Effect of steaming process on the structural characteristics and antioxidant activities of polysaccharides from Polygonatum sibiricum rhizomes

Polygonatum sibiricum (P. sibiricum) rhizomes are widely used as a tonic and functional food, and are often processed to enhance their tonic function by repeated steaming and drying. As the most important constituent, the polysaccharide from P. sibiricum rhizomes (PSP) has demonstrated various activities, but the alteration of structural characteristics and activities of the purified PSPs during steaming process was rarely investigated. To well understand the effect of steaming process on the polysaccharides of P. sibiricum, neutral polysaccharides from P. sibiricum rhizomes (PSP0?~?PSP9) after steaming were first isolated and purified, and then the chemical properties and antioxidant activities were determined. The results showed that the molar ratios of monosaccharides in PSPs were different. The molecular weights of PSPs were increased significantly after the fourth steaming. Morphological studies showed that the surface of PSPs became much tighter during the steaming process. Fourier transform infrared spectroscopy spectra displayed the polysaccharides had similar backbones and chemical groups. Furthermore, the antioxidant activity of PSPs was measured through radical scavenging tests. It was found that the radical scavenging activity of PSPs was elevated strikingly after steaming, and increased gradually with numbers of steaming process. The biological and chemical variance of PSPs revealed considerable segregation of PSP0, PSP1?~?PSP4 and PSP5?~?PSP9. In conclusion, our results proposed the fourth time as the optimal number of steaming to extract functional polysaccharide from P. sibiricum rhizomes.

     

In situ measurements of bioluminescence response of Gonyaulax spinifera to various mechanical stimuli

A conspicuous bioluminescence during nighttime was reported in an aquaculture farm in the Cochin estuary due to Gonyaulax spinifera bloom on March 20, 2020. In situ measurements on bioluminescence was carried out during nighttime to quantify the response of G. spinifera to various mechanical stimuli. The bioluminescence intensity (BI) was measured using Glowtracka, an advanced single channel sensor, attached to a Conductivity–Temperature–Depth Profiler. In steady environment, without any external stimuli, the bioluminescence generated due to the movement of fishes and shrimps in the water column was not detected by the sensor. However, stimuli such as a hand splash, oar and swimming movements, and a mixer could generate measurable bioluminescence responses. An abundance of?~?2.7?×?10⁶ cells L^?1 of G. spinifera with exceptionally high chlorophyll a of 25 mg m^?3 was recorded. The BI in response to hand splash was recorded as high as 1.6?×?10¹¹ photons cm^?2 s^?1. Similarly, BI of?~?1–6?×?10¹⁰ photons cm^?2 s^?1 with a cumulative bioluminescence of?~?2.51?×?10¹² photons cm^?2 (for 35 s) was recorded when there is a mixer with a constant force of 494 N/800 rpm min^?1. The response of G. spinifera was spontaneous with no time lapse between application of stimuli and the bioluminescence response. Interestingly, in natural environment, application of stimulus for longer time periods (10 min) does not lower the bioluminescence intensity due to the replenishment of water thrusted in by the mixer from surrounding areas. We also demonstrated that the bioluminescence intensity decreases with increase in distance from the source of stimuli (mixer) (av. 1.84?×?10¹⁰ photons cm^?2 s^?1 at 0.2 m to av. 0.05?×?10¹⁰ photons cm^?2 s^?1 at 1 m). The BI was highest in the periphery of the turbulent wake generated by the stimuli (av. 3.1?×?10¹⁰ photons cm^?2 s^?1) compared to the center (av. 1.8?×?10¹⁰ photons cm^?2 s^?1). When the stimuli was applied vertically down, the BI decreased from 0.2 m (0.3?×?10¹⁰ photons cm^?2 s^?1) to 0.5 m (0.10?×?10¹⁰ photons cm^?2 s^?1). Our study demonstrates that the BI of G. spinifera increases with increase in mechanical stimuli and decreases with increase in distance from the stimuli.

     

Remediation of Explosive-Contaminated Soils: Alkaline Hydrolysis and Subcritical Water Degradation

Alkaline hydrolysis and subcritical water degradation were investigated as ex-situ remediation processes to treat explosive-contaminated soils from military training sites in South Korea. The addition of NaOH solution to the contaminated soils resulted in rapid degradation of the explosives. The degradation of explosives via alkaline hydrolysis was greatly enhanced at pH ≥12. Estimated pseudo-first-order rate constants for the alkaline hydrolysis of 2,4-dinitrotoluene (DNT), 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in contaminated soil at pH 13 were (9.6?±?0.1)×10^?2, (2.2?±?0.1)×10^?1, and (1.7?±?0.2)×10^?2 min^?1, respectively. In the case of subcritical water degradation, the three explosives were completely removed at 200–300°C due to oxidation at high temperatures and pressures. The degradation rate increased as temperature increased. The pseudo-first-order rate constants for DNT, TNT, and RDX at 300°C were (9.4?±?0.8)×10^?2, (22.8?±?0.3)×10^?2, and (16.4?±?1.0)×10^?2, respectively. When the soil-to-water ratio was more than 1:5, the extent of alkaline hydrolysis and subcritical water degradation was significantly inhibited.     

Immunochemistry of the Lewis-blood-group system: Partial characterization of Lea-, Leb-, and H-type 1 (LedH)-blood-group active glycosphingolipids from human plasma

Four different H-type 1 (Le^dH) blood-group-active glycosphingolipids (Le^dH-I–IV) have been isolated from the plasma of blood-group O Le(a?b?) secretors. The agglutination of O Le(a?b?) erythrocytes from secretors by 50 μl of 4 hemagglutinating units of caprine anti-Le^dH (anti-H-type 1) serum was inhibited by 0.02 μg of each of all four glycolipids. No Le^a or Le^b activities or reaction against Ulex europaeus lectin could be found. Le^dH-I, -II, -III, and -IV at 0.05, 0.01, 0.01, and 0.02 μg each are sufficient for incubation in order to convert 9 × 10⁷ O Le(a?b?) erythrocytes from nonsecretors into H-type 1 (Le^dH)-positive cells. Structural analysis of the H-type 1 glycolipids was performed in comparison to that of Le^a- and Le^b-blood-group-active glycolipids from human plasma isolated previously: Gas chromatography of peracetylated alditols revealed sugar composition. Combined gas chromatography-mass spectrometry established the glycosidic linkages. Together with the results obtained by direct inlet mass spectrometry of permethylated glycosphingolipids and by 360-MHz ¹H nuclear magnetic resonance spectroscopy (Egge, H., and Hanfland, P., 1981, Arch. Biochem. Biophys., 210, 396–404; Dabrowski, J., Hanfland, P., Egge, H., and Dabrowski, U., 1981, Arch. Biochem. Biophys., 210, 405–411) the complete structures of the oligosaccharide chains of the Le^a-, Le^b-, and H-type 1-active glycolipids were established: Galβ1 → 3GlcNAc(4 ← 1αFuc)β1 → 3Galβ1 → 4Glcβ1 → 1 Cer for the Le^a antigens; Fucα1 → 2Galβ1 → 3GlcNAc(4 ← 1αFuc)β1 → 3Galβ1 → 4Glcβ1 → 1 Cer for the Le^b antigens; and Fucα1 → 2Galβ1 → 3GlcNAcβ1 → 3Galβ1 → 4Glcβ1 → 1 Cer for the H-type 1 (Le^dH) glycolipids. The diverse antigens of the same blood-group specificity obviously differ from one another in their lipid residue. In addition, plasmatic neolactotetraosylceramide could be identified, differing from that of human erythrocytes by a slower migration behavior in thin-layer chromatography.     

Biochemical characterization of recombinant β-carbonic anhydrase (PgiCAb) identified in the genome of the oral pathogenic bacterium Porphyromonas gingivalis

Abstract

Carbonic anhydrases (CAs, EC 4.2.1.1) belonging to the α-, β-, γ-, δ- and ζ-CAs are ubiquitous metalloenzymes present in prokaryotes and eukaryotes. CAs started to be investigated in detail only recently in pathogenic bacteria, in the search for antibiotics with a novel mechanism of action, since it has been demonstrated that in many such organisms they are essential for the life cycle of the organism. CA inhibition leads to growth impairment or growth defects in several pathogenic bacteria. The microbiota of the human oral mucosa consists of a myriad of bacterial species, Porphyromonas gingivalis being one of them and the major pathogen responsible for the development of chronic periodontitis. The genome of P. gingivalis encodes for a β- and a γ-CAs. Recently, our group purified the recombinant γ-CA (named PgiCA) which was shown to possess a significant catalytic activity for the reaction that converts CO₂ to bicarbonate and protons, with a k_cat of 4.1?×?10^5?s^?1 and a k_cat/K_m of 5.4?×?10^7?M^?1?×?s^?1. We have also investigated its inhibition profile with a range of inorganic anions such as thiocyanate, cyanide, azide, hydrogen sulfide, sulfamate and trithiocarbonate. Here, we describe the cloning, purification and kinetic parameters of the other class of CA identified in the genome of P. gingivalis, the β-CA, named PgiCAb. This enzyme has a good catalytic activity, with a k_cat of 2.8?×?10^5?s^?1 and a k_cat/K_m of 1.5?×?10^7?M^?1?×?s^?1. PgiCAb was also inhibited by the clinically used sulfonamide acetazolamide, with an inhibition constant of 214?nM. The role of CAs as possible virulence factors of P. gingivalis is poorly understood at the moment but their good catalytic activity and the fact that they might be inhibited by a large number of compounds, which may pave the way for finding inhibitors with antibacterial activity that may elucidate these phenomena and lead to novel antibiotics.     

ON THE STRUCTURE OF A NEW, FUCOSE CONTAINING GANGLIOSIDE FROM PIG CEREBELLUM

A new ganglioside, provisionally named GLIVa, was isolated in pure form from pig cerebellum. Ganglioside GLIVa is a disialoganglioside containing fucose. Its basic neutral glycosphingolipid core is the gangliotetraose ceramide: Gal, β 1 → 3 GalNAc, β 1 → 4 Gal, β 1 → 4 Glc, β 1 → Cer. Fucose is α-glycosidically linked to the 2-position of external galactose and one N-acetylneuraminic acid is linked to the other one by an α, 2 → 8 linkage. Thus the total structure of ganglioside GLIVa is the following: Fuc, α 1 → 2 Gal, β 1 → 3 GalNAc, β 1 → 4 (NeuAc, α 2 48 NeuAc, α 2 → 3) Gal, β 1 → 4 Glc, β 1 → Ceramide. According to the IUPAC-IUB Commission on Biochemical Nomenclature is indicated as II³α(NeuAc)₂ IV²αFuc-GgOse₄Cer.     

Temporal variability of algicidal and growth-inhibiting bacteria at an eelgrass bed in the Ariake Sea,Japan

Abstract

Temporal changes of algicidal and growth-inhibiting bacteria on the fish-killing raphidophyte flagellate, Chattonella antiqua, at an eelgrass (Zostera marina) bed in southern Ariake Sea, Japan in 2011 was investigated. The maximum value (5.1?×?10⁷ CFU g^?1 wet leaf) of algicidal bacteria (AB) was detected from a biofilm formed on Z. marina on August 1 when AB in the adjacent seawater had also peaked (1.2?×?10⁴ CFU mL^?1). Two causative bacteria isolated from the biofilm and seawater on August 1 were both identified to be of the genus Alteromonas (γ-proteobacteria). AB and growth-inhibiting bacteria (GIB) were present from the beginning of sampling (May 20) to August 26, fluctuating between 8.6?×?10² and 1.2?×?10⁴, 1.2?×?10³ and 9.3?×?10³ CFU mL^?1, respectively. The highest phytoplankton density observed was 6423 cells mL^?1 on September 29 and was comprised of centric diatoms such as Chaetoceros, Skeletonema, and Thalassiosira and coincided with the absence of AB and GIB where the decline of Z. marina was also observed. These findings provide a new ecological insight on AB and GIB associated with Z. marina beds, indicating eelgrass beds have the important role as the nursery of those bacteria that can be utilized as mitigation measures of harmful algal blooms (HABs) in the future.     

Histone-induced conformational changes in DNA as probed by quasi-elastic light scattering.

Quasi-elastic light scattering as measured by intensity fluctuation (self-beat) spectroscopy in the time domain can be profitably used to follow both the translational diffusion D and the dominant internal flexing mode τ_int of DNA and its complexes with various histones in aqueous salt solutions. Without histones, DNA is found to have D = 1.6 × 10^?8 cm²/sec and τ_int ? 5 × 10^?4 sec in 0.8 M NaCl, 2 M urea at 20°C. Total histone as well as fraction F2A induce supercoiling (D = 2.6 × 10^?8 cm²/sec, τ_int ? 2.8 × 10^?4 sec) whereas fraction F1 induces uncoiling (D = 1.0 × 10^?8 cm²/sec, τ_int ? 9.4 × 10^?4 sec). Upon increasing the salt concentration to 1.5 M the DNA–histone complex dissociates (D = 1.8 × 10^?8 cm²/sec). Upon decreasing the salt concentration to far below 0.8 M, the DNA–histone complex eventually precipitates as a chromatin gel.     

Utilization of oligo‐ and polysaccharides at microgram‐per‐litre levels in freshwater by Flavobacterium johnsoniae

Aims: To obtain a bacterial strain that can be used to quantify biodegradable polysaccharides at concentrations of a few micrograms per litre in freshwater. Methods and Results: Flavobacterium johnsoniae strain A3 was isolated from tap water supplemented with laminarin, pectin or amylopectin at 100 μg C l^?1 and river Rhine water. The organism utilized 14 of 23 oligo‐ and polysaccharides, and 1 of 9 monosaccharides, but none of the sugar acids, sugar alcohols, carboxylic acids or aromatic acids tested at 10 μg C l^?1. Amino acids promoted growth of strain A3, but not in coculture with assimilable organic carbon (AOC) test strain Pseudomonas fluorescens P17, which utilized these compounds more rapidly than strain A3. Compounds released by strain P17 and AOC test strain Spirillum sp. NOX grown on acetate promoted the growth of strain A3 at N_max values of ≥ 2 × 10⁵ CFU ml^?1 of strain P17 and ≥ 5 × 10⁵ CFU ml^?1 of strain NOX. Significant growth of strain A3 was observed in surface water and in tap water in the presence of strain P17 (N_max P17 < 2 × 10⁵ CFU ml^?1). Conclusions: Strain A3 utilizes oligo‐ and polysaccharides at microgram‐per‐litre levels. In surface water and in tap water, the organism was able to utilize compounds that were not utilized by strain P17. These compounds may include oligo‐ and/or polysaccharides. Significance and Impact of the Study: Phytoplanktonic and bacterial polysaccharides can constitute an important biodegradable fraction of natural organic matter in water and may promote growth of heterotrophic bacteria during water treatment and drinking water distribution. Strain A3 can be used to quantify a group of compounds that includes oligo‐ and polysaccharides at microgram‐per‐litre levels in freshwater.     

Evolution of 1, 3, 5-trisubstituted bipyrazole scaffold based platinum(II) complexes as a biological active agent

Abstract

Square planar mononuclear platinum(II) complexes having general formula [Pt(Lⁿ)Cl₂], (where, Lⁿ?=?L^1–4) were synthesized with neutral bidentate heterocyclic 1,3,5-trisubstituted bipyrazole based ligands. The synthesized compounds were characterized by physicochemical method such as TGA, molar conductance, micro-elemental analysis and magnetic moment, and spectroscopic method such as, FT-IR, UV–vis, ¹H NMR, ¹³C NMR and mass spectrometry. Biological applications of the compounds were carried out using in vitro brine shrimp lethality bioassay, in vitro antimicrobial study against five different pathogens, and cellular level cytotoxicity against Schizosaccharomyces pombe (S. Pombe) cells. Pt(II) complexes were tested for DNA interaction activities using electronic absorption titration, viscosity measurements study, fluorescence quenching technique and molecular docking assay. Binding constants (K_b) of ligands and complexes were observed in the range of 0.23–1.07?×?10⁵?M^?1 and 0.51–3.13?×?10⁵?M^?1, respectively. Pt(II) complexes (I–IV) display an excellent binding tendency to biomolecule (DNA) and possess comparatively high binding constant (K_b) values than the ligands. The DNA binding study indicate partial intercalative mode of binding in complex-DNA. The gel electrophoresis activity was carried out to examine DNA nuclease property of pUC19 plasmid DNA.     

Equilibrium studies on neutral red-DNA binding.

Spectrophotometric binding studies were undertaken on the interaction of neutral red with native and heat-denatured, sonicated, calf thymus DNA in a 0.2M ionic strength buffer containing Tris–sodium acetate–potassium chloride at 25°C. The pK_A of neutral red was found to be 6.81. At pH 5 the binding of protonated neutral red was complicated even at low concentration ratios of dye to DNA. In the pH range 7.5–8.5 the tight binding process could be studied and it was found that both protonated and free base species of neutral red significantly bind with DNA having association constants (in terms of polynucleotide phosphate) of 5.99 × 10³ M^?1 and 0.136 × 10³ M^?1, respectively, for native DNA and 7.48 × 10³ M^?1 and 0.938 × 10³ M^?1, respectively, for denatured DNA. The pK_A value of the neutral red–DNA complexes were 8.46 for native DNA and 7.72 for denatured DNA. These results are discussed in terms of possible binding mechanisms.     

Development of an experimental laboratory fertilization protocol for the South African abalone,Haliotis midae (Linnaeus 1758)

In this study, effective gamete concentrations, egg viability, and fertilization volumes were evaluated for Haliotis midae (L.). Sperm concentrations between 5?×?10³ and 5?×?10⁴?mL^?1 (p?>?0.05) consistently resulted in high hatch-out rates (96?±?1%). At concentrations higher than 5?×?10⁵?mL^?1, hatch-out rates decreased to 69?±?7% (p??1 resulted in high fertilization rates, with 50?eggs?mL^?1 being the ideal concentration for fertilization in H. midae. Egg viability was consistently high up to 100?min post-spawning, with a decrease in hatch-out success, when eggs were fertilized 120?min post-spawning. Fertilization volumes did not affect successful hatch-out. The results from this study can be implemented by South African abalone farms to increase hatch-out rates and subsequent culture. It can also be used as basis for the development of fertilization protocols in other marine invertebrate species.     

The molecular basis for cold agglutination: effect of receptor density upon thermal amplitude of a cold agglutinin.

Cold agglutinin MKV is a Waldenström macroglobulin that agglutinates human erythrocytes in the cold by binding $\text{N}$-acetylneuraminosyl-containing carbohydrate chains on their surfaces. Neuraminidase-treated cells are not agglutinated but their reactivity can be restored by allowing them to adsorb hematoside (NeuNAcα2-3Galβ1-4Glcβ1-ceramide). When between 7 × 10⁴ and 10⁶ molecules are adsorbed per cell, the cells are agglutinated at 0° but not at 37°. When over 10⁶ molecules of hematoside are adsorbed, they are agglutinated at both 0° and 37°. The density of receptors on the erythrocyte surface can thus influence the thermal amplitude of cold agglutinins.     

Chemical characterization of milk oligosaccharides of the koala (Phascolarctos cinereus)

Previous structural characterizations of marsupial milk oligosaccharides had been performed in only two macropod species, the tammar wallaby and the red kangaroo. To clarify the homology and heterogeneity of milk oligosaccharides among marsupial species, which could provide information on their evolution, the oligosaccharides of the koala milk carbohydrate fraction were characterized in this study. Neutral and acidic oligosaccharides were separated from the carbohydrate fraction of milk of the koala, a non-macropod marsupial, and characterized by ¹H-nuclear magnetic resonance spectroscopy. The structures of the neutral saccharides were found to be Gal(β1-4)Glc (lactose), Gal(β1-3)Gal(β1-4)Glc (3′-galactosyllactose), Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc (3′,3″-digalactosyllactose), Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (lacto-N-novopentaose I) and Gal(β1-3){Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6)}Gal(β1-4)Glc (fucosyl lacto-N-novopentaose I), while those of the acidic saccharides were Neu5Ac(α2-3)Gal(β1-4)Glc (3′-SL), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-4)Gal (sialyl 3′-galactosyllactose), Neu5Ac(α2-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose a), Gal(β1-3)[Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose b), Gal(β1-3)[Neu5Ac(α2-3)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose c), and Neu5Ac(α2-3)Gal(β1-3){Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6)}Gal(β1-4)Glc (fucosyl sialyl lacto-N-novopentaose a). The neutral oligosaccharides, other than fucosyl lacto-N-novopentaose I, a novel hexasaccharide, had been found in milk of the tammar wallaby, a macropod marsupial, while the acidic oligosaccharides, other than fucosyl sialyl lacto-N-novopentaose a had been identified in milk carbohydrate of the red kangaroo. The presence of fucosyl oligosaccharides is a significant feature of koala milk, in which it differs from milk of the tammar wallaby and the red kangaroo.     

Characterization of the Galactose-Specific Binding Activity of a Purified Soluble Entamoeba histolytica Adherence Lectin

ABSTRACT. We studied galactose (Gal)-specific binding of the soluble purified 260-kDa Entamoeba histolytica adherence protein to glycosylation deficient Chinese hamster ovary (CHO) cell mutants. Our goal was to further define the lectin's functional activity and carbohydrate receptor specificity. The adherence protein was purified by acid elution from an immunoaffnity column; however, exposure of the surface membrane lectin on viable trophozoites to identical acid pH conditions had no effect on carbohydrate binding activity. Saturable Gal-specific binding of soluble lectin to parental CHO cells was demonstrated at 4°C by radioimmunoassay; the dissociation coefficient (Kd was 2.39 × 10^?8 M^?1 with 5.97 × 10⁴ lectin receptors present per CHO cell. Gal-specific binding of lectin to Lec2 CHO cell mutants, which have increased N- and O-linked terminal Gal residues on cell surface carbohydrates, was increased due to an enhanced number of receptors (2.41 × 10⁵/cell) rather than a significantly reduced dissociation constant (4.93 × 10^?8 M^?1). At 4°C, there was no measurable Gal-specific binding of the adherence protein to the Lec and IdlD.Lecl CHO mutants, which contain surface carbohydrates deficient in terminal Gal residues. Binding of lectin (20 μg/ml) to CHO cells was equivalent at 4°C and 37°C and unaltered by adding the microfilament inhibitor, Cytochalasin D (10 μg/ml). Gal-specific binding of the lectin at 4°C was calcium independent and reduced by 81% following adsorption of only 0.2% of the lectin to CHO cells. In summary, these findings indicate that the purified E. histolytica adherence lectin demonstrates saturable Gal-specific binding to 1–6 branched-N-linked and not O-linked galactose terminal cell surface carbohydrates; however, apparently only a small percentage of purified amebic lectin molecules actually possess galactose binding activity.     

Hydrochemical,bacteriological assessment,and classification of groundwater quality in Thulamela Municipality,South Africa: potential health risk

Abstract

Potential effects of anthropogenic activities on different boreholes around Thulamela Municipality, South Africa, were evaluated by quantifying the bacteria indicators and physicochemical parameters during summer, autumn, and winter. The purpose was to determine whether the borehole water in this region is safe for domestic use across the seasons. The concentrations of physicochemical (Temperature, pH, electrical conductivity (EC), turbidity, and nitrate) and bacteriological (both Escherichia coli and Enterococcus faecalis) contaminants in the borehole water samples were determined using standard microbiology methods. The mean concentration of NO₃ for most of the boreholes failed to comply with the recommended guidelines throughout the season. High microbial load of E. coli (2.0?×?10¹ – 4.6?×?10³ CFU/100?ml) and E. faecalis (2.0?×?10° – 6.0?×?10² CFU/100?ml) was recorded in the wet season than in the dry season (0.0 – 7.0?×?10² and 0.0 – 1.0?×?10¹ CFU/100?ml, respectively). Sanitary inspection and water source classification showed that most boreholes are prone to chemical contaminants during summer and autumn due to rainfall and this correlates with the measured microbial contamination. The result is significantly important because water from these boreholes is used for domestic purposes without treatment and could pose major public health risks to the consumers.     

Understanding nucleosomal histone and DNA interactions: a biophysical study

Histones are associated with DNA to form nucleosome essential for chromatin structure and major nuclear processes like gene regulation and expression. Histones consist of H1, H2A, H2B and H3, H4 type proteins. In the present study, combined histones from calf thymus were complexed with ct DNA and their binding affinities were measured fluorimetrically. All the five histones were resolved on SDS page and their binding with DNA was visualized. The values of biding affinities varied with pH and salt concentration. Highest affinity (4.0?×?10⁵ M^?1) was recorded at pH 6.5 in 50 mM phosphate buffer and 1.5?×?10⁴ M^?1 in 2 M NaCl at pH 7.0. The CD spectra support the highest binding affinity with maximum conformational changes at pH 7.0. The time-resolved fluorescence data recorded two life times for histone tyrosine residues at 300 nm emission in phosphate buffer pH 6.5. These life times did not show much change upon binding with DNA in buffer as well as in 2 M NaCl. The isothermal calorimetric studies yielded thermodynamic parameters ΔG, ΔH and ΔS as ?1.6?×?10⁵ cal/mol, ?1.13?×?10³ cal/mol and ?3.80 cal/mol/deg, respectively, evidencing a spontaneous exothermic reaction. The dominant binding forces in building the nucleosome are electrostatic interactions.     

Unraveling the stability of plasma proteins upon interaction of synthesized uridine products: biophysical and molecular dynamics approach

Abstract

Most of the drugs binding to human serum albumin (HSA) are transported to various parts of the body. Here, we have studied the molecular interaction between HSA and synthesized uridine derivatives, 1-[(3R, 4S, 5?R)-2-methyl-3, 4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidine-2,4-dion.)(C-MU); [(2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxy-4-methyl-tetrahydrofuran-2-yl] methyl methyl phosphochloridate (CM-MU) and [(2R,3S,4R,5R)-5-(2,4-dioxopyrimidin-1-yl)-2-methyl-3,4-dihydroxyoxolan-2-yl] methyl dihydrogen phosphate (P-MU). Cytotoxic studies of these synthesized compounds with mouse macrophages (RAW 246.7) and HeLa cells (human cervical cancer cells) and binding mechanism of these uridine derivatives with HSA were performed. Subsequently, fluorescence quenching was observed upon titration of uridine derivatives with HSA via static mode of quenching, and the binding constants (K_2-C-MU = 4?±?0.03?×?10⁴M^?1, K_5-CM-MU = 1.95?±?0.03?×?10⁴ M^?1 and K_5-P-MU =1.56?±?0.03?×?10⁴ M^?1) were found to be in sync with the computational results. Further, molecular displacement and molecular docking data revealed that all the derivatives are binding in the subdomain IIA and IIB regions of HSA. The protein secondary structure of complexes was determined by circular dichroism, indicating partial unfolding of the protein upon addition of the uridine derivatives. Furthermore, atomic force microscopy data reveal the change in topology upon binding of 2-C-MU, 5-CM-MU and 5-P-MU with HSA, indicating change in the microenvironment around tryptophan region. Additionally, cytotoxicity studies on HeLa and Raw Cell lines suggested that these molecules have significant anti-proliferative and anti-inflammatory properties. Hence, the study may be of help for development of new drugs based on uridine derivatives which may be helpful for combating various potential diseases.

Communicated by Ramaswamy H. Sarma     

Evaluation of the pathogenicity of Aschersonia aleyrodis on Bemisia tabaci in the laboratory and greenhouse

The entomopathogenic fungus Aschersonia aleyrodis (Webber) is a promising fungal species against whiteflies. In this work, the pathogenicity of A. aleyrodis isolate Aa005 against MEAM1 Bemisia tabaci (Gennadius) was evaluated under laboratory and greenhouse conditions. The bioassay results indicated that the percentage of larval mortalities was concentration and age dependent. A. aleyrodis showed high pathogenicity against second and third instars and pupae with LC₅₀ values of 7.93?×?10⁶, 1.08?×?10⁷, and 1.56?×?10^7?conidia?mL^?1, respectively. The median lethal time (LT₅₀) was lower (4.60 days) for second instars and was the highest (6.17 days) for pupae when inoculated with a concentration of 1?×?10^7?conidia?mL^?1. Weekly sampling of immatures showed that the per cent mortality caused by A. aleyrodis at a conidial concentration of 1?×?10^7?conidia?mL^?1 was 71.21% in small nymphs, 69.31% in large nymphs and 53.36% in pupae. The dispersion index (DI) and Lloyd’s Index of Patchiness (LIP) values indicated that the infected immatures had a tendency to aggregate. The study demonstrated that A. aleyrodis isolate A005 is an effective biocontrol agent for B. tabaci control under laboratory and greenhouse conditions.     

On the impact of low concentrations of chlorophos in the period of early ontogenesis on digestive carbohydrases of underyearlings of roach Rutilus rutilus

The remote effects of brief exposure to low doses of chlorophos (1 × 10^?6 ?1 × 10^?2 mg/l) in the period of early ontogenesis of roach Rutilus rutilus have been studied. Diverse changes of the activity of carbohydrases and of the kinetic characteristics of the hydrolysis of di-and polysaccharides in experimental under-yearlings are revealed. Especially interesting are the data on the greater effect of superlow concentrations on changes of the kinetic characteristics; in some cases adaptive changes in the affinity of enzymes to the substrate are recorded.