In sturgeon, the acquisition of the potential for motility activation called spermatozoon maturation takes place outside testes. This process can be accomplished in vitro by pre-incubation of immature testicular spermatozoa in seminal fluid collected from fully mature Wolffian duct sperm. Addition of trypsin inhibitor to the pre-incubation medium disrupts spermatozoon maturation. There are no available data for the role of proteolysis regulators in fish spermatozoon maturation, while their role is recognized in mammalian sperm maturation. The present study evaluated the involvement of seminal fluid proteases and anti-proteolytic activity in the sterlet spermatozoon maturation process. Casein and gelatin zymography and quantification of amidase and anti-proteolytic activity were conducted in sturgeon seminal fluid from Wolffian duct sperm and seminal fluid from testicular sperm, along with spermatozoon extracts from Wolffian duct spermatozoa, testicular spermatozoa, and testicular spermatozoa after in vitro maturation. We did not find significant differences in proteolytic profiles of seminal fluids from Wolffian duct sperm and ones from testicular sperm. Zymography revealed differences in spermatozoon extracts: Wolffian duct spermatozoon extracts were characterized by the presence of a broad proteolytic band ranging from 48 to 41 kDa, while testicular spermatozoon extracts did not show such activity until after in vitro maturation. The differences in amidase activity coincided with these results. It may not be the levels of proteolytic and anti-proteolytic activity per se, but the alterations in their interactions triggering a cascade of signaling events, that is crucial to the maturation process.

• Klíčová slova
• Amidase activity, Anti-proteolytic activity, Casein and gelatin zymography, Siberian sturgeon sperm, Spermatozoon maturation, Sterlet sperm,
• MeSH
• amidohydrolasy metabolismus   MeSH
• motilita spermií MeSH
• proteolýza MeSH
• ryby fyziologie   MeSH
• spermie fyziologie   MeSH
• testis cytologie   MeSH
• Wolffovy vývody cytologie   MeSH
• zrání spermie * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amidase MeSH Prohlížeč
• amidohydrolasy MeSH

A new procedure for the determination of effective proteolytic activity in biodetergents has been developed. Effective enzyme activity is defined as the activity exhibited by the tested enzyme under real washing conditions, i.e., in the water suspensions of the biodetergent tested at the working temperature (usually 40 degrees C). Two insoluble chromolytic substrates, namely black gelatin (gelatin cross-linked with glutaraldehyde in the presence of black drawing ink) and the protease substrate based on the immobilization of dyed casein in the structure of polyacrylamide gel (blue casein-PAAG) were successfully used. It was found that there is a great difference in effective proteolytic activity among various biodetergents. The proposed procedure is suitable both for the manufacturers for a quick control of their products and for the quality control laboratories.

• MeSH
• detergenty analýza   chemie   MeSH
• proteasy chemie   MeSH
• rozpustnost MeSH
• spektrofotometrie MeSH
• substrátová specifita MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• detergenty MeSH
• proteasy MeSH

A simple procedure for spectrophotometric determination of proteolytic activity in coloured solutions is described. Gelatin cross-linked with glutaric dialdehyde or formaldehyde in the presence of black drawing ink is used as an insoluble chromolytic substrate. The absorbances of reaction mixture filtrates are read in the near infra-red region (at 800-900 nm) where the majority of coloured substances does not absorb; on the contrary black drawing ink released from the substrate during the proteolysis absorbs strongly even at these wavelengths.

• MeSH
• hydrolýza MeSH
• indikátory a reagencie MeSH
• proteasy analýza   MeSH
• roztoky MeSH
• spektrofotometrie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• indikátory a reagencie MeSH
• proteasy MeSH
• roztoky MeSH

• Klíčová slova
• HEPATITIS, INFECTIONS/blood *, PROTEASES/blood *,
• MeSH
• hepatitida A krev   MeSH
• hepatitida * MeSH
• hydrolasy * MeSH
• lidé MeSH
• proteasy krev   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hydrolasy * MeSH
• proteasy MeSH

• MeSH
• Ascaris * MeSH
• houby enzymologie   MeSH
• proteasy metabolismus   MeSH
• vaječné proteiny metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• proteasy MeSH
• vaječné proteiny MeSH

A new insoluble chromolytic substrate for the determination of proteolytic activity was prepared by immobilization of dyed casein into the structure of polyacrylamide gel. The prepared substrate could detect approximately 0.1 microgram of trypsin per ml. The spontaneous leakage of dyed casein molecules in water solutions was negligible.

• MeSH
• akrylové pryskyřice MeSH
• barvení a značení MeSH
• enzymy imobilizované * MeSH
• imobilizace MeSH
• indikátory a reagencie * MeSH
• kaseiny MeSH
• proteasy metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akrylové pryskyřice MeSH
• enzymy imobilizované * MeSH
• indikátory a reagencie * MeSH
• kaseiny MeSH
• polyacrylamide gels MeSH Prohlížeč
• proteasy MeSH

The present study deals with the effect of hypergravity (2xg) on the pH and on the proteolytic activity in the digesta of the gastrointestinal tract of Japanese quails during intense growth. The birds were raised on a semisynthetic diet containing free amino acids (A) and a commercial diet (B). During days 35 till 40 post-hatching the quails were exposed to hypergravity (2xg) using a specially designed centrifuge. On days 40 (experimental group, 2xg) and 41 (control group, 1xg) the animals were sacrificed. The pH of the digesta in various segments of the gastrointestinal tract was measured by means of a semi-microelectrode. Total proteolytic activity was determined by means of azo-dye-modified proteins serving as general proteolytic substrates. Hypergravity leads in general to an alkalization of digesta in various parts of the gastrointestinal tract. In case of the gizzard and duodenum (diet A) and also in the distal jejunum (diet B) the differences are significant. With both diets, hypergravity leads to a considerable decrease in the total proteolytic activity. The reduction is most expressed in the duodenum and jejunum. Changes in the pH of digesta compensate for the decrease in the proteolytic activity. This may explain why hypergravity per se does not seem to impair growth of the Japanese quails.

• MeSH
• aminokyseliny metabolismus   MeSH
• Coturnix metabolismus   MeSH
• gravitace * MeSH
• koncentrace vodíkových iontů MeSH
• mikroelektrody MeSH
• proteasy metabolismus   MeSH
• trávicí systém enzymologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aminokyseliny MeSH
• proteasy MeSH

Dye-stained gelatin microcarriers were used as insoluble chromolytic substrates for the determination of proteolytic activity. Trypsin at concentration 100 ng/ml could be easily detected. The spontaneous leakage of dyed fragments into water solutions was negligible.

• MeSH
• barvicí látky * MeSH
• micely MeSH
• proteasy analýza   MeSH
• želatina * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• barvicí látky * MeSH
• micely MeSH
• proteasy MeSH
• želatina * MeSH

An insoluble chromogenic substrate for the determination of proteolytic activity was prepared by heating azocasein in a thin layer at 200 degrees C for 4 h in a hot-air thermostat. The activity of an extracellular bacterial proteinase produced by Bacillus sp. was determined with this new substrate.

• MeSH
• kaseiny * MeSH
• kinetika MeSH
• proteasy metabolismus   MeSH
• spektrofotometrie metody   MeSH
• vysoká teplota MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• azocasein MeSH Prohlížeč
• kaseiny * MeSH
• proteasy MeSH

• Klíčová slova
• HEPARIN/pharmacology *, PROTEASES/blood *,
• MeSH
• endopeptidasy * MeSH
• heparin farmakologie   MeSH
• hydrolasy * MeSH
• injekce intravenózní * MeSH
• lidé MeSH
• proteasy krev   MeSH
• proteolýza * MeSH
• sérum * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• endopeptidasy * MeSH
• heparin MeSH
• hydrolasy * MeSH
• proteasy MeSH