Mutation detection analysis of a region of 16S-like ribosomal RNA gene of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii

<p>Abstract</p> <p>Background</p> <p>The level of intra-species genetic variation in <it>Entamoeba histolytica, Entamoeba dispar </it>and <it>Entamoeba moshkovskii </it>populations in a localized geographic area, like Puducherry, India, remains unknown.</p> <p>Methods</p> <p>In the present study the existence of genetic variation in the nested multiplex polymerase chain reaction (NM-PCR) amplified region of the 16S-like ribosomal RNA genes of <it>E. histolytica, E. dispar </it>and <it>E. moshkovskii </it>was investigated by riboprinting and single strand conformation polymorphism (SSCP) analysis.</p> <p>Results</p> <p>We found that 70 stool specimens were positive for <it>E. histolytica</it>, 171 stool specimens were positive for <it>E. dispar</it>, and 37 stool specimens were positive for <it>E. moshkovskii </it>by NM-PCR. Ninety liver abscess pus specimens, 21 urine specimens, and 8 saliva specimens were positive for <it>E. histolytica </it>by NM-PCR. Riboprinting analysis detected a mutation in the PCR product of only one <it>E. histolytica </it>isolate from a stool specimen. However, SSCP analysis detected mutations in the PCR products of five <it>E. histolytica </it>isolates and three <it>E. moshkovskii </it>isolates from stool specimens, and one <it>E. histolytica </it>isolate from a saliva specimen. The mutations detected by riboprinting and SSCP analysis were confirmed by sequencing. All the nucleotide sequences showing mutations in this study have already been deposited into the NCBI GenBank database under accession numbers [GenBank: <ext-link ext-link-type="gen" ext-link-id="EF682200">EF682200</ext-link> to GenBank: <ext-link ext-link-type="gen" ext-link-id="EF682208">EF682208</ext-link>].</p> <p>Conclusion</p> <p>The present study has revealed the subsistence of mutations in the ribosomal RNA genes of <it>E. histolytica </it>and <it>E. moshkovskii</it>, which points towards the existence of intra-species genetic variation in <it>E. histolytica </it>and <it>E. moshkovskii </it>isolates infecting humans.</p

Dyeing of wool and silk with tea

Silk and wool fabrics were coloured employing aqueous tea extract in absence and presence of magnesium sulphate, aluminium sulphate and ferrous sulphate as the mordanting agents. Colour uptake for wool was found to be more than that for silk under all conditions studied. Colouring biochemical components of tea revealed highest affinity for both the protein fibres at pH 2 to 4 in presence and absence of such mordanting agents. Use of ferrous sulphate and aluminium sulphate produced significant improvement in colour uptake following a pre- or post mordanting methods. Coloured protein fibres in general produced a light-fastness rating of 5 on a scale of 1- 5 and a wash-fastness rating of 4 on a scale of 1-5. Ferrous sulphate and aluminium sulphate improved colour retention on washing and the fastness properties further. Coloured protein fibres became blackish, when ferrous sulphate was employed as mordanting agent.This item was scanned with a HP 4850 Scanjet at 300 dpi and consists of 9 pages

Dyeing of wool and silk with <i>Punica granatum</i>

559-564The application of dye obtained from Punica granatum fruit rind on wool and silk fabric in the presence and absence of environment-friendly mordanting agents has been studied. The dyeing of silk and wool with pomegranate solution is found to be effectively accomplished at pH 4.0. Pre- and post-mordanting employing ferrous sulphate and aluminium sulphate improve the colour uptake, light fastness and colour retention on repeated washing. The use of such mordants, however, does not improve wash fastness property of dyed substrates

Colouration of wool and silk with Rheum emodi

163-170Silk and wool fabrics have been dyed with colourant extracted from Rheum emodi in the absence and presence of magnesium sulphate, aluminium sulphate and ferrous sulphate mordants for producing shades of different colours, ranging from yellow to olive green. Colouring component of Rheum emodi has close resemblance with a typical anthraquinonoid type disperse dye; the uptake of such colouring component by the protein fibres remains unaffected within a wide pH range of 4 – 8. Colour uptake, rate of dyeing and affinity of colour are found to be more for silk than that for wool under all the conditions studied. The dyeing mechanism corresponds to the partition mechanism, confirming that this anthraquinonoid-based colourant is adsorbed by silk and wool fibres as a disperse dye and the dyeing process is endothermic. The use of ferrous sulphate and aluminium sulphate produces significant improvement in depth of shade, when both the substrates are treated with such salts prior to application of the colourant. Coloured protein fibres, in general, show a common light fastness and wash fastness ratings of 4 and 3 respectively. Ferrous sulphate, however, improves the colour fastness properties and colour retention on washing of both wool and silk fabrics further

Active site mapping of porcine liver esterase (PLE) and porcine pancreatic lipase (PPL): Stereo and regiochemical outcome of hydrolysis of conformationally constrained esters†

974-980The regio and stereoselectivity of PLE and PPL-catalyzed hydrolysis of a number of conformationally constrained substrates have been studied. The results are discussed based on the existing active site models for the two enzymes

Exploring the interaction of phenothiazinium dyes methylene blue, new methylene blue,azure A and azure B with tRNA Phe: spectroscopic, thermodynamic, voltammetric and molecular modeling approach

This study focuses on the understanding of the interaction of phenothiazinium dyes methylene blue (MB), new methylene blue (NMB), azure A (AZA) and azure B (AZB) with tRNAPhe with particular emphasis on deciphering the mode and energetics of the binding. Strong intercalative binding to tRNAPhe was observed for MB, NMB and AZB, bound by a partial intercalative mode. AZA has shown groove binding characteristics. From spectroscopic studies binding affinity values of the order of 105 M�1 were deduced for these dyes; the trend varied as MB 4 NMB 4 AZB 4 AZA. The binding was characterized by an increase of thermal melting temperatures and perturbation in the circular dichroism spectrum of tRNA. All the dyes acquired optical activity upon binding to tRNA. The binding was predominantly entropy driven with a favorable enthalpy term that increased with temperature in all the cases. Dissection of the Gibbs energy to polyelectrolytic and non-polyelectrolytic terms revealed a major role of the nonelectrostatic forces in the binding. The small but significant heat capacity changes and the observed enthalpy–entropy compensation phenomenon confirmed the involvement of multiple weak noncovalent forces driving the interaction. The mode of binding was confirmed from quenching, viscosity and cyclic voltammetric results. Using density functional theory, ground state optimized structures of the dyes were calculated to provide insight into theoretical docking studies to correlate the experimental approaches. The modeling results verified the binding location as well as the binding energy of complexation. The results may provide new insights into the structure–activity relationship useful in the design of effective RNA targeted therapeutic agents

Single step aqueous synthesis of pure rare earth nanoparticles in biocompatible polymer matrices

The room temperature synthesis of water soluble, stable rare earth (RE) metal nanoparticles (MNPs) with controlled size is a long standing interest. In the present work, we have established a synthetic strategy for the preparation of pure europium (Eu-0) metal nanoparticles (NPs) in aqueous solution employing a gamma-radiolytic reduction technique. Since radiolysis is the cleanest method amongst all other chemical routes, we preferentially choose this technique for the reduction of precursor Eu3+ ions to nanoscale metals in our work. This has been possible as hydrated electrons (e(aq)(-)) having a very high reduction potential (E-0(H2O/e(aq)(-)) = -2.87 V-NHE) produced in situ can efficiently reduce Eu3+ to Eu-0. Synthesized Eu-0 MNPs were stabilised within the matrices of biocompatible polymers, polyvinyl alcohol (PVA) and polyvinylpyrrolidone (PVP). Reduction of the metal ion has been conducted at different irradiation doses with a maximum dose of 83.88 kGy. The irradiated solution shows an absorption maximum at 266 +/- 2 nm and an emission maximum at 394 +/- 5 nm. Analysis of transmission electron microscopy (TEM) images shows that the average sizes of PVA and PVP encapsulated Eu-0 NPs are 13 +/- 0.6 nm and 17 +/- 1.01 nm, respectively ([Eu3+] = 5.0 x 10(-3) mol dm(-3), [polymer] = 1.0%). Formation of monodisperse pure Eu-0 MNPs was further characterised by dynamic light scattering (DLS), energy dispersive X-ray (EDX) as well as Fourier transformed infrared (FTIR) spectroscopy and cyclic voltammetry (CV) studies

Aggregation-Induced Fabrication of Fluorescent Organic Nanorings: Selective Biosensing of Cysteine and Application to Molecular Logic Gate

Self-aggregation behavior in aqueous medium of four naphthalimide derivatives has exhibited substitution-dependent, unusual, aggregation induced emission enhancement (AIEE) phenomena. Absorption, emission, and time-resolved study initially indicated the formation of J-type fluorescent organic nanoaggregates (FONs). Simultaneous applications of infrared spectroscopy, theoretical studies, and dynamic light scattering (DLS) measurements explored the underlying mechanism of such substitution-selective aggregation of a chloro-naphthalimide organic molecule. Furthermore, transmission electron microscopy (TEM) visually confirmed the formation of ring like FONs with average size of 7.5–9.5 nm. Additionally, naphthalimide FONs also exhibited selective and specific cysteine amino acid sensing property. The specific behavior of NPCl aggregation toward amino acids was also employed as a molecular logic gate in information technology (IT)

Deciphering the Role of the Length of the Corona in Controlled NSET within Triblock Copolymers

Nanometal surface energy transfer (NSET) from 2-anthracene sulfonate (2-AS) to gold nanoparticles in water-soluble triblock copolymers (TBP) P123 (PEO₁₉PPO₆₉PEO₁₉) and F127 (PEO₁₀₀PPO₆₅PEO₁₀₀) is systematically investigated. Fluorescence lifetime and anisotropy experiments provide thorough information on the locations of the 2-AS within these micelles. Variation in the size of the micellar corona region of the TBP is found to be the prime factor for different positions of 2-AS in them. Of late, nanometal surface energy transfer (NSET) from the donor probe to the surface of the metal nanoparticles has emerged as a potential tool for sensing and biolabeling. In the present work, the quenching of emission of the water-soluble 2-AS confined in the two different triblock copolymers in the proximity of a monolayer of the gold nanoparticles has been explored. Closer agreement between the experimental and theoretical characteristic distances has been found across the full wavelength range by the NSET approach. Understanding the location of the water-soluble dye in the vicinity of a polymeric drug delivery system is of significant importance, and how altered locations can trigger different controlled energy transfer efficiency from the 2-AS to the surfaces of gold nanoparticles (GNPs) has been discussed. This strategy could offer a new prospect in designing novel optical materials for chemical sensing and light harvesting endeavors