Revisiting marine redox conditions during the Ediacaran Shuram carbon isotope excursion

The Neoproterozoic carbonate record contains multiple carbon isotope anomalies, which are the subject of intense debate. The largest of these anomalies, the Shuram excursion (SE), occurred in the mid-Ediacaran (~574–567 Ma). Accurately reconstructing marine redox landscape is a clear path toward making sense of the mechanism that drives this δ¹³C anomaly. Here, we report new uranium isotopic data from the shallow-marine carbonates of the Wonoka Formation, Flinders Ranges, South Australia, where the SE is well preserved. Our data indicate that the δ²³⁸U trend during the SE is highly reproducible across globally disparate sections from different depositional settings. Previously, it was proposed that the positive shift of δ²³⁸U values during the SE suggests an extensive, near-modern level of marine oxygenation. However, recent publications suggest that the fractionation of uranium isotopes in ferruginous and anoxic conditions is comparable, opening up the possibility of non-unique interpretations of the carbonate uranium isotopic record. Here, we build on this idea by investigating the SE in conjunction with additional geochemical proxies. Using a revised uranium isotope mass balance model and an inverse stochastic carbon cycle model, we reevaluate models for δ¹³C and δ²³⁸U trends during the SE. We suggest that global seawater δ²³⁸U values during the SE could be explained by an expansion of ferruginous conditions and do not require a near-modern level of oxygenation during the mid-Ediacaran.     

Preparation of Some New Pyrazolo[1,5-a]pyrimidines and Evaluation of Their Antioxidant,Antibacterial (MIC and ZOI) Activities,and Cytotoxic Effect on MCF-7 Cell Lines

This study aims to synthesize some novel pyrazolo[1,5-a]pyrimidine derivatives, and investigate their biological activities. These compounds exhibited good to high antioxidant activities [2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capabilities]. Among them, Ethyl 5-(2-ethoxy-2-oxoethyl)-7-hydroxy-2-methylpyrazolo[1,5-a]pyrimidine-3-carboxylate ( 3h ) showed the highest antioxidant activity [Half-maximal Inhibitory Concentration (IC₅₀)=15.34 μM] compared to ascorbic acid (IC₅₀=13.53 μM) as a standard compound. Their antibacterial activities were investigated against two Gram-positive bacteria (Bacillus subtilis, and Staphylococcus aureus) and two Gram-negative bacteria (Pseudomonas aeruginosa, and Escherichia coli). The results showed that Ethyl 7-hydroxy-5-phenylpyrazolo[1,5-a]pyrimidine-3-carboxylate ( 3i ) has the best antibacterial activity against Gram-positive B. subtilis [Zone of Inhibition (ZOI)=23.0±1.4 mm, Minimum Inhibitory Concentration (MIC)=312 μM]. Also, the cytotoxicity of these compounds was assessed against breast cancer cell lines [human breast adenocarcinoma (MCF-7)], which 7-Hydroxy-2-methyl-5-phenylpyrazolo[1,5-a]pyrimidine-3-carbonitrile ( 3f ) displayed the most cytotoxicity (IC₅₀=55.97 μg/mL), in contrast with Lapatinib (IC₅₀=79.38 μg/mL) as a known drug.     

Comparison of destructive and nondestructive DNA extraction methods for the metabarcoding of arthropod bulk samples

DNA metabarcoding is routinely used for biodiversity assessment, in particular targeting highly diverse groups for which limited taxonomic expertise is available. Various protocols are currently in use, although standardization is key to its application in large-scale monitoring. DNA metabarcoding of arthropod bulk samples can be conducted either destructively from sample tissue, or nondestructively from sample fixative or lysis buffer. Nondestructive methods are highly desirable for the preservation of sample integrity but have yet to be experimentally evaluated in detail. Here, we compare diversity estimates from 14 size-sorted Malaise trap samples processed consecutively with three nondestructive approaches (one using fixative ethanol and two using lysis buffers) and one destructive approach (using homogenized tissue). Extraction from commercial lysis buffer yielded comparable species richness and high overlap in species composition to the ground tissue extracts. A significantly divergent community was detected from preservative ethanol-based DNA extraction. No consistent trend in species richness was found with increasing incubation time in lysis buffer. These results indicate that nondestructive DNA extraction from incubation in lysis buffer could provide a comparable alternative to destructive approaches with the added advantage of preserving the specimens for postmetabarcoding taxonomic work but at a higher cost per sample.     

Lipid-protein interactions in membranes: Effect of lipid composition on mobility of spin-labeled cysteine residues in yeast plasma membrane

In order to gain direct evidence for lipid-dependent protein conformation in membrane, effects of modification of lipid composition on mobility of spin-labeled cysteine residues were investigated in the plasma membrane of the yeast Saccharomyces cerevisiae. Conversion of the bulk of phospholipids to diglycerides by treatment of the membrane with phospholipase C substantially enhanced spectral anisotropy. However, alteration of the viscosity of the lipid-bilayer by enriching the membrane with palmitelaidic or oleic acid had no effect on mobility of spin-labeled cysteine residues. These observations indicate that while the spin-labeled residues are not in direct contact with the lipid core of the membrane, there are lipid-protein interactions to the extent that removal of polar portion of the bulk of phospholipids induces conformational changes in proteins, which in turn restrict mobility of these residues. It is concluded that conformation of membrane proteins depends on lipid structure and that phospholipids have a role in preserving the native conformation of proteins.     

Effect of exogenous fatty acids on growth,membrane fluidity,and phospholipid fatty acid composition in yeast

The growth response of a double-mutant fatty acid auxotroph of yeast Saccharomyces cerevisiae to exogenous saturated fatty acids of a homologous series from 12:0 to 16:0, each supplied with oleate, linoleate, linolenate, or cis-Δ¹¹- eicosenoate, cannot be explained in terms of the efficiency of incorporation of the fatty acids into phospholipids or alteration of membrane fluidity. There is, however, a negative correlation between growth and levels of 12:0 plus 13:0 in phospholipids, as well as a positive correlation between growth and levels of 14:0, 1 5:0, and 1 6:0. We, therefore, conclude that the predominant factor in these phospholipid fatty acyl chain modifications is maintenance of an optimal concentration of C14:0 through C16:0 in phospholipids of this organism.     

Complexes of Cu with mixed-donor phenanthroline-containing macrocycles: analysis of their structural, redox and spectral properties in the context of Type-1 blue copper proteins biomimetic models

The macrocycles L¹-L³ having N₂S₂O-, N₂S₂-, and N₂S₃-donor sets, respectively, and incorporating the 1,10-phenanthroline unit interact in EtOH and MeCN solutions with Cu^II to give 1:1 [M(L)]²⁺ complex species. The compounds [Cu(L¹)(ClO₄)]ClO₄ (1), [Cu(L²)(ClO₄)]ClO₄ ·  (2) and [Cu(L³)](ClO₄)₂ (3) were isolated at the solid state and the first two also characterised by X-ray diffraction studies. The conformation adopted by L¹ and L² in the cation complexes reveals the aliphatic portion of the rings folded over the plane containing the heteroaromatic moiety with the ligands encapsulating the metal centre within their cavity by imposing, respectively, a square-based pyramidal and a square planar geometry. In both complexes, the metal ion completes its coordination sphere by interacting with a ClO₄⁻ ligand. The compound [Cu(L³)₂](PF₆)₂ (4) containing a 1:2 cation complex was also isolated at the solid state: EPR spectroscopy measurements suggest the presence of a CuN₄ chromophore in this complex. The EPR and electronic spectral features of 1-4 have been studied and their redox properties examined in comparison with those observed for Type-1 blue copper proteins.The reactivity of L¹-L³ has also been tested toward stoichiometric amounts of the Cu^I salt [CuCl(PPh₃)₃].     

Predation by Allothrombium pulvinum on the spider mites Tetranychus urticae and Amphitetranychus viennensis: Predation Rate, Prey Preference and Functional Response

The deutonymphs of Allothrombium pulvinum Ewing (Acari: Trombidiidae) are among the most important natural enemies of spider mites in North, North East and West Iran. In this study, maximum predation rate and preference experiments were conducted with A. pulvinum deutonymphs on apple leaf discs, to determine their preference for either of two spider mite species: Amphitetranychus viennensis (Zacher) and Tetranychus urticae Koch (Acari: Tetranychidae). Maximum predation rate tests showed that the predatory mite consumed more eggs and females of T. urticae than of A. viennensis. Furthermore, the Manly’s preference index for eggs and females of T. urticae confirmed that T. urticae were the preferred prey. The functional response of A. pulvinum deutonymphs on females of T. urticae was examined over a 24-h period. Predation of A. pulvinum deutonymphs presented with females of T. urticae followed a type III functional response. Estimated handling time for the predatory mites was 4.51 h and attack coefficient b, which describes the changes in attack rate with prey densities in a type III functional response, was 0.021.     

Investigation on the surface hydrophobicity and aggregation kinetics of human calprotectin in the presence of calcium

Calcium and zinc binding protein, calprotectin is a multifunctional protein with broad spectrum antimicrobial and antitumoural activity. It was purified from human neutrophil, using a two-step ion exchange chromatography. Since surface hydrophobicity of calprotectin may be important in membrane anchoring, membrane penetration, subunits oligomerization and some biological roles of protein, in this study attempted to explore the effect of calcium in physiological range on the calprotectin lipophilicity. Incubation of human calprotectin (50 microg/ml) with different calcium concentrations showed that 1-anilino-8-naphthalene sulfonic acid (ANS) fluorescence intensity of the protein significantly elevates with calcium in a dose dependent manner, suggesting an increase in calprotectin surface hydrophobicity upon calcium binding. Our study also indicates that calcium at higher concentrations (6, 8 and 10 mM) induces aggregation of human calprotectin. Our finding demonstrates that the starting time and the rate constant of calprotectin aggregation depend on the calcium concentration.     

The Bt gene <Emphasis Type="Italic">cry2Aa2</Emphasis> driven by a tissue specific ST-LS1 promoter from potato effectively controls <Emphasis Type="Italic">Heliothis virescens</Emphasis>

Expression of the Cry2Aa2 protein was targeted specifically to the green tissues of transgenic tobacco Nicotiana tabacum cv. Xanthi plants. This deployment was achieved by using the promoter region of the gene encoding the Solanum tuberosum leaf and stem specific (ST-LS1) protein. The accumulated levels of toxin in the leaves were found to be effective in achieving 100 mortality of Heliothis virescens larvae. The levels of Cry2Aa2 expression in the leaves of these transgenic plants were up to 0.21 of the total soluble proteins. Bioassays with R₁ transgenic plants indicated the inheritance of cry2Aa2 in the progeny plants. Tissue-specific expression of the Bt toxin in transgenic plants may help in controlling the potential occurrence of insect resistance by limiting the amount of toxin to only predated tissues. The results reported here validate the use of the ST-LS1 gene promoter for a targeted expression of Bt toxins in green tissues of plants.     

Inter-insertional distance is a poor correlate for ligament load: Analysis from in vivo gait kinetics data

In many analytic models of the knee joint, inter-insertional distance is used as the measure to define the load in a ligament. In addition, the direction of the load is taken to be the direction between the two insertions. Our in vivo data on the ovine ligament loads during gait, however, indicate that a wide range of forces is possible in the ligament for any specified inter-insertional distance. To understand the complex relationship between the bone orientations and ligament load better, an artificial neural network (ANN) model was developed. The six degree-of-freedom (6-DOF) in vivo kinematics of femur relative to tibia (joint kinematics) was used as input, and the magnitude of the anterior cruciate ligament (ACL) load was used as output/target. While the trained network was able to predict peak ligament loads with remarkable accuracy (R-square=0.98), an explicit relationship between joint kinematics and ACL load could not be determined. To examine the experimental and ANN observations further, a finite element (FE) model of the ACL was created. The geometry of the FE model was reconstructed from magnetic resonance images (MRI) of an ACL, and an isotropic, hyperelastic, nearly incompressible constitutive model was implemented for the ACL. The FE simulation results also indicate that a range of loads is possible in the ACL for a given inter-insertional distance, in concordance with the experimental/ANN observations. This study provides new insights for models of the knee joint; a simple force–length relationship for the ligament is not exact, nor is a single point to single point direction. More detailed microstructure-function data is required.