International Veterinary Epilepsy Task Force recommendations for systematic sampling and processing of brains from epileptic dogs and cats

Traditionally, histological investigations of the epileptic brain are required to identify epileptogenic brain lesions, to evaluate the impact of seizure activity, to search for mechanisms of drug-resistance and to look for comorbidities. For many instances, however, neuropathological studies fail to add substantial data on patients with complete clinical work-up. This may be due to sparse training in epilepsy pathology and or due to lack of neuropathological guidelines for companion animals. The protocols introduced herein shall facilitate systematic sampling and processing of epileptic brains and therefore increase the efficacy, reliability and reproducibility of morphological studies in animals suffering from seizures. Brain dissection protocols of two neuropathological centres with research focus in epilepsy have been optimised with regards to their diagnostic yield and accuracy, their practicability and their feasibility concerning clinical research requirements. The recommended guidelines allow for easy, standardised and ubiquitous collection of brain regions, relevant for seizure generation. Tissues harvested the prescribed way will increase the diagnostic efficacy and provide reliable material for scientific investigations

Alkaline sulfite thermomechanical pretreatment of sugarcane bagasse

Chemithermomechanical (CTMP) processing of lignocellulosic materials can be used as an efficient pretreatment of lignocellulosic materials. This type of process has been largely used in the mechanical pulping industry with currently available equipment and robust technology. The use of alkaline sulfite in this process provides selective lignin removal up to 50% delignification, which has been proved efficient to decrease recalcitrance of the pretreated materials. The current work evaluates sugarcane bagasse pretreatment by alkaline sulfite CTMP processing. Solids yield after processing with increasingly chemical loads (from 1.25% to 10% sulfite) were in the range of 87% to 82%, respectively. The most severe pretreated sample was completely hydrolyzed by 10 FPU cellulases (Cellic CtecII)/g of substrate within 24 h hydrolysis. Lower chemical doses in the pretreatment provided substrates with higher recalcitrance. Mass balance of sugarcane bagasse components at the most severe condition indicated 49% and 28% of lignin and xylan removal, respectively. Under this condition, the glucan loss was limited to 4%. Besides delignification, the residual lignins appeared sulfonated after CTMP pretreatment. Both delignification and sulfonation of residual lignins resulted in significant increases in the water retention values of the pretreated material (from 134% to 244% in the untreated and severely pretreated material, respectively). 2D-NMR evaluation of the pretreated materials corroborated intense delignification with increasingly sulfite loads. Xylan side groups were progressively removed, whereas part of the original p-coumaric acid (PCA), acylating gama-OH of the lignin side chain, resisted to the most severe pretreatment condition. Direct correlations were observed for lignin removal, WRV, sulfonation of residual lignins and digestibility of the polysaccharides fraction.FAPESP 2014/06923-6Peer Reviewe

Fate of p-hydroxycinnamates and structural characteristics of residual hemicelluloses and lignin during alkaline-sulfite chemithermomechanical pretreatment of sugarcane bagasse

12 páginas.-- 2 figuras.-- 4 tablas.-- 39 referencias.-- The online version of this article ( https://doi.org/10.1186/s13068-018-1155-3) contains supplementary material, which is available to authorized users.Background: Preparing multiple products from lignocellulosic biomass feedstock enhances the profit and sustainability of future biorefineries. Grasses are suitable feedstocks for biorefineries as they permit a variety of possible by-products due to their particular chemical characteristics and morphology. Elucidating the fate of p-hydroxycinnamates (ferulates—FAs and p-coumarates—pCAs) and major structural components during bioprocessing helps to discriminate the sources of recalcitrance in grasses and paves the way for the recovery of p-hydroxycinnamates, which have multiple applications. To address these subjects, we assessed sugarcane bagasse biorefining under alkalinesulfite chemithermomechanical (AS-CTM) pretreatment and enzymatic saccharification. Results: The mass balances of the major bagasse components were combined with 2D-NMR structural evaluation of process solids to advance our understanding of sugarcane bagasse changes during biorefining. AS-CTM pretreatment provided a high yield and thoroughly digestible substrates. The pretreated material was depleted in acetyl groups, but retained 62 and 79% of the original lignin and xylan, respectively. Forty percent of the total FAs and pCAs were also retained in pretreated material. After pretreatment and enzymatic hydrolysis, the residual solids contained mostly lignin and ester-linked pCAs, with minor amounts of FAs and non-digested polysaccharides. Saponification of the residual solids, at a higher alkali load, cleaved all the ester linkages in the pCAs; nevertheless, a significant fraction of the pCAs remained attached to the saponified solids, probably to lignin, through 4-O ether-linkages. Conclusion: AS-CTM pretreatment provided soundly digestible substrates, which retain substantial amounts of xylans and lignin. Acetyl groups were depleted, but 40% of the total FAs and pCAs remained in pretreated material. Ester-linked pCAs detected in pretreated material also resisted to the enzymatic hydrolysis step. Only a more severe saponification reaction cleaved ester linkages of pCAs from residual solids; nevertheless, pCAs remained attached to the core lignin through 4-O ether-linkages, suggesting the occurrence of an alkali-stable fraction of pCAs in sugarcane bagasse.This work was supported by FAPESP (Contract Number 14/06923-6), CNPq, and CAPES. Felipe AM Reinoso thanks CAPES for his student fellowships. JCdR, JR, and AG were funded by the Spanish Projects AGL2014-53730-R, CTQ2014-60764-JIN and AGL2017-83036-R (financed by Agencia Estatal de Investigación, AEI, and Fondo Europeo de Desarrollo Regional, FEDER), and the CSIC Project 2014-40E-097Peer reviewe