The cytopoxic effect of RL2 lactaptin (the recombinant analog of proteolytic fragment of human kappa-casein) toward tumor cells in vitro and in vivo presents it as a novel promising antitumor drug. The binding of any drug with serum proteins can affect their activity, distribution, rate of excretion and toxicity in the human body. Here, we studied the ability of RL2 to bind to various blood serum proteins. Using magnetic microparticles bearing by RL2 as an affinity matrix, in combination with mass spectrometry and western blot analysis, we found a number of blood serum proteins possessing affinity for RL2. Among them IgA, IgM and IgG subclasses of immunoglobulins, apolipoprotein A1 and various cortactin isoforms were identified. This data suggests that in the bloodstream RL2 lactaptin takes part in complicate protein-protein interactions, which can affect its activity.
- MeSH
- antitumorózní látky metabolismus MeSH
- chromatografie afinitní metody MeSH
- kaseiny metabolismus MeSH
- krevní proteiny analýza izolace a purifikace metabolismus MeSH
- kyseliny polymethakrylové chemie MeSH
- lidé MeSH
- magnety chemie MeSH
- mikrosféry MeSH
- rekombinantní proteiny metabolismus MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Proteome modifications in a biological fluid can potentially indicate the occurrence of pathologies, even if the identification of a proteome fingerprint correlated to a specific disease represents a very difficult task. When a nanomaterial is introduced into a biological fluid, macromolecules compete to form a protein corona on the nanoparticle surface, and depending on the specific proteome, different patterns of proteins will form the final protein corona shell depending on their affinity for the nanoparticle surface. Novel surface active maghemite nanoparticles (SAMNs) display a remarkable selectivity toward protein corona formation, and they are able to concentrate proteins and peptides presenting high affinities for their surface even if they are present in very low amounts. Thus, SAMNs may confer visibility to hidden biomarkers correlated to the occurrence of a pathology. In the present report, SAMNs were introduced into milk samples from healthy cows and from animals affected by mastitis, and the selectively bound protein corona shell was easily analyzed and quantified by gel electrophoresis and characterized by mass spectrometry. Upon incubation in mastitic milk, SAMNs were able to selectively bind αs2-casein fragments containing the FALPQYLK sequence, as part of the larger casocidin-1 peptide with strong antibacterial activity, which were not present in healthy samples. Thus, SAMNs can be used as a future candidate for the rapid diagnosis of mastitis in bovine milk. The present report proposes protein competition for SAMN protein corona formation as a means of mirroring proteome modifications. Thus, the selected protein shell on the nanoparticles results in a fingerprint of the specific pathology.
- MeSH
- biologické markery chemie metabolismus MeSH
- hmotnostní spektrometrie metody MeSH
- kaseiny chemie metabolismus MeSH
- kovové nanočástice chemie MeSH
- mastitida diagnóza metabolismus MeSH
- mléčné bílkoviny chemie metabolismus MeSH
- mléko chemie metabolismus MeSH
- povrchové vlastnosti MeSH
- proteinová korona chemie metabolismus MeSH
- proteom chemie metabolismus MeSH
- proteomika metody MeSH
- reprodukovatelnost výsledků MeSH
- sekvence aminokyselin MeSH
- senzitivita a specificita MeSH
- skot MeSH
- železité sloučeniny chemie MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Přehled nových přídavků do kojeneckých formulí.
New additions to infant formulas.
- Klíčová slova
- nové přídavky do kojeneckých formulí,
- MeSH
- alergie na mléko dietoterapie imunologie MeSH
- aminokyseliny imunologie terapeutické užití MeSH
- dietní proteiny metabolismus normy terapeutické užití MeSH
- hydrolýza účinky léků MeSH
- kaseiny metabolismus terapeutické užití MeSH
- kojenec MeSH
- lidé MeSH
- mateřské mléko MeSH
- náhražky mateřského mléka * metody normy MeSH
- nenasycené mastné kyseliny metabolismus terapeutické užití MeSH
- novorozenec MeSH
- nukleotidy metabolismus normy terapeutické užití MeSH
- potrava pro kojence * normy využití MeSH
- prebiotika využití MeSH
- probiotika metabolismus terapeutické užití MeSH
- Check Tag
- kojenec MeSH
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- přehledy MeSH
Birth is an event that exposes the newborn baby to a high concentration of free radicals (reactive oxygen species (ROS)/reactive nitrogen species (RNS)), which can contribute to several diseases. The balanced equilibrium between ROS/RNS and the antioxidant system of the newborn is a key factor in preventing a plethora of diseases. The antioxidant system involves endogenous and exogenous molecules, from vitamins (A, E, C) to enzymes (glutathione peroxidase, superoxide dismutase), metals (copper, zinc, and selenium) and other molecules (coenzyme Q10, melatonin) that can act in a synergistic manner to deactivate free radicals. A competent antioxidant system of the baby is strongly dependent on the intake of free radical deactivating molecules from feeding, either maternal or formula milk, with several studies pointing that breast milk has more powerful antioxidant effects on lowering the infant's oxidative status. An improved understanding of the antioxidant molecules, their mechanism of action, and the relationships between them, are key factors to comprehend all the potential benefits of human breastfeeding in this matter. The purpose of this review is to describe different research efforts and methodologies of evaluation of TAS in human milk, and to evaluate and summarize the contribution of different antioxidant molecules.
- Klíčová slova
- TAS/TAC (total antioxidant system/total antioxidant capacity) - (celková antioxidační kapacita lidského mléka),
- MeSH
- antioxidancia farmakologie metabolismus MeSH
- ceruloplasmin imunologie izolace a purifikace metabolismus MeSH
- farmakologické účinky - molekulární mechanismy MeSH
- glutathionperoxidasa imunologie izolace a purifikace metabolismus MeSH
- kaseiny imunologie izolace a purifikace metabolismus MeSH
- katalasa imunologie izolace a purifikace metabolismus MeSH
- kojenec MeSH
- kojení * MeSH
- kovy imunologie izolace a purifikace metabolismus MeSH
- kyselina askorbová MeSH
- laktoferrin imunologie izolace a purifikace metabolismus MeSH
- lidé MeSH
- mateřské mléko * enzymologie imunologie metabolismus MeSH
- melatonin imunologie izolace a purifikace metabolismus MeSH
- mléčné bílkoviny imunologie izolace a purifikace metabolismus MeSH
- ochranné látky farmakologie metabolismus MeSH
- oxidační stres fyziologie imunologie MeSH
- proteiny imunologie izolace a purifikace metabolismus MeSH
- statistika jako téma MeSH
- stopové prvky imunologie izolace a purifikace metabolismus MeSH
- superoxiddismutasa imunologie izolace a purifikace metabolismus MeSH
- ubichinon imunologie izolace a purifikace metabolismus MeSH
- vitamin A MeSH
- vitamin E MeSH
- volné radikály antagonisté a inhibitory metabolismus škodlivé účinky MeSH
- Check Tag
- kojenec MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
Kozí mléko je ve srovnání s mlékem kravským svým složením bližší mléku mateřskému. Jelikož je při konzumaci kravského mléka či z něho vyráběných potravin v populaci zjišťována relativně často alergie, může být kozí mléko jeho alternativou. V článku je diskutován především jeho vztah k alergii z hlediska patofyziologie a poukázáno je i na další benefity týkající se např. zažívacího traktu.
Goat milk, when compared to cow milk, is more similar to maternal milk with regard to its composition. Since the feeding with cow milk or diary products might be relatively often associated with allergy, goat milk can be an interesting alternative. In this article, its relation to allergy is mostly discussed, especially from the pathophysiologic point of view, and other possible benefits on gastrointestinal tract are outlined.
- Klíčová slova
- Kozí kaše,
- MeSH
- alergeny MeSH
- alergie etiologie prevence a kontrola MeSH
- dítě * MeSH
- kaseiny imunologie metabolismus MeSH
- kozy * MeSH
- lidé MeSH
- mléčné výrobky * MeSH
- mléko škodlivé účinky MeSH
- potravinová alergie etiologie prevence a kontrola MeSH
- zácpa * etiologie prevence a kontrola MeSH
- Check Tag
- dítě * MeSH
- lidé MeSH
Microplate assays with 96 wells were optimized to screen proteolytic activities in mite homogenates. Whole-mite extracts of Acarus siro, Aleuroglyphus ovatus, Tyrophagus putrescentiae, Tyroborus lini, Carpoglyphus lactis, Lepidoglyphus destructor, and Dermatophagoides farinae exhibited non-specific proteolytic activity in buffers from pH 2 to 12, and three peaks of highest activity at pH 3, 5-6, and 10 were distinguished. The reducing agent Tris(2-carboxyethyl)phosphine hydrochloride decreased general proteolytic activity on azocasein at pH 5 and 6. The results obtained on two non-specific substrates, azocasein and azoalbumin, showed highly different ranks of the species at pH 5 and 6. Proteolytic activities toward N(α)-Benzoyl-D,L-arginine 4-nitroanilide hydrochloride, N-Succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanine 4-nitroanilide, N-Succinyl-L-alanyl-L-alanyl-L-alanine 4-nitroanilide, Benzyloxycarbonyl-L-arginine-L-arginyl 4-nitroanilide, and N-Methoxysuccinyl-L-alanyl-L-alanyl-L-prolyl-L-methionine 4-nitroanilide (MAAPMpNA) were highest at alkaline pH, but the activity toward MAAPMpNA was also high at pH 5 and 6. In contrast, N-Succinyl-L-alanyl-L-alanyl-L-phenylalanine 4-nitroanilide (AAPpNA) and L-arginyl 4-nitroanilide (ArgpNA) had the highest activity recorded at pH 6. The high activities observed on AAPpNA, ArgpNA, and MAAPMpNA at digestive pH suggest that enzymes present in these extracts could have the majority of proteolysis in the mite gut. Evidence of the presence of proteolytic activities on all tested substrates and in all the tested mite homogenates suggests that the proteolytic activities may contribute to allergenicity. Poor or undetected hydrolytic activities of mite extracts toward substrates for keratin and collagen at digestive pH underline the importance of ecological interactions between mites and microorganisms in the utilization of such substrates.
- MeSH
- Acaridae enzymologie MeSH
- albuminy metabolismus MeSH
- druhová specificita MeSH
- fosfiny MeSH
- kaseiny metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- oligopeptidy metabolismus MeSH
- proteasy metabolismus MeSH
- substrátová specifita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Comparison of cell-wall-bound extracellular proteinases (CEPs) from Lactobacillus paracasei (LBP) ssp. paracasei natural isolates BGHN14, BGAR75 and BGAR76 with Lactococcus lactis (LCL) ssp. cremoris Wg2, in their action on alpha(S1)-, beta- and kappa-casein was done. The CEPs of LBP strains were able to degrade alpha(S1)- and beta-caseins and their caseinolytic specificity depended on the type of buffer used. These CEPs, compared with LCL Wg2, differ in four amino acid residues in small segments predicted to be involved in substrate binding. The most striking features of this comparison are the presence of Ala instead of Ser(329) and the presence of Thr instead of Asn(256) and Ala(299), in the subtilisin-like region of the CEP in LBP natural isolates. Additional conservative amino acid substitution Leu to Ile(364) was found.
- MeSH
- bakteriální geny genetika MeSH
- bakteriální proteiny genetika metabolismus MeSH
- cysteinové endopeptidasy genetika metabolismus MeSH
- financování organizované MeSH
- kaseiny metabolismus MeSH
- katalytická doména genetika MeSH
- Lactobacillus enzymologie genetika MeSH
- Lactococcus lactis enzymologie genetika MeSH
- potravinářská mikrobiologie MeSH
- substituce aminokyselin MeSH
- substrátová specifita MeSH
- subtilisin genetika MeSH
- sýr mikrobiologie MeSH
- Publikační typ
- srovnávací studie MeSH
