The ocular microcirculation represents an important target to treat inflammatory diseases of eye, where impairment of microvascular blood flow plays key role as, for example, in anterior uveitis. To evaluate novel interventions targeting the microcirculation, appropriate and reliable tools to study this particular microvascular bed are needed. Intravital microscopy (IVM) belongs to several methods allowing evaluation of microcirculation experimentally, even in small animals. The aim of our study was to examine the iridial microcirculation (IMIC) in uveitis induced by local or systemic endotoxin administration in rats and mice by IVM and to propose new parameters to quantify the changes within the IMIC. Systemic inflammation was induced in rats by intravenous endotoxin administration, control group received normal saline intravenously. Local inflammation was induced in mice by intravitreal endotoxin administration, the control group received normal saline intravitreally. IVM of IMIC was performed in animals receiving systemic endotoxin prior injection and 1 and 2 h afterwards, respectively, in animals receiving intravitreal endotoxin/saline prior local injection and 5 h afterwards. Obtained video recordings were analyzed off-line. Functional capillary density (FCD) and dysfunctional capillary density (DCD) were evaluated for description of IMIC, and calculation of FCD/DCD ratio was performed. In systemic inflammation, FCD was significantly decreased compared to control animals. In local inflammation, the number of functional capillaries in the IMIC was significantly reduced following the endotoxin challenge. Analysis of the DCD revealed a significant increase in capillaries with reduced perfusion after intravitreal endotoxin administration and right shift of the FCD/DCD ratio was observed after endotoxin local injection. Detecting and quantifying changes in IMIC during systemic or local inflammation in experimental animals by IVM was feasible. Therefore, IVM of the IMIC represents a valuable tool to evaluate and quantify inflammatory changes in experimental eye disease.
- MeSH
- endotoxiny aplikace a dávkování toxicita MeSH
- intravitální mikroskopie metody MeSH
- kapiláry patologie MeSH
- krysa rodu rattus MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- uveitida chemicky indukované patologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Approximately 40% of hormone receptor-positive, HER2-negative breast cancers (BCs) are associated with activating mutations of the phosphatidylinositol 3-kinase (PI3K) pathway. Pictilisib, a potent and highly specific class I pan-PI3K inhibitor, demonstrated preclinical activity in BC cell lines and may potentiate the effect of taxanes, benefiting patients with or without aberrant activation of the PI3K pathway. PEGGY (NCT01740336), a randomised, placebo-controlled phase II trial, examined whether pictilisib augments the anti-tumour activity of paclitaxel in patients with hormone receptor-positive, HER2-negative locally recurrent or metastatic BC (mBC). We report results from the protocol-specified interim analysis. PATIENTS AND METHODS: One hundred and eighty-three eligible patients were randomised (1:1) to receive paclitaxel (90 mg/m2 weekly for 3 weeks in every 28-day cycle) with either 260 mg pictilisib or placebo (daily on days 1-5 every week). The primary end point was progression-free survival (PFS) in the intention-to-treat (ITT) population and patients with PIK3CA-mutated tumours. Secondary end points included overall response rate (ORR), duration of response, and safety. RESULTS: In the ITT population, the median PFS was 8.2 months with pictilisib (n = 91) versus 7.8 months with placebo (n = 92) [hazard ratio (HR) for progression or death, 0.95; 95% confidence interval (CI) 0.62-1.46; P = 0.83]. In patients with PIK3CA-mutated tumours, the median PFS was 7.3 months for pictilisib (n = 32) versus 5.8 months with placebo (n = 30) (HR, 1.06; 95% CI 0.52-2.12; P = 0.88). ORR was similar between treatment arms. The safety profile of pictilisib was consistent with previous reports, with no new safety signals. Proportions of patients with grade ≥3 adverse events (AEs), serious AEs, and dose reductions/discontinuations due to AEs were higher with pictilisib. CONCLUSIONS: PEGGY did not meet its primary end point, revealing no significant benefit from adding pictilisib to paclitaxel for patients with hormone receptor-positive, HER2-negative locally recurrent or mBC. CLINICAL TRIAL NUMBER: NCT01740336.
- MeSH
- dospělí MeSH
- fosfatidylinositol-3-kinasy třídy I antagonisté a inhibitory genetika MeSH
- humanizované monoklonální protilátky aplikace a dávkování MeSH
- indazoly aplikace a dávkování MeSH
- lidé středního věku MeSH
- lidé MeSH
- lokální recidiva nádoru farmakoterapie genetika patologie MeSH
- metastázy nádorů MeSH
- nádory prsu farmakoterapie genetika patologie MeSH
- paclitaxel aplikace a dávkování MeSH
- přežití po terapii bez příznaků nemoci MeSH
- protokoly antitumorózní kombinované chemoterapie aplikace a dávkování MeSH
- receptor erbB-2 genetika MeSH
- senioři MeSH
- sulfonamidy aplikace a dávkování MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky, fáze II MeSH
- multicentrická studie MeSH
- práce podpořená grantem MeSH
- randomizované kontrolované studie MeSH
Generation of reactive oxygen species significantly contributes to the pathogenesis of acute renal failure (ARF) induced by myoglobin release. Ginsenosides (GS), the principal active ingredients of ginseng, is considered as an extremely good antioxidative composition of Chinese traditional and herbal drugs. The purpose of the present study was to investigate the protective effect of ginsenoside in rats with ARF on the changes of cholinergic nervous system in the kidney as well as on the involvement of mitogen-activated protein kinases (MAPK) in the hypothalamic paraventricular nuclei (PVN). In our assay, glycerol-induced acute renal failure in rats was employed to study the protective effects of ginsenoside. Our results indicated that the treatment of ARF rats with ginsenosides for 48 h significantly reduced lipid peroxidation, restored the superoxide dismutase (SOD) level. Meanwhile, the obvious increase of choline acetyltransferase-immunoreactivity (ChAT-IR) in the proximal convoluted tubular cells (PCT) was observed by immunohistochemistry in ARF+GS group. The same effect was also observed in the changes of p-ERK1/2-IR in the hypothalamic paraventricular nuclei. Our results suggest that ginsenoside administered orally may have a strong renal protective effect against glycerol-induced ARF, reduce the renal oxidative stress, and ginsenoside can also activate the cholinergic system in PCT, simultaneously MAPK signal pathway in the PVN was also activated.
- MeSH
- akutní poškození ledvin chemicky indukované enzymologie patologie prevence a kontrola MeSH
- antioxidancia aplikace a dávkování farmakologie MeSH
- aplikace orální MeSH
- časové faktory MeSH
- cholin-O-acetyltransferasa metabolismus MeSH
- cytoprotekce MeSH
- fosforylace MeSH
- ginsenosidy aplikace a dávkování farmakologie MeSH
- glycerol MeSH
- malondialdehyd metabolismus MeSH
- mitogenem aktivovaná proteinkinasa 1 metabolismus MeSH
- mitogenem aktivovaná proteinkinasa 3 metabolismus MeSH
- modely nemocí na zvířatech MeSH
- nucleus paraventricularis hypothalami účinky léků enzymologie MeSH
- oxidační stres účinky léků MeSH
- peroxidace lipidů účinky léků MeSH
- potkani Sprague-Dawley MeSH
- proximální tubuly ledvin účinky léků enzymologie patologie MeSH
- signální transdukce účinky léků MeSH
- superoxiddismutasa metabolismus MeSH
- upregulace MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Sidestream dark field imaging represents a novel, noninvasive method to study the microcirculation in humans and animals. To-date, it has been used extensively in various peripheral tissues (e.g. sublingual area, intestinal mucosa), however no data for the ocular vasculature, including the iridial microcirculation, are currently available. Therefore, the aim of this study was to examine the reliability and reproducibility of sidestream dark field imaging within the iridial microcirculation in experimental animals. Male Lewis rats were anaesthetized and the iris microvasculature was observed using an sidestream dark field probe gently placed against a cover slip covering the right eye. All video sequences recorded were analysed off-line by using AVA 3.0 software (MicroVision Medical, Amsterdam, The Netherlands). Results are expressed as mean (±SE) or median (interquartile range). Clear images were recorded from each animal and the total number of analysable video sequences was 50. All raw data for selected vessel density parameters passed normality test. The total all and small vessel density (in mm mm(-2) ) were 22,6 (±0,58) and 19,6 (±0,68), respectively. The perfused all and small vessel density were 20,9 (±0,61) and 19,1 (±0,65), respectively. The mean values of all iris vessel density parameters are shown in Figure 4. The DeBacker Score (n/mm) was 15,2 (±0,45), the proportion of perfused vessel was 94,5% (89,8-99,1%), and the MFI was 3 points (3-3). Taken together, these results indicate that SDF imaging provides a reliable and noninvasive method to examine the iridial microvascular bed in vivo and, thus, may provide unique opportunities for the study of the iridial vascular network in various experimental and clinical settings and disease models.
- MeSH
- iris anatomie a histologie fyziologie MeSH
- krysa rodu rattus MeSH
- mikrocévy anatomie a histologie fyziologie MeSH
- počítačové zpracování obrazu metody MeSH
- videomikroskopie metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Disturbance of capillary perfusions due to leukocyte adhesion, disseminated intravascular coagulation, tissue edema is critical components in the pathophysiology of sepsis. Alterations in brain microcirculation during sepsis are not clearly understood. The aim of this study is to gain an improved understanding of alterations through direct visualization of brain microcirculations in an experimental endotoxemia using intravital microscopy (IVM). Endotoxemia was induced in Lewis rats with Lipopolysaccharide (LPS, 15 mg/kg i.v.). The dura mater was removed via a cranial window to expose the pial vessels on the brain surface. Using fluorescence dyes, plasma extravasation of pial venous vessels and leukocyte-endothelial interaction were visualized by intravital microscopy 4 h after LPS administration. Plasma cytokine levels of IL1-β, IL-6, IFN-γ, TNF-α and KC/GRO were evaluated after IVM. A significant plasma extravasation of the pial venous vessels was found in endotoxemia rats compared to control animals. In addition, a significantly increased number of leukocytes adherent to the pial venous endothelium was observed in septic animals. Endotoxemia also induced a significant elevation of plasma cytokine levels of IL1-β, IL-6, IFN-γ, TNF-α and KC/GRO. Endotoxemia increased permeability in the brain pial vessels accompanied by an increase of leukocyte-endothelium interactions and an increase of inflammatory cytokines in the plasma.
- MeSH
- buněčná adheze MeSH
- cévní endotel metabolismus patofyziologie MeSH
- endotoxemie metabolismus patofyziologie MeSH
- interferon gama krev MeSH
- interleukin-1beta krev MeSH
- interleukin-6 krev MeSH
- kapilární permeabilita MeSH
- krysa rodu rattus MeSH
- leukocyty fyziologie MeSH
- mikrocévy metabolismus patofyziologie MeSH
- mikrocirkulace fyziologie MeSH
- mozek krevní zásobení MeSH
- mozkové žíly metabolismus MeSH
- potkani inbrední LEW MeSH
- TNF-alfa krev MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Whole cell patch-clamp recordings from GABAergic cells of thalamic reticular nucleus (RTN) in thalamocortical slices made from postnatal day 6 (P6) to 10 (P10) were used to investigate the pattern of rebound bursts (RBs) triggered by an injection of hyperpolarizing current into RTN cells. The number of RBs in the RTN and the overlying Na+/K+ spikes changed in an agedependent manner. The generation of RBs depended largely on the amplitude of the after-hyperpolarizations (AHPs). RB patterns in response to hyperpolarizing current injection into relay cells were markedly different from RB patterns in RTN cells with an after-depolarization. GABAA receptor antagonist bicuculline methiodide (BMI) changed burst firing patterns, increasing the duration of RB and decreasing the amplitude of AHP in RTN cells. Furthermore, local puffs of NMDA in the presence of BMI induced RBs. K+ channel blocker 4-aminopyridine partially mimicked the effect of BMI on AHPs. The shapes of RBs were altered by a selective CaMKII inhibitor KN-62, but not by an inactive analog KN-04.
- MeSH
- 1-(5-isochinolinsulfonyl)-2-methylpiperazin analogy a deriváty farmakologie MeSH
- 4-aminopyridin farmakologie MeSH
- akční potenciály fyziologie účinky léků MeSH
- bikukulin analogy a deriváty farmakologie MeSH
- blokátory draslíkových kanálů farmakologie MeSH
- financování organizované MeSH
- GABA antagonisté farmakologie MeSH
- GABA fyziologie MeSH
- inhibitory proteinkinas farmakologie MeSH
- jádra thalamu cytologie fyziologie MeSH
- myši inbrední ICR MeSH
- myši MeSH
- nervový útlum fyziologie účinky léků MeSH
- neurony fyziologie MeSH
- novorozená zvířata MeSH
- orgánové kultury - kultivační techniky MeSH
- proteinkinasa závislá na vápníku a kalmodulinu typ 2 antagonisté a inhibitory metabolismus MeSH
- receptory GABA-A - antagonisté MeSH
- receptory GABA-A fyziologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
Acute renal failure (ARF) is mainly characterized by acute tubular necrosis. No significant change was found for mortality rates over the past few decades despite significant advances in supportive care. In recent years, great effort has been focused on traditional and herbal medicine, which is much less toxic than those agents conventionally used and which is nowadays considered as a novel therapeutic agent for ARF. However, the effect of ginsenosides (GS) administered orally on ARF has not been reported yet and little is known about its cellular and molecular mechanism. The purpose of the study is to investigate the protective effect of ginsenoside in rats with ARF on the changes of tyrosine hydroxylase immunoreactivity (TH-IR) as well as on the involvement of mitogen-activated protein kinases (MAPK) in the locus coeruleus. In our assay, glycerol-induced acute renal failure in rats was employed to study the protective effects of ginsenoside. Our results indicated that the treatment of ARF rats with ginsenosides for 48 h significantly reduced the serum blood urea nitrogen, creatinine level, and lipid peroxidation, restored the GSH level and the normal renal morphology. Immunohistochemistry showed that an obvious increase of TH-IR was further enhanced in ARF+GS group. The same effect was also observed in the changes of p-ERK1/2-IR in the locus coeruleus. Our results suggest that ginsenoside administered orally may have a strong renal protective effect against glycerol-induced ARF, and ginsenoside can also activate the brain catecholaminergic neurons in the locus coeruleus. Our future attention will be focused to the question whether there is a correlation between the renal protective effect of ginsenosides against acute renal failure and the activation of tyrosine hydroxylase in the locus coeruleus.
- MeSH
- akutní poškození ledvin MeSH
- aplikace orální MeSH
- biologické markery krev MeSH
- časové faktory MeSH
- deprivace z nedostatku vody MeSH
- dusík močoviny v krvi MeSH
- financování organizované MeSH
- fosforylace MeSH
- ginsenosidy aplikace a dávkování farmakologie MeSH
- glutathion metabolismus MeSH
- glycerol MeSH
- imunohistochemie MeSH
- kreatinin krev MeSH
- krysa rodu rattus MeSH
- ledviny MeSH
- locus coeruleus enzymologie účinky léků MeSH
- malondialdehyd metabolismus MeSH
- mitogenem aktivovaná proteinkinasa 1 metabolismus MeSH
- mitogenem aktivovaná proteinkinasa 3 metabolismus MeSH
- modely nemocí na zvířatech MeSH
- ochranné látky aplikace a dávkování farmakologie MeSH
- peroxidace lipidů účinky léků MeSH
- potkani Sprague-Dawley MeSH
- tyrosin-3-monooxygenasa metabolismus MeSH
- upregulace MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
