Novel carbohydrate specificity of the 16-kDa galectin from Caenorhabditis elegans: binding to blood group precursor oligosaccharides (type 1, type 2, Talpha,and Tbeta) and gangliosides

Galectins, a family of soluble beta-galactosyl-binding lectins, are believed to mediate cell-cell and cell-extracellular matrix interactions during development, inflammation, apoptosis, and tumor metastasis. However, neither the detailed mechanisms of their function(s) nor the identities of their natural ligands have been unequivocally elucidated. Of the several galectins present in the nematode Caenorhabditis elegans, the 16-kDa "proto" type and the 32-kDa "tandem-repeat" type are the best characterized so far, but their carbohydrate specificities have not been examined in detail. Here, we report the carbohydrate-binding specificity of the recombinant C. elegans 16-kDa galectin and the structural analysis of its binding site by homology modeling. Our results indicate that unlike the galectins characterized so far, the C. elegans 16-kDa galectin interacts with most blood group precursor oligosaccharides (type 1, Galbeta1,3GlcNAc, and type 2, Galbeta1,4GlcNAc; Talpha, Galbeta1,3GalNAcalpha; Tbeta, Galbeta1,3GalNAcbeta) and gangliosides containing the Tbeta structure. Homology modeling of the C. elegans 16-kDa galectin CRD revealed that a shorter loop containing residues 66-69, which enables interactions of Glu(67) with both axial and equatorial -OH at C-3 of GlcNAc (in Galbeta1,4GlcNAc) or at C-4 of GalNAc (in Galbeta1,3GalNAc), provides the structural basis for this novel carbohydrate specificity.     

Nutrient use preferences among soil Streptomyces suggest greater resource competition in monoculture than polyculture plant communities

Background an aims

Nutrient use overlap among sympatric Streptomyces populations is correlated with pathogen inhibitory capacity, yet there is little information on either the factors that influence nutrient use overlap among coexisting populations or the diversity of nutrient use among soil Streptomyces.

Methods

We examined the effects of plant host and plant species richness on nutrient use of Streptomyces isolated from the rhizosphere of Andropogon gerardii (Ag) and Lespedeza capitata (Lc) growing in communities of 1 (monoculture) or 16 (polyculture) plant species. Growth on 95 carbon sources was assessed over 5d.

Results

Cumulative growth was significantly greater for polyculture vs. monoculture isolates, and for Lc vs. Ag isolates. Isolates from monocultures, but not polycultures, exhibited a drop in growth rates between 24 h and 72 h post-inoculation, suggesting resource allocation to non-growth functions. Isolates from high-carbon (polyculture) or high-nitrogen (Lc) soils had larger niche widths than isolates from low-C (monocultures) or low-N (Ag) soils. Sympatric isolates from polycultures were significantly more differentiated from one another in preferred nutrients for growth than sympatric isolates from monocultures.

Conclusions

These results suggest that Streptomyces populations respond to selection imposed by plant host and plant community richness and that populations from polyculture but not from monoculture, mediate resource competition via niche differentiation.

     

Effect of herbal melanin on IL-8: a possible role of Toll-like receptor 4 (TLR4)

The production of IL-8 can be induced by LPS via TLR4 signaling pathway. In this study, we tested the effect of a herbal melanin (HM) extract, from black cumin seeds (Nigella sativa L.), on IL-8 production. We used HM and LPS in parallel to induce IL-8 production by THP-I, PBMCs, and TLR4-transfected HEK293 cells. Both HM and LPS induced IL-8 mRNA expression and protein production in THP-1 and PBMCs. On applying similar treatment to HEK293 cells that express TLR4, MD2, and CD14, both HM and LPS significantly induced IL-8 protein production. We have also demonstrated that HM and LPS had identical effects in terms of IL-8 stimulation by HEK293 transfected with either TLR4 or MD2-CD14. Melanin extracted from N. sativa L. mimics the action of LPS in the induction of IL-8 by PBMC and the other used cell lines. Our results suggest that HM may share a signaling pathway with LPS that involves TLR4.     

Fab fragments imprinted SPR biosensor for real-time human immunoglobulin G detection

F(ab) fragments imprinted surface plasmon resonance (SPR) chip was prepared for the real-time detection of human immunoglobulin G (IgG). In order to attach polymerization precursor on SPR chip, the SPR chip surface was modified with allyl mercaptan. F(ab) fragments of the IgG molecules were prepared by papain digestion procedure and collected by fast protein liquid chromatography (FPLC) system using Hi-Trap_r Protein A FF column. The collected F(ab) fragments were complexed with histidine containing specific monomer, N-methacryloyl-l-histidine methyl ester (MAH). Molecular imprinted polymeric nanofilm was prepared on SPR chip in the presence of ethylene glycol dimethacrylate and 2-hydroxyethylmethacrylate. The template molecules, F(ab) fragments, were removed from the polymeric nanofilm using 1M NaCl solution (pH: 7.4, phosphate buffer system). The molecular imprinted SPR chip was characterized by contact angle, atomic force microscopy and Fourier transform infrared spectroscopy. By the real-time IgG detection studies carried out using aqueous IgG solutions in different concentrations, the kinetics and isotherm parameters of the molecular imprinted SPR chip-IgG system were calculated. To show selectivity and specificity of the molecular imprinted SPR chip, competitive kinetic analyses were performed using bovine serum albumin (BSA), IgG, F(ab) and F(c) fragments in singular and competitive manner. As last step, IgG detection studies from human plasma were performed and the measured IgG concentrations were well matched with the results determined by enzyme-linked immunosorbent assay (ELISA). The results obtained with the molecular imprinted SPR chip were well fitted to Langmuir isotherm and the detection limit was found as 56 ng/mL. In the light of the results, we can conclude that the proposed molecular imprinted SPR chip can detect IgG molecules from both aqueous solutions and complex natural samples.     

Rational MD simulations for improvement the affinity of nanobody against PlGF (placenta growth factor): mutagenesis based on electrostatic interactions

Abbreviations COM center of mass distance

MD molecular dynamics

MM-PBSA Molecular Mechanics Poisson–Boltzmann Surface Area

Nb nanobody

PlGF placenta growth factor

Rg radius of gyration

RMSD root mean-square deviation

SASA solvent-accessible surface area

VEGF vascular endothelial growth factor

     

Physiological insights into sulfate and selenium interaction to improve drought tolerance in mung bean

The present study involved two pot experiments to investigate the response of mung bean to the individual or combined SO₄²⁻ and selenate application under drought stress. A marked increment in biomass and NPK accumulation was recorded in mung bean seedlings fertilized with various SO₄²⁻ sources, except for CuSO₄. Compared to other SO₄²⁻ fertilizers, ZnSO₄ application resulted in the highest increase in growth attributes and shoot nutrient content. Further, the combined S and Se application (S + Se) significantly enhanced relative water content (16%), SPAD value (72%), photosynthetic rate (80%) and activities of catalase (79%), guaiacol peroxidase (53%) and superoxide dismutase (58%) in the leaves of water-stressed mung bean plants. Consequently, the grain yield of mung bean was markedly increased by 105% under water stress conditions. Furthermore, S + Se application considerably increased the concentrations of P (47%), K (75%), S (80%), Zn (160%), and Fe (15%) in mung bean seeds under drought stress conditions. These findings indicate that S + Se application potentially increases the nutritional quality of grain legumes by stimulating photosynthetic apparatus and antioxidative machinery under water deficit conditions. Our results could provide the basis for further experiments on cross-talk between S and Se regulatory pathways to improve the nutritional quality of food crops.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-00992-6.     

The Effects of Intermittent Fasting during the Month of Ramadan in Chronic Haemodialysis Patients in a Tropical Climate Country

Background

Chronic kidney disease is an emerging problem in the majority Muslim countries. Despite the uncertainties of the risks involved, some Muslim patients undergoing chronic haemodialysis choose to observe intermittent fasting during the month of Ramadan. This study aims to investigate the effect of Ramadan fasting in haemodialysis patients residing in a tropical climate country.

Methods

This prospective cross sectional study recruited Muslim patients on regular haemodialysis from three haemodialysis centres in Kuala Lumpur from 15^th July 2011 to 29^th August 2011. Patients who fasted for any number of days were included (n = 35, 54% female, age 54±11 years). 89% of patients fasted for more than 15 days and 49% were diabetics. Dialysis parameters and blood samples were obtained one week prior to Ramadan and during the last week of Ramadan. The differences in dialysis parameters and biochemical values pre- and end-Ramadan were examined using paired t-test.

Results

Both pre- and post-dialysis weight were significantly decreased during Ramadan fasting compared to the month prior (p = <0.001). There was a significant decrease in the amount of ultrafiltration (p = 0.002). There were no significant differences in dry weight, inter-dialytic weight gain, mean urea reduction ratio or blood pressure measurements comparing pre- and end of Ramadan fasting. There was a significant increase in serum albumin level (p = 0.006) and decrease in serum phosphate level (p = 0.02) at the end of Ramadan.

Conclusion

Ramadan fasting is associated with reduced weight, improved serum albumin and phosphate level in our population of haemodialysis patients. A larger multi-centre study will allow us to understand more about the effects of fasting in this population.     

VdNop12, containing two tandem RNA recognition motif domains,is a crucial factor for pathogenicity and cold adaption in Verticillium dahliae

Previous studies have reported the ability of fungi to overwinter in soil or on crop debris under different environmental conditions, but how fungi adapt to chilling is still largely unknown. In this study, we have identified and characterized the RNA binding protein (RBP) (VdNop12) by screening an Agrobacterium tumefaciens-mediated transformation-mediated insertional mutational library of Verticillium dahliae. We determined that this protein was essential to the pathogen for virulence on cotton plants. VdNop12 contains two tandem RNA recognition motif domains, and its orthologs are widely distributed in filamentous fungi. Mutants produced by disruption of VdNop12 showed defects in vegetative growth, conidiation and cell wall integrity. The mutant also showed an increase in sensitivity to low temperature, as compared to the wildtype and complementation strains. Yeast complementation assay showed that VdNop12 could functionally restore the growth phenotype of ΔScNop12 mutant of Saccharomyces cerevisiae at 15°C. We demonstrated that the VdNop12 is localized in the nucleus, and its loss resulted in the downregulated expression of several genes related to cAMP-PKA and MAPK pathways in V. dahliae. Our results demonstrated a crucial role of RBPs in the regulation of morphology, cold adaption, and pathogenic development in V. dahliae.     

Synthesis of tentacle-type magnetic beads as immobilized metal-chelate affinity support for cytochrome c adsorption

Magnetic poly(2-hydroxyethylmethacrylate) (mPHEMA) beads with an average diameter of 100-140 microm were produced by suspension polymerization in the presence of magnetite particles (i.e. Fe3O4). Specific surface area and average pore size of the magnetic beads was found to be 50 m2/g and 819 nm, respectively. Ester groups in the mPHEMA structure were converted to imine groups by reacting with poly(ethyleneimine) (PEI) in the presence of NaH. Amino (-NH2) content of PEI-attached mPHEMA beads was determined as 102 mg PEI/g. Then, Cu2+ ions were chelated on the magnetic beads in the range of 20-793 micromol Cu2+/g. Cytochrome c (cyt c) adsorption was performed on the metal chelating beads from aqueous solutions containing different amounts of cyt c at different pHs, Cu2+ loadings and temperatures. Cyt c adsorption on the mPHEMA/PEI beads was 4.6 mg/g. Cu2+ chelation increased the cyt c adsorption significantly (40.1 mg/g). Adsorption capacity increased with Cu2+ loading and then reached a saturation value. Cyt c adsorption decreased with increasing temperature. Cyt c molecules could be reversibly adsorbed and eluted ten times with the magnetic adsorbents without noticeable loss in their cyt c adsorption capacity. The applicability of two kinetic models including pseudo-first order and pseudo-second order model was estimated on the basis of comparative analysis of the corresponding rate parameters, equilibrium capacity and correlation coefficients. Results suggest that chemisorption processes could be the rate-limiting step in the adsorption process. In the last part of this article, cyt c adsorption experiments were performed in a magnetically stabilized fluidized bed (MSFB) system at optimum conditions determined from the batch experiments. The adsorption capacity decreased significantly from 46.8 to 15.4 mg/g polymer with the increase of the flow-rate from 0.5 to 4.0 ml/min. The resulting magnetic chelator beads possessed excellent long-term storage stability.