BACKGROUND: Human colostrum and milk contain components that influence development. Our aim was to use a protein array to determine the cytokine profile of human lacteal secretions and changes that occur during the early postpartum period. METHODS: We collected 17 samples of colostrum during the first 2 days postpartum and a 2nd group of 5 sets of 2 to 3 sequential colostrum or milk samples (at 20- to 30-h intervals). We analyzed the samples with array membranes consisting of 42 or 79 antibodies directed against cytokines. RESULTS: In most samples, we detected the previously described cytokines interleukin-8 (IL-8)/CXCL8, epidermal growth factor (EGF), growth-related oncoprotein (GRO)/CXCL1-3, angiogenin, transforming growth factor beta-2, and monocyte chemotactic protein 1 (MCP-1/CCL2). In addition, we found 32 cytokines that have not been described before in colostrum. Cytokine concentrations differed among mothers, and the spectrum of cytokines changed with time after delivery. A significant decrease occurred in IL-12 and macrophage inflammatory protein-1delta/CCL15 and a significant increase in MCP-1/CCL2. The production of angiogenin, vascular endothelial growth factor, GRO/CXCL1-3, EGF, and IL-8/CXCL8 remained high throughout. The concentrations of 2 selected cytokines measured with the array technique and ELISA showed moderate to strong correlation (r = 0.63 for EGF and r = 0.84 for IL-8/CXCL8). CONCLUSION: Despite the lack of precise quantification, the protein array might be suitable for cytokine screening. It allows simultaneous detection of a broad spectrum of cytokines (including those not described before) in lacteal secretions.

• MeSH
• časové faktory MeSH
• chemokiny analýza   MeSH
• čipová analýza proteinů MeSH
• cytokiny analýza   MeSH
• dospělí MeSH
• ELISA MeSH
• kolostrum chemie   MeSH
• lidé MeSH
• mateřské mléko chemie   MeSH
• mezibuněčné signální peptidy a proteiny analýza   MeSH
• poporodní období MeSH
• proteomika MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• chemokiny MeSH
• cytokiny MeSH
• mezibuněčné signální peptidy a proteiny MeSH

Intrinsically disordered regions (IDRs) are protein regions that lack persistent secondary or tertiary structure under native conditions. IDRs represent >40% of the eukaryotic proteome and play a crucial role in protein-protein interactions. The classical approach for identification of these interaction interfaces is based on mutagenesis combined with biochemical techniques such as coimmunoprecipitation or yeast two-hybrid screening. This approach either provides information of low resolution (large deletions) or very laboriously tries to precisely define the binding epitope via single amino acid substitutions. Here, we report the use of a peptide microarray based on the human scaffold protein AXIN1 for high-throughput and -resolution mapping of binding sites for several AXIN1 interaction partners in vitro For each of the AXIN1-binding partners tested, i.e. casein kinase 1 ϵ (CK1ϵ); c-Myc; peptidyl-prolyl cis/trans isomerase, NIMA-interacting 1 (Pin1); and p53, we found at least three different epitopes, predominantly in the central IDR of AXIN1. We functionally validated the specific AXIN1-CK1ϵ interaction identified here with epitope-mimicking peptides and with AXIN1 variants having deletions of short binding epitopes. On the basis of these results, we propose a model in which AXIN1 competes with dishevelled (DVL) for CK1ϵ and regulates CK1ϵ-induced phosphorylation of DVL and activation of Wnt/β-catenin signaling.

• Klíčová slova
• Myc (c-Myc), Wnt pathway, axin, casein kinase 1ϵ, dishevelled, intrinsically disordered region, p53, peptide array, scaffold protein, serine/threonine protein kinase,
• MeSH
• axin protein metabolismus   MeSH
• beta-katenin metabolismus   MeSH
• čipová analýza proteinů metody   MeSH
• fosforylace MeSH
• interakční proteinové domény a motivy * MeSH
• kaseinkinasa Iepsilon metabolismus   MeSH
• kompetitivní vazba MeSH
• lidé MeSH
• peptidy metabolismus   MeSH
• protein dishevelled metabolismus   MeSH
• proteiny Wnt metabolismus   MeSH
• signální dráha Wnt MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• axin protein MeSH
• AXIN1 protein, human MeSH Prohlížeč
• beta-katenin MeSH
• kaseinkinasa Iepsilon MeSH
• peptidy MeSH
• protein dishevelled MeSH
• proteiny Wnt MeSH

Overexpression of procathepsin D (pCD) is reported to occur in numerous types of cancer and is associated with increased growth and metastasis. It has been established that pCD affects multiple stages of tumor progression including proliferation, angiogenesis and metastasis. Previously, we showed that the mitogenic effect of pCD on cancer cells is mediated by interaction of its activation peptide (AP) with yet unidentified cell surface receptor. In this investigation, gene expression profiles were compared between AP-treated and control human breast cancer ZR-75-1 cells to elucidate the mechanism of AP mitogenicity. Several differentially expressed genes involved in signal transduction, regulation of cell cycle, apoptosis, tumor invasion and metastasis were identified using microarray technology. These findings, including overexpression of NF-kappaB2, were confirmed in breast cancer cell lines by reverse-transcriptase PCR (RT-PCR). Understanding the mechanism of pCDs effect on breast cancer cells could extend possibilities of breast cancer treatment in the future.

• MeSH
• kathepsin D farmakologie   MeSH
• lidé MeSH
• nádorové buňky kultivované MeSH
• nádorové proteiny genetika   MeSH
• nádory prsu genetika   MeSH
• NF-kappa B - podjednotka p52 genetika   metabolismus   MeSH
• peptidové fragmenty farmakologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prekurzory enzymů farmakologie   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• signální transdukce MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• kathepsin D MeSH
• nádorové proteiny MeSH
• NF-kappa B - podjednotka p52 MeSH
• peptidové fragmenty MeSH
• prekurzory enzymů MeSH
• procathepsin D MeSH Prohlížeč

Jumonji (JMJ, Jarid2), a prototypical member of the jumonji domain-containing protein family, plays a major role in embryonic cardiac development, but its role in the developed heart is unclear. Cardiomyocytes from neonatal mouse heart were treated in culture with NO donor SIN-1, 500 microM, for 2, 4, and 20 h. SIN-1 treatment was associated with a significant and 6.9 +/- 2.5 fold increase in jmj gene expression over all time points. The expression of jmj increased markedly and significantly 4.2 +/- 1.1 fold, 16.6 +/- 4.1 fold, and 2.7 +/- 0.3 fold, respectively, at time points 2 h, 4 h, and 20 h after treatment. The ability of the increase in gene expression to translate into an increase in cellular protein expression was ascertained by Western blotting, which showed an increase in the JMJ protein in whole-cell lysates. Because of the relationship of JMJ to Rb and ANP in the heart, gene expression of these proteins was also examined. SIN-1 produced a small but significant increase in Rb2, but not Rb1 or Rb-binding proteins 4, 6, or 7. In contrast, SIN-1 produced a marked and significant reduction in natriuretic peptide precursor type B but not type C to 0.24 +/- 0.09 fold of the control. These data suggest that JMJ may be a critical, previously unrecognized factor that mediates some of the cellular effects of NO, that NO may be able to increase JMJ in diseases associated with reduced JMJ expression.

• MeSH
• donory oxidu dusnatého farmakologie   MeSH
• kardiomyocyty účinky léků   metabolismus   MeSH
• kultivované buňky MeSH
• molsidomin analogy a deriváty   farmakologie   MeSH
• myši MeSH
• natriuretický peptid typu B genetika   metabolismus   MeSH
• oxid dusnatý metabolismus   MeSH
• PRC2 MeSH
• protein p130 podobný retinoblastomu genetika   MeSH
• proteiny nervové tkáně genetika   metabolismus   MeSH
• regulace genové exprese MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• srdce MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• donory oxidu dusnatého MeSH
• Jarid2 protein, mouse MeSH Prohlížeč
• linsidomine MeSH Prohlížeč
• molsidomin MeSH
• natriuretický peptid typu B MeSH
• oxid dusnatý MeSH
• PRC2 MeSH
• protein p130 podobný retinoblastomu MeSH
• proteiny nervové tkáně MeSH
• RBL2 protein, human MeSH Prohlížeč

AIM: Evaluation of serum levels of 17 cytokines and 5 adhesion molecules in patients with newly diagnosed acute myeloid leukemia (AML) and in healthy subjects using biochip array technology. METHODS: A total of 15 AML patients and 15 healthy subjects (blood donors) were studied. Serum samples were analyzed by biochip based immunoassays on the Evidence Investigator analyzer. This approach allows multi-analytical determination from a single sample. T-tests were used for statistical analysis. RESULTS: In newly diagnosed AML patients, we found significant increase (p < 0.01) in serum VCAM-1, ICAM-1, E-selectin, L-selectin, and significant increase (p < 0.05) in serum IL-6, IL-8. No significant differences were found in the levels of other evaluated cytokines and adhesion molecules. CONCLUSION: Our results indicate that serum levels of specific cytokines and adhesion molecules (VCAM-1, ICAM-1, E-selectin, L-selectin, IL-6, IL-8) are significantly altered in patients with newly diagnosed AML, showing activity of the disease. Whether these alterations could serve as a prognostic marker for AML is not known. Further studies will be needed to define the potential role of these and additional markers in the risk stratification of AML.

• MeSH
• akutní myeloidní leukemie krev   genetika   patologie   MeSH
• cévní buněčněadhezivní molekula-1 krev   MeSH
• čipová analýza proteinů metody   MeSH
• cytokiny krev   MeSH
• dospělí MeSH
• E-selektin krev   MeSH
• interleukin-6 krev   MeSH
• interleukin-8 krev   MeSH
• L-selektin krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mezibuněčná adhezivní molekula-1 krev   MeSH
• molekuly buněčné adheze krev   MeSH
• nádorové biomarkery krev   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cévní buněčněadhezivní molekula-1 MeSH
• cytokiny MeSH
• E-selektin MeSH
• interleukin-6 MeSH
• interleukin-8 MeSH
• L-selektin MeSH
• mezibuněčná adhezivní molekula-1 MeSH
• molekuly buněčné adheze MeSH
• nádorové biomarkery MeSH

A combined approach to signal enhancement in fluorescence affinity biosensors and assays is reported. It is based on the compaction of specifically captured target molecules at the sensor surface followed by optical probing with a tightly confined surface plasmon (SP) field. This concept is utilized by using a thermoresponsive hydrogel (HG) binding matrix that is prepared from a terpolymer derived from poly(N-isopropylacrylamide) (pNIPAAm) and attached to a metallic sensor surface. Epi-illumination fluorescence and SP-enhanced total internal reflection fluorescence readouts of affinity binding events are performed to spatially interrogate the fluorescent signal in the direction parallel and perpendicular to the sensor surface. The pNIPAAm-based HG binding matrix is arranged in arrays of sensing spots and employed for the specific detection of human IgG antibodies against the Epstein-Barr virus (EBV). The detection is performed in diluted human plasma or with isolated human IgG by using a set of peptide ligands mapping the epitope of the EBV nuclear antigen. Alkyne-terminated peptides were covalently coupled to the pNIPAAm-based HG carrying azide moieties. Importantly, using such low-molecular-weight ligands allowed preserving the thermoresponsive properties of the pNIPAAm-based architecture, which was not possible for amine coupling of regular antibodies that have a higher molecular weight.

• Klíčová slova
• biomarkers, click chemistry, microarrays, pNIPAAm, peptide, plasmon-enhanced fluorescence, serotesting, thermoresponsive hydrogel,
• MeSH
• akrylové pryskyřice chemie   MeSH
• biosenzitivní techniky metody   MeSH
• fluorescence MeSH
• hydrogely chemie   metabolismus   MeSH
• imunoglobulin G analýza   imunologie   MeSH
• infekce virem Epsteina-Barrové diagnóza   imunologie   metabolismus   virologie   MeSH
• lidé MeSH
• peptidové fragmenty imunologie   metabolismus   MeSH
• polymery chemie   MeSH
• virus Epsteinův-Barrové - jaderné antigeny imunologie   MeSH
• virus Epsteinův-Barrové imunologie   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akrylové pryskyřice MeSH
• hydrogely MeSH
• imunoglobulin G MeSH
• peptidové fragmenty MeSH
• poly-N-isopropylacrylamide MeSH Prohlížeč
• polymery MeSH
• virus Epsteinův-Barrové - jaderné antigeny MeSH

The emergence of communicable and non-communicable diseases has posed a health challenge for millions of people worldwide and is a major threat to the economic and social development in the coming century. The occurrence of the recent pandemic, SARS-CoV-2, caused by lethal severe acute respiratory syndrome coronavirus 2, is one such example. Rapid research and development of drugs for the treatment and management of these diseases have become an incredibly challenging task for the pharmaceutical industry. Although, substantial attention has been paid to the discovery of therapeutic compounds from natural sources having significant medicinal potential, their synthesis has made a slow progress. Hence, the discovery of new targets by the application of the latest biotechnological and synthetic biology approaches is very much the need of the hour. Polyketides (PKs) and non-ribosomal peptides (NRPs) found in bacteria, fungi and plants are a diverse family of natural products synthesized by two classes of enzymes: polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS). These enzymes possess immense biomedical potential due to their simple architecture, catalytic capacity, as well as diversity. With the advent of the latest in-silico and in-vitro strategies, these enzymes and their related metabolic pathways, if targeted, can contribute highly towards the biosynthesis of an array of potentially natural drug leads that have antagonist effects on biopolymers associated with various human diseases. In the face of the rising threat from multidrug-resistant pathogens, this will further open new avenues for the discovery of novel and improved drugs by combining natural and synthetic approaches. This review discusses the relevance of polyketides and non-ribosomal peptides and the improvement strategies for the development of their derivatives and scaffolds, and how they will be beneficial for future bioprospecting and drug discovery.

• Klíčová slova
• NRPS, Polyketides, bioactive molecules, natural products, secondary metabolites,
• MeSH
• farmakoterapie COVID-19 * MeSH
• lidé MeSH
• peptidy farmakologie   terapeutické užití   MeSH
• polyketidy * chemie   metabolismus   farmakologie   MeSH
• SARS-CoV-2 MeSH
• vyvíjení léků MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• peptidy MeSH
• polyketidy * MeSH

Capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC) were used for the analysis of new synthetic derivatives of hypophysis neurohormones--vasopressin and oxytocin, and pancreatic hormone--human insulin (HI) and its octapeptide fragment, derivatized by fluorescent probe, 4-chloro-7-nitrobenzo[1,2,5]oxadiazol (NBD). The suitable composition of background electrolytes (BGEs) was selected on the basis of calculated pH dependence of effective charge of analyzed peptides. Basic ionogenic peptides were analyzed by CZE in the acidic BGE composed of 100 mM H3PO4, 50 mM Tris, pH 2.25. The ionogenic peptides with fluorescent label, NBD, were analyzed in 0.5 M acetic acid, pH 2.5. The best MEKC separation of non-ionogenic peptides was achieved in alkaline BGE, 20 mM Tris, 5 mM H3PO4, with micellar pseudophase formed by 50 mM sodium dodecylsulfate (SDS), pH 8.8. Selected characteristics (noise, detectability of substance, sensitivity of detector) of the UV-absorption detectors (single wavelength detector, multiple-wavelength photodiode array detector (PDA), both of them operating at constant wavelength 206 nm) and laser-induced fluorescence (LIF) detector (excitation/emission wavelength 488/520 nm) were determined. The detectability of peptides in the single wavelength detector was 1.3-6.0 micromol dm(-3) and in the PDA detector 1.6-3.1 micromol dm(-3). The LIF detection was more sensitive, the applied concentration of NBD derivative of insulin fragment in CZE analysis with LIF detection was three orders lower than in CZE with UV-absorption detector, and the detectability of this peptide was improved to 15.8 nmol dm(-3).

• MeSH
• chromatografie micelární elektrokinetická kapilární metody   MeSH
• elektroforéza kapilární metody   MeSH
• fluorescenční spektrometrie metody   MeSH
• hormony analýza   MeSH
• lasery MeSH
• peptidy analýza   MeSH
• sekvence aminokyselin MeSH
• senzitivita a specificita MeSH
• spektrofotometrie ultrafialová metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• hormony MeSH
• peptidy MeSH

Several species of the genus Liposcelis are common insect pests that cause serious qualitative and quantitative losses to various stored grains and processed grain products. They also can contaminate foods, transmit pathogenic microorganisms and cause allergies in humans. The common occurrence of multi-species infestations and the fact that it is difficult to identify and discriminate Liposcelis spp. make accurate, rapid detection and discriminatory tools absolutely necessary for confirmation of their identity. In this study, PCR primers and probes specific to different Liposcelis spp. were designed based on nucleotide sequences of the cytochrome oxidase 1 (CO1) gene. Primer sets ObsCo13F/13R, PeaCo15F/14R, BosCO7F/7R, BruCo5F/5R, and DecCo11F/11R were used to specifically detect Liposcelis obscura Broadhead, Liposcelis pearmani Lienhard, Liposcelis bostrychophila Badonnel, Liposcelis brunnea Motschulsky and Liposcelis decolor (Pearman) in multiplex endpoint PCRs, which amplified products of 438-, 351-, 191-, 140-, and 87-bp, respectively. In multiplex TaqMan qPCR assays, orange, yellow, red, crimson and green channels corresponding to reporter dyes 6-ROXN, HEX, Cy5, Quasar705 and 6-FAM specifically detected L. obscura, L. brunnea, L. bostrychophila, L. pearmani and L. decolor, respectively. All developed primer and probe sets allowed specific amplification of corresponding targeted Liposcelis species. The development of multiplex endpoint PCR and multiplex TaqMan qPCR will greatly facilitate psocid identification and their management. The use of APCs will streamline and standardize PCR assays. APC will also provide the opportunity to have all positive controls in a single tube, which reduces maintenance cost and labor, but increases the accuracy and reliability of the assays. These novel methods from our study will have applications in pest management, biosecurity, quarantine, food safety, and routine diagnostics.

• MeSH
• dezinsekce MeSH
• DNA genetika   izolace a purifikace   MeSH
• hmyz genetika   MeSH
• hmyzí proteiny genetika   MeSH
• oligonukleotidové sondy chemie   genetika   MeSH
• oligonukleotidy chemie   genetika   MeSH
• polymerázová řetězová reakce metody   MeSH
• respirační komplex IV genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH
• hmyzí proteiny MeSH
• oligonukleotidové sondy MeSH
• oligonukleotidy MeSH
• respirační komplex IV MeSH

Some cyclic peptides and depsipeptides are synthesized in microorganisms by large multienzymes called nonribosomal peptide synthetases. The structures of peptide products originating in this way are complex and diverse and are microorganism-specific. This work proposes the use of fungal cyclic peptides and depsipeptides as extremely specific markers of fungal infections. Since a reliable molecular tool for diagnosing fungal infections at an early stage is still missing, we present mass spectrometry as a new, modern, broadband (with respect to fungal strain) and specific tool for clinical mycologists. More than 40 different fungal species can be rapidly characterized according to specific families of cyclic peptides, and in some cases, a particular fungal strain can be identified on the basis of its cyclopeptide profile. This paper is also aimed at initiating discussion on the biological role of these secondary metabolites, especially of those synthesized by medically important strains. Proven cytotoxic, anti-inflammatory or immunosuppressive activities of some cyclic peptides indicate that these molecules may contribute to the synergistic array of fungal virulence factors and support microbial invasion during fungal infection. In addition to an overview on recent mass spectrometric protocols for cyclic peptide sequencing, the structures of new peptides from Paecilomyces and Pseudallescheria are presented.

• MeSH
• biologické markery analýza   chemie   metabolismus   MeSH
• biosyntéza peptidů nezávislá na nukleových kyselinách * MeSH
• fungální proteiny analýza   chemie   metabolismus   MeSH
• hmotnostní spektrometrie * MeSH
• lidé MeSH
• mikrobiologické techniky MeSH
• mykózy diagnóza   metabolismus   mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• biologické markery MeSH
• fungální proteiny MeSH