From fix to fit into the autoptic human brains.

Formalin-fixed, paraffinembedded (FFPE) human brain tissues are very often stored in formalin for long time. Formalin fixation reduces immunostaining, and the DNA/RNA extraction from FFPE brain tissue becomes suboptimal. At present, there are different protocols of fixation and several procedures and kits to extract DNA/RNA from paraffin embedding tissue, but a gold standard protocol remains distant. In this study, we analyzed four types of fixation systems and compared histo and immuno-staining. Based on our results, we propose a modified method of combined fixation in formalin and formic acid for the autoptic adult brain to obtain easy, fast, safe and efficient immunolabelling of long-stored FFPE tissue. In particular, we have achieved an improved preservation of cellular morphology and obtained success in postmortem immunostaining for NeuN. This nuclear antigen is an important marker for mapping neurons, for example, to evaluate the histopathology of temporal lobe epilepsy or to draw the topography of cardiorespiratory brainstem nuclei in sudden infant death syndrome (SIDS). However, NeuN staining is frequently faint or lost in postmortem human brain tissues. In addition, we attained Fluoro Jade C staining, a marker of neurodegeneration, and immunofluorescent staining for stem cell antigens in the postnatal human brain, utilizing custom fit fixation procedures

Pasta-making properties of the new durum wheat variety biensur suitable for the northern mediterranean environment

Industrial pasta is commonly made from mixtures of semolina from different durum wheat varieties, and there is a very low market presence of mono-varietal pasta from local, short supply chains. In this work, dough rheological properties and pasta quality traits of the new durum wheat cv. Biensur, which has a high HMW/LMW-GS ratio, were evaluated with a view to developing short-chain, mono-varietal pasta production in NE Italy. Chemical and sensory analyses on short-cut pasta, viz. tubetti, made with semolina from cv. Biensur at two drying temperatures revealed that it has good technological characteristics and stability, excellent cooking and sensory properties, and is comparable to the high-quality commercial reference cv. Aureo. We conclude that Biensur provides farmers and traders with new market opportunities and offers improvements to the environmental and economic sustainability of the durum wheat chain

Investigaciones estratigráfico-tectónicas en el curso medio del arroyo Picún-Leufú (Gobernación del Neuquén)

Tesis presentada para optar al Grado de Doctor en Ciencias NaturalesFil: Simonato, Italo B.. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo; Argentin

Correlating Noble Rot Infection of Garganega Withered Grapes with Key Molecules and Odorants of Botrytized Passito Wine

Experimental passito wines with dierent percentages of naturally noble-rotten grapes of the Garganega variety were analyzed to evaluate key molecules and odorants related to the typical aroma and sensory profile of botrytized passito wine. Remarkable changes in the concentration of 1-octen-3-ol, 4-terpineol, benzaldehyde, N-(3-methylbutyl)acetamide, and sherry lactone 1 and 2 were observed between sound and noble-rotten wines. Wines were perceived to be dierent for floral, honey, figs, apricot, and caramel scents. By partial least square regression these descriptors were well correlated to samples. An important positive contribution of sherry lactones, N-(3-methylbutyl)acetamide, vanillin, benzaldehyde, and -butyrolactone to honey, apricot, and caramel was observed. It is conceivable that oxidative eects of Botrytis cinerea infection play an important role in the genesis of these chemical and sensory aroma markers. This study provides a predictive tool for winemakers that use natural grape withering to produce wines whose aroma profile is not standardized due to the seasonal variation of noble rot incidence

Development and evaluation of a methodology to integrate technical and sensorial properties in materials selection.

In the materials selection process, the use of different tools, languages and perspectives frequently causes disagreement between engineers and industrial designers. The aim of the paper is to define an integrated method for materials selection that provides industrial designers with measurable data to support and explain aesthetic decisions on materials. A new method for materials selection consisting of multiple tools structured in a two-step framework is presented. The method is tested through a case study of professional kitchen appliances where metal components are replaced with polymers. The first step involved the application of an established technique to identify polymeric bulk solutions, based on their technical properties. The second step employed a sensory analysis test to choose suitable finishes. Thirty-seven individuals performed the test: the subjects highlighted their main perceptions of metal and metal-look polymer finishes. The research demonstrates that the proposed method is suitable for the evaluation of both technical and sensorial properties of materials. In particular, Mapping test represents a rapid, low cost and effective tool to help industrial designers justify Colour Materials and Finish (CMF) choices with quantifiable information

322. Benign Herpes Simplex Virus Vector Design for Efficient Delivery of Large or Multiple Transgenes To a Diversity of Cells

Viral vectors derived from herpes simplex virus (HSV) have the potential to revolutionize gene therapy due to their ability to accommodate large and multiple therapeutic transgenes. However, current HSV gene therapy vectors express toxic levels of an immediate-early (IE) protein, ICP0, whose function is required for robust and sustained transgene expression. Here we report the development of a new generation of HSV vectors that are IE-gene independent and non-toxic, yet capable of persistent transgene expression in a variety of human primary non-neuronal cell types. We identified a CTCF motif cluster upstream of the latency promoter and a known long-term regulatory region as key elements for the protection of transgene expression cassettes from global silencing of the viral genome in the absence of all viral IE gene products. Using this new HSV vector system, we have observed vigorous expression of full-length dystrophin cDNA (14 kb) for several weeks in a dystrophin-deficient muscle cell line. We further tested our vectors for transgene expression in rodent brain. While we detected variable persistence of gene expression from the latency locus, we were surprised to observe vigorous long-term reporter gene expression from one other locus despite the absence of gene expression from this locus in non-neuronal cells. These findings demonstrate that transgene expression in neurons is operatively different from that in non-neuronal cells and suggest that multiple loci can be used for expression of foreign genes in the nervous system. In addition, our data raise the prospect that our highly defective HSV vector system will be applicable as a safe delivery tool for large and multiple therapeutic genes to a wide range of non-neuronal tissues

A comprehensive overview on Kratom

Kratom (Mitragyna speciosa Korth) is a tropical tree, indigenous to South East Asia. Historically, the plant is locally used as a stimulant, a remedy in traditional medicine and in social context. Imported to Western countries, Kratom is classified as a novel psychoactive substance (NPS). A systematic review of the literature on Mitragyna speciosa and its main constituents was carried by our international multidisciplinary group. Results were qualitatively analysed in three main areas of interest: in-vitro and preclinical data on pharmacology and behavioral effects, laboratoristic techniques for identification/characterization, epidemiological/toxicological reports on humans. At present, there is no systematic data on the prevalence of Kratom use in all the native countries, but it seems to be considerable. In South-East Asia, Kratom, even if banned, might be still considered a better option than other illicit drugs, an alternative opioid treatment, a “natural” remedy with no real social stigma attached to its consumption. In parallel, this ethno-drug seems to be popular in Western countries, largely unregulated, easily available on the Internet. Kratom pharmacology appears to be complex, with many alkaloids involved. The subjective effects in humans are very peculiar and seem to be dose-dependent, ranging from psycho-stimulant to sedative-narcotic. Available data on Kratom suggest caution: this psychoactive plant could exhibit a serious harmful potential. Kratom use seems to be associated with drug dependency, development of withdrawal symptoms, craving, serious adverse effects and life-threatening effects in a multidrug-intoxicating scenario. On the other hand, its anxyiolitic, antidepressant and analgesic properties deserve to be further studied

Endoparasites and vector-borne pathogens in dogs from greek islands: Pathogen distribution and zoonotic implications

The present study investigated the presence of endo-and ecto-parasites, and vector-borne pathogens, in dogs from four islands of Greece. A total of 200 (123 owned and 77 sheltered) dogs were examined with different microscopic, serological and molecular methods. Of the examined dogs, 130 (65%) were positive for one or more parasites and/or vector-borne pathogens. The most common zoonotic intestinal helminths recorded were Ancylos-tomatidae (12.5%) and Toxocara canis (3.5%). Ninety-three dogs (46.5%) seroreacted to Rickettsia conorii. Twenty-two (11%) of them were also PCR positive and 7 (3.5%) showed corpuscles suggestive of Rickettsia spp. on the blood smears. Nineteen dogs (9.5%) were seropositive for Ehrlichia canis, three of them being also PCR positive. Dogs positive for Anaplasma phagocytophilum-Anaplasma platys (1%), Dirofilaria immitis (0.5%) and Babesia canis (0.5%) were also found. Fleas and ticks were recorded in 53 (26.5%) and 50 (25%) dogs, respectively, and all specimens were identified as Ctenocephalides felis felis and Rhipicephalus sanguineus sensu lato. Binary multiple univariate Generalized Linear Models were used to investigate factors and clinical signs related to the recorded positivity, while the association of specific signs with the pathogens was evaluated using tests of independence. Knowledge of occurrence and impact of zoonotic parasites and vector-borne pathogens in dog populations is crucial to prevent the infection in animals and people, and to control the risk of spreading of these pathogens in endemic and non-endemic areas

Deletion of the Virion Host Shut-off Gene Enhances Neuronal-Selective Transgene Expression from an HSV Vector Lacking Functional IE Genes

The ability of herpes simplex virus (HSV) to establish lifelong latency in neurons suggests that HSV-derived vectors hold promise for gene delivery to the nervous system. However, vector toxicity and transgene silencing have created significant barriers to vector applications to the brain. Recently, we described a vector defective for all immediate-early gene expression and deleted for the joint region between the two unique genome segments that proved capable of extended transgene expression in non-neuronal cells. Sustained expression required the proximity of boundary elements from the latency locus. As confirmed here, we have also found that a transgene cassette introduced into the ICP4 locus is highly active in neurons but silent in primary fibroblasts. Remarkably, we observed that removal of the virion host shutoff (vhs) gene further improved transgene expression in neurons without inducing expression of viral genes. In rat hippocampus, the vhs-deleted vector showed robust transgene expression exclusively in neurons for at least 1 month without evidence of toxicity or inflammation. This HSV vector design holds promise for gene delivery to the brain, including durable expression of large or complex transgene cassettes

Distilled grape pomace as a functional ingredient in vegan muffins: effect on physicochemical, nutritional, rheological and sensory aspects

Wheat-based vegan muffins were formulated with increasing levels (0, 5 and 10 g per 100 g substitutes to flour) of dried grape pomace powder obtained after distillation (DGPP). The DGPP-muffins showed a greater (P < 0.05) total dietary fibre and a lower (P < 0.05) total starch content to the control. The inclusion of DGPP influenced (P < 0.05) both the batter's rheological properties and the baked muffins' technological characteristics, including the baking loss, the volume, the firmness, the spread ratio and the colour. The in vitro antioxidant activity and the phenolic content increased (P < 0.05) following DGPP inclusion. The untargeted phenolic profiling by UHPLC-HRMS revealed an (P < 0.05) increase of several phenolic classes (i.e. free and bound fractions) following the inclusion of DGPP in the recipe. Flavonoids, mainly anthocyanins, were the most abundant compounds. A quantitative descriptive sensory analysis detected the DGPP-vegan muffins showed good sensory acceptability. The vegan muffin with 5 g per 100 g of DGPP obtained the highest overall acceptability score by the panellists