BACKGROUND: CD16 was previously suggested to be a new marker of basophils that is subject to downregulation by FcεRI crosslinking. Certain compounds, including supraoptimal concentrations of the PKC inhibitors, bisindolylmaleimides, decouple the release of granules containing CD203c, CD63 and histamine, and may thus help to identify the mechanisms related to the CD16 externalization. OBJECTIVE: We hypothesized that CD16 is differentially expressed on the surface of basophils in patients with birch pollen or insect venom allergy and is subject to a regulation in response to allergens. We also employed CD203c and CD63 externalization decoupling by bisindolylmaleimides. METHODS: We performed a basophil activation test coupled with CD16 and histamine detection using cells isolated from patients with allergy to birch pollen or insect venom and negative controls. We employed two PKC inhibitors, bisindolylmaleimide II and Ro 31-8220 at their supraoptimal concentrations and, after difficulties reproducing previously published data, we analyzed the fluorescence of these inhibitors alone. We identified the CD16 isoforms by sequencing nested RT-PCR amplicons from flow cytometry sorted basophils and by cleaving the CD16b GPI anchor using a phospholipase C. RESULTS: We provide the first evidence that CD16a is expressed as a surface antigen on a small subpopulation of human basophils in patients with respiratory and insect venom allergy, and this antigen shows increased surface expression following allergen challenge or FcεRI crosslinking. We rejected the apparent decoupling of the surface expression of basophil activation markers following the administration of bisindolylmaleimides. CONCLUSIONS & CLINICAL RELEVANCE: The inclusion of αCD16 in negative selection cocktails selects against a subset of basophils that are CD16+ or CD16dim . Using CD16dim basophils and unstained leucocytes, we show that previous studies with supraoptimal concentrations of bisindolylmaleimides are likely flawed and are not associated with the differential expression of CD203c and CD63.
- MeSH
- alergie imunologie patologie MeSH
- antigeny CD63 imunologie MeSH
- bazofily imunologie patologie MeSH
- dospělí MeSH
- fosfodiesterasy imunologie MeSH
- GPI-vázané proteiny imunologie MeSH
- indoly chemie MeSH
- jedy členovců toxicita MeSH
- kousnutí a bodnutí hmyzem imunologie patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- maleimidy chemie MeSH
- pyrofosfatasy imunologie MeSH
- receptory IgG imunologie MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Over the last decades, multidrug-resistant bacteria have emerged and spread, increasing the number of bacteria, against which commonly used antibiotics are no longer effective. It has become a serious public health problem whose solution requires medical research in order to explore novel effective antimicrobial molecules. On the one hand, antimicrobial peptides (AMPs) are regarded as good alternatives because of their generally broad-spectrum activities, but sometimes they can be easily degraded by the organism or be toxic to animal cells. On the other hand, cationic carbosilane dendrons, whose focal point can be functionalized in many different ways, have also shown good antimicrobial activity. In this work, we synthetized first- and second-generation cationic carbosilane dendrons with a maleimide molecule on their focal point, enabling their functionalization with three different AMPs. After different microbiology studies, we found an additive effect between first-generation dendron and AMP3 whose study reveals three interesting effects: (i) bacteria aggregation due to AMP3, which could facilitate bacteria detection or even contribute to antibacterial activity by preventing host cell attack, (ii) bacteria disaggregation capability of second-generation cationic dendrons, and (iii) a higher AMP3 aggregation ability when dendrons were added previously to peptide treatment. These compounds and their different effects observed over bacteria constitute an interesting system for further mechanism studies.
Directing the organization of cells into a tissue with defined architectures is one use of biomaterials for regenerative medicine. To this end, hydrogels are widely investigated as they have mechanical properties similar to native soft tissues and can be formed in situ to conform to a defect. Herein, we describe the development of porous hydrogel tubes fabricated through a two-step polymerization process with an intermediate microsphere phase that provides macroscale porosity (66.5%) for cell infiltration. These tubes were investigated in a spinal cord injury model, with the tubes assembled to conform to the injury and to provide an orientation that guides axons through the injury. Implanted tubes had good apposition and were integrated with the host tissue due to cell infiltration, with a transient increase in immune cell infiltration at 1 week that resolved by 2 weeks post injury compared to a gelfoam control. The glial scar was significantly reduced relative to control, which enabled robust axon growth along the inner and outer surface of the tubes. Axon density within the hydrogel tubes (1744 axons/mm2) was significantly increased more than 3-fold compared to the control (456 axons/mm2), with approximately 30% of axons within the tube myelinated. Furthermore, implantation of hydrogel tubes enhanced functional recovery relative to control. This modular assembly of porous tubes to fill a defect and directionally orient tissue growth could be extended beyond spinal cord injury to other tissues, such as vascular or musculoskeletal tissue. STATEMENT OF SIGNIFICANCE: Tissue engineering approaches that mimic the native architecture of healthy tissue are needed following injury. Traditionally, pre-molded scaffolds have been implemented but require a priori knowledge of wound geometries. Conversely, hydrogels can conform to any injury, but do not guide bi-directional regeneration. In this work, we investigate the feasibility of a system of modular hydrogel tubes to promote bi-directional regeneration after spinal cord injury. This system allows for tubes to be cut to size during surgery and implanted one-by-one to fill any injury, while providing bi-directional guidance. Moreover, this system of tubes can be broadly applied to tissue engineering approaches that require a modular guidance system, such as repair to vascular or musculoskeletal tissues.
- MeSH
- axony účinky léků patologie MeSH
- hydrogely farmakologie MeSH
- jizva patologie MeSH
- lokomoce účinky léků MeSH
- maleimidy chemie MeSH
- mikrosféry MeSH
- myelinová pochva účinky léků metabolismus MeSH
- myši inbrední C57BL MeSH
- neuroglie patologie MeSH
- polyethylenglykoly chemie MeSH
- polymerizace MeSH
- poranění míchy patologie patofyziologie MeSH
- poréznost MeSH
- reagencia zkříženě vázaná chemie MeSH
- regenerace nervu účinky léků MeSH
- tkáňové podpůrné struktury chemie MeSH
- zadní končetina účinky léků fyziologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, N.I.H., Extramural MeSH
The paper is focused on pilot study of effects of novel synthetic protein kinase inhibitors-maleimide derivatives in different concentrations on normal, transformed and multipotent cell lines. Influence on cell proliferation and morphological characteristics has been demonstrated. The chosen agents cause antiproliferative effect on transformed cells and are not cytotoxic to normal cell lines. Moreover, different maleimide derivatives' effects on multipotent cells in attached and floating states has been shown. Described results can be used for further research of the maleimide derivatives as antitumor agents.
- MeSH
- buňky cytologie účinky léků MeSH
- endoteliální buňky cytologie účinky léků MeSH
- fibroblasty cytologie účinky léků MeSH
- HEK293 buňky MeSH
- HeLa buňky MeSH
- inhibiční koncentrace 50 MeSH
- kultivované buňky MeSH
- lidé MeSH
- maleimidy chemie farmakologie MeSH
- mezenchymální kmenové buňky cytologie účinky léků MeSH
- MFC-7 buňky MeSH
- multipotentní kmenové buňky cytologie účinky léků MeSH
- pupečník cytologie MeSH
- transformované buněčné linie MeSH
- tvar buňky účinky léků MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
INTRODUCTION AND OBJECTIVE: In Italy, rabbits are frequently reared for meat production. The aim of the study was to find the seroprevalence of T. gondii and N. caninum parasites, and risk factors of infection in rabbit farms. MATERIAL AND METHODS: Blood samples from 260 apparently healthy breeding rabbits were collected on 13 commercial farms in Northern Italy. Rabbits were divided into categories according to age, number of births, breed, province and size of farm. Samples were tested for antibodies to T. gondii and N. caninum using the indirect fluorescence antibody test (IFAT); samples with a titre ≥ 50 were considered positive. RESULTS: Antibodies to T. gondii and N. caninum were found in 38 (14.6 %) and 3 (1.2 %) rabbits, respectively. A statistically significant difference (p-value ≤ 0.05) was found only in T. gondii prevalence among different rabbit breeds and provinces. CONCLUSION: Rabbits from Northern Italy are at risk of T. gondii and N. caninum infection; however, it is lower compared to seroprevalence noted in other animal species or in humans.
- MeSH
- kokcidióza epidemiologie parazitologie přenos veterinární MeSH
- králíci * MeSH
- maleimidy MeSH
- Neospora fyziologie MeSH
- prevalence MeSH
- rizikové faktory MeSH
- séroepidemiologické studie MeSH
- Toxoplasma fyziologie MeSH
- toxoplazmóza zvířat epidemiologie parazitologie přenos MeSH
- železnaté sloučeniny MeSH
- zvířata MeSH
- Check Tag
- králíci * MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Itálie MeSH
The WNT receptors of the Frizzled family comprise ten mammalian isoforms, bind WNT proteins and mediate downstream signaling to regulate stem cell fate, neuronal differentiation, cell survival and more. WNT-induced signaling pathways are either β-catenin-dependent or -independent, thereby dividing the 19 mammalian WNT proteins into two groups. So far hardly any quantitative, pharmacological information is available about WNT-FZD interaction profiles, affinities or mechanisms of signaling specification through distinct WNT/FZD pairings. This lack of knowledge originates from difficulties with WNT purification and a lack of suitable assays, such as ligand binding assays and FZD activity readouts. In order to minimize this gap, we employ fluorescence recovery after photobleaching (FRAP) to investigate WNT effects on the lateral mobility of FZD6-GFP in living cells. Pharmacological uncoupling of heterotrimeric G proteins by pertussis toxin and N-ethylmaleimide argues that changes in FZD6 mobility are related to putative precoupling of heterotrimeric Gi/o proteins to FZD6. We show that recombinant WNT-1, -2, 3A, -4, -5A, -7A, -9B and -10B affect FZD6 surface mobility and thus act on this receptor. WNT-5B and WNT-11, on the other hand, have no effect on FZD6 mobility and we conclude that they do not act through FZD6. We introduce here a novel way to assess WNT-FZD interaction by live cell imaging allowing further mapping of WNT-FZD interactions and challenging previous experimental limitations. Increased understanding of WNT-FZD selectivity provides important insight into the biological function of this crucial signaling system with importance in developmental biology, stem cell regulation oncogenesis, and human disease.
- MeSH
- buněčná membrána metabolismus MeSH
- ethylmaleimid farmakologie MeSH
- FRAP * MeSH
- frizzled receptory agonisté genetika metabolismus MeSH
- HEK293 buňky MeSH
- heterotrimerní G-proteiny metabolismus MeSH
- lidé MeSH
- pertusový toxin toxicita MeSH
- proteiny vázající GTP - alfa-podjednotky Gi-Go metabolismus MeSH
- proteiny Wnt genetika metabolismus MeSH
- protoonkogenní proteiny genetika metabolismus MeSH
- rekombinantní proteiny genetika metabolismus MeSH
- signální transdukce účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Význam PET/CT s 18F fluorodeoxyglukózou (FDG) je dobre dokumentovaný pre viaceré klinické situácie mnohých malignít. Včasné zhodnotenie terapeutickej odpovede pomocou funkčných zobrazovacích metód poskytujúce významnú prediktívnu a prognostickú informáciu je osobitne zaujímavé. Niektoré protirakovinové látky vykazujú významnú terapeutickú špecifickosť pre niektoré typy malignít a včasné hodnotenie liečby môže slúžiť ako náhrada za histológiu („metabolická biopsia“). Gastrointestinálne stromálne tumory (GIST) sú FDG avídne mezenchýmové nádory, ktoré vo väčšine prípadov dobre reagujú na liečbu tyrozínkinázovými inhibítormi (imatinib mesylát, sunitinib maleát). PET/CT s FDG môže potenciálne slúžiť ako „metabolická biopsia“ v terapeutickom teste s imatinib mesylátom u pacientov s nádorom evokujúcim GIST. Článok ilustruje potenciálne miesto „metabolickej biopsie“ v rutinnom manažmente pacientov s abdominálnou masou imponujúcou ako GIST.
x
- MeSH
- diferenciální diagnóza MeSH
- gastrointestinální stromální tumory * diagnóza radiografie MeSH
- lidé MeSH
- maleimidy * diagnostické užití MeSH
- multimodální zobrazování * metody využití MeSH
- nádory podle histologického typu MeSH
- nádory trávicího systému diagnóza metabolismus radiografie terapie MeSH
- piperaziny aplikace a dávkování metabolismus terapeutické užití MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- stupeň nádoru MeSH
- tyrosinkinasy antagonisté a inhibitory aplikace a dávkování terapeutické užití MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Current therapies proven to slow the progression of diabetic nephropathy include blockade of the renin-angiotensin system (RAS) with either angiotensin-converting enzyme inhibitors or angiotensin receptor blockers. Given our better understanding of the pathophysiology of diabetic nephropathy, newer therapies to treat this condition are slowly emerging. RECENT FINDINGS: Animal studies and a single clinical trial demonstrate efficacy of the renin inhibitor, aliskiren, to decrease a marker of nephropathy progression, that is albuminuria. On the basis of animal study results, pyridoxamine, an inhibitor of advanced glycation and ruboxistaurin, a protein kinase C inhibitor showed promise as new agent to treat nephropathy. The clinical trial results were less than gratifying, however. Sulodexide, a glycosaminoglycan, works to reduce proteinuria presumably by restoring the already reduced glycoproteins present in the glomerular basement membrane. Like other agents, sulodexide also looked promising in animal studies, but failed to demonstrate albuminuria reduction in a large multicentre clinical trial (SUN-Micro-Trial). SUMMARY: This review summarizes newer therapies for slowing the progression of diabetic nephropathy. Aliskiren shows promise from small clinical studies, but we await the results of the multicentre, international ALTITUDE trial in about 2012. On the basis of the results of trials only, pyridoxamine may have a chance at further evaluation, but that is also unclear.
- MeSH
- amidy terapeutické užití MeSH
- diabetické nefropatie etiologie farmakoterapie metabolismus MeSH
- fumaráty terapeutické užití MeSH
- indoly terapeutické užití MeSH
- lidé MeSH
- maleimidy terapeutické užití MeSH
- pyridoxamin terapeutické užití MeSH
- superoxidy metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
Hypotonic solution alters ion channel activity, but little attention has been paid to voltage-dependent sodium channels. The aim of this study was to investigate the effects of hypotonic solution on transient sodium currents (INaT) and persistent sodium currents (INaP). We also explored whether the intracellular signal transduction systems participated in the hypotonic modifications of sodium currents. INaT and INaP were recorded by means of whole-cell patch-clamp technique in isolated rat ventricular myocytes. Our results revealed that hypotonic solution reduced INaT and simultaneously augmented INaP with the occurrence of interconversion between INaT and INaP. Hypotonic solution shifted steady-state inactivation to a more negative potential, prolonged the time of recovery from inactivation, and enhanced intermediate inactivation (IIM). Ruthenium red (RR, inhibitor of TRPV4), bisindolylmaleimide VI (BIM, inhibitor of PKC), Kn-93 (inhibitor of Ca/CaMKII) and BAPTA (Ca2+-chelator) inhibited the effects of hypotonic solution on INaT and INaP. Therefore we conclude that hypotonic solution inhibits INaT, enhances INaP and IIM with the effects being reversible. TRPV4 and intracellular Ca2+, PKC and Ca/CaMKII participate in the hypotonic modifications of sodium currents.
- MeSH
- benzylaminy farmakologie MeSH
- chelátory farmakologie MeSH
- EGTA analogy a deriváty farmakologie MeSH
- financování organizované MeSH
- hypotonické roztoky MeSH
- indoly farmakologie MeSH
- inhibitory proteinkinas farmakologie MeSH
- kardiomyocyty metabolismus účinky léků MeSH
- kationtové kanály TRPV antagonisté a inhibitory metabolismus MeSH
- kinetika MeSH
- krysa rodu rattus MeSH
- maleimidy farmakologie MeSH
- membránové potenciály MeSH
- metoda terčíkového zámku MeSH
- proteinkinasa C antagonisté a inhibitory metabolismus MeSH
- proteinkinasa závislá na vápníku a kalmodulinu typ 2 antagonisté a inhibitory metabolismus MeSH
- rutheniová červeň farmakologie MeSH
- signální transdukce účinky léků MeSH
- sodík metabolismus MeSH
- srdeční komory cytologie metabolismus účinky léků MeSH
- sulfonamidy farmakologie MeSH
- vápník metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
The vitamin E analogue alpha-tocopheryl succinate (alpha-TOS) is an efficient anti-cancer drug. Improved efficacy was achieved through the synthesis of alpha-tocopheryl maleamide (alpha-TAM), an esterase-resistant analogue of alpha-tocopheryl maleate. In vitro tests demonstrated significantly higher cytotoxicity of alpha-TAM towards cancer cells (MCF-7, B16F10) compared to alpha-TOS and other analogues prone to esterase-catalyzed hydrolysis. However, in vitro models demonstrated that alpha-TAM was cytotoxic to non-malignant cells (e.g. lymphocytes and bone marrow progenitors). Thus we developed lyophilized liposomal formulations of both alpha-TOS and alpha-TAM to solve the problem with cytotoxicity of free alpha-TAM (neurotoxicity and anaphylaxis), as well as the low solubility of both drugs. Remarkably, neither acute toxicity nor immunotoxicity implicated by in vitro tests was detected in vivo after application of liposomal alpha-TAM, which significantly reduced the growth of cancer cells in hollow fiber implants. Moreover, liposomal formulation of alpha-TAM and alpha-TOS each prevented the growth of tumours in transgenic FVB/N c-neu mice bearing spontaneous breast carcinomas. Liposomal formulation of alpha-TAM demonstrated anti-cancer activity at levels 10-fold lower than those of alpha-TOS. Thus, the liposomal formulation of alpha-TAM preserved its strong anti-cancer efficacy while eliminating the in vivo toxicity found of the free drug applied in DMSO. Liposome-based targeted delivery systems for analogues of vitamin E are of interest for further development of efficient and safe drug formulations for clinical trials.
- MeSH
- alfa-tokoferol analogy a deriváty aplikace a dávkování farmakologie MeSH
- antitumorózní látky aplikace a dávkování farmakologie MeSH
- chemie farmaceutická MeSH
- lidé MeSH
- liposomy MeSH
- maleimidy aplikace a dávkování farmakologie MeSH
- melanom experimentální farmakoterapie patologie MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši transgenní MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory prsu farmakoterapie patologie MeSH
- polyethylenglykoly aplikace a dávkování farmakologie MeSH
- vitamin E analogy a deriváty aplikace a dávkování farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
