Detection of VAMP Proteolysis by Tetanus and Botulinum Neurotoxin Type B In Vivo with a Cleavage-Specific Antibody

Tetanus and Botulinum type B neurotoxins are bacterial metalloproteases that specifically cleave the vesicle-associated membrane protein VAMP at an identical peptide bond, resulting in inhibition of neuroexocytosis. The minute amounts of these neurotoxins commonly used in experimental animals are not detectable, nor is detection of their VAMP substrate sensitive enough. The immune detection of the cleaved substrate is much more sensitive, as we have previously shown for botulinum neurotoxin type A. Here, we describe the production in rabbit of a polyclonal antibody raised versus a peptide encompassing the 13 residues C-terminal with respect to the neurotoxin cleavage site. The antibody was affinity purified and found to recognize, with high specificity and selectivity, the novel N-terminus of VAMP that becomes exposed after cleavage by tetanus toxin and botulinum toxin type B. This antibody recognizes the neoepitope not only in native and denatured VAMP but also in cultured neurons and in neurons in vivo in neurotoxin-treated mice or rats, suggesting the great potential of this novel tool to elucidate tetanus and botulinum B toxin activity in vivo.     

Agronomic performance of experimental fertilizers on spinach (<Emphasis Type="Italic">Spinacia oleracea</Emphasis> L.) in organic farming

In organic farming, soil application of processed agro-industrial by-products could sustain soil fertility for vegetables, which have short cropping cycles. Therefore, the objectives of this 2-year research on organic spinach crop were to assess the productive performance of different experimental fertilizers, the effects on soil fertility, and investigate the dynamics of some soil properties and the N balance. Two types of olive pomace mixtures, with a different initial C/N ratio, were composted and both stopped at the active phase (A1 and B1) and processed until maturation (A2 and B2). Also an anaerobic digestate (DA), and the B2 applied as amendment (B2A) were studied. The four composts, DA, and B2A were compared with a commercial organic fertilizer (Org), and a control (N0). The Org resulted as not sustainable in maintaining soil fertility in the long-term, mainly due to reduction in the soil of total organic carbon by 32 %, compared to the average of the other treatments. Conversely, choosing stage of maturity and adequate C/N of starting mixtures was among the best practices for compost use in spinach crop. The great content of nutrients (N and K higher by 102 and 86 % than Org, respectively), and N surplus (1431 kg ha^?1) in the B2A plots would suggest that they could accumulate after subsequent soil applications, with the risk of losses in the environment. The DA appeared to be the most suitable fertilizer to get a favorable trade-off among yield, quality and N-use efficiency, when applied according to best agronomic practices.     

Urinary Metabolic Phenotyping Reveals Differences in the Metabolic Status of Healthy and Inflammatory Bowel Disease (IBD) Children in Relation to Growth and Disease Activity

Background: Growth failure and delayed puberty are well known features of children and adolescents with inflammatory bowel disease (IBD), in addition to the chronic course of the disease. Urinary metabonomics was applied in order to better understand metabolic changes between healthy and IBD children. Methods: 21 Pediatric patients with IBD (mean age 14.8 years, 8 males) were enrolled from the Pediatric Gastroenterology Outpatient Clinic over two years. Clinical and biological data were collected at baseline, 6, and 12 months. 27 healthy children (mean age 12.9 years, 16 males) were assessed at baseline. Urine samples were collected at each visit and subjected to ¹H Nuclear Magnetic Resonance (NMR) spectroscopy. Results: Using ¹H NMR metabonomics, we determined that urine metabolic profiles of IBD children differ significantly from healthy controls. Metabolic differences include central energy metabolism, amino acid, and gut microbial metabolic pathways. The analysis described that combined urinary urea and phenylacetylglutamine—two readouts of nitrogen metabolism—may be relevant to monitor metabolic status in the course of disease. Conclusion: Non-invasive sampling of urine followed by metabonomic profiling can elucidate and monitor the metabolic status of children in relation to disease status. Further developments of omic-approaches in pediatric research might deliver novel nutritional and metabolic hypotheses.     

The effect of lighting changes on simultaneous colour contrast: a new aspect of colour rendition

Among the many metrics used to compute the colour rendition of a light source which have been investigated in recent years, most consider only the spectral power distribution of the light source being tested. To highlight aspects not usually considered in the development of colour rendition metrics, and to propose future directions for alternative approaches consistent with human visual perception, an experiment was conducted to examine colour appearance variations on certain spatial visual content configurations, in other words the effects of illusions under different light sources. Results indicate that stronger simultaneous contrast configurations have a more unpredictable variation in appearance. Therefore, it is proposed that colour spatial distribution should be considered for the future generation of rendering metrics.     

Helena’s Many Daughters: More Mitogenome Diversity behind the Most Common West Eurasian mtDNA Control Region Haplotype in an Extended Italian Population Sample

The high number of matching haplotypes of the most common mitochondrial (mt)DNA lineages are considered to be the greatest limitation for forensic applications. This study investigates the potential to solve this constraint by massively parallel sequencing a large number of mitogenomes that share the most common West Eurasian mtDNA control region (CR) haplotype motif (263G 315.1C 16519C). We augmented a pilot study on 29 to a total of 216 Italian mitogenomes that represents the largest set of the most common CR haplotype compiled from a single country. The extended population sample confirmed and extended the huge coding region diversity behind the most common CR motif. Complete mitogenome sequencing allowed for the detection of 163 distinct haplotypes, raising the power of discrimination from 0 (CR) to 99.6% (mitogenome). The mtDNAs were clustered into 61 named clades of haplogroup H and did not reveal phylogeographic trends within Italy. Rapid individualization approaches for investigative purposes are limited to the most frequent H clades of the dataset, viz. H1, H3, and H7.     

Surface modification of activated carbon fabric with ozone. Part 3: Thermochemical aspects and electron spin resonance

It is shown that the differential scanning calorimetry (DSC) analysis of water desorption from activated carbon fabric (ACF) surfaces having different degrees of oxidation, is a valid alternative to immersion calorimetry in determining the desorption/adsorption energy of an adsorbate on the oxidized ACF surfaces. Processing the DSC endotherm relative to water desorption through the Clausius-Clapeyron equation it is possible to calculate either the water desorption energy from the oxidized ACF surfaces and also the hydrogen bond energy through which the water molecules are bound to the oxidized ACF surface. The DSC analysis of the ozonized ACF has revealed that secondary ozonides and peroxides are formed together with other oxygenated groups on the ACF surface, and were detected through an exothermal transition with two peaks at 118°C and 142°C with the measured decomposition enthalpy directly proportional to the degree of oxidation of the ACF surface. The ESR analysis on pristine ACF has revealed the presence of clustered paramagnetic centers. The reaction of the ACF with ozone greatly enhances the ESR signal since more free radicals are formed on the surface mainly in the form of oxygen-centered peroxide-type.     

Graphene nanoribbons produced by the oxidative unzipping of single-wall carbon nanotubes

Graphene nanoribbons were synthesized by oxidative unzipping of single-wall carbon nanotubes (SWCNTs). The nanoribbons produced from SWCNTs were characterized using FT-IR, Raman and X-ray photoelectron spectroscopy. For the morphological study of the product obtained from the SWCNT unzipping reaction, transmission electron microscopy and atomic force microscopy were used, confirming the typical graphene nanoribbon structure.     

Extensive Placental Methylation Profiling in Normal Pregnancies

The placental methylation pattern is crucial for the regulation of genes involved in trophoblast invasion and placental development, both key events for fetal growth. We investigated LINE-1 methylation and methylome profiling using a methylation EPIC array and the targeted methylation sequencing of 154 normal, full-term pregnancies, stratified by birth weight percentiles. LINE-1 methylation showed evidence of a more pronounced hypomethylation in small neonates compared with normal and large for gestational age. Genome-wide methylation, performed in two subsets of pregnancies, showed very similar methylation profiles among cord blood samples while placentae from different pregnancies appeared very variable. A unique methylation profile emerged in each placenta, which could represent the sum of adjustments that the placenta made during the pregnancy to preserve the epigenetic homeostasis of the fetus. Investigations into the 1000 most variable sites between cord blood and the placenta showed that promoters and gene bodies that are hypermethylated in the placenta are associated with blood-specific functions, whereas those that are hypomethylated belong mainly to pathways involved in cancer. These features support the functional analogies between a placenta and cancer. Our results, which provide a comprehensive analysis of DNA methylation profiling in the human placenta, suggest that its peculiar dynamicity can be relevant for understanding placental plasticity in response to the environment.     

Impact of waste cooking oil in biodiesel blends on particle size distributions from a city-car engine

Using biodiesel as a blending component in diesel engine has demonstrated to reduce hydrocarbon and particulate matter emissions. Literature showed that biodiesel type, engine architecture and test conditions deeply affect performance and emission characteristics. Among suitable biodiesel fuels, waste cooking oil (WCO) is considered very attractive due to the reduced environmental impact without sacrificing engine performance.This paper aims at investigating how mixing ratio of biodiesel from WCO and mineral diesel affects the particle size distributions of a current state of art small displacement diesel engine.Experimental tests have been performed on an up-to date light common rail diesel engine. Its complete operative field has been investigated. The results obtained show that the use of biodiesel blends from WCO reduces the total number of particles emitted from the engine with respect to the diesel fuel; the reduction is more evident as the percentage of biodiesel in the blend increases. The number of particles in WCO biodiesel soot with diameter smaller than 10 nm is reduced as compared to diesel fuel; the same trend is observed for diameters larger than 200 nm; comparable particle numbers were obtained in the ultrafine range (Dp < 100 nm).