scholarly journals Action of cardosin A from Cynara humilis on ovine and caprine caseinates

2000 ◽  
Vol 67 (3) ◽  
pp. 449-454 ◽  
Author(s):  
SOFIA V. SILVA ◽  
F. XAVIER MALCATA
Keyword(s):  
Proteolytic Activity ◽  
Small Ruminants ◽  
Sodium Caseinate ◽  
Cheese Ripening ◽  
Caprine Milk ◽  
Proteolytic Action ◽  
Milk Coagulation ◽  
Cardosin A ◽  
Intermediate System

In the Iberian Peninsula, the proteinases present in the flowers of members of the Cynara genus, C. cardunculus, C. humilis and C. scolymus, have for many years been successfully used in the manufacture of traditional cheeses from ovine and/or caprine milk on individual farms (Vieira de Sá & Barbosa, 1972; Trujillo et al. 1994). In Portugal, C. cardunculus is the species most frequently employed. Although commercial thistle was tentatively assumed to be pure in taxonomic terms, accurate analyses have shown that the flowers of C. cardunculus are often mixed with flowers of C. humilis (Pires et al. 1994). The clotting activity of C. humilis is due to an aspartic proteinase, currently designated cardosin A and similar to another enzyme obtained from C. cardunculus. This enzyme is similar in specificity and activity to chymosin (Pires et al. 1994).The action of cardosin A from C. cardunculus upon ovine and caprine caseins has been reported recently (Ramalho-Santos et al. 1996; Simo4es, 1998; Sousa & Malcata, 1998), but as yet there is no information on the proteolytic activity of the proteinase from C. humilis upon caseins from milks other than bovine. Caseins from small ruminants' milks are the usual substrates of cardosin during milk coagulation and cheese ripening, and sodium caseinate represents an intermediate system between isolated caseins and the cheese matrix that is free from interference by fat. Thus ovine and caprine caseinates may be useful substrates for investigating the proteolytic activity of cardosin.The aim of the present study was to compare the action of pure cardosin A, obtained from C. humilis, on caprine and ovine caseinates, and to assess the in vitro contribution of this enzyme to the overall proteolytic action of thistle rennet.

In Vitro Studies on a Proteolytic Enzyme from Aspergillus Oryzae (Protease I)

1968 ◽  
Vol 19 (01/02) ◽  
pp. 136-144 ◽  
Author(s):  
D Ogston ◽  
C. M Ogston
Keyword(s):  
Proteolytic Activity ◽  
Fibrinolytic Activity ◽  
Proteolytic Enzyme ◽  
Thrombolytic Agent ◽  
Inhibitory Effect ◽  
In Vitro Studies ◽  
Clotting Time ◽  
Proteolytic Action ◽  
Fibrinolytic Action

Summary1. Protease I was found to have potent fibrinolytic activity in concentrations which exceeded the blood inhibitory capacity when tested on fibrin plates and artificial thrombi.2. Plasma inhibited the proteolytic activity of protease I to a greater extent than serum; serum had a greater inhibitory effect on protease I than on plasmin. Trasylol did not inhibit the proteolytic action of protease I.3. Protease I caused the slow formation of fibrin in plasma in concentrations which did not produce fibrinogenolysis; this effect was seen in Al(OH)3-adsorbed plasma, and was not inhibited by heparin. Protease I also shortened the recalcified plasma clotting time.4. The fibrinogenolytic action of protease I was more rapid than its fibrinolytic action both in the presence and absence of plasma inhibitors. No concentration of protease I lysed fibrin in plasma without prior destruction or conversion to fibrin of the surrounding plasma fibrinogen.5. It is concluded from these in vitro studies that protease I does not have the properties necessary for a satisfactory thrombolytic agent.

Effect of Jatropha curcas Latex on L3 Haemonchus contortus Larval Motility

2017 ◽  
Vol 9 (3) ◽  
Author(s):  
Noorzaid Muhamad ◽  
Syahirah Sazeli ◽  
Resni Mona ◽  
Jannathul Firdous
Keyword(s):  
Larvicidal Activity ◽  
Jatropha Curcas ◽  
Haemonchus Contortus ◽  
Anthelmintic Resistance ◽  
Small Ruminants ◽  
Gastrointestinal Nematodes ◽  
Dose Dependent ◽  
Plants Extract

The anthelmintic resistance has limited the control of gastrointestinal nematodes of small ruminants and thus has awakened interest in the study of plants extract as a source of anthelmintics. These experiments were carried out to evaluate the in vitro efficacy of Jatrophacurcas latex extract against Haemonchuscontortus larval motility. To evaluate the larvicidal activity, H.contortus L3 were incubated with the extracts with varying concentration of 5 mg/mL, 10 mg/mL, 15 mg/mL and 20 mg/mL at 27°C for 48, 72 and 96 hrs. The results were subjected to the Kruskal-Wallis test (P less than 0.05). The extracts showed dose-dependent larvicidal effects. These results suggest that J.curcas can be used to control gastrointestinal nematodes of small ruminants.

scholarly journals Chromatographic Characterization and In Vitro Bioactivity Evaluation of Lactobacillus helveticus Hydrolysates upon Fermentation of Different Substrates

2021 ◽  
Vol 11 (2) ◽  
pp. 811
Author(s):  
Federica Ianni ◽  
Alessandra Anna Altomare ◽  
Beniamino T. Cenci-Goga ◽  
Francesca Blasi ◽  
Luca Grispoldi ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Proteolytic Activity ◽  
Bioactive Peptides ◽  
Biological Activities ◽  
Skim Milk ◽  
Brain Heart Infusion ◽  
Starter Cultures ◽  
Lactobacillus Helveticus ◽  
Food Sources

Among various food sources, milk proteins remain the major vector for functional peptides endowed with several biological activities. Particularly, the proteolytic activity of lactic acid bacteria during milk fermentation has been one of the most followed strategies to produce bioactive peptides. In the present study, the exploration of the activity of several starter cultures, at different fermentation times, was firstly investigated by reversed phase-high performance liquid chromatography. Among the tested strains, Lactobacillus helveticus showed a higher proteolytic activity and it was submitted to further investigations by changing the fermentation substrate (skim milk, brain heart infusion, peptone water) as well as the extraction strategy (trichloroacetic acid vs. glass beads). The chromatographic analyses and the in vitro antioxidant and antihypertensive assays highlighted considerable differences for L. helveticus hydrolysates from different substrates, while a negligible impact by the two extraction protocols emerged. Furthermore, nano-high pressure liquid chromatography coupled with a high resolution mass spectrometry analyzer allowed the preliminary discrimination of fractions from fermented skim milk, likely responsible for the found activity. The obtained results suggest the possibility of varying the fermentation parameters in order to maximize the functional effects of the bioactive peptides.

scholarly journals FURTHER STUDIES ON THE ROLE OF PROTEASES IN THE ALLERGIC REACTION

1961 ◽  
Vol 113 (2) ◽  
pp. 359-380 ◽  
Author(s):  
Georges Ungar ◽  
Takuso Yamura ◽  
Jacqueline B. Isola ◽  
Sidney Kobrin
Keyword(s):  
Amino Acid ◽  
Guinea Pig ◽  
Proteolytic Activity ◽  
Guinea Pigs ◽  
Protease Activity ◽  
Amino Acid Esters ◽  
Protein Substrates ◽  
Protease Activation ◽  
Acid Esters

Protease activity was measured through the hydrolysis of synthetic amino acid esters in body fluids and tissues of guinea pigs, rats, mice, and humans. Significant in vitro activation was observed in serum and lung slices of sensitized guinea pigs on addition of the specific antigen. Increased proteolytic activity was also seen in reverse anaphylaxis. More marked activation occurred when guinea pig serum was treated with peptone and guinea pig or rat serum was treated with agar. Protease activation was demonstrated in specimens of human skin under the influence of a poison ivy extract or croton oil added in vitro. Urinary protease activity of guinea pigs increased significantly during the first hours of anaphylactic shock and very markedly in peptone shock. Peptone shock, elicited in mice pretreated with H. pertussis, was accompanied by a considerable increase in protease activity in the peritoneal fluid as compared with non-pretreated mice which were insensitive to peptone. Proteolytic activity resulting from the activation procedures was due to a number of proteases. The dominant substrate affinity and inhibition patterns suggest that serum and urine proteases are similar to but not identical with plasmin. Anaphylactic activation exhibited patterns different from those resulting from the action of anaphylactoid agents. Tissue enzymes are either of cathepsin- or chymotrypsin-type or mixtures of both. Some of the activated enzymes, although remarkably effective in hydrolyzing amino acid esters, show no activity on protein substrates. This does not justify, however, their designation as "esterases." They probably belong to the class of specific proteases acting only on a single or a small number of functionally significant protein substrates. There is at present sufficient evidence to prove not only that protease activation does occur in anaphylaxis and anaphylactoid conditions but also that it is an important component of the chain of reactions leading to the allergic response.

scholarly journals Isolation of autophagic vacuoles from rat liver: morphological and biochemical characterization.

1982 ◽  
Vol 93 (1) ◽  
pp. 144-154 ◽  
Author(s):  
L Marzella ◽  
J Ahlberg ◽  
H Glaumann
Keyword(s):  
Proteolytic Activity ◽  
Rat Liver ◽  
Biochemical Characterization ◽  
Secretory Granules ◽  
Density Lipoprotein ◽  
Marker Enzymes ◽  
Cell Organelles ◽  
Protein Ratio ◽  
Autophagic Vacuoles

The induction of autophagy caused by vinblastine (VBL) has been found to be concomitant with a stimulation of proteolysis in a mitochondrial-lysosomal (ML) fraction from the rat liver (Marzella and Glaumann, 1980, Lab. Invest., 42: 8-17. Marzella and Glaumann, 1980, Lab. Invest., 42:18-27). In this fraction the enhanced proteolysis is associated with a threefold increase in the relative fractional volume of autophagic vacuoles (AVs). In an attempt to isolate the AVs, we subfractionated the ML suspension at different intervals after the induction of autophagy by VBL by centrifugation on a discontinuous Metrizamide gradient ranging from 50% to 15%. The material banding at the 24 to 20% and the 20 to 15% interphases was collected. Morphological analysis reveals that 3 h after induction of autophagy these fractions consist predominantly (approximately 90%) of intact autophagic vacuoles. These autophagic vacuoles contain cytosol, mitochondria, portions of endoplasmic reticulum, and occasional very low density lipoprotein, particles either free or in Golgi apparatus derivatives, in particular secretory granules. The sequestered materials show ultrastructural signs of ongoing degradation. In addition to containing typical autophagic vacuoles, the isolated fractions consist of lysosomes lacking morphologically recognizable cellular components. Contamination from nonlysosomal material is only a few percent as judged from morphometric analysis. Typical lysosomal "marker" enzymes are enriched 15-fold, whereas the proteolytic activity is enriched 10- to 20-fold in the isolated AV fraction as compared to the homogenate. Initially, the yield of nonlysosomal mitochondrial and microsomal enzyme activities increases in parallel with the induction of autophagy but, later on, decreases with advanced degradation of the sequestered cell organelles. Therefore, in the case of AVs the presence of nonlysosomal marker enzymes cannot be used for calculation of fraction purity, since newly sequestered organelles are enzymatically active. Isolated autophagic vacuoles show proteolytic activity when incubated in vitro. The comparatively high phospholipid/protein ratio (0.5) of the AV fraction suggests that phospholipids are degraded more slow than proteins. Is it concluded that AVs can be isolated into a pure fraction and are the subcellular site of enhanced protein degradation in the rat liver after induction of autophagy.

Abstract 1770: Proteasome Switching is Associated with Adverse Cardiac Remodeling in a Transgenic Mouse Model of Sustained Inflammatory Signaling

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Wei Wang ◽  
Gabor Szalai ◽  
Natarajan Sivasubramanian ◽  
Douglas L Mann
Keyword(s):  
Proteolytic Activity ◽  
Cardiac Remodeling ◽  
Fold Increase ◽  
Left Ventricular ◽  
26S Proteasome ◽  
Myocardial Inflammation ◽  
Fluorescent Substrate ◽  
Lv Remodeling ◽  
Apoptotic Proteins

The 26S proteasome possess proteolytic activity and deubiquitinating (DUB) activity of ubiquitin tagged proteins. Whereas the proteolytic activity of the 26S proteasome facilitates protein degradation, proteasome DUB activity spares proteins from degradation by shortening the length of the ubiquitin chains, thereby preventing proteins from being degraded by the 26S proteasome. In yeast, increased DUB activity is beneficial by preventing depletion of ubiquitin pools critical for cell signaling. However, the role of DUB activity in mammalian systems is not known. We have shown that mice with cardiac restricted overexpression of tumor necrosis factor (sTNF mice) develop a heart failure phenotype characterized by progressive left ventricular (LV) remodeling and accumulation of pro-apoptotic proteins, including Smac/Diablo. To determine whether the adverse LV remodeling in sTNF mice was related to alterations in DUB activity we measured the cleavage of ubiquitin-AMC, an in vitro fluorescent substrate for DUBs, in purified preparations of the 26S proteasome obtained from hearts of 4 week old sTNF and littermate (LM) control mice. Compared to LM controls we observed a significant (p < 0.001) 60.8% decrease in activity of the 26S proteasome and a significant (p < 0.01) 24.2% increase in DUB activity in sTNF mouse hearts. There was also a significant (p < 0.01) 11-fold increase myocardial protein levels of USP14, a critical DUB associated with the 26S proteasome in sTNF mouse hearts. The decrease in 26S proteasome activity and increased DUB activity in sTNF mouse hearts was accompanied by an increase in myocardial levels of ubiquitinated SMAC/Diablo. Taken together these results show for the first time that sustained myocardial inflammation leads to switch in the function of the proteasome from a proteolytic function to a protein sparing function. Although this “proteasome switching” may provide a short-term adaptive benefit by preventing depletion of critical ubiquitin pools, it may lead to long-term maladaptive consequences by allowing the progressive accumulation of potentially harmful pro-apoptotic proteins in the cytosol, which may in turn promote programmed cell death and adverse cardiac remodeling.

scholarly journals Treatment of Toxoplasmosis and Neosporosis in Farm Ruminants: State of Knowledge and Future Trends

2018 ◽  
Vol 18 (15) ◽  
pp. 1304-1323 ◽  
Author(s):  
Roberto Sánchez-Sánchez ◽  
Patricia Vázquez ◽  
Ignacio Ferre ◽  
Luis Miguel Ortega-Mora
Keyword(s):  
Vaccine Development ◽  
Neospora Caninum ◽  
Treatment Options ◽  
Small Ruminants ◽  
Additional Treatment ◽  
Current Status ◽  
Caninum Infection ◽  
Zoonotic Parasites ◽  
Toxoplasma Gondii Infection

Toxoplasmosis and neosporosis are closely related protozoan diseases that lead to important economic impacts in farm ruminants. Toxoplasma gondii infection mainly causes reproductive failure in small ruminants and is a widespread zoonosis, whereas Neospora caninum infection is one of the most important causes of abortion in cattle worldwide. Vaccination has been considered the most economic measure for controlling these diseases. However, despite vaccine development efforts, only a liveattenuated T. gondii vaccine has been licensed for veterinary use, and no promising vaccines against neosporosis have been developed; therefore, vaccine development remains a key goal. Additionally, drug therapy could be a valuable strategy for disease control in farm ruminants, as several drugs that limit T. gondii and N. caninum proliferation and dissemination have been evaluated. This approach may also be relevant to performing an initial drug screening for potential human therapy for zoonotic parasites. Treatments can be applied against infections in adult ruminants to minimize the outcomes of a primo-infection or the reactivation of a chronic infection during gestation or in newborn ruminants to avoid infection chronification. In this review, the current status of drug development against toxoplasmosis and neosporosis in farm ruminants is presented, and in an effort to promote additional treatment options, prospective drugs that have shown efficacy in vitro and in laboratory animal models of toxoplasmosis and neosporosis are examined.

Diet effects on methane production by goats and a comparison between measurement methodologies

2008 ◽  
Vol 146 (6) ◽  
pp. 705-715 ◽  
Author(s):  
R. BHATTA ◽  
O. ENISHI ◽  
N. TAKUSARI ◽  
K. HIGUCHI ◽  
I. NONAKA ◽  
...  
Keyword(s):  
Small Ruminants ◽  
National Research Council ◽  
Mean Value ◽  
Open Circuit ◽  
Gas Production ◽  
Mitigation Strategies ◽  
Production Test ◽  
Gross Energy ◽  
Nutritive Composition

SUMMARYA series of studies were carried out to measure the methane (CH4) production by Japanese goats fed 19 different diets (D1–D19) varying in nutritive composition in the open circuit respiration chamber (RC) and to compare them with CH4 estimated by the in vitro gas production test (IVGPT). Adult Japanese goats (>2 years old) with a mean body weight of 26±5·4 kg were used in these experiments. Each diet was fed to four randomly selected goats and feeding was carried out at 1·1 maintenance (M) as per National Research Council (NRC) (1981) for goats. Average CH4 emission by goats in the RC ranged from 0·23 to 0·39 (mean value 31 ml/g dry matter intake (DMI)); when it was expressed as a proportion of gross energy or, with methane conversion rate (MCR), it ranged from 5·0 to 8·2, with an average of 6·6. Incorporation of by-products like sweet potato vine silage (SPVS) (P=0·016), dried pumpkin (P=0·052) and brewers' grain in the diet suppressed (P<0·01) methanogenesis in goats, when compared with that of standard farm diet (D1). The CH4 output measured in the RC was very close to that estimated from the gas collected after 24 h and higher after 48 h of in vitro incubation. Although composition of the diets' acid detergent fibre (ADF) had a significant effect on methane emission, methane output estimated by IVGPT was very close to that measured in the RC demonstrating that this system could be used to estimate the CH4 production potential from diets in preparing a database and also in the planning of mitigation strategies in small ruminants to improve their performance as well as to reduce greenhouse gas emissions.

Sign in / Sign up

Export Citation Format

Share Document