Possibilities of the diagnostic utilization of determined thermostable portion of the ALP activity in bovine serum were studied. The work is based on the finding that the total ALP activity is formed by two fractions with different sensitivity to the temperature of 56 degrees C. Dynamics of the loss of ALP activity during serum heating was of an equal character in all four tested groups. The loss was always highest during the first minutes of heating, after 15 to 20 minutes it became equal. A period of 15 minutes of the serum heating is sufficient for obtaining information on the size of the ALP thermostable fraction. The ALP activity and its residue after 15-minute thermal inhibition (thermostable portion) were tested in the sera of 163 head of cattle divided into seven groups. The size of this residue was related to the age of animals, degree of gravidity did not affect the residue. The thermostable portion in 6 to 10 months old bullocks was 14.86 +/- 4.93%, in 2.5 to 5 years old heifers 41.20 +/- 20.37% and in clinically healthy dairy cows 54.35 +/- 18.26%. In a group of calved breeding cows 44.13 +/- 19.74% of the initial activity remained. Determination of the thermostable ALP fraction can be applied with certain limitation in pre-clinical diagnostics of disorders of mineral metabolism in bones of cattle. In herds of healthy dairy cows the value of thermostable portion should not be lower than 45% of the original ALP activity. In 2.5 to 3 years old heifers this level is considered to be 35%.

• MeSH
• alkalická fosfatasa krev   MeSH
• klinické enzymatické testy MeSH
• nemoci skotu diagnóza   MeSH
• skot krev   MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• skot krev   MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• alkalická fosfatasa MeSH

Alkaline proteases were produced by a virulent strain of Aspergillus fumigatus during growth on media containing glucose and proteins or peptides. After isoelectric focusing, six bands of proteolytic activity were detected with synthetic substrates after blotting on nitrocellulose membranes. The main protease (pI = 8.6) corresponded to the known subtilisin-like protease Alp of A. fumigatus and five minor components had lower isoelectric points (8.1 to 5.2). All proteases were produced on different media and in various phases of growth with only small quantitative variations. They also had identical pH optima, were denatured above 45 degrees C and stabilized by Ca2+ ions, were affected by the inhibitors of serine proteases only and had nearly identical substrate specificity against 13 synthetic substrates. On gel chromatography the three most acidic components had higher molecular weights than the main enzyme Alp. It remains to be determined if the enzymes under study arise through posttranslational processing of the main protease or are true isoenzymes, products of a gene family.

• MeSH
• Aspergillus fumigatus enzymologie   růst a vývoj   MeSH
• aspergilóza mikrobiologie   veterinární   MeSH
• elektroforéza v polyakrylamidovém gelu metody   MeSH
• imunoblotting MeSH
• isoelektrická fokusace metody   MeSH
• izoenzymy metabolismus   MeSH
• kur domácí MeSH
• nemoci drůbeže mikrobiologie   MeSH
• plicní mykózy mikrobiologie   veterinární   MeSH
• serinové endopeptidasy chemie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• izoenzymy MeSH
• oryzin MeSH Prohlížeč
• serinové endopeptidasy MeSH

Biofabrication and maturation of bone constructs is a long-term task that requires a high degree of specialization. This specialization falls onto the hierarchy complexity of the bone tissue that limits the transfer of this technology to the clinic. This work studied the effects of the short-term cryopreservation on biofabricated osteoblast-containing structures, with the final aim to make them steadily available in biobanks. The biological responses studied include the osteoblast post-thawing metabolic activity and the recovery of the osteoblastic function of 3D-bioprinted osteoblastic structures and beta tricalcium phosphate (β-TCP) scaffolds infiltrated with osteoblasts encapsulated in a hydrogel. The obtained structures were cryopreserved at -80 °C for 7 days using dimethyl sulfoxide (DMSO) as cryoprotectant additive. After thawing the structures were cultured up to 14 days. The results revealed fundamental biological aspects for the successful cryopreservation of osteoblast constructs. In summary, immature osteoblasts take longer to recover than mature osteoblasts. The pre-cryopreservation culture period had an important effect on the metabolic activity and function maintain, faster recovering normal values when cryopreserved after longer-term culture (7 days). The use of β-TCP scaffolds further improved the osteoblast survival after cryopreservation, resulting in similar levels of alkaline phosphatase activity in comparison with the non-preserved structures. These results contribute to the understanding of the biology of cryopreserved osteoblast constructs, approaching biofabrication to the clinical practice.

• Klíčová slova
• ALP activity, biofabrication, bioprinting, bone construct, cryopreservation, metabolic activity, osteoblast,
• Publikační typ
• časopisecké články MeSH

Injectable hydrogels for bone regeneration consisting of chitosan, sodium beta-glycerophosphate (Na-β-GP) and alkaline phosphatase (ALP) were enriched with the polyphenols phloroglucinol (PG) and gallic acid (GA) and characterized physicochemically and biologically with respect to properties relevant for applications in bone regeneration, namely gelation kinetics, mineralizability, antioxidant properties, antibacterial activity, cytocompatibility and ability to support adhesion and growth of human osteoblast-like MG63 cells. Enrichment with PG and GA had no negative effect on gelation kinetics and mineralizability. PG and GA both enhanced antioxidant activity of unmineralized hydrogels. Mineralization reduced antioxidant activity of hydrogels containing GA. Hydrogels containing GA, PG and without polyphenols reduced colony forming ability of Escherichia coli after 1h, 3h and 6h incubation and slowed E. coli growth in liquid culture for 150min. Hydrogels containing GA were cytotoxic and supported cell growth more poorly than polyphenol-free hydrogels. PG had no negative effect on cell adhesion and growth.

• Klíčová slova
• Antibacterial, Antioxidant, Cytocompatibility, Hydrogel, Polyphenol,
• MeSH
• alkalická fosfatasa metabolismus   MeSH
• antibakteriální látky farmakologie   MeSH
• buněčná adheze účinky léků   MeSH
• buněčné linie MeSH
• chitosan farmakologie   MeSH
• hydrogely farmakologie   MeSH
• kinetika MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• minerály chemie   MeSH
• osteoblasty cytologie   účinky léků   MeSH
• polyfenoly farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• spektroskopie infračervená s Fourierovou transformací MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alkalická fosfatasa MeSH
• antibakteriální látky MeSH
• chitosan MeSH
• hydrogely MeSH
• minerály MeSH
• polyfenoly MeSH

• Klíčová slova
• PHOSPHATASES/blood *,
• MeSH
• alkalická fosfatasa * MeSH
• fosfatasy krev   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alkalická fosfatasa * MeSH
• fosfatasy MeSH

Biochemical properties of a termostable alkaline phosphatase obtained from the mycelium extract of A. caespitosus were described. The enzyme was purified 42-fold with 32% recovery by DEAE-cellulose and concanavalin A-Sepharose chromatography. The molar mass estimated by Sephacryl S-200 or by 7% SDS-PAGE was 138 kDa and 71 kDa, respectively, indicating a homodimer. Temperature and pH optima were 80 degrees C and pH 9.0. This enzyme was highly glycosylated (approximately 74% saccharide content). The activity was enhanced by Mg2+ (19-139%), NH4+ (64%), Na+ (51%) and Mn2+ (38%). 4-Nitrophenyl phosphate (4-NPP) was preferentially hydrolyzed, but glucose 1-phosphate (93%), UTP (67%) and O-phosphoamino acids also acted as substrates. V(lim) and K(m) were 3.78 nkat per mg protein and 270 micromol/L in the absence of Mg2+ and 7.35 nkat per mg protein and 410 micromol/L in the presence of Mg2+, using 4-NPP as substrate. The purified alkaline phosphatase removed the 5'-phosphate group of a linearized plasmid without showing DNAase activity, indicating its potential for recombinant DNA technology.

• MeSH
• alkalická fosfatasa chemie   izolace a purifikace   metabolismus   MeSH
• Aspergillus enzymologie   MeSH
• deoxyribonukleasy metabolismus   MeSH
• kinetika MeSH
• mycelium enzymologie   MeSH
• substrátová specifita MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alkalická fosfatasa MeSH
• deoxyribonukleasy MeSH

Alkaline phosphatase (EC 3.1.3.1) is synthesized in media with a low phosphate concentration (0.37 mM of total and 19 microM of inorganic phosphate, respectively) already during the exponential phase of growth of Bacillus cereus. The enzyme is repressed by higher phosphate concentrations (3.7 mM) during the whole growth period; during sporogenesis the enzyme activity in cells slightly increases even under these conditions. During growth the enzyme is not secreted into the medium, a minor amount being released after cessation of growth. The enzyme activity can be increased by adding Zn2+ ions (10 microM). When during growth without phosphate the pH of the medium decreases below 5.0, the enzyme activity temporarily decreases and growth is slowed down, followed by a subsequent increase of the enzyme activity. In this case the onset of sporulation is also delayed.

• MeSH
• alkalická fosfatasa metabolismus   MeSH
• Bacillus cereus enzymologie   růst a vývoj   fyziologie   MeSH
• draslík farmakologie   MeSH
• fosfáty farmakologie   MeSH
• koncentrace vodíkových iontů MeSH
• kultivační média MeSH
• sloučeniny draslíku * MeSH
• spory bakteriální fyziologie   MeSH
• zinek farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alkalická fosfatasa MeSH
• draslík MeSH
• fosfáty MeSH
• kultivační média MeSH
• potassium phosphate MeSH Prohlížeč
• sloučeniny draslíku * MeSH
• zinek MeSH

The influence of static homogeneous magnetic field on alkali and acid phosphatase in guinea pigs (regarding the twenty four hours rhythm) was studied. The increase of acid phosphatase activity was determined by the time of exposure to magnetic field. No changes in alkali phosphatase activity were observed.

• MeSH
• alkalická fosfatasa krev   MeSH
• časové faktory MeSH
• kyselá fosfatasa krev   MeSH
• magnetismus * MeSH
• morčata MeSH
• vystavení vlivu životního prostředí MeSH
• zvířata MeSH
• Check Tag
• morčata MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alkalická fosfatasa MeSH
• kyselá fosfatasa MeSH

The activities were studied in five kinds of enzymes (aspartate aminotransferase - AST, alanine aminotransferase - ALT, lactate dehydrogenase - LD, the thermally stable fraction of lactate dehydrogenase - LD-1, and alkaline phosphatase - ALP) of 30 male dogs. The dogs, divided into two age categories, were studied during a long-continued training (130 days). Both transaminases exhibit characteristic changes in the activity, with a depression at the beginning between the 30th and 40th days of training, followed by a slow increase in AST and by a rapid increase in ALT, continuing until the end of the training period. A statistically significant activity pattern was recorded in LD: the activity declined continuously in both age groups of dogs. LD-1 exhibited an activity depression continuing until the 70th day of training, followed by an increase which reached statistical significance towards the end of the training. ALP activity varied regularly, but always remained significantly below the starting values. The enzymatic activities can be used as partial tests during the scientific management of the training of dogs in relation to the physiological and pathophysiological processes in the bodies of the dogs subjected to the training stress.

• MeSH
• alanintransaminasa krev   MeSH
• alkalická fosfatasa krev   MeSH
• aspartátaminotransferasy krev   MeSH
• enzymy krev   MeSH
• fyziologický stres krev   veterinární   MeSH
• L-laktátdehydrogenasa krev   MeSH
• psi metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• psi metabolismus   MeSH
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• alanintransaminasa MeSH
• alkalická fosfatasa MeSH
• aspartátaminotransferasy MeSH
• enzymy MeSH
• L-laktátdehydrogenasa MeSH

• MeSH
• alkalická fosfatasa metabolismus   MeSH
• arterioskleróza metabolismus   MeSH
• infarkt myokardu diagnóza   MeSH
• klinické enzymatické testy MeSH
• leukocyty enzymologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• neutrofily enzymologie   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alkalická fosfatasa MeSH