Evidence for antibody-catalyzed ozone formation in bacterial killing and inflammation

Recently, we showed that antibodies catalyze the generation of hydrogen peroxide (H2O2) from singlet molecular oxygen (1O2*) and water. Here, we show that this process can lead to efficient killing of bacteria, regardless of the antigen specificity of the antibody. H2O2 production by antibodies alone was found to be not sufficient for bacterial killing. Our studies suggested that the antibody-catalyzed water-oxidation pathway produced an additional molecular species with a chemical signature similar to that of ozone. This species is also generated during the oxidative burst of activated human neutrophils and during inflammation. These observations suggest that alternative pathways may exist for biological killing of bacteria that are mediated by potent oxidants previously unknown to biology.     

Development of simple and co-dominant PCR markers to genotype puroindoline a and b alleles for grain hardness in bread wheat (Triticum aestivum L.)

Grain hardness is one of the most important quality characteristics of cultivated bread wheat (Triticum aestivum L.). A large deletion in the puroindoline a (Pina) gene or single nucleotide polymorphisms (SNPs) in the puroindoline b (Pinb) gene results in hard grain texture. So far, nine Pina alleles (Pina-D1a–Pina-D1b, Pina-D1k–Pina-D1q) and seventeen Pinb alleles (Pinb-D1a–Pinb-D1g, Pinb-D1p–Pinb-D1ab) have been identified in bread wheat. The major Pina and Pinb alleles identified in hard wheat cultivars are Pina-D1b, Pinb-D1b, Pinb-D1c and Pinb-D1d. In this study, a three-primer PCR system was employed to develop nine co-dominant STS markers for genotyping Pina-D1a and Pina-D1b, whereas temperature-switch (TS) PCR was used to develop six co-dominant SNP markers for genotyping the Pinb-D1a, Pinb-D1b, Pinb-D1c and Pinb-D1d alleles. These STS and TS-PCR markers were used to verify the grain hardness genotype of 100 wheat cultivars. The reliability and genotyping accuracy of TS-PCR markers were confirmed through sequencing of PCR products and a comparison with previously published results. Therefore, STS and TS-PCR markers offer a simple, cost-effective and reliable method for high-throughput genotyping Pina and Pinb alleles to select grain hardness in wheat quality breeding programs and for wheat market classification.     

Cracks in bread crust cause longer crispness retention

Crispness is among the most important factors that the consumer uses to assess the quality of crispy bread. However, this quality attribute is rapidly lost after baking. It is known that crispness retention can be increased more than eight times by enhancing the water vapor permeability of the crust. Current methods to achieve this, i.e., puncturing the bread before baking, require an extra process step. We hypothesize that cracks that appear spontaneously on the crust surface after baking can also enhance water vapor permeability and therefore improve crispness retention. We were able to confirm this hypothesis by preparing composite breads containing the same crumb but different crusts, with crust recipes of varying starch/protein ratios. Crusts systems that were generally high in gelatinized starch content and poor in evenly distributed gluten were more prone to crack after the whole process of part-baking, freezing, and baking off. These cracks led to an increased water vapor permeability of the crust and an eight times longer instrumental crispness retention compared to standard bread. In this paper we also discuss possible causes for crack formation in the crust. We hypothesize that effective cracks are caused by thermal shock in materials with a low ability to dissipate energy.     

The feeding ecology of elaterid larvae in central European arable land: New perspectives based on naturally occurring stable isotopes

To understand soil food webs, empirically generated data on the trophic connections and the feeding ecology of the major below-ground animal taxa are needed. Here we used stable isotope analysis to assess the trophic ecology of wireworms, the larvae of click beetles, in Central European arable land. Wireworms are amongst the major soil macroinvertebrates and are of practical importance in arable soils. Besides feeding on crops, they are thought to feed on weeds, soil organic matter (SOM), and even animal prey, but their feeding ecology is poorly studied under natural conditions. Elaterid larvae and their putative feeding substrates—plant roots, SOM, and litter—were sampled at 17 locations in Austria, Germany, and Italy and their isotope ratios of carbon (¹²C/¹³C) and nitrogen (¹⁴N/¹⁵N) measured to determine the wireworms’ trophic level, the importance of SOM and weeds within the diet of Agriotes larvae, as well as the individual diet variation in Agriotes obscurus larvae. δ¹⁵N signatures suggested that Agriotes larvae are predominately herbivorous, whereas the other wireworm species primarily fed on animal prey. In contrast to SOM, weeds were readily eaten by Agriotes larvae: their dietary contribution ranged between 28% and 67% in weedy maize fields. Most A. obscurus larvae fed on a mixed diet of weeds and maize, although ～15% of the larvae fed primarily on one of the two food sources only. δ¹⁵N signatures indicated that ～10% of the “herbivorous” A. obscurus larvae fed primarily on animal prey, revealing high intraspecific trophic plasticity in these soil insects. Wireworm feeding behaviour is apparently complex at the individual level: the population consists of types A and B generalists, a phenomenon which needs further assessment.     

Diversity of arbuscular mycorrhizal fungi associated with the rhizosphere of tea growing in ‘natural’ and ‘cultivated’ ecosites

The colonization and diversity of arbuscular mycorrhizal fungi (AMF) associated with the rhizosphere of tea [Camellia sinensis (L.) O. Kuntze] growing under ‘natural’ as well as ‘cultivated’ conditions in the Kumaun region of Uttaranchal Himalaya (India), during the periods of active growth and dormancy were investigated. Root and rhizosphere soil samples, collected from both the ecosites (natural and cultivated), were monitored for root colonization. While the percent root colonization was quite high (77.66 ± 4.40 and 86.40 ± 3.02%, in the natural and cultivated tea, respectively) during the period of active growth in both the ecosites, relatively higher colonization (97.33 ± 0.78 and 98.13 ± 0.80%, in the natural and cultivated tea, respectively) was recorded during the period of dormancy. The rhizosphere of cultivated tea bushes was found to be dominated by Glomus morhpotypes (88.89% of the total isolates) along with three morphotypes of Acaulospora; occurrence of 35 morphotypes belonging to four genera viz. Acaulospora (11.43%), Gigaspora (11.43%), Glomus (68.57%) and Scutellospora (8.57%) was recorded in the rhizosphere of tea plants from the natural ecosite. A total of 51 AMF morphotypes were detected. Shannon–Weaver index of diversity was higher (1.80 ± 0.13 and 2.05 ± 0.10 during periods of active growth and dormancy, respectively) at the species level for the natural ecosite over its counterparts from the cultivated ecosite. Values for the diversity indices of natural and cultivated ecosites did not show much variation in the period of dormancy. These data suggest that collectively, various cultural practices negatively affect AMF diversity at the genus level in tea plantations of the colder regions.     

Oocyte Diameter and Plasma Vitellogenin as Predictive Factors to Identify Potential Channel Catfish,Ictalurus punctatus,Suitable for Induced Spawning

This study monitors the progression of oocyte size and plasma hormone profiles of female channel catfish, Ictalurus punctatus, at monthly intervals to stage expectant ovulating, females for strip spawning. The “critical minimum” diameter of an oocyte to reach threshold maturity for channel catfish was 2.5 ± 0.21 mm. Monthly increases of oocyte diameters and plasma vitellogenin concentrations were linear until spawning. The reproductive performance of cannulated and noncannulated catfish did not differ, negating adverse effects of routine cannulation. This study suggests oocyte diameter in channel catfish can be used as a predictive factor to determine and stage potential broodfish suitable for hormone‐induced spawning.     

The structure of a pH-sensing mycobacterial adenylyl cyclase holoenzyme

Class III adenylyl cyclases contain catalytic and regulatory domains, yet structural insight into their interactions is missing. We show that the mycobacterial adenylyl cyclase Rv1264 is rendered a pH sensor by its N-terminal domain. In the structure of the inhibited state, catalytic and regulatory domains share a large interface involving catalytic residues. In the structure of the active state, the two catalytic domains rotate by 55 degrees to form two catalytic sites at their interface. Two alpha helices serve as molecular switches. Mutagenesis is consistent with a regulatory role of the structural transition, and we suggest that the transition is regulated by pH.     

Automated counting of nucleated cells in equine synovial fluid without and with hyaluronidase pretreatment

Background: Microscopy is usually used to obtain manual total and differential cell counts in equine synovial fluid. A faster, more precise method is desirable. Objectives: The objectives were to compare an automated impedance method with a manual method for obtaining total and differential cell counts in equine synovial fluid and to evaluate the effect of pretreatment with hyaluronidase on automated results. Methods: Synovial fluid samples (n=48) were collected into EDTA and analyzed within 48 hours. Automated total and differential cell counts were evaluated using a Medonic CA620‐VET hematology analyzer before and after pretreatment for 5–30 minutes with hyaluronidase (final concentration 0.01 mg/mL). A hemacytometer count and microscopic evaluation of a direct smear were used as the reference method. Intra‐assay coefficients of variation (CV) were determined. Results: Thirty‐one of 46 untreated samples and 0/46 hyaluronidase‐treated samples were error‐flagged by the analyzer. Correlation between automated (ANCC) and manual (MNCC) nucleated cell counts in untreated samples (n=15; R²=0.93) and pretreated samples (n=46; R²=0.94) was high, and pseudomedian difference was low. Intra‐assay CVs for samples with medium and high cellularity were significantly lower for ANCC (1.5–2.7%) compared with MNCC (6.1–15.7%) (P<.01). Valid automated differential cell counts were not obtained. Conclusions: Automated total cell counts obtained on the Medonic analyzer correlate well with manual counts in equine synovial fluid; however, pretreatment with hyaluronidase is required to minimize error flags. Automated differential counts are not accurate for synovial fluid.     

Efficiency of mannose-binding plant lectins in controlling a homopteran insect,the red cotton bug

Yield losses of different crops due to the attack of various classes of insects are a worldwide problem. Sucking type homopteran pests causing damage to many crop species are not controlled by commonly known insecticidal proteins, namely, Bacillus thuringiensis delta-endotoxin (Bt). This study describes the purification of mannose-binding lectins from three different monocotyledonous plants (Allium sativum, Colocasia esculenta, and Diffenbachia sequina) and their effects on a homopteran insect, the red cotton bug. All of them had a detrimental effect on the growth and development of the insect, where A. sativum bulb lectin showed the highest mortality of all, in particular. The same bulb lectin not only affected the growth and fecundity of the insect but also imparted drastic changes in the color, weight, and size, even on the second generation of the insects which have been reared on artificial diet supplemented with a sublethal dose of the lectin. Thus, this finding opens up a possibility of using this lectin as an important component in crop management.