Identification of CRISP2 from human sperm as PSP94‐binding protein and generation of CRISP2‐specific anti‐peptide antibodies

Cysteine‐rich secretory proteins (CRISPs) are mainly found in the mammalian male reproductive tract and reported to be involved at different stages of fertilization. CRISPs have been shown to interact with prostate secretory protein of 94 amino acids (PSP94) from diverse sources, and the binding of these evolutionarily conserved proteins across species is proposed to be of functional significance. Of the three mammalian CRISPs, PSP94–CRISP3 interaction is well characterized, and specific binding sites have been identified; whereas, CRISP2 has been shown to interact with PSP94 in vitro. Interestingly, human CRISP3 and CRISP2 proteins are closely related showing 71.4% identity. In this study, we identified CRISP2 as a potential binding protein of PSP94 from human sperm. Further, we generated antisera capable of specifically detecting CRISP2 and not CRISP3. In this direction, specific peptides corresponding to the least conserved ion channel regulatory region were synthesized, and polyclonal antibodies were generated against the peptide in rabbits. The binding characteristics of the anti‐CRISP2 peptide antibody were evaluated using competitive ELISA. Immunoblotting experiments also confirmed that the peptide was able to generate antibodies capable of detecting the mature CRISP2 protein present in human sperm lysate. Furthermore, this anti‐CRISP2 peptide antibody also detected the presence of native CRISP2 on sperm.Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.     

Tracking Economic and Environmental Indicators of Exported Wood Pellets to the United Kingdom from the Southern United States: Lessons for Policy?

This study estimates the abatement cost of greenhouse gas (GHG) emissions for a unit of electricity generated in the UK from wood pellets imported from Southern USA. We assumed that only pulpwood obtained from loblolly pine (Pinus taeda) plantations was used for manufacturing exported wood pellets. The use of imported wood pellets for electricity generation could save at least 69.9 % of GHG emissions relative to coal-based electricity in the UK. The average unit production cost of electricity generated from imported wood pellets (US$222.3 MWh^?1) was higher by 30.0 % than the unit production cost of electricity generated from coal (US$171.0 MWh^?1) without any price support. In the presence of payments from the established price support mechanisms of Renewable Obligation Certificates (ROCs) and Levy Exemption Certificates (LECs), the unit production cost of electricity generated from imported wood pellets (US$142.9 MWh^?1) was lower by about 16.0 % than the unit production cost of electricity generated from coal. Policy makers should consider 1 MWh of electricity generated from imported wood pellets equivalent to 0.58 ROCs or 0.71 ROCs in presence and absence of payments from LECs, respectively. This will ensure zero abatement cost and lead to economic efficiency in reducing GHG emissions. However, a more in-depth analysis focusing on the market risks for power-generating companies and other wood pellet supply chains is required before modifying existing equivalency factors for ensuring continuous use of imported wood pellets for displacing coal-based electricity in the UK.     

Immunochemical Studies on Lipopolysaccharide from Agglutinable and Non-Agglutinable Vibrios

Lipopolysaccharide (LPS) prepared from Vibrio cholerae and a non-agglutinable (NAG; not agglutinable with O-group I serum according to Gardner and Venkatraman [13]) vibrio strain, isolated from a patient with cholera-like clinical symptoms, have been compared with respect to their chemical composition and immunological behavior. In addition to a significant difference in the chemical composition between the two lipopolysaccharides, the LPS from V. cholerae, unlike that from the NAG vibrio, requires prior treatment with alkali for it to be an effective antigen in the indirect hemagglutination test with sheep cells. It has been suggested that the alkali acts by removing excess O-acetyl group from LPS of agglutinable vibrios. LPS from the NAG vibrio consistently showed a lower antibody response in rabbits in terms of agglutinin and vibriocidal titer. Also, the class of agglutinin antibody elicited by LPS of the NAG vibrio was predominantly immunoglobin M, and that from V. cholerae was immunoglobulin G under comparable conditions.     

Mycoparasitism ofFusarium SPP. onRhizoctonia solani Kühn

Summary Experiments on nutrient and staled agar were carried out to investigate the mycoparasitic activity of some fusaria againstRhizoctonia solani Kühn. Penetration and coiling byFusarium oxysporum Sch.,F. semitectum Berk & Rav. andF. udum Butler in and around theR. solani hyphae was observed. Lysis ofF. udum hyphae was observed inside theR. solani hyphae showing the reverse of the normal direction of necrotrophic mycoparasitic relationships. The mycoparasitic activity ofFusarium spp. was much affected in staled agar plates.     

Ultrastructure of meristematic cells of dormant and released buds in Tradescantia paludosa

The lateral bud meristems of Tradescantia paludosa show a characteristic cytohistological zonation during dormancy. The cells comprising this so called ‘zone of inhibition’, which is located at the extreme tip of the bud apex, rarely synthesize nuclear DNA or undergo mitotic division. These nuclei are as large as prophase nuclei, yet contain only telophase (2C) amounts of DNA and significantly lower amounts of histone as compared to the 2C nuclei of the actively dividing cells.Ultrastructural observations of the nuclei in the ‘zone of inhibition’ show that a large proportion of the chromatin is organized as less condensed, diffuse, euchromatin fibrils; however, the chromatin of the actively dividing nuclei of the cells outside the ‘zone of inhibition’ or in the released bud meristems is organized to a greater extent as condensed clumps of heterochromatin. When the dormancy is released, the nuclei in the ‘zone of inhibition’ synthesize DNA and histone and undergo cell division in approx. 4 days. Striking changes in the organization of chromatin fibrils take place during this transition period. The diffuse chromatin fibrils of the nuclei in the ‘zone of inhibition’ progressively become more and more condensed as the cell prepares to undergo the first mitotic division after the release of dormancy. This change which is coupled with the synthesis of histones in the nuclei of the ‘zone of inhibition’ suggests a prominent structural role of these basic proteins in the organization of the chromatin. The large volume of 2C nuclei of the ‘zone of inhibition’ seems, therefore, to result not from a great nuclear mass, but probably from a relatively small degree of condensation of chromatin.     

Selection and characterization of DNA aptamers with binding selectivity to Campylobacter jejuni using whole-cell SELEX

The need for pre-analytical sample processing prior to the application of rapid molecular-based detection of pathogens in food and environmental samples is well established. Although immunocapture has been applied in this regard, alternative ligands such as nucleic acid aptamers have advantages over antibodies such as low cost, ease of production and modification, and comparable stability. To identify DNA aptamers demonstrating binding specificity to Campylobacter jejuni cells, a whole-cell Systemic Evolution of Ligands by EXponential enrichment (SELEX) method was applied to a combinatorial library of FAM-labeled single-stranded DNA molecules. FAM-labeled aptamer sequences with high binding affinity to C. jejuni A9a as determined by flow cytometric analysis were identified. Aptamer ONS-23, which showed particularly high binding affinity in preliminary studies, was chosen for further characterization. This aptamer displayed a dissociation constant (K _d value) of 292.8 ± 53.1 nM with 47.27 ± 5.58% cells fluorescent (bound) in a 1.48-μM aptamer solution. Binding assays to assess the specificity of aptamer ONS-23 showed high binding affinity (25–36%) for all other C. jejuni strains screened (inclusivity) and low apparent binding affinity (1–5%) with non-C. jejuni strains (exclusivity). Whole-cell SELEX is a promising technique to design aptamer-based molecular probes for microbial pathogens without tedious isolation and purification of complex markers or targets.     

Interpreting Quantifier Scope Ambiguity: Evidence of Heuristic First,Algorithmic Second Processing

The present work suggests that sentence processing requires both heuristic and algorithmic processing streams, where the heuristic processing strategy precedes the algorithmic phase. This conclusion is based on three self-paced reading experiments in which the processing of two-sentence discourses was investigated, where context sentences exhibited quantifier scope ambiguity. Experiment 1 demonstrates that such sentences are processed in a shallow manner. Experiment 2 uses the same stimuli as Experiment 1 but adds questions to ensure deeper processing. Results indicate that reading times are consistent with a lexical-pragmatic interpretation of number associated with context sentences, but responses to questions are consistent with the algorithmic computation of quantifier scope. Experiment 3 shows the same pattern of results as Experiment 2, despite using stimuli with different lexical-pragmatic biases. These effects suggest that language processing can be superficial, and that deeper processing, which is sensitive to structure, only occurs if required. Implications for recent studies of quantifier scope ambiguity are discussed.