scholarly journals The Effects of pH and Temperature on Cysteine Protease (Cathepsin B) Activity in Miracidia and Eggs of Fasciola hepatica

2020 ◽  
Author(s):  
Leila KIANIFARD ◽  
Mohammad YAKHCHALI ◽  
Mehdi IMANI
Keyword(s):  
Cysteine Protease ◽  
Cathepsin B ◽  
Fasciola Hepatica ◽  
Distilled Water ◽  
Acidic Ph ◽  
Optimum Temperature ◽  
Egg Hatching ◽  
Ph Values ◽  
Zoonotic Parasite ◽  
Effects Of Ph

Abstract Background: Fascioliasis is a worldwide zoonotic disease caused by the trematodes Fasciola hepatica in humans and animals. Proteases are essential for the survival of parasites and have important activities such as penetration, tissue migration, and egg hatching. This study was conducted to analyze cysteine protease of the miracidia and eggs of F. hepatica, and to assess the effects of pH and temperature on the proteases activity and stability. Methods: Adults F. hepatica were isolated from infected livers and were morphologically identified in 2018. Eggs collected from the adults and incubated in distilled water at 28 °C for 16 d to produce miracidia. The extract was collected from miracidia and eggs. A substrate for cathepsin B (Z-Arg-Arg-Pna) was used to assess the enzyme activity at different (2-12) pH levels. After homogenization, protein level was measured with Bradford method. Estimation of optimum temperature and pH was performed in the temperature range of 10-90 ° C and pH values from 2-12. Results: The highest activity of the miracidia and eggs enzyme extracts for Z-ArgArg-pNA was at pH 4. The miracidia extract was most stable at neutral pH and the eggs extract was most stable in acidic pH. The optimum temperature activity for both stages was 40 °C. These proteases were stable up to 40 °C. Conclusion: Upon the importance of pH and temperature in the life cycle of F. hepatica, the current findings can be used for induction of some modifications in pH and preventing the activity of the enzymes for decrement of the efficacy of miracidia penetration into the intermediate snails and egg hatching of this zoonotic parasite.

scholarly journals Activity of serine proteases from Fasciola hepatica eggs in relation to pH and temperature

2020 ◽  
Vol 23 (3) ◽  
pp. 350-358
Author(s):  
L. Kianifard ◽  
M. Yakhchali ◽  
M. Imani
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Proteolytic Activity ◽  
Serine Proteases ◽  
Fasciola Hepatica ◽  
Ethylenediaminetetraacetic Acid ◽  
Optimum Temperature ◽  
Egg Hatching ◽  
Protein Levels ◽  
Ph Values

This study was conducted to analyse the serine protease of Fasciola hepatica eggs by specific substrates and inhibitors, and investigation of the effects of pH and temperature on proteases’ activity and stability. Adult worms were isolated from infected livers. After homogenisation, their protein levels were measured with the Bradford method. Total proteolytic activity of the Fasciola hepatica extract was evaluated with azocasein substrate at pH values from 2 to 12. N-benzoyl–arginine–p-nitroanilide (BApNA) trypsin and N-succinyl-alanine-alanine-prolin-phenylalanine-p-nitroanilide (SAAPFpNA) chymotrypsin substrates were used to measure specific protease activities. The effect of protease inhibitors phenylmethane sulfonyl fluoride (PMSF), pepsin, and ethylenediaminetetraacetic acid (EDTA) on these enzymes was evaluated. Estimation of optimum temperature and pH was performed in the temperature range of 10–90 °C and pH values from 2–12. The optimum pH activities for trypsin and chymotrypsin were at alkaline pH and for total proteolytic activity at acidic pH. The results using protease inhibitors showed that the eggs had serine protease activity. The optimum temperature activity of trypsin and chymotrypsin was 50 °C. These proteases were stable up to 40 °C. Due to the importance of pH and temperature in life cycle of Fasciola hepatica, these findings can be used for induction of some modifications in pH and preventing the activity of the enzyme for decrement of the efficacy of embryonic development and egg hatching of this zoonotic parasite.

scholarly journals Mechanisms of Heat-induced Antigen Retrieval: Does pH or Ionic Strength of the Solution Play a Role for Refolding Antigens?

2005 ◽  
Vol 53 (11) ◽  
pp. 1311-1321 ◽  
Author(s):  
Katsura Emoto ◽  
Shuji Yamashita ◽  
Yasunori Okada
Keyword(s):  
Ionic Strength ◽  
The Other ◽  
Antigen Retrieval ◽  
Antigenic Determinants ◽  
Acidic Ph ◽  
Cooling Process ◽  
Ph Values ◽  
Ph Dependency ◽  
Nacl Concentration ◽  
Effects Of Ph

We investigated the effects of pH and ionic strength of solutions used for antigen retrieval to elucidate the mechanism of heat-induced antigen retrieval (HIAR) in immunohistochemistry. The immunostaining intensity of nuclear, cytoplasmic, cell membrane, and extracellular matrix antigens with 17 different antibodies was evaluated in formaldehyde-fixed and paraffin-embedded mouse and human tissues. Deparaffinized sections were autoclaved for 10 min in buffers with different pH values ranging from 3.0 to 10.5. To test the influence of ionic strength on immunoreactions, the sections were autoclaved for 10 min in 20 mM Tris-HCl buffers (TB) at pH 9.0 and 10.5 with or without 25, 50, and 100 mM NaCl. There were two immunostaining patterns for pH dependency of HIAR. First, the majority of antibodies recovered their antigenicity when heated in the buffers with both acidic pH (pH 3.0) and basic pH (pH 9.0 and 10.5). Second, some antibodies showed strong immunostaining only at basic pH values (pH 9.0 and 10.5). When the sections were autoclaved in TB at pH 9.0, immunostaining of all eight antibodies examined decreased as the NaCl concentration increased. On the other hand, when the sections were treated with TB at pH 10.5, all antibodies yielded stronger reactions in the buffer containing NaCl than in the buffer without NaCl; five antibodies exhibited the strongest immunoreaction at concentrations from 25 to 50 mM. These results suggest that the extended polypeptides by heating are charged negatively or positively at basic or acidic pH, and that an electrostatic repulsion force acts to prevent random entangling of polypeptides caused by hydrophobic attractive force and to expose antigenic determinants, during cooling process of HIAR solution.

scholarly journals Effects of pH on Nanofibrillation of TEMPO-Oxidized Paper Mulberry Bast Fibers

Polymers ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 414 ◽  
Author(s):  
Jung Park ◽  
Chan-Woo Park ◽  
Song-Yi Han ◽  
Gu-Joong Kwon ◽  
Nam-Hun Kim ◽  
...  
Keyword(s):  
Tensile Strength ◽  
Cellulose Nanofibrils ◽  
Crystallinity Index ◽  
Electrostatic Repulsion ◽  
Acidic Ph ◽  
Tempo Oxidation ◽  
Bast Fibers ◽  
Ph Values ◽  
Ph Dependency ◽  
Effects Of Ph

TEMPO oxidation was conducted as a pretreatment to achieve efficient nanofibrillation of long paper mulberry bast fibers (PMBFs). The pH dependency of nanofibrillation efficiency and the characteristics of the resulting cellulose nanofibrils (CNFs) were investigated. As the pH increased, the negative value of the zeta potential of TEMPO-oxidized fibers increased. The increase in electrostatic repulsion at pH values of greater than 9 prevented the entanglement of long PMBFs, which was a drawback for defibrillation at acidic pH. With increasing pH, the CNF production yield was increased. The crystallinity index of TEMPO-oxidized CNFs from PMBFs was 83.5%, which was higher than that of TEMPO-oxidized CNFs from softwood fibers in the same conditions. The tensile strength of nanopaper from TEMPO-oxidized PMBF CNFs was 110.18 MPa, which was approximately 30% higher than that (84.19 MPa) of the TEMPO-oxidized CNFs from softwood fibers.

Is the Acidic pH of the External Auditory Canal Playing a Significant Role in the Treatment of Acute Otitis Externa?

2018 ◽  
Vol 69 (8) ◽  
pp. 2304-2305
Author(s):  
Oana Ruxandra Iana ◽  
Dragos Cristian Stefanescu ◽  
Viorel Zainea ◽  
Razvan Hainarosie
Keyword(s):  
External Auditory Canal ◽  
Otitis Externa ◽  
Proton Pumps ◽  
Acidic Ph ◽  
External Ear ◽  
Ph Values ◽  
The World ◽  
Subcutaneous Tissues ◽  
Low Levels ◽  
Value Changes

Variable pH values for skin have been reported in the literature, all within the acidic range, varying from 4.0 up to 7. 0. The origin of the acidic pH remains conjectural, and several factors have been incriminated with this role, such as eccrine and sebaceous secretions as well as proton pumps. Keeping low levels of pH prevents microbial dispersal as well as multiplication. The skin in the external auditory canal is also covered with this acidic mantle with antimicrobial value. Changes of pH in the external ear can lead to acute otitis externa. This condition is defined by the inflammation and infection of the cutaneous and subcutaneous tissues of the external auditory canal. 10% of the world�s population may suffer from acute otitis externa at least once in their lifetime. This paper aims to consolidate the relevance of an acidic pH in the healthy external ear and its relation to the pathogenesis and treatment of otitis externa through a prospective and interventional clinical study on 80 patients who presented to the outpatient department at Prof. Dr D. Hociota ENT Institute in Buch

Nitrogen Removal from Anaerobically-Treated Leachate

1984 ◽  
Vol 19 (1) ◽  
pp. 87-100
Author(s):  
D. Prasad ◽  
J.G. Henry ◽  
P. Elefsiniotis
Keyword(s):  
Nitrogen Removal ◽  
Air Flow ◽  
Operating Conditions ◽  
Air Flow Rate ◽  
Flow Rates ◽  
Anaerobic Filter ◽  
Ph Values ◽  
Wide Range ◽  
Ammonia Desorption ◽  
Effects Of Ph

Abstract Laboratory studies were conducted to demonstrate the effectiveness of diffused aeration for the removal of ammonia from the effluent of an anaerobic filter treating leachate. The effects of pH, temperature and air flow on the process were studied. The coefficient of desorption of ammonia, KD for the anaerobic filter effluent (TKN 75 mg/L with NH3-N 88%) was determined at pH values of 9, 10 and 11, temperatures of 10, 15, 20, 30 and 35°C, and air flow rates of 50, 120, and 190 cm3/sec/L. Results indicated that nitrogen removal from the effluent of anaerobic filters by ammonia desorption was feasible. Removals exceeding 90% were obtained with 8 hours aeration at pH of 10, a temperature of 20°C, and an air flow rate of 190 cm3/sec/L. Ammonia desorption coefficients, KD, determined at other temperatures and air flow rates can be used to predict ammonia removals under a wide range of operating conditions.

scholarly journals First report on the prevalence of Fasciola hepatica in the endangered Père David’s deer (Elaphurus davidianus) in China

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Si-Yang Huang ◽  
Jing-Zhi Gong ◽  
Yi-Jun Ren ◽  
Ming Pan ◽  
Wei-Min Cai ◽  
...  
Keyword(s):  
Fasciola Hepatica ◽  
Potential Threat ◽  
Prevalence Data ◽  
First Report ◽  
Natural Reserve ◽  
Zoonotic Parasite ◽  
The World ◽  
Père David’S Deer ◽  
Microscopy Examination ◽  
Elaphurus Davidianus

Abstract Background Fasciola hepatica is an important zoonotic parasite that causes fasciolosis in a broad range of animals. No information is available about the prevalence of F. hepatica in Père David’s deer (Elaphurus davidianus), an endangered species in the world. Therefore, the purpose of the study was to evaluate the prevalence of fasciolosis in Père David’s deer in the Dafeng Elk National Natural Reserve, Jiangsu province, China. Results In this study, 142 fecal samples from Père David’s deer were analyzed for F. hepatica by microscopy and nest-PCR. Only one sample was positive for F. hepatica according to microscopy examination, while 18 of 142 (12.68, 95%CI: 2.841–22.45%) samples were positive for F. hepatica according to nest-PCR results. Conclusions This is the first report of prevalence of F. hepatica in Père David’s deer. The prevalence data indicated that F. hepatica was also present in this endangered animal, which may cause a potential threat to this precious species.

scholarly journals In Vitro Incorporation of Helicobacter pylori into Candida albicans Caused by Acidic pH Stress

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 489 ◽  
Author(s):  
Kimberly Sánchez-Alonzo ◽  
Cristian Parra-Sepúlveda ◽  
Samuel Vega ◽  
Humberto Bernasconi ◽  
Víctor L. Campos ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Candida Albicans ◽  
Pcr Amplification ◽  
Growth Curves ◽  
Acidic Ph ◽  
Ph Values ◽  
16S Gene ◽  
Wide Range ◽  
H Pylori

Yeasts can adapt to a wide range of pH fluctuations (2 to 10), while Helicobacter pylori, a facultative intracellular bacterium, can adapt to a range from pH 6 to 8. This work analyzed if H. pylori J99 can protect itself from acidic pH by entering into Candida albicans ATCC 90028. Growth curves were determined for H. pylori and C. albicans at pH 3, 4, and 7. Both microorganisms were co-incubated at the same pH values, and the presence of intra-yeast bacteria was evaluated. Intra-yeast bacteria-like bodies were detected using wet mounting, and intra-yeast binding of anti-H. pylori antibodies was detected using immunofluorescence. The presence of the H. pylori rDNA 16S gene in total DNA from yeasts was demonstrated after PCR amplification. H. pylori showed larger death percentages at pH 3 and 4 than at pH 7. On the contrary, the viability of the yeast was not affected by any of the pHs evaluated. H. pylori entered into C. albicans at all the pH values assayed but to a greater extent at unfavorable pH values (pH 3 or 4, p = 0.014 and p = 0.001, respectively). In conclusion, it is possible to suggest that H. pylori can shelter itself within C. albicans under unfavorable pH conditions.

Effects of pH and ionic strength on the cation exchange capacity of soils with variable charge

Soil Research ◽  
1981 ◽  
Vol 19 (1) ◽  
pp. 93 ◽  
Author(s):  
GP Gillman
Keyword(s):  
Ionic Strength ◽  
Cation Exchange ◽  
Cation Exchange Capacity ◽  
Exchange Capacity ◽  
Surface Soils ◽  
Ph Values ◽  
Variable Charge ◽  
Effects Of Ph ◽  
Variable Charge Soils

The cation exchange capacity of six surface soils from north Queensland and Hawaii has been measured over a range of pH values (4-6) and ionic strength values (0.003-0.05). The results show that for variable charge soils, modest changes in electrolyte ionic strength are as important in their effect on caton exchange capacity as are changes in pH values.

scholarly journals Characterization of cathepsins in cartilage

1967 ◽  
Vol 105 (2) ◽  
pp. 549-557 ◽  
Author(s):  
S. Y. Ali ◽  
L. Evans ◽  
E. Stainthorpe ◽  
C. H. Lack
Keyword(s):  
Cathepsin B ◽  
Cartilage Matrix ◽  
Heat Inactivation ◽  
Ph Values ◽  
Fixed Weight ◽  
Exogenous Substrate ◽  
Nitrophenyl Phosphate ◽  
Rabbit Ear ◽  
Ear Cartilage

The presence of a cathepsin B-like enzyme in rabbit ear cartilage was established by the use of the synthetic substrates benzoyl-l-arginine amide and benzoyl-dl-arginine 2-naphthylamide. This was facilitated by using a technique that permits the incubation of a fixed weight of thin (18μ) cartilage sections with an appropriate exogenous substrate. The enzymic properties of cathepsin B in cartilage have been compared with an endogenous enzyme that liberates chondromucopeptide by degrading the cartilage matrix autocatalytically at pH5. Besides being maximally active at pH4·7, these cartilage enzymes are enhanced in activity by cysteine and inhibited by arginine analogues, iodoacetamide, chloroquine and mercuric chloride. They are not inhibited by EDTA, di-isopropyl phosphorofluoridate and diethyl p-nitrophenyl phosphate. When inhibiting the release of chondromucopeptide from cartilage at pH5, the arginine-containing synthetic substrates are hydrolysed simultaneously. These enzymes also share the same heat-inactivation characteristics at various pH values, being stable at acid pH and unstable at neutral and alkaline pH. The experimental evidence indicates that a cathepsin B-like enzyme may be partly responsible for the autolytic degradation of cartilage matrix at pH5.

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