Structures from anomalous diffraction of native biological macromolecules

Crystal structure analyses for biological macromolecules without known structural relatives entail solving the crystallographic phase problem. Typical de novo phase evaluations depend on incorporating heavier atoms than those found natively; most commonly, multi- or single-wavelength anomalous diffraction (MAD or SAD) experiments exploit selenomethionyl proteins. Here, we realize routine structure determination using intrinsic anomalous scattering from native macromolecules. We devised robust procedures for enhancing the signal-to-noise ratio in the slight anomalous scattering from generic native structures by combining data measured from multiple crystals at lower-than-usual x-ray energy. Using this multicrystal SAD method (5 to 13 equivalent crystals), we determined structures at modest resolution (2.8 to 2.3 angstroms) for native proteins varying in size (127 to 1148 unique residues) and number of sulfur sites (3 to 28). With no requirement for heavy-atom incorporation, such experiments provide an attractive alternative to selenomethionyl SAD experiments.     

Short‐term potassium release and fixation in some calcareous soils

Potassium (K⁺) directly released from primary K‐bearing minerals can contribute to plant nutrition. The objective of this research was to assess short‐term K⁺ release and fixation on a range of intensively cropped calcareous soils. Potassium sorption and desorption properties and the contributions of exchangeable‐K⁺ (EK) and nonexchangeable‐K⁺ (NEK) pools to K⁺ dynamics of the soil‐solution system was measured using a modified quantity‐to‐intensity (Q : I) experiment. Release and fixation of K⁺ were varied among soils. The relation between the change in the amount of NEK during the experiment and the initial constrain was linear, and soil ability for K⁺ release and fixation (β) for all soils varied from 0.041 to 0.183, indicating that 4% to 18% of added K⁺ converted to NEK when fixation occurred. The equilibrium potential buffering capacity (PBC) for K⁺ derived from Q : I experiments had significant correlation (r = 0.75, p < 0.01) with β, indicating that PBC depends not only on exchange properties but also on release and fixation properties. The depleted soils showed higher β value than the other soils, indicating much of the added K⁺ was converted to NEK in case of positive constraint. The range of the amount of EK which was not in exchange equilibrium with Ca (E_min) in the experimental conditions was large and varied from 0.68 to 9.00 mmol kg^–1. On average, E_min amounted to 64% of EK. This fraction of EK may not be available to the plant. The parameters obtained from these short‐term K⁺ release and fixation experiments can be used in plant nutrition.     

Effect of nitrogen and potassium fertilizer on growth,yield and chemical composition of sweet fennel

Abstract

In this study, the effect of different levels of nitrogen (N) and potassium (K) fertilizers on vegetative fresh bulb yield, quality and biochemical attributes of sweet fennel were investigated in spring 2017 and 2018. Treatments consisted of 0, 50, 100, or 150?kg ha^?1 of N and K in a randomized complete block design with three replications. Results showed that, the bulb fresh yield increased with application of N and K in both years, while the higher level of K had significant adverse effect in first year. Results also showed that sweet fennel bulb treated by N and K exhibited higher vitamin C, total phenols and flavonoid contents, and antioxidant capacity, especially in first year. In both years, alone or combined application of N and K increased total soluble solids (TSS) content as compared to control, and this effect was more pronounced in 2017 season. Macronutrient contents of bulb including magnesium (Mg), phosphorus (P), K, N, and calcium (Ca) were significantly enhanced by NK fertilizer, however an alone application of K decreased Ca and Mg nutrient contents. Therefore, to improve fresh bulb yield, quality and antioxidant capacity in sweet fennel, 150?kg ha^?1 K and 100?kg ha^?1 N is recommended.     

Functional involvement of Ca(2+) and Ca(2+)-activated K(+) channels in anethol-induced changes in Ca(2+) dependent excitability of F1 neurons in Helix aspersa

The effects of anethol, the major component of anise oil, on the Ca²⁺-dependent excitability and afterhyperpolarization (AHP) in snail neurons were examined using intracellular recording. Anethol (0.5%) significantly broadened the spike, reduced the firing frequency and enhanced the AHP amplitude. In contrast, anethol (2%) significantly increased the firing frequency and decreased the AHP. Blockade of Ca²⁺ channels after anethol application depolarized the membrane potential and significantly reduced the firing rate. Furthermore, in the presence of anethol (0.5%) a significant decrease in the AHP was observed by Ca²⁺ channels blockage. Here, anethol-induced functional modification of Ca²⁺ and Ca²⁺-activated K⁺ channels is suggested.     

Morphometric study of the testis in sheep embryos using unbiased design-based stereology

Sheep have been used as translational models of human postnatal testicular development. However, the morphometric features of the normal developing testis in sheep embryos have not been previously investigated using stereology. The objective of the present work was to establish normal quantitative parameters for fetal testicular tissue components in sheep, using unbiased design-based stereological methods. Twenty-four sheep embryos were divided into four gestational age groups (9–11, 12–14, 15–17 and 18–20 weeks of gestation) on the basis of the embryos’ crown-rump length. Isotropic, systematic uniform random sections of the left testes were obtained by employing the orientator method. Testicular total volume, the absolute and proportional volumes occupied by the seminiferous tubules and interstitial tissue, as well as the seminiferous tubule length, were estimated using the point-counting system and the unbiased counting frame principle. All the parameters, with the exception of the interstitial tissue's fractional volume, gradually increased along with gestational age, with the maximum increase especially seen in the late fetal stages. The proportional volume of the interstitial tissue, on the other hand, showed a decreasing trend along with increasing gestational age. The absolute volume of the testes, of the seminiferous tubules and of the interstitial tissue, and the length of the seminiferous tubules showed a significant (p< 0.05) positive linear correlation with gestational age. Several similarities were observed with human testicular embryogenesis. The stereological data emerging from the present study might prove useful as basic contribution to the fields of andrology and embryology and stimulate further research in these areas.     

Enantiomeric discrimination and quantification of the chiral organophosphorus pesticide fenamiphos in aqueous samples by a novel and selective ³¹P nuclear magnetic resonance spectroscopic method using cyclodextrins as chiral selector

A rapid, selective, and accurate quantitative 31P nuclear magnetic resonance (31P NMR) spectroscopy method was used for the chiral recognition of the racemic organophosphorus pesticide fenamiphos using chiral solvating agents (CSAs). Six neutral cyclodextrins (CDs) (α-CD, β-CD, methyl-β-CD, hydroxyethyl-β-CD, hydroxypropyl-β-CD, and hydroxypropyl-γ-CD) and two anionic CDs (carboxymethyl-β-CD and carboxyethyl-β-CD) were selected for these experiments. The shift displacement values (Δδ), after addition of each of the eight CDs in the highest possible molar ratio to a guest, were recorded. The results showed that β-CD and hydroxypropyl-β-CD were the best chiral solvating agents for the enantiomeric discrimination of fenamiphos. Two-dimension rotating frame nuclear Overhauser spectroscopy (ROESY) was used to investigate the structure of the β-CD-fenamiphos inclusion complex in aqueous solution. To determine the fenamiphos enantiomers, a calibration curve was drawn for two enantiomers over the range of 0.05-0.25 mg mL?1. The limits of detection (S/N = 3) were obtained as 0.0068 and 0.0060 mg mL?1 for fenamiphos enantiomers. The recovery studies were performed on aqueous real samples ranging from 94 to 107% with coefficients of variation of ≤ 9%.     

Gut Organoid as a New Platform to Study Alginate and Chitosan Mediated PLGA Nanoparticles for Drug Delivery

Intestinal organoids can be used as an ex vivo epithelial model to study different drug delivery effects on epithelial cells’ luminal surface. In this study, the impact of surface charge on the delivery of 5-ASA loaded PLGA nanoparticles into the lumen of organoids was investigated. Alginate and chitosan were used to coat the nanoparticles and provide negative and positive charges on the particles, respectively. The organoid growth and viability were not affected by the presence of either alginate- or chitosan-coated nanoparticles. It was shown that nanoparticles could be transported from the serosal side of the organoids to the lumen as the dye gradually accumulated in the lumen by day 2–3 after adding the nanoparticles to the Matrigel. By day 5, the dye was eliminated from the lumen of the organoids. It was concluded that the positively charged nanoparticles were more readily transported across the epithelium into the lumen. It may be attributed to the affinity of epithelial cells to the positive charge. Thus, the organoid can be utilized as an appropriate model to mimic the functions of the intestinal epithelium and can be used as a model to evaluate the benefits of nanoparticle-based drug delivery.     

The response of drought-tolerant sugar beet to salinity stress under field and controlled environmental conditions

Abstract

Thirty-four sugar beet drought-tolerant half-sib families were screened in greenhouse experiment under saline (16?dSm^?1) and normal conditions, and a subset of 10 tolerant genotypes were selected for evaluating of yield-related traits, Na⁺, K⁺, and α-amino-N nitrogen contents under saline field condition (12?dSm^?1). Drought-tolerant genotypes showed a different tolerance level under salt stress. Significant correlation was observed between EP2_s and STIEP2 in greenhouse with root yield (RY) in saline field condition. It should be possible that a simple measurement at 16?dSm^?1 in the greenhouse can be used as main part of sugar beet salt-tolerant breeding program. In total, genotypes SBSI-DR I-HSF14-P.23 (# 16) and 436B-HSF9-P.27 (# 26) were recognized as salt- and drought-tolerant genotypes with low Na⁺ and K⁺ contents in root and high white sugar content (WSC) and RY in field. A larger panel of genotypes to validate this kind of association is necessary.