BACKGROUND: Vascular endothelial growth factor (VEGF) is not only a potent angiogenic factor but it also promotes axonal outgrowth and proliferation of Schwann cells. The aim of the present study was to quantitatively assess reinnervation of musculocutaneous nerve (MCN) stumps using motor and primary sensory neurons after plasmid phVEGF transfection and end-to-end (ETE) or end-to-side (ETS) neurorrhaphy. The distal stump of rat transected MCN, was transfected with plasmid phVEGF, plasmid alone or treated with vehiculum and reinnervated following ETE or ETS neurorrhaphy for 2 months. The number of motor and dorsal root ganglia neurons reinnervating the MCN stump was estimated following their retrograde labeling with Fluoro-Ruby and Fluoro-Emerald. Reinnervation of the MCN stumps was assessed based on density, diameter and myelin sheath thickness of regenerated axons, grooming test and the wet weight index of the biceps brachii muscles. RESULTS: Immunohistochemical detection under the same conditions revealed increased VEGF in the Schwann cells of the MCN stumps transfected with the plasmid phVEGF, as opposed to control stumps transfected with only the plasmid or treated with vehiculum. The MCN stumps transfected with the plasmid phVEGF were reinnervated by moderately higher numbers of motor and sensory neurons after ETE neurorrhaphy compared with control stumps. However, morphometric quality of myelinated axons, grooming test and the wet weight index were significantly better in the MCN plasmid phVEGF transfected stumps. The ETS neurorrhaphy of the MCN plasmid phVEGF transfected stumps in comparison with control stumps resulted in significant elevation of motor and sensory neurons that reinnervated the MCN. Especially noteworthy was the increased numbers of neurons that sent out collateral sprouts into the MCN stumps. Similarly to ETE neurorrhaphy, phVEGF transfection resulted in significantly higher morphometric quality of myelinated axons, behavioral test and the wet weight index of the biceps brachii muscles. CONCLUSION: Our results showed that plasmid phVEGF transfection of MCN stumps could induce an increase in VEGF protein in Schwann cells, which resulted in higher quality axon reinnervation after both ETE and ETS neurorrhaphy. This was also associated with a better wet weight biceps brachii muscle index and functional tests than in control rats.

• MeSH
• dextrany MeSH
• fluoresceiny MeSH
• genetická terapie metody   MeSH
• krysa rodu Rattus MeSH
• mícha patologie   MeSH
• modely nemocí na zvířatech MeSH
• nemoci periferního nervového systému patologie   terapie   MeSH
• nervová vlákna myelinizovaná patologie   MeSH
• nervus musculocutaneus metabolismus   patologie   fyziologie   MeSH
• neurologické vyšetření MeSH
• neurony metabolismus   patologie   MeSH
• potkani Wistar MeSH
• přední končetina patofyziologie   MeSH
• regenerace nervu genetika   fyziologie   MeSH
• rhodaminy MeSH
• vaskulární endoteliální růstový faktor A biosyntéza   metabolismus   terapeutické užití   MeSH
• velikost orgánu fyziologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dextrany MeSH
• fluoresceiny MeSH
• fluoro-emerald MeSH Prohlížeč
• Fluoro-Ruby MeSH Prohlížeč
• rhodaminy MeSH
• vaskulární endoteliální růstový faktor A MeSH

In spite of the inhibitory effects of ethanol (EtOH) on platelet function, soft blood clots are often observed in cadaveric blood in cases of sudden death after alcohol ingestion. In order to resolve this discrepancy, we have focused on the role of vascular endothelial cells. We tried to investigate the effects of EtOH and LPS on endothelial cells from various perspectives; thrombogenic factor (Von Willebrand factor, VWF), fibrinolytic factor (tissue plasminogen activator, tPA) and inflammatory factor (Interleukin-6, IL-6). Human umbilical vein endothelial cells (HUVECs) were incubated with various concentrations of EtOH (0-160 mM) with or without LPS. Treatment with EtOH and LPS increased VWF release from HUVECs without enhancement mRNA expression. Treatment with 40 mM of EtOH also increased IL-6 release from HUVECs without enhancement mRNA expression. Although EtOH inhibited LPS-induced IL-6 mRNA expression, 20 mM of EtOH still had an increasing effect on the release of IL-6. These doses of EtOH are consistent with a moderate drunkenness level in a normal person. On the other hand, mRNA expression and release reaction of tPA were not affected by EtOH and LPS addition. In conclusion, EtOH enhances procoagulant status via VWF release and IL-6 production cooperation with LPS and may contribute to soft blood clot formation in cadaveric blood.

• MeSH
• buněčné linie MeSH
• cévní endotel účinky léků   metabolismus   MeSH
• ethanol farmakologie   MeSH
• hemokoagulace MeSH
• interleukin-6 metabolismus   MeSH
• koagulační faktory metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• lipopolysacharidy farmakologie   MeSH
• posmrtné změny MeSH
• tkáňový aktivátor plazminogenu metabolismus   MeSH
• venae umbilicales účinky léků   MeSH
• von Willebrandův faktor metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ethanol MeSH
• interleukin-6 MeSH
• koagulační faktory MeSH
• lipopolysacharidy MeSH
• tkáňový aktivátor plazminogenu MeSH
• von Willebrandův faktor MeSH

Tumor necrosis factor-α (TNF-α) is a pleiotropic cytokine playing critical roles in host defense and acute and chronic inflammation. It has been described in fish, amphibians, and mammals but was considered to be absent in the avian genomes. Here, we report on the identification and functional characterization of the avian ortholog. The chicken TNF-α (chTNF-α) is encoded by a highly GC-rich gene, whose product shares with its mammalian counterpart 45% homology in the extracellular part displaying the characteristic TNF homology domain. Orthologs of chTNF-α were identified in the genomes of 12 additional avian species including Palaeognathae and Neognathae, and the synteny of the closely adjacent loci with mammalian TNF-α orthologs was demonstrated in the crow (Corvus cornix) genome. In addition to chTNF-α, we obtained full sequences for homologs of TNF-α receptors 1 and 2 (TNFR1, TNFR2). chTNF-α mRNA is strongly induced by lipopolysaccharide (LPS) stimulation of monocyte derived, splenic and bone marrow macrophages, and significantly upregulated in splenic tissue in response to i.v. LPS treatment. Activation of T-lymphocytes by TCR crosslinking induces chTNF-α expression in CD4+ but not in CD8+ cells. To gain insights into its biological activity, we generated recombinant chTNF-α in eukaryotic and prokaryotic expression systems. Both, the full-length cytokine and the extracellular domain rapidly induced an NFκB-luciferase reporter in stably transfected CEC-32 reporter cells. Collectively, these data provide strong evidence for the existence of a fully functional TNF-α/TNF-α receptor system in birds thus filling a gap in our understanding of the evolution of cytokine systems.

• Klíčová slova
• avian, biological activity, chicken, missing gene, tumor necrosis factor-α, tumor necrosis factor-α receptors,
• MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• GC bohatá sekvence genetika   MeSH
• klonování DNA MeSH
• kultivované buňky MeSH
• kur domácí imunologie   MeSH
• lidé MeSH
• makrofágy imunologie   MeSH
• NF-kappa B metabolismus   MeSH
• Palaeognathae imunologie   MeSH
• ptačí proteiny genetika   metabolismus   MeSH
• receptory TNF genetika   metabolismus   MeSH
• savci imunologie   MeSH
• sekvenční seřazení MeSH
• TNF-alfa genetika   MeSH
• vrány imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• NF-kappa B MeSH
• ptačí proteiny MeSH
• receptory TNF MeSH
• TNF-alfa MeSH

Exercise order is one of the significant factors modulating training effects. Therefore, the aim of this study was to compare the effectiveness of an 8-week complex (CPX) training program utilizing intra-CPX active recovery with compound training (CMP) on bilateral and single-leg jumping performance, change of direction test time (shuttle test), and the post-activation performance enhancement (PAPE) response in a group of basketball players. Thirteen participants were performing CPX bi-weekly combined with regular pre-season basketball practice, while eleven participants were performing CMP for 8 weeks. Before and after the interventions, the following fitness tests were assessed: (i) bilateral countermovement jump, (ii) single-leg countermovement jump, (iii) shuttle run test. All tests were performed pre- and post-conditioning activity (CA-three sets of five drop jumps). The results showed a statistically significant increase in non-dominant (p = 0.019) and dominant single-leg jump relative peak power (p = 0.001), and in non-dominant single-leg jump height (p = 0.022) post-training compared to pre-training. The CA was significantly and similarly effective in eliciting a PAPE response in all tests before and after each intervention (p < 0.039; for all). However, the magnitude of improvement in CMJ and shuttle test time was trivial to small and did not reach statistical significance. Both 8 weeks of CPX and CMP training led to significant improvements in the SLJ power output of both the dominant and non-dominant limbs as well as the height of the non-dominant SLJ. Neither of the training methods had significant impacts on the magnitude of the PAPE response.

• Klíčová slova
• exercise, fatigue, post-activation performance enhancement, resistance training, sport performance,
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Sigma factors are one of the components of RNA polymerase holoenzymes, and an essential factor of transcription initiation in bacteria. Corynebacterium glutamicum possesses seven genes coding for sigma factors, most of which have been studied to some detail; however, the role of SigD in transcriptional regulation in C. glutamicum has been mostly unknown. RESULTS: In this work, pleiotropic effects of sigD overexpression at the level of phenotype, transcripts, proteins and metabolites were investigated. Overexpression of sigD decreased the growth rate of C. glutamicum cultures, and induced several physiological effects such as reduced culture foaming, turbid supernatant and cell aggregation. Upon overexpression of sigD, the level of Cmt1 (corynomycolyl transferase) in the supernatant was notably enhanced, and carbohydrate-containing compounds were excreted to the supernatant. The real-time PCR analysis revealed that sigD overexpression increased the expression of genes related to corynomycolic acid synthesis (fadD2, pks), genes encoding corynomycolyl transferases (cop1, cmt1, cmt2, cmt3), L, D-transpeptidase (lppS), a subunit of the major cell wall channel (porH), and the envelope lipid regulation factor (elrF). Furthermore, overexpression of sigD resulted in trehalose dicorynomycolate accumulation in the cell envelope. CONCLUSIONS: This study demonstrated that SigD regulates the synthesis of corynomycolate and related compounds, and expanded the knowledge of regulatory functions of sigma factors in C. glutamicum.

• Klíčová slova
• Corynebacterium glutamicum, Mycomembrane, SigD, Sigma factor, Trehalose dicorynomycolate,
• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• Corynebacterium glutamicum genetika   růst a vývoj   metabolismus   MeSH
• kyseliny mykolové metabolismus   MeSH
• regulace genové exprese u bakterií MeSH
• sigma faktor genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bakteriální proteiny MeSH
• corynomycolic acid MeSH Prohlížeč
• kyseliny mykolové MeSH
• sigma faktor MeSH

The transcription factor PU.1 (Purine-rich DNA binding, SPI1) is a key regulator of hematopoiesis, whose level is influenced by transcription through its enhancers and its post-transcriptional degradation via microRNA-155 (miR-155). The degree of transcriptional regulation of the PU.1 gene is influenced by repression via DNA methylation, as well as other epigenetic factors, such as those related to progenitor maturation status, which is modulated by the transcription factor Myeloblastosis oncogene (MYB). In this work, we show that combinatorial treatment of acute myeloid leukemia (AML) cells with DNA methylation inhibitors (5-Azacytidine), MYB inhibitors (Celastrol), and anti-miR-155 (AM155) ideally leads to overproduction of PU.1. We also show that PU.1 reactivation can be compensated by miR-155 and that only a combined approach leads to sustained PU.1 derepression, even at the protein level. The triple effect on increasing PU.1 levels in myeloblasts stimulates the myeloid transcriptional program while inhibiting cell survival and proliferation, leading to partial leukemic differentiation.

• Klíčová slova
• 5-Azacytidine, Celastrol, microRNA miR-155, transcription factor PU.1,
• MeSH
• akutní myeloidní leukemie * farmakoterapie   genetika   MeSH
• buněčná diferenciace genetika   MeSH
• lidé MeSH
• mikro RNA * genetika   metabolismus   MeSH
• protoonkogenní proteiny genetika   metabolismus   MeSH
• regulace genové exprese u leukemie MeSH
• trans-aktivátory metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• mikro RNA * MeSH
• MIRN155 microRNA, human MeSH Prohlížeč
• proto-oncogene protein Spi-1 MeSH Prohlížeč
• protoonkogenní proteiny MeSH
• trans-aktivátory MeSH

Secondary diversification of the Ig repertoire occurs through somatic hypermutation (SHM), gene conversion (GCV), and class switch recombination (CSR)-three processes that are initiated by activation-induced cytidine deaminase (AID). AID targets Ig genes at orders of magnitude higher than the rest of the genome, but the basis for this specificity is poorly understood. We have previously demonstrated that enhancers and enhancer-like sequences from Ig genes are capable of stimulating SHM of neighboring genes in a capacity distinct from their roles in increasing transcription. Here, we use an in vitro proteomics approach to identify E-box, MEF2, Ets, and Ikaros transcription factor family members as potential binders of these enhancers. ChIP assays in the hypermutating Ramos B cell line confirmed that many of these factors bound the endogenous Igλ enhancer and/or the IgH intronic enhancer (Eμ) in vivo. Further investigation using SHM reporter assays identified binding sites for E2A and MEF2B in Eμ and demonstrated an association between loss of factor binding and decreases in the SHM stimulating activity of Eμ mutants. Our results provide novel insights into trans-acting factors that dictate SHM targeting and link their activity to specific DNA binding sites within Ig enhancers.

• Klíčová slova
• AID, E2A, MEF2B, Ramos B cell line, Somatic hypermutation,
• MeSH
• geny pro imunoglobuliny MeSH
• kur domácí MeSH
• lidé MeSH
• somatická hypermutace imunoglobulinových genů fyziologie   MeSH
• transkripční faktory genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• transkripční faktory MeSH

In an attempt to compare effects of different neurotrophic factors on impaired memory function, young adult naive rats were trained to find the hidden platform in the Morris water maze (3 consecutive days, eight trials/day). The fimbria-fornix was unilaterally removed by aspiration and nerve growth factor (NGF) (11 micrograms/ml and 0.5 microgram/ml; groups NGF and ngf, respectively) or basic fibroblast growth factor (bFGF) (0.2 microgram/ml, group FGF) were applied via intra-cerebroventricular infusion by the osmotic minipump (flow rate 0.5 microliter/h, 14 days). Nootropic drug Cerebrolysin (EBEWE Arzneitmittel; 2.5 ml/kg/day, group CER) was applied via intraperitoneal injection (14 days). One group was formed by the rats treated with NGF (11 micrograms/ml) and Cerebrolysin (group NGFCER). Non-lesioned and lesioned only rats served as controls (groups INT and LES). After a 14-day treatment, rats were tested using the retention test (1 day, four trials). On the next day, the rats were tested using transfer test (3 days, eight trials/day). Escape latency and length of trajectory was recorded. Groups NGF, ngf, FGF and LES were similarly impaired in their ability to retrieve the old position of the platform (retention test), as well as in their ability to navigate to the new position of the platform (transfer test). In the latter, NGF group significantly differed from lesioned animals. Groups CER and NGFCER were comparable to group INT in the retention or transfer test. It is concluded that anterograde amnesia elicited by fimbria-fornix lesion can be abbreviated by NGF and/or CER, while retrograde amnesia is absent only in rats treated by CER. No short-term influence of bFGF was found. It is suggested that biochemical systems other than the cholinergic one are involved.

• MeSH
• aminokyseliny farmakologie   MeSH
• bludiště - učení účinky léků   MeSH
• fibroblastový růstový faktor 2 farmakologie   MeSH
• hipokampus cytologie   fyziologie   MeSH
• krysa rodu Rattus MeSH
• neurotrofní faktory farmakologie   MeSH
• nootropní látky farmakologie   MeSH
• parasympatický nervový systém cytologie   účinky léků   MeSH
• poruchy paměti farmakoterapie   psychologie   MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aminokyseliny MeSH
• cerebrolysin MeSH Prohlížeč
• fibroblastový růstový faktor 2 MeSH
• neurotrofní faktory MeSH
• nootropní látky MeSH

Intestinal homeostasis is precisely regulated by a number of endogenous regulatory molecules but significantly influenced by dietary compounds. Malfunction of this system may result in chronic inflammation and cancer. Dietary essential n-3 polyunsaturated fatty acids (PUFAs) and short-chain fatty acid butyrate produced from fibre display anti-inflammatory and anticancer activities. Both compounds were shown to modulate the production and activities of TNF family cytokines. Cytokines from the TNF family (TNF- α, TRAIL, and FasL) have potent inflammatory activities and can also regulate apoptosis, which plays an important role in cancer development. The results of our own research showed enhancement of apoptosis in colon cancer cells by a combination of either docosahexaenoic acid (DHA) or butyrate with TNF family cytokines, especially by promotion of the mitochondrial apoptotic pathway and modulation of NF κ B activity. This review is focused mainly on the interaction of dietary PUFAs and butyrate with these cytokines during colon inflammation and cancer development. We summarised recent knowledge about the cellular and molecular mechanisms involved in such effects and outcomes for intestinal cell behaviour and pathologies. Finally, the possible application for the prevention and therapy of colon inflammation and cancer is also outlined.

• MeSH
• apoptóza MeSH
• butyráty metabolismus   MeSH
• cytokiny metabolismus   MeSH
• dieta MeSH
• kolon patologie   MeSH
• kyseliny dokosahexaenové metabolismus   MeSH
• lidé MeSH
• mitochondrie patologie   MeSH
• myši MeSH
• nádory metabolismus   MeSH
• nenasycené mastné kyseliny metabolismus   MeSH
• NF-kappa B metabolismus   MeSH
• střevní sliznice metabolismus   MeSH
• tumor nekrotizující faktory metabolismus   MeSH
• zánět metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• butyráty MeSH
• cytokiny MeSH
• kyseliny dokosahexaenové MeSH
• nenasycené mastné kyseliny MeSH
• NF-kappa B MeSH
• tumor nekrotizující faktory MeSH

We investigated the role of the 5-lipoxygenase (5-LOX) pathway of arachidonic acid metabolism in tumour necrosis factor-alpha (TNF-alpha)-induced differentiation of human leukemic HL-60 cells using MK-886, an inhibitor of 5-LOX activating protein. MK-886 augmented cell cycle arrest and differentiation induced by TNF-alpha; however, both effects were probably 5-LOX-independent, because a general LOX inhibitor, NDGA, had no effect. Apoptosis was significantly elevated after combined TNF-alpha and MK-886 treatment, which could be partially associated with changes of Mcl-1 protein expression. NF-kappaB signalling or activation of JNKs were not modulated by MK-886. Thus, in addition to apoptosis, MK-886 can enhance TNF-alpha-induced differentiation.

• MeSH
• apoptóza * MeSH
• arachidonát-5-lipoxygenasa metabolismus   MeSH
• buněčná diferenciace MeSH
• buněčný cyklus MeSH
• časové faktory MeSH
• HL-60 buňky MeSH
• indoly farmakologie   MeSH
• inhibitory lipoxygenas farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• signální transdukce MeSH
• TNF-alfa metabolismus   MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• arachidonát-5-lipoxygenasa MeSH
• indoly MeSH
• inhibitory lipoxygenas MeSH
• MK-886 MeSH Prohlížeč
• TNF-alfa MeSH