backbone and demonstrate experimentally the advantage of this method. In addition, we add a theoretical dimension to our work, performing full quantum chemical (DFT) calculations of monomeric-, dimeric-, and trimeric-pectic compounds, in various states of partial methylesterification. These extensive calculations not only confirm the identity of the proposed methyl-band and illustrate its scaling with DM; but also demonstrate the success of the theoretical approach. Thus, DFT calculations are expected to be a valuable tool in the interpretation of IR spectra obtained from more complex systems such as polysaccharide conjugates.     

Exploring wild genetic resources of Musa acuminata Colla distributed in the humid forests of southern Western Ghats of peninsular India using ISSR markers

Musa acuminata ssp. burmannica, one of the wild progenitors contributing 'A genome' to the present-day dessert bananas, has a long evolutionary history intervened by human activities. In this study, ISSR markers were used to analyze the pattern of genetic variation and differentiation in 32 individuals along with two reference samples (viz., Musa acuminata ssp. burmannicoides, var. Calcutta 4 and Musa balbisiana) of wild Musa, which corresponded to three populations across the biodiversity-rich hot spot of southern Western Ghats of India. High levels of genetic diversity were revealed both at the species and population levels, using Nei's diversity indices. The hierarchical analysis of molecular variance showed pronounced genetic differentiation, as 96?% of the total variance was fixed within population and only 4?% among populations. Nei's genetic differentiation coefficient (G (ST)?=?0.1823) and low gene flow (Nm?=?1.18) further confirmed this. The positive correlation (Mantel test) between geographic distance and genetic distance (r?=?0.338 P?相似文献   

C-tail mediated modulation of somatostatin receptor type-4 homo- and heterodimerizations and signaling

Somatostatin receptors show great diversity in response to agonist mediated receptor-specific homo- and heterodimerizations. Here, using photobleaching-fluorescence resonance energy transfer, immunocytochemistry, western blot and co-immunoprecipitation, we investigated dimerization, trafficking, coupling to adenylyl cyclase and signaling of human somatostatin receptor-4 (hSSTR4) in HEK-293 cells. We also determined the role of the C-tail of hSSTR4 on physiological responses of the cells. wt-hSSTR4 exogenously expressed in HEK-293 cells exhibits constitutive dimerization, inhibits forskolin-stimulated cAMP, and displays agonist dependent changes in pERK1/2 and pERK5 expressions. Upon C-tail deletion, the receptor loses membrane expression and ability to dimerize and inhibition of cAMP and pERK5 however, displays several-fold increases in the expression of pERK1/2. Chimeric hSSTR4 with the C-tail of hSSTR5 functions like wt-hSSTR4, in contrast, with the C-tail of hSSTR1 functions like C-tail deleted hSSTR4. hSSTR4 dimerization and signaling are associated with increased cyclin-dependent-kinase p27^kip1 expression and inhibition of the cell proliferation. We also report heterodimerization between hSSTR4/hSSTR5, but not between hSSTR4/hSSTR1, with significant changes in receptor functions. Taken together, these data define a novel mechanism for the role of hSSTR4 in cell proliferation and modulation of signaling pathways.     

The Effect of Tensile Stress on the Conformational Free Energy Landscape of Disulfide Bonds

Disulfide bridges are no longer considered to merely stabilize protein structure, but are increasingly recognized to play a functional role in many regulatory biomolecular processes. Recent studies have uncovered that the redox activity of native disulfides depends on their C–C–S–S dihedrals, and . Moreover, the interplay of chemical reactivity and mechanical stress of disulfide switches has been recently elucidated using force–clamp spectroscopy and computer simulation. The and angles have been found to change from conformations that are open to nucleophilic attack to sterically hindered, so–called closed states upon exerting tensile stress. In view of the growing evidence of the importance of C–C–S–S dihedrals in tuning the reactivity of disulfides, here we present a systematic study of the conformational diversity of disulfides as a function of tensile stress. With the help of force-clamp metadynamics simulations, we show that tensile stress brings about a large stabilization of the closed conformers, thereby giving rise to drastic changes in the conformational free energy landscape of disulfides. Statistical analysis shows that native TDi, DO and interchain Ig protein disulfides prefer open conformations, whereas the intrachain disulfide bridges in Ig proteins favor closed conformations. Correlating mechanical stress with the distance between the two –carbons of the disulfide moiety reveals that the strain of intrachain Ig protein disulfides corresponds to a mechanical activation of about 100 pN. Such mechanical activation leads to a severalfold increase of the rate of the elementary redox reaction step. All these findings constitute a step forward towards achieving a full understanding of functional disulfides.     

Transcranial Near-Infrared Laser Transmission (NILT) Profiles (800 nm): Systematic Comparison in Four Common Research Species

Background and Purpose

Transcranial near-infrared laser therapy (TLT) is a promising and novel method to promote neuroprotection and clinical improvement in both acute and chronic neurodegenerative diseases such as acute ischemic stroke (AIS), traumatic brain injury (TBI), and Alzheimer’s disease (AD) patients based upon efficacy in translational animal models. However, there is limited information in the peer-reviewed literature pertaining to transcranial near-infrared laser transmission (NILT) profiles in various species. Thus, in the present study we systematically evaluated NILT characteristics through the skull of 4 different species: mouse, rat, rabbit and human.

Results

Using dehydrated skulls from 3 animal species, using a wavelength of 800nm and a surface power density of 700 mW/cm2, NILT decreased from 40.10% (mouse) to 21.24% (rat) to 11.36% (rabbit) as skull thickness measured at bregma increased from 0.44 mm in mouse to 0.83 mm in rat and then 2.11 mm in rabbit. NILT also significantly increased (p<0.05) when animal skulls were hydrated (i.e. compared to dehydrated); but there was no measurable change in thickness due to hydration.In human calvaria, where mean thickness ranged from 7.19 mm at bregma to 5.91 mm in the parietal skull, only 4.18% and 4.24% of applied near-infrared light was transmitted through the skull. There was a slight (9.2-13.4%), but insignificant effect of hydration state on NILT transmission of human skulls, but there was a significant positive correlation between NILT and thickness at bregma and parietal skull, in both hydrated and dehydrated states.

Conclusion

This is the first systematic study to demonstrate differential NILT through the skulls of 4 different species; with an inverse relationship between NILT and skull thickness. With animal skulls, transmission profiles are dependent upon the hydration state of the skull, with significantly greater penetration through hydrated skulls compared to dehydrated skulls. Using human skulls, we demonstrate a significant correlation between thickness and penetration, but there was no correlation with skull density. The results suggest that TLT should be optimized in animals using novel approaches incorporating human skull characteristics, because of significant variance of NILT profiles directly related to skull thickness.     

Aldehyde oxidoreductase as a biocatalyst: Reductions of vanillic acid

Aldehyde oxidoreductase (carboxylic acid reductase) catalyzes the Mg(2+), ATP and NADPH dependent reduction of carboxylic acids to their corresponding aldehydes. The identification of the gene from Nocardia sp. NRRL 5646 and its expression in E. coli BL21-CodonPlus(?)(DE3)-RP/pHAT305 provided an avenue to develop a biocatalyst for reduction of carboxylic acids. In addition to aromatic acids, the recombinant carboxylic acid reductase also accepts several aliphatic mono, di and tri carboxylic acids as substrates. A recently identified Nocardia sp., phosphopantetheinyl transferase gene (npt) enhanced the activity of carboxylic acid reductase. Coexpression of car and npt in E. coli BL21-CodonPlus(?)(DE3)-RP/pPV2.83 resulted in a purified recombinant carboxylic acid reductase with improved specific activity of 2.2U/mg protein. The utility of the recombinant carboxylic acid reductase as a biocatalyst has been demonstrated using vanillic acid as substrate. E. coli BL21-CodonPlus(?)(DE3)-RP/pHAT305 expressing Car reduced 50% of vanillic acid to vanillin in 10h. E. coli BL21-CodonPlus(?)(DE3)-RP/pPV2.83 resting cells expressing Car and Npt reduced 90% of vanillic acid to vanillin in 6h. Enhanced, in vivo cofactor NADPH regeneration by glucose dehydrogenase (gdh) was accomplished using E. coli BL21-CodonPlus(?)(DE3)-RP/pPV2.85, that carried car, npt, and gdh. Resting cell reactions using E. coli BL21-CodonPlus(?)(DE3)-RP/pPV2.85 with in situ product removal by XAD-2 resin efficiently reduced 5g/L of vanillic and benzoic acids within 2h.     

Evidence for Methylerythritol Pathway (MEP) Contributions to Zerumbone Biosynthesis as Revealed by Expression Analysis of Regulatory Genes and Metabolic Inhibitors Studies

Plant Molecular Biology Reporter - Sesquiterpenoid Zerumbone, the principal secondary metabolite in Zingiber zerumbet Smith, has been identified as the putative molecule conferring resistance...     

Estimation of genetic diversity in varieties of Mucuna pruriens using RAPD

Genetic diversity was estimated in 13 accessions of the otherwise self pollinated Mucuna pruriens (L.) DC. (velvetbean) comprising varieties pruriens and utilis collected from tropical humid forest using 15 RAPD primers. Similarity index value of 0.68 based on Nei and Li's similarity coefficient indicated high degree of genetic variability. Analysis of various genetic diversity indices like total heterozygosity, Nei's gene diversity, percentage of polymorphic loci, expected and observed number of alleles and Shannon index strongly suggests that variety pruriens is genetically more diverse than variety utilis. Chemical analysis with respect to 3,4-dihydroxy-L-phenylalanine (L-DOPA) content showed uniform distribution. Cluster analysis showed grouping of accessions into two major clusters and tendency of accessions of variety pruriens to group according to their geographical locations. Bootstrap analysis confirmed the robustness of the phenogram. The putative hybrid MMP6 with relatively low similarity value index and low L-DOPA content was promising as food or fodder.