Eurythrematosis as a developmental model of the Diabetes Mellitus type 1 pathological condition: pathophysiological parameters and oxidative stress / Eurytrematose como modelo de desenvolvimento da patologia da Diabetes Mellitus tipo 1: parâmetros fisiopatológicos e stress oxidativo

Eurytrematosis is a helminthic disease caused by trematodes belonging to the genus Eurytrema spp. that parasitize the pancreas of many animals and humans. This parasitosis causes chronic fibrosing pancreatitis, fat infiltration in the pancreatic parenchyma, besides damaging the exocrine pancreas, which is similar to that found in patients with Diabetes Mellitus type 1 (DM1). The current work aimed to evaluate the use of bovine pancreas infected with E. coelomaticum as a model to study DM1 pathophysiology. It was carried out macroscopic analyses, parasite identification, total pancreatic lipid determination and oxidative damage biomarkers levels of pancreas naturally infected with E. coelomaticum. Macroscopically, we observed that the infected pancreas had duct obstruction, organ stiffness due to the visible presence of fibrosis, increased adipose tissue deposition, increased protein and lipid damage, as well as increased antioxidant biomarkers (GSH, CAT and VIT C). Thus, it is possible to show that DM1 may have pancreatic parasitism as a possible primary origin. However, more studies are needed to better investigate this possible primary origin; the results obtained here suggest that the use of pancreas parasitized by E. coelomaticum could be a model to investigate DM1 pathophysiology.

Comparative Studies on the Susceptibility of (R)-2,3-Dipalmitoyloxypropylphosphonocholine (DPPnC) and Its Phospholipid Analogues to the Hydrolysis or Ethanolysis Catalyzed by Selected Lipases and Phospholipases

Susceptibility of soybean phosphatidylcholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and its phosphono analogue (R)-2,3-dipalmitoyloxypropylphosphonocholine (DPPnC) towards selected lipases and phospholipases was compared. The ethanolysis of substrates at sn-1 position was carried out by lipase from Mucor miehei (Lipozyme®) and lipase B from Candida antarctica (Novozym 435) in 95% ethanol at 30 °C, and the hydrolysis with LecitaseTM Ultra was carried out in hexane/water at 50 °C. Hydrolysis at sn-2 position was carried out in isooctane/Tris-HCl/AOT system at 40 °C using phospholipase A2 (PLA2) from porcine pancreas and PLA2 from bovine pancreas or 25 °C using PLA2 from bee venom. Hydrolysis in the polar part of the studied compounds was carried out at 30 °C in acetate buffer/ethyl acetate system using phospholipase D (PLD) from Streptococcus sp. and PLD from white cabbage or in Tris-HCl buffer/methylene chloride system at 35 °C using PLD from Streptomyces chromofuscus. The results showed that the presence of C-P bond between glycerol and phosphoric acid residue in DPPnC increases the rate of enzymatic hydrolysis or ethanolysis of ester bonds at the sn-1 and sn-2 position and decreases the rate of hydrolysis in the polar head of the molecule. The most significant changes in the reaction rates were observed for reaction with PLD from Streptococcus sp. and PLD from Streptomyces chromofuscus that hydrolyzed DPPnC approximately two times slower than DPPC and soybean PC. The lower susceptibility of DPPnC towards enzymatic hydrolysis by phospholipases D gives hope for the possibility of using DPPnC-like phosphonolipids as the carriers of bioactive molecules that, instead of choline, can be bounded with diacylpropylphosphonic acids (DPPnA).

Efficient Biofilms Eradication by Enzymatic-Cocktail of Pancreatic Protease Type-I and Bacterial α-Amylase

Removal of biofilms is extremely pivotal in environmental and medicinal fields. Therefore, reporting the new-enzymes and their combinations for dispersal of infectious biofilms can be extremely critical. Herein, for the first time, we accessed the enzyme “protease from bovine pancreas type-I (PtI)” for anti-biofilm properties. We further investigated the anti-biofilm potential of PtI in combination with α-amylase from Bacillus sp. (αA). PtI showed a very significant biofilm inhibition effect (86.5%, 88.4%, and 67%) and biofilm prevention effect (66%, 64%, and 70%), against the E. coli, S. aureus, and MRSA, respectively. However, the new enzyme combination (Ec-PtI+αA) exhibited biofilm inhibition effect (78%, 90%, and 93%) and a biofilm prevention effect (44%, 51%, and 77%) against E. coli, S. aureus, and MRSA, respectively. The studied enzymes were found not to be anti-bacterial against the E. coli, S. aureus, and MRSA. In summary, the PtI exhibited significant anti-biofilm effects against S. aureus, MRSA, and E. coli. Ec-PtI+αA exhibited enhancement of the anti-biofilm effects against S. aureus and MRSA biofilms. Therefore, this study revealed that this Ec-PtI+αA enzymatic system can be extremely vital for the treatment of biofilm complications resulting from E. coli, S. aureus, and MRSA.

Application of modified silica gel in the process of trypsin immobilization

The paper presents the use of modified silica gel for the production of immobilized trypsin from bovine pancreas. Silica gel was modified with 3-aminopropyltriethoxysilane, followed by glutaraldehyde. The influence of stirring time on activity of the prepared biocatalyst was determined for individual stages of the modification. Activity of both native and immobilized trypsin was measured using Kunitz method. At the temperature of 55℃ and pH 7.6 native and immobilized trypsin onto modified silica gel indicate optimum activity. The influence of multiple recycling and storage time on activity of immobilized trypsin was tested. After fourteen days of storage at the temperature of 4℃ immobilized trypsin exhibits 75% of its initial activity.

The α-chymotrypsin-catalyzed Povarov reaction: one-pot synthesis of tetrahydroquinoline derivatives

The α-chymotrypsin from bovine pancreas (BPC) catalyzed three-component one-pot Povarov reaction for the synthesis of tetrahydroquinoline derivatives is reported.

Molecular characterization of Eurytrema coelomaticum in cattle from Paraná, Brazil

This study investigated the occurrence of Eurytremaspp. in cattle by analysis of the partial 18S rRNA gene sequence. Trematodes from 44 bovine pancreas were collected and classified based on typical morphological features. PCR assay and sequence analyses of amplified products confirmed that the trematodes classified as Eurytrema coelomaticum were phylogenetically distinct from those identified as E. pancreaticum. The results of this study represent the first molecular characterization of E. coelomaticum within the Americas, and provide an efficient method to differentiate digenean trematodes of domestic animals.