Immunoadsorbents with synthetic oligosaccharide hapten representing blood group A substances

Immunoadsorbents with a synthetic oligosaccharide hapten representing human blood group A specific substances are prepared. The synthetic hapten, known as A-trisaccharide, which carries a space arm, is chemically attached to various solid supports, either directly through a suitable functional group at the end of the spacer arm or indirectly via a protein conjugated to the hapten. The preparation involves simple and mild procedures for the activation and/or derivatization of the supports. The latter includes naturally occurring polyhydroxy materials such as agarose, cellulose, or cellulose derivatives, and other particulate materials such as inorganic diatomites and a synthetic organic copolymer. The methods used for the coupling concern specifically the preparation of controlled-capacity and high-efficiency immunoadsorbents, with limited incorporations, which may be prepared easily and used for the selective removal, or affinity chromatographic separation, of specific antibodies from plasma environment or blood. It has been found that while hapten incorporation to the support may be varied rather easily, the physical nature of the support as well as the form of the hapten is important in determining the efficiency of an immunoadsorbent.     

Development of enzyme-linked immunosorbent assay for prostaglandin D2 using the stable isosteric analogue as a hapten mimic and its application

For the determination of prostaglandin (PG) D(2) produced by cultured cells in response to external stimuli, immunological methods would be convenient and useful. However, PGD(2) is unstable under the physiological conditions, so that it has been difficult to get a specific antibody for the parent PGD(2). In an attempt to get a specific antibody for PGD(2), we tried to prepare monoclonal antibodies for 11-deoxy-11-methylene-PGD(2), a novel, chemically stable, isosteric analogue of PGD(2). We successfully cloned a hybridoma cell line secreting a monoclonal antibody reacting specifically with the parent PGD(2). To develop the enzyme-linked immunosorbent assay (ELISA) for PGD(2), the immobilized antigen using the stable PGD(2) derivative was immunoreacted in a competitive manner with the monoclonal antibody in presence of free PGD(2). The optimization of the assay provided a sensitive calibration curve for PGD(2) from 0.32 pg to 0.18 ng with a value of 7.6 pg at 50% displacement. PGD(2) was almost stable during the ELISA condition. The developed assay method was useful for applying to the direct determination of PGD(2) in the culture medium of mouse 3T3-L1 adipocytes. The incubation of PGD(2) in the maturation medium of adipocytes at 37 degrees C caused the chemical conversion into PGJ(2) derivatives. The conversion became more evident after 6 h of the incubation. These findings indicate the importance of considering the optimal time for collecting the samples to be determined for PGD(2) before the conversion starts to occur.     

Yield Loss Caused by Meloidogyne graminicola on Lowland Rainfed Rice in Bangladesh

The impact of Meloidogyne graminicola on growth and yield of lowland rainfed rice was assessed with and without carbofuran in a rice-wheat rotation area of northwestern Bangladesh. The experiment was conducted on farmer fields and at a research station, with experimental plots arranged in a randomized complete block design. Prior to transplanting, rice seedling height and dry weight were greater (P ≤ 0.05) and soil levels of M. graminicola were lower (P ≤ 0.05) in the treated seedbed plots compared to the nontreated control plots. Nematicide application to the field at transplanting had a greater effect (P ≤ 0.05) on mid-season plant growth than did nematicide application to the seedbed at sowing, and rice yield increased by 1.0 t/ha where carbofuran was applied to the seedbed and field-both at the research station (P ≤ 0.05) and on farmer fields (P ≤ 0.10)- compared to a nontreated control. This is the first report of a negative impact of M. graminicola on growth and yield of lowland rainfed rice in production fields in Bangladesh.     

Cholinesterase inhibitory activity of tinosporide and 8-hydroxytinosporide isolated from Tinospora cordifolia: In vitro and in silico studies targeting management of Alzheimer’s disease

Tinosporide and 8-hydroxytinosporide isolated from Tinospora cordifolia were evaluated for acetylcholinesterase (AChE) and butylcholinesterase (BuChE) inhibitory activities. The structure of the compound was confirmed by spectroscopic analysis, whereas cholinesterase inhibition was investigated by Ellman method using donepezil as standard drug and the data were presented as IC₅₀ (μg/ml ± SEM). Furthermore, donepezil, tinosporide and 8-hydroxytinosporide were executed for docking analysis. The results from the isolated compounds TC-16R confirmed as tinosporide promisingly inhibited AChE with IC₅₀ value of 13.45 ± 0.144, whereas TC-19R confirmed as 8-hydroxytinosporide moderately inhibited AChE with IC₅₀ value of 46.71 ± 0.511. In case of BuChE inhibition, the IC₅₀ values were found to be 408.50 ± 17.197 and 317.26 ± 6.918 for tinosporide and 8-hydroxytinosporide, respectively. The in silico studies revealed that the ligand tinosporide fit with the binding sites and inhibited AChE. Overall, the study findings suggested that tinosporide would be a complementary noble molecule of donepezil which is correlated with its pharmacological activity through in vitro studies, while 8-hydroxytinosporide modestly inhibited BuChE and the results are very close to the standard donepezil.     

Genetic variation,heritability, divergence and biomass accumulation of rice genotypes resistant to bacterial blight revealed by quantitative traits and ISSR markers

A field experiment was carried out in order to evaluate genetic diversity of 41 rice genotypes using physiological traits and molecular markers. All the genotypes unveiled variations for crop growth rate (CGR), relative growth rate (RGR), net assimilation rate (NAR), yield per hill (Yhill^?1), total dry matter (TDM), harvest index (HI), photosynthetic rate (PR), leaf area index (LAI), chlorophyll‐a and chlorophyll‐b at maximum tillering stage. The CGR values varied from 0.23 to 0.76 gm cm^?2 day^?1. The Yhill^?1 ranged from 15.91 to 92.26 g, while TDM value was in the range of 7.49 to 20.45 g hill^?1. PR was found to vary from 9.40 to 22.34 µmol m^?2 s^?1. PR expressed positive relation with Yhill^?1. Significant positive relation was found between CGR and TDM (r = 0.61**), NAR and CGR (r = 0.62**) and between TDM and NAR (r = 0.31**). High heritability was found in RGR and Yhill^?1. Cluster analysis based on the traits grouped 41 rice genotypes into seven clusters. A total of 310 polymorphic loci were detected across the 20 inter‐simple sequence repeats (ISSR) markers. The UPGMA dendrogram grouped 41 rice genotypes into 11 clusters including several sub‐clusters. The Mantel test revealed positive correlation between quantitative traits and molecular markers (r = 0.41). On the basis of quantitative traits and molecular marker analyses parental genotypes, IRBB54 with MR84, IRBB60 with MR84, Purbachi with MR263, IRBB65 with BR29, IRBB65 with Pulut Siding and MRQ74 with Purbachi could be hybridized for future breeding program.     

Endogenous 15-deoxy-Delta(12,14)-prostaglandin J(2) synthesized by adipocytes during maturation phase contributes to upregulation of fat storage

15-Deoxy-Δ^12,14-prostaglandin J₂ (15d-PGJ₂) has been identified as a natural ligand for peroxisome proliferator-activated receptor (PPAR) γ to promote adipogenesis. However, it remains elusive about the ability of PPARγ-expressing adipocytes to produce PGJ₂ series and the role in the life cycle of adipocytes. Here, we developed an enzyme-linked immunosorbent assay specific for 15d-PGJ₂. The analysis using this method revealed the increase in the endogenous synthesis of immunoreactive 15d-PGJ₂ in cultured adipocytes during the maturation phase. Further studies using cyclooxygenase inhibitors clarified the contribution of endogeous 15d-PGJ₂ produced by mature adipocytes to upregulation of fat storage in an autocrine manner.     

Kinetics of yeast alcohol dehydrogenase and its coenzyme coimmobilized in a tubular flow reactor

Yeast alcohol dehydrogenase and nicotinamide adenine dinucleotide (NAD) were coimmobilized, with covalent attachment, to the interior surface of a nylon tube. The NAD was attahed at the N(6) group of the adenine moiety; an NAD derivative was prepared and attached to free carboxyl groups at a partially hydrolyzed nylon surface. The enzyme was attached, through glutaraldehyde residues, to free amino groups on the surface. Kinetic studies were carried out in which the reduced NAD was recycled by means of phenazine ethosulfate and 2,6-dichlorophenol indophenol. The reaction was studied over a range of flow rates and ethanol concentrations. The variation of rate with flow rate suggested that there was little diffusion control with respect to ethanol and that there was no observable inhibition by the reaction products. These conclusions were confirmed by evidence based on dimensionless parameters for the reaction and by direct inhibition experiments. The apparent Michaelis constant was lower than when only the enzyme was immobilized, suggesting that the immobilized enzyme-coenzyme system is of high efficiency. Overall rates of reaction were lower than when there was saturation with NAD. The tube showed no measurable loss of catalytic activity over a period of one month.     

Hi-Bi Photonic Crystal Fiber for Broadband Filter Realization Using Copper Microwires

This paper presents a highly birefringence (Hi-Bi) photonic crystal fiber (PCF)-based single-polarization filter, which consists of copper microwires. Copper is chemically stable and the use of microwires is benefit to fabricate than any metal-coated PCF. The filter characteristics are inspected by the full-vector finite element method (FEM). The proposed filter can filter out y-polarized mode, while the x-polarized mode can be guided. The confinement loss of the y-polarized mode at the wavelength of 1.31 μm is achieved of 696.79 dB/cm, while the x-polarized loss is only 4.34 dB/cm. According to numerical results, 20 dB bandwidth of the proposed filter with a maximum value of crosstalk of 601.37 dB is achieved of 650 nm that range from 1.1 to 1.75 μm. Furthermore, the insertion loss of the guided mode (x-polarization) is as low as 0.142 dB for 1 mm of fiber length. Moreover, by optimizing the structural parameters, it has shown that the proposed filter can be effective at any wavelength at the optical communication window.

     

Zinc Oxide Nanoparticles Promote the Aggregation of Concanavalin A

The ability of nanoparticles to influence protein folding and aggregation is interesting, not only because of the potential beneficial applications, but also the potential risks to human health and the environment. The interactions of concanavalin A (Con A) with zinc oxide nanoparticles (ZnO-NPs) were investigated by using fluorescence, fourier transform infrared spectroscopy, circular dichroism (CD) and dynamic light scattering techniques. ANS fluorescence and CD spectroscopy authenticated the formation of molten globule state of Con A after its incubation with ZnO-NPs for 36 h. Further incubation of 48 h resulted in the aggregation of unadsorbed Con A, proved by decrease in ANS fluorescence while an increase in thioflavin T fluorescence, characteristic of an aggregates. Moreover, Fourier transform-infrared spectroscopy confirmed the aggregation of unadsorbed Con A. The aggregated products were negligible genotoxic as analyzed by pUC19 plasmid degradation and comet assay. It is clear that ZnO-NPs morphology affect unadsorbed proteins structure. A better understanding of these differences will be essential to engineer fully functional nanobioconjugates and NPs which do not damage the proteins present in the biological system.     

Functional Divergence and Evolutionary Turnover in Mammalian Phosphoproteomes

Protein phosphorylation is a key mechanism to regulate protein functions. However, the contribution of this protein modification to species divergence is still largely unknown. Here, we studied the evolution of mammalian phosphoregulation by comparing the human and mouse phosphoproteomes. We found that 84% of the positions that are phosphorylated in one species or the other are conserved at the residue level. Twenty percent of these conserved sites are phosphorylated in both species. This proportion is 2.5 times more than expected by chance alone, suggesting that purifying selection is preserving phosphoregulation. However, we show that the majority of the sites that are conserved at the residue level are differentially phosphorylated between species. These sites likely result from false-negative identifications due to incomplete experimental coverage, false-positive identifications and non-functional sites. In addition, our results suggest that at least 5% of them are likely to be true differentially phosphorylated sites and may thus contribute to the divergence in phosphorylation networks between mouse and humans and this, despite residue conservation between orthologous proteins. We also showed that evolutionary turnover of phosphosites at adjacent positions (in a distance range of up to 40 amino acids) in human or mouse leads to an over estimation of the divergence in phosphoregulation between these two species. These sites tend to be phosphorylated by the same kinases, supporting the hypothesis that they are functionally redundant. Our results support the hypothesis that the evolutionary turnover of phosphorylation sites contributes to the divergence in phosphorylation profiles while preserving phosphoregulation. Overall, our study provides advanced analyses of mammalian phosphoproteomes and a framework for the study of their contribution to phenotypic evolution.