Decrease In Vascular Permeability Research Articles

AGE FEATURES OF REMODELING OF ARTERIES OF THE PROSTATE GLAND AT ETHANOL INTOXICATION

Chronic alcohol intoxication leads to damage of almost all organs and systems, the degree of structural and functional disorders of which in this pathology is different and depends on the duration and severity of intoxication. Organs of the reproductive system are always involved in the pathological process with long-term effects of alcohol on the body. Age-related remodeling of prostate arteries in ethanol intoxication has not been studied enough.
 The aim of the study: morphologically study the age-related features of prostate artery remodeling under conditions of ethanol intoxication.
 Materials and methods. The prostate arteries of 60 white rats were morphologically studied, which were divided into 4 groups: 1 group – 15 intact animals aged 8 months; group 2 – 15 rats aged 24 months; group 3 – 15 8-month-old animals with ethanol intoxication; Group 4 – 15 24-month-old rats with the indicated simulated pathology. A 30% ethanol solution was injected intragastrically at the rate of 2 ml per 100 g of animal weight for 28 days once a day. Animals were euthanized by bloodletting under anesthesia. The external and internal diameters of small-caliber arteries, the thickness of the intima, media, adventitia, intimo-medial, intimo-adventitous, adventitio-medial, Vogenvort and Kernogan indices, and the relative volumes of damaged endotheliocytes were determined on the micropreparations of the prostate. Quantitative morphological indicators were processed statistically.
 Research results. The wall of arteries thickens, their lumen narrows, the thickness of the media and adventitia increases, and the studied indices change significantly in ethanol intoxication in the prostate. The thickness of the media in young animals with a pronounced statistically significant difference increased by 49.2 %, in old animals by 50.2 % (p<0.001), and the Wogenvoort index changed by 1.56 and 1.6 times, respectively (p <0.001). The lumen of the studied vessels in young rats was statistically significantly (p<0.001) reduced by 16.6%, in old rats by 35.0%, the Kernoghan index by 12.7 % and 13.0 % (p<0.001), which indicated a marked decrease in vascular permeability and deterioration of blood supply to the organ. The relative volume of damaged endotheliocytes in the small-caliber arteries of the prostate gland during long-term ethanol intoxication in young rats was equal to (24.80±0.18) %, and in animals of the older age group – (47.60±0.21) %. Light-optically, pronounced vascular disorders, hemoptysis, expansion of mainly venous vessels, perivisceral and stromal edema, foci of dystrophically, necrobiotically, apoptically altered endotheliocytes, epitheliocytes of glandular structures, focal infiltrates and growth of connective tissue were observed in the prostate gland during chronic alcohol intoxication. Swelling of endotheliocytes, their dystrophy, necrobiosis, desquamation and proliferation were also noted.
 Conclusions. Ethanol intoxication leads to thickening of walls of arteries of the prostate gland, narrowing of their lumen, atrophic processes in the intima, an increase of thickness of media, adventitia, disruption of the relationship between them, apoptotic, dystrophic and necrobiotic changes in endotheliocytes, endothelial dysfunction, hypoxia, dystrophy, necrobiosis cells and stromal structures, infiltration and sclerosis. The degree of structural rearrangement of prostate arteries dominates in experimental animals of the older age group.

Comparative study of the effectiveness of phlebotonic drugs in the experiment

Introduction. Chronic venous diseases are a common group of diseases with a significant risk of complications requiring timely correction. As a rule, phlebotonic drugs based on flavonoid complexes are used for the treatment and prevention of venous diseases.Aim. To evaluate the effectiveness of the use of various phlebotonic drugs in the conditions of experimental chronic venous insufficiency.Materials and methods. Varicose veins were modeled in Wistar rats by partial stricture of the deep femoral vein. The studied medicines were administered orally in a course of 30 days from the moment of surgery. During the work, the change in the following parameters was evaluated: the rate oflocal blood flow in the skin in dynamics, the degree of vascular permeability, the concentration of proinflammatory cytokines (TNF-α, IL-6) and matrix metalloproteinase 9 (MMP9) in the vascular wall. The rate of local blood flow was assessed by ultrasound Dopplerography. The change of vascular permeability was studied by the degree of extravasation of the Evans blue dye in the Miles test. The content of proinflammatory cytokines and MMR9 was determined by enzyme-linked immunoassay. The results were statistically processed.Results. The study showed that the course oral administration of all the studied venotonizing drugs led to the restoration of hemodynamics and a significant (p˂ 0.05) decrease in the degree of vascular permeability in relation to untreated animals. It is worth noting that the use of a micronised purified flavonoid fraction 1 contributed to the development of a more pronounced vasal effect, which was reflected in an increase in blood flow velocity and a decrease in vascular permeability compared to the rest of the studied drugs. At the same time, the administration of micronised purified flavonoid fraction 1 to animals led to a statistically significant decrease in the concentration of proinflammatory cytokines, which was not observed when using other drugs.Conclusion. The course administration of the micronised purified flavonoid fraction 1leads to the development of a pronounced phlebotonic effect, expressed in the restoration of microcirculation, a decrease of the inflammation in the vascular wall.

Targeting Pdzrn3 maintains adult blood-brain barrier and central nervous system homeostasis

Blood brain barrier (BBB) disruption is a critical component of the pathophysiology of cognitive impairment of vascular etiology (VCI) and associated with Alzheimer’s disease (AD). The Wnt pathway plays a crucial role in BBB maintenance, but there is limited data on its role in cognitive pathologies. The E3 ubiquitin ligase PDZRN3 is a regulator of the Wnt pathway. In a murine model of VCI, overexpressing Pdzrn3 in endothelial cell (EC) exacerbated BBB hyperpermeability and accelerated cognitive decline. We extended these observations, in both VCI and AD models, showing that EC-specific depletion of Pdzrn3, reinforced the BBB, with a decrease in vascular permeability and a subsequent spare in cognitive decline. We found that in cerebral vessels, Pdzrn3 depletion protects against AD-induced Wnt target gene alterations and enhances endothelial tight junctional proteins. Our results provide evidence that Wnt signaling could be a molecular link regulating BBB integrity and cognitive decline under VCI and AD pathologies.

Effect of GnRH-antagonist, mifepristone and letrozole on preventing ovarian hyperstimulation syndrome in rat model

Research questionCan luteolysis-targeted drugs, gonadotrophin-releasing hormone antagonist (GnRH-ant), mifepristone and letrozole, administered separately or in combination, prevent the progression of ovarian hyperstimulation syndrome (OHSS) in a rat model? DesignThirty-six female Wistar rats were randomly divided into six groups, including control group (OHSS group, ovarian hyperstimulation-induced OHSS); GnRH-ant group (OHSS with GnRH-ant treatment); mifepristone group (OHSS with mifepristone treatment); letrozole group (OHSS with letrozole treatment); combination group (OHSS with GnRH-ant, mifepristone and letrozole treatment in combination). The main outcomes were the alterations in OHSS-related indices, including ovarian weight, vascular permeability, serum oestradiol and progesterone levels, corpus luteum proportion and diameter, ovarian vascular endothelial growth factor (VEGF), interleukin 6 (IL-6), caspase-3 and cleaved caspase-3 levels. ResultsNo significant difference was found in body weight gain among the six groups. Compared with the control group, the OHSS group showed significant increases in all OHSS-related indices. GnRH-ant treatment showed decreases in vascular permeability, serum oestradiol level, corpus luteum diameter, ovarian VEGF /IL-6 mRNA levels, and increases in ovarian caspase-3 and cleaved caspase-3 levels. Mifepristone treatment demonstrated reduction in serum progesterone level and corpus luteum diameter, and elevation in ovarian caspase-3 and cleaved caspase-3 levels. Letrozole treatment displayed a decline in serum oestradiol level and corpus luteum diameter, and up-regulation in ovarian caspase-3 and cleaved caspase-3 levels. The combination treatment by GnRH-ant, mifepristone and letrozole showed enhanced synergistic effect on reducing OHSS-related indices. ConclusionsGnRH-ant, mifepristone and letrozole are beneficial in preventing the progression of OHSS through different luteolytic mechanisms. Cocktail style treatment shows enhanced synergistic effect on preventing the progression of OHSS.

Identification and quantification of oleanane triterpenoid saponins and potential analgesic and anti-inflammatory activities from the roots and rhizomes of Panax stipuleanatus

BackgroundPanax stipuleanatus represents a folk medicine for treatment of inflammation. However, lack of experimental data does not confirm its function. This article aims to investigate the analgesic and anti-inflammatory activities of triterpenoid saponins isolated from P. stipuleanatus. MethodsThe chemical characterization of P. stipuleanatus allowed the identification and quantitation of two major compounds. Analgesic effects of triterpenoid saponins were evaluated in two models of thermal- and chemical-stimulated acute pain. Anti-inflammatory effects of triterpenoid saponins were also evaluated using four models of acetic acid–induced vascular permeability, xylene-induced ear edema, carrageenan-induced paw edema, and cotton pellet–induced granuloma in mice. ResultsTwo triterpenoid saponins of stipuleanosides R1 (SP-R1) and R2 (SP-R2) were isolated and identified from P. stipuleanatus. The results showed that SP-R1 and SP-R2 significantly increased the latency time to thermal pain in the hot plate test and reduced the writhing response in the acetic acid–induced writhing test. SP-R1 and SP-R2 caused a significant decrease in vascular permeability, ear edema, paw edema, and granuloma formation in inflammatory models. Further studies showed that the levels of inflammatory mediators, nitric oxide, malondialdehyde, tumor necrosis factor-α, and interleukin 6 in paw tissues were downregulated by SP-R1 and SP-R2. In addition, the rational harvest of three- to five-year-old P. stipuleanatus was preferable to obtain a higher level of triterpenoid saponins. SP-R2 showed the highest content in P. stipuleanatus, which had potential as a chemical marker for quality control of P. stipuleanatus. ConclusionThis study provides important basic information about utilization of P. stipuleanatus resources for production of active triterpenoid saponins.

Visceral adipogenesis inhibited by Pref-1 is associated with peritoneal angiogenesis.

Studies showed an increased visceral adipose tissue and peritoneal angiogenesis in peritoneal dialysis (PD) patients. However, the relationship between the visceral adipose expands and peritoneal angiogenesis remains unclear. Pref-1 (preadipocyte factor-1) recombinant adeno-associated virus (AAV) and control AAV were constructed. Mice were divided into four groups, mice in control and PD group were injected intraperitoneally with PBS, mice in control-AAV-PD were injected intraperitoneally with plaque-forming unit (PFU) control AAV and mice in pref-1-AAV-PD group were injected with PFU recombinant AAV. Two weeks later, control group was injected intraperitoneally with normal saline while other groups were injected intraperitoneally with 4.25% peritoneal dialysis fluid (PDF). Thirty days later, viscerall adipose tissue was collected and weighed. Pref-1 protein expression was measured by Western blot, and peritoneal permeability was measured by Evans blue. Cluster of differentiation 31(CD31) immunohistochemical staining was used to detect mesenteric blood vessel number, and vascular endothelial growth factor (VEGF) in serum were measured by enzyme-linked immunosorbent assay. Pref-1 protein expression increased in pref-1-AAV-PD group. Visceral adipose expanded in PD and control-AAV-PD group while decreased in pref-1-AAV-PD group, which approves PD fluid enhance visceral adipogensis, and the process could be inhibited by Pref-1 recombinant AAV. The reduction of peritoneal vessel number and the decrease of vascular permeability as well as down-regulation of serum vascular endothelial growth factor observed in pref-1-AAV-PD group suggested peritoneal angiogenesis could be inhibited following visceral adipose tissue reduction. Visceral adipose expands is associated with peritoneal angiogenesis in PD treatment, and prevention of visceral adipogenesis may be an alternative way to protect the validity of peritoneum. Copyright © 2019 John Wiley & Sons, Ltd.

Final analysis of a phase II study of MRI based functional imaging of bone metastases in men with metastatic castrate-resistant prostate cancer (mCRPC) receiving cabozantinib.

213 Background: Cabozantinib (C) is a small molecule inhibitor of receptor tyrosine kinases including VEGFR-2, c-MET. C trials have shown significant improvements in bone pain and bone scintigraphy in mCRPC patients (pts). We hypothesized that functional imaging using MRI could elucidate underlying biological processes by demonstrating an early decrease in vascular permeability (decrease in transfer consant - Ktrans) and subsequent decrease in cell density (increase in apparent diffusion coefficient - ADC) within bone metastases. Methods: mCRPC pts received C 60 mg daily. The primary endpoint was change in Ktrans at 2 weeks (wks) of treatment. Secondary endpoints included Ktrans and ADC longitudinal changes, and correlation with bone scan, PSA, RECIST, and changes in reported pain. All pts underwent MRI at baseline, day 0, day 15 and every 12 wks. Results: 17 pts were treated at two sites. Median age: 68 yrs (range:51-83), baseline PSA 94.78 ng/mL (7.4-2971), number of prior CRPC therapies 2 (1-8). Median progression free survival was 5.1 months; 5 pts discontinued therapy for adverse events, and 12 for progressive disease. The most common grades 3/4 toxicities were fatigue (24%) and palmarplantar erythrodysesthesia (12%). 14 pts were evaluable for the primary endpoint. At 2 wks, Ktrans decreased an average 35%, 0.074 to 0.048 min-1 (SD=0.016, p<0.0001). There was no change in Ktrans between wk 2 and end of study. There was an increase in median ADC of 150 at wk 12. There were no RECIST or 50% PSA responses. Conclusions: Ktrans decreased significantly after 2 wks of treatment, consistent with antiangiogenic properties of C. Compared to wk 2, K trans at disease progression is unchanged, which could signify that adaptive vascular change is not the primary mechanism of tumor resistance to the drug. Contrary to our hypothesis, ADC increased during the trial, which perhaps correlates with the short median time to progression. Still, MRI of bone metastases could benefit drug development for other agents in mCRPC or interrogation of bone metastases, notoriously challenging for disease response analysis, in other cancers. Clinical trial information: NCT01599793.

Comparison of dynamic contrast-enhanced magnetic resonance imaging and contrast-enhanced ultrasound for evaluation of the effects of sorafenib in a rat model of hepatocellular carcinoma

ObjectivesTo compare the accuracy of contrast-enhanced ultrasound (CEUS) and Dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) for the assessment of changes in tissue vascularization as result of sorafenib treatment in a rat model of hepatocellular carcinoma (HCC). MethodsMale Buffalo rats with orthotopic liver tumors treated daily with 7.5 mg/kg sorafenib via oral gavage for 2 weeks (n = 9) were subject to DCE-MRI and CEUS 2 weeks after tumor implantation - right before treatment initiation - and also after treatment completion - right before tumor harvest. Untreated animals (n = 10) were used as control. Tumor tissue sections were stained for hematoxylin-eosin, pimonidazole, and CD34 for quantitative assessment of necrosis, hypoxia, and microvessel density (MVD), respectively. ResultsOf all the DCE-MRI parameters that were evaluated, only volume transfer constant (Ktrans) measurements were significantly lower in sorafenib-treated tumors (0.18 vs 0.33 min−1, p < 0.01), indicating a substantial decrease in vascular permeability caused by the therapy. This reduction was associated with decreased MVD (3.9 vs 10.8% CD34+ cells, p < 0.01), higher tumor necrosis (31.9 vs 21.8%, p < 0.001) and hypoxia (19.7 vs 10.5% pimonidazole binding, p < 0.01). Moreover, statistical analysis demonstrate significant correlation of DCE-MRI Ktrans with histopathologic tissue necrosis (r = −0.537, p < 0.05) and MVD (r = 0.599, p < 0.05). Interestingly, none of the CEUS measurements were significantly different between the control and treatment groups, and did not show statistical correlation with any of the histopathological parameters assessed (p > 0.05). ConclusionsSorafenib-induced reduction in vascular permeability in this preclinical model of HCC is detected more accurately through DCE-MRI than CEUS, and DCE-MRI parameters strongly correlate with histopathological changes in tissue vascularization and tissue necrosis.

Angiopoietin-Like 4 Attenuates Brain Edema and Neurological Deficits in a Mouse Model of Experimental Intracerebral Hemorrhage.

BackgroundAngiopoietin-like 4 (ANGPTL4) is neuroprotective when administered acutely for the treatment of cerebral ischemia. The aim of the present study was to evaluate the preventive effects of ANGPTL4 on the formation of brain edema and to determine whether it promotes the recovery of neurological function following intracerebral hemorrhage (ICH).Material/MethodsRecombinant human ANGPTL4 (rhANGPTL4; 40 μg/kg) or a vehicle was administered intraperitoneally 5 min prior to bacterial collagenase-induced ICH in male C57/B6J mice. Behavioral tests were performed prior to ICH and at days 1, 3, 7, 14, 21, and 28 after ICH. Brain edema and hematoma volume were examined separately using the wet weight/dry weight method and hematoxylin-eosin staining. The integrity of the tight and adherens junctions was quantified via immunofluorescence. The ultrastructure of the blood-brain barrier (BBB) was examined using transmission electron microscopy. Vascular endothelial (VE)-cadherin, claudin-5, Src, and phospho-Src in the ipsilateral and contralateral striatum were detected by Western blot analysis.ResultsRhANGPTL4 reduced brain edema and hematoma volume and improved neurological functional recovery over the subsequent 4 weeks when compared with the control group. rhANGPTL4 significantly increased VE-cadherin and claudin-5-positive areas and relative amounts in the peri-hematoma region compared with the control group. In addition, ANGPTL4 significantly reduced the ratio of phospho-Src to Src. The significant reduction of Src kinase activity in the peri-hematoma region of ANGPTL-treated mice was paralleled by a decrease in vascular permeability and edema formation.ConclusionsThese results suggest that ANGPTL4 is a relevant target for vasculoprotection and cerebral protection during stroke.

Effect of oral tranexamic acid on macular edema associated with retinal vein occlusion or diabetes.

PurposeTranexamic acid (TXA) is a widely used antifibrinolytic agent that can also cause a decrease in vascular permeability. We hypothesized that TXA could improve macular edema (ME) that is caused by an increase in retinal vascular permeability. The aim of this study is to evaluate the efficacy of oral TXA for ME associated with retinal vein occlusion (RVO) or diabetic ME (DME).Patients and methodsOral TXA (1,500 mg daily for 2 weeks) was administered to patients with persistent ME secondary to RVO (7 eyes) and DME (7 eyes). After 2 weeks (ie, the final day of administration) and 6 weeks (ie, 4 weeks after the final administration), best-corrected visual acuity and central macular thickness (CMT) were measured and compared with baseline. Analyses were performed for RVO and DME cases. No other treatment was performed during the study period.ResultsIn RVO cases, significant improvement in CMT was found between baseline (467.7±121.4 μm) and 2-week measurements after treatment (428.7±110.5 μm, p=0.024). No significant change was found in CMT between measurements taken at baseline and 6 weeks after treatment. In DME cases, no significant change was found in CMT between measurements taken at baseline and 2 or 6 weeks after treatment. In all analyses of best-corrected visual acuity, no significant change was observed.ConclusionThe results support the hypothesis that plasmin plays a role in the development of ME associated with RVO, and oral TXA administration may be useful as an adjuvant treatment when combined with other agents such as anti-vascular endothelial growth factor.

Abstract 2873: Imaging tumor heterogeneity after multikinase inhibitor therapy in rat hepatocellular carcinoma

Abstract Purpose To assess tumor tissue perfusion and vascular permeability through multiparametric functional imaging, and to evaluate their correspondence with histopathologic parameters of necrosis and hypoxia in a rat model of hepatocellular carcinoma treated with a multikinase inhibitor. Materials and Methods Rat hepatoma McA-RH7777 cells were implanted in the left liver lobe of nineteen male Buffalo rats. Exactly 2 weeks after tumor inoculation, the animals were randomly assigned to remain untreated (n=10) or to receive a daily dose of 7.5 mg/kg sorafenib by oral gavage (n=9) for 2 additional weeks. T2-weighted spin-echo magnetic resonance imaging (MRI), dynamic contrast-enhanced (DCE) and contrast-enhanced ultrasound (CEUS) were performed weekly. All tumors were harvested 4 weeks post-implantation &amp;gt;90 minutes after injecting 60mg/kg pimonidazole. Tissue sections were stained for hematoxylin-eosin and pimonidazole for quantitative assessment of necrosis and hypoxia, respectively. Differences between treatment groups were assessed using the Mann-Whitney test, and the correlation of imaging and histopathology parameters was determined by Spearman correlation analysis. Results In spite of the relatively low dose and short treatment duration, the response to sorafenib therapy was characterized by a significantly higher median tumor necrosis (60 vs 15%, P &amp;lt; 0.0001) and increased tumor tissue hypoxia (35 vs 18.5% pimonidazole binding, P &amp;lt; 0.0001). Similarly, the sorafenib regimen caused a substantial decrease in vascular permeability, measured by DCE-MRI Ktrans (P = 0.002) as well as reduction in tumor perfusion evaluated by CEUS peak enhancement (PE) (P =0.043). Our findings show a strong correlation between the two histopathologic parameters we assessed, tumor tissue necrosis and hypoxia (r = 0.645, P=0.002). Furthermore, our results demonstrate significant correlation of functional imaging parameters of vascular permeability, namely DCE-MRI Ktrans and DCE-MRI normalized initial area under the curve (NIAUC), with histopathologic tissue hypoxia (r=-0.663, P= 0.002, and r= -0.512, P=0.029, respectively). In contrast, histopathologic tumor necrosis appears more strongly correlated with a functional imaging parameter of tumor perfusion, specifically, CEUS PE (r = -0.547, P = 0.028). Conclusion Tumors exhibited wide heterogeneity in vascular perfusion and hypoxia after treatment. Interestingly, functional MRI parameters appear to correlate more strongly with tissue oxygenation, whereas functional CEUS parameters correlate with tumor viability. Novel predictive imaging biomarkers of treatment response may be developed through further analyses of spatial tumor heterogeneity in our animal model. Citation Format: Nina M. Muñoz, Adeeb A. Minhaj, Kiersten L. Maldonado, Charles Kingsley, Hideyuki Nishiofuku, Keith A. Michel, Andrea C. Cortes, James A. Bankson, Asif Rashid, Rony Avritscher. Imaging tumor heterogeneity after multikinase inhibitor therapy in rat hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2873. doi:10.1158/1538-7445.AM2017-2873

2-Methoxyestradiol protects against IgG immune complex-induced acute lung injury by blocking NF-κB and CCAAT/enhancer-binding protein β activities.

Increasing evidences indicate that 2-Methoxyestradiol (2ME2) plays an essential role in protecting against inflammatory responses. However, its effect on IgG immune complex (IC)-induced acute lung injury (ALI) remains enigmatic. In the study, by using i.p. administration of 2ME2, we evaluated its influence on IgG IC-induced pulmonary injury in mice. We found that during IgG IC-induced ALI, mice treated by 2ME2 displayed a substantial decrease in vascular permeability and neutrophil influx (represented by myeloperoxidase activity) when compared with their counterparts receiving vehicle treatment. Furthermore, 2ME2 treatment significantly decreased pro-inflammatory mediator production and inflammatory cell, especially neutrophil accumulation in bronchoalveolar lavage fluids (BALFs) upon IgG IC stimulation. In vitro, IgG IC-triggered inflammatory mediator production was markedly down-regulated by 2ME2 in macrophages. Moreover, we verified that the activation of the transcription factors, NF-κB and CCAAT/enhancer-binding protein (C/EBP) β, were inhibited by 2ME2 in IgG IC-challenged macrophages. We demonstrated that alleviation of NF-κB-dependent transcription might be associated with reduced phosphorylation of NF-κB p65, and reduction of C/EBP activation was directly linked to its expression. In addition, we discovered that IgG IC-stimulated phosphorylation of both p38 MAPK and ERK1/2 was alleviated by 2ME2. These data indicated a novel strategy for blockade of IgG IC-induced inflammatory activities.

Association of Heme Oxygenase 1 with Lung Protection in Malaria-Associated ALI/ARDS.

Malaria is a serious disease, caused by the parasite of the genus Plasmodium, which was responsible for 440,000 deaths in 2015. Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is one of the main clinical complications in severe malaria. The murine model DBA/2 reproduces the clinical signs of ALI/ARDS in humans, when infected with Plasmodium berghei ANKA. High levels of HO-1 were reported in cases of severe malaria. Our data indicated that the HO-1 mRNA and protein expression are increased in mice that develop malaria-associated ALI/ARDS (MA-ALI/ARDS). Additionally, the hemin, a HO-1 inducing drug, prevented mice from developing MA-ALI/ARDS when administered prior to the development of MA-ALI/ARDS in this model. Also, hemin treatment showed an amelioration of respiratory parameters in mice, high VEGF levels in the sera, and a decrease in vascular permeability in the lung, which are signs of ALI/ARDS. Therefore, the induction of HO-1 before the development of MA-ALI/ARDS could be protective. However, the increased expression of HO-1 on the onset of MA-ALI/ARDS development may represent an effort to revert the phenotype of this syndrome by the host. We therefore confirm that HO-1 inducing drugs could be used for prevention of MA-ALI/ARDS in humans.

The study of the action of the thick extract from the leaves of Corylus avellana on the vascular tissue permeability

Research of the action of the thick extract from the leaves of Corylus avellana on the vascular-tissue permeability of the rats skin has been conducted. Effect of the thick extract from the leaves of Corylus avellana on vascular tissue permeability in rats has been assessed by the time of papules coloring (animal skin at the injection side of flohohen substances) in seconds. We used formalin, histamine, protein, which have different mechanisms of action on capillary permeability. In the group, which has been administrated with the studied extract at its conditional therapeutic dose of 60 mg/kg we have observed the decrease of vascular permeability induced by flohogene substances. It was determined that the most pronounced vessel strengthening effect during the experiment with histamine, where papules coloring has been 1.9 times slower compared with the control group. In comparison with eskuvit the data rates has been approaching to 1. Taking into consideration the mechanism of the action of the applied flohogenes we can suggest that an extract from the leaves of Corylus avellana detects an expressed validity of capillaries permeability and mikrocirculations. Extract showed a moderate effect against the background of formalin and protein inflammation, but this index has exceeded the index of the eskuvit group by 170 seconds against the background of formalin inflammation. The obtained results indicate the presence of capillary resistance activity that is cause for further research of the efficiency of the extract in experimental pathology.

NI-62 * DCE-MRI DISCRIMINATES BETWEEN RESPONDER AND NON-RESPONDER IN PROGRESSIVE HIGH GRADE GLIOMA PATIENTS UNDERGOING ANTI-ANGIOGENIC TREATMENT

OBJECTIVES: In this prospective phase II clinical study we analyzed whether dynamic contrast enhanced (DCE) - MRI may distinguish between anti-vascular and anti-tumor effects in the course of anti-angiogenic treatment of progressive high-grade glioma patients (pHGG). METHODS: Twenty-two patients with pHGG (16 glioblastoma, 5 anaplastic astrocytoma WHO III, 1 anaplastic oligoastrocytoma) were treated biweekly with bevacizumab/irinotecan. DCE-MRI scans were performed at baseline and eight weeks after treatment commencement. DCE-MRI analysis included four semiquantitative parameters, describing gadolinium contrast medium behavior; 1) initial slope of signal increase (marker for vessel permeability; slope), 2) maximum enhancement (marker for extravascular volume; ME), 3) time from starting of curve increase until ME (time-to-peak; TTP), 4) area under the curve until ME (IAUC). Responders to anti-angiogenic therapy were defined as having a progression free survival (PFS) greater than six months (PFS6). DCE-MRI parameters were compared to PFS6 by Wilcoxon rank test. RESULTS: Mean PFS was 4.0 ± 2.2 months and 4 patients showed a PFS greater than 6 months and were classified as responders. At baseline there was no difference in ME, TTP, IAUC and slope between responder and non-respopnder. At follow-up responders showed a significant decrease in ME, TTP and IAUC, whereas slope did not change (mean ME 60.03% vs. 23.0%, mean TTP 579.5sec vs 367.4sec, mean IAUC 55040 vs 12150 and slope 1.03 vs 1.04, p < 0.001). In contrary non-responder did not show a change in ME, TTP and IAUC but an increase in slope (mean ME 55.45% vs 50.21%, mean sTTP 539.0sec vs 523.4sec, mean IAUC 53440 vs 50320 and slope 1.49 vs 1.74, p < 0.001). CONCLUSION: By semiquantitative DCE-MRI analysis we could show that responders to anti-angiogenic therapy show a significant decrease in extravascular volume despite no decrease in vascular permeability. These results might provide insights into anti-angiogenic treatment effects in responding patients.

NI-57 * DYNAMIC CONTRAST-ENHANCED MAGNETIC RESONANCE PERFUSION WEIGHTED IMAGING (DCE-MRI) AND DIFFUSION WEIGHTED IMAGING (DWI) FOR PHARMACODYNAMIC EVALUATION OF CARBOXYAMIDOTRIAZOLE OROTATE (CTO) AND TEMOZOLOMIDE IN MALIGNANT GLIOMA

BACKGROUND: Advanced imaging techniques have been proposed as potentially useful tools for evaluation of biological effects of new targeted agents. CTO is an oral inhibitor of non-voltage-dependent calcium signaling that modulates several pathways, including EGFR, MEK, RAS, HDAC, HSP90, WNT/B-catenin, Akt, ERK, VEGF and Bcr-Abl. We prospectively evaluated advanced MRI parameters in a phase I trial of combined CTO and adjuvant temozolomide for temozolomide-refractory recurrent/progressive malignant glioma, aiming at characterizing early pharmacodynamic effects after drug exposure. METHODS: Within an exploratory imaging correlative study, a subset of patients enrolled in this trial underwent DCE-MRI and DWI at baseline and after two 28-day cycles. Changes in imaging parameters were evaluated utilizing volumetric histogram analysis of the entire tumor (NordicICE v2.3; NordicNeuroLab, Bergen, Norway), including blood perfusion (relative plasma volume [Vp]), vascular permeability (Ktrans) and diffusion (apparent diffusion coefficient [ADC]). Normalized results were analyzed with Mann-Whitney u tests. RESULTS: Among 17 enrolled patients, 7 were included in the imaging component (glioblastoma: 5; grade III: 2; MGMT methylated: 3, unmethylated: 2, unknown: 2; bevacizumab failures: 3), with CTO dose range of 219-625mg/m2/day. All patients displayed a reduction in blood perfusion, with mean Vp = 10.65 at baseline vs 3.82 after 2 cycles, p = 0.001. There were no statistically significant differences in vascular permeability parameters, such as Ktrans (mean 11.07 vs 5.14, p = 0.38) or ADC (mean 1.54 vs 1.54, p = 0.81). CONCLUSIONS: DCE-MRI depicted decreases in blood perfusion, demonstrating biological effects of CTO in all evaluated patients after two cycles. Interestingly, such changes were not accompanied by significant decreases in vascular permeability, suggesting an effect distinct from that observed with bevacizumab and VEGF blockade, and likely reflecting modulation of other signaling pathways. This study provides insight into the mechanism of action of CTO, and highlights the usefulness of advanced neuroimaging in early drug development in brain tumors.

Outcome of discontinuing bevacizumab prior to malignant glioma progression

Bevacizumab (Avastin , Genentech Inc., San Francisco, CA) was granted accelerated approval from the Food and Drug Administration as single agent treatment of recurrent glioblastoma (GB) based on the high radiologic response rate demonstrated in two phase 2 clinical trials. The approval of bevacizumab has been controversial despite the impressive radiographic response. It is argued that neutralization of vascular endothelial growth factor results in stabilization of the blood brain barrier (BBB) with resultant decrease in vascular permeability, leading to an improvement in edema and a decrease in enhancement on imaging, rather than a true anti-tumor effect [1, 2]. In addition, the optimal dose, schedule, and duration of bevacizumab treatment has not been determined. Given its moderate toxicity and the treatment unresponsive, invasive ‘‘gliomatosis’’ pattern of recurrence that occurs in a subset of patients [3] it may be favorable to discontinue bevacizumab prior to tumor progression. Contrarily, early discontinuation of bevacizumab could lead to a ‘‘rebound’’ in edema and tumor progression, compromising patient function and survival. The outcome of discontinuing therapy in bevacizumab responders has not been described, but many clinicians fear rebound cerebral edema and rapid clinical decline. We evaluated whether our bevacizumab responders who discontinued therapy prior to tumor progression experienced a radiographic ‘‘rebound’’ effect, early tumor recurrence and clinical decline, and a second radiographic response following re-challenge with bevacizumab at subsequent tumor progression.

Role of leukotrienes in NSAID induced gastric ulceration and inflammation in wistar rats

ObjectiveTo evaluate the effects of Montelukast and Curcumin against indomethacin induced gastric damage in rats in order to assess the role of leukotriene (LTs) if any, in non steroidal anti-inflammatory drug (NSAID) induced gastroinflammation. MethodsThe effects of Montelukast (10 mg/kg) and Curcumin (100 mg/kg) were observed on gastric lesion induced by Indomethacin. The blood samples were analyzed for neutrophil adhesion and lipid peroxide levels in gastric tissue measured spectrophotometrically. The skin vascular permeability study was performed by using compound 48/80 induced vascular permeability model. ResultsMontelukast and Curcumin significantly reduced Indomethacin induced gastric lesion score. Pretreatment with Montelukast and Curcumin significantly counteracted Indomethacin induced gastropathy by a combination of its effect on inhibition of neutrophil adherence, through decrease in related production of free radicals that disrupts integrity of stomach mucosa and decrease in vascular permeability as compared to Indomethacin group. The results of the present study further indicates the role of 5-LOX metabolites in NSAIDs induced gastro inflammation and suggests that Montelukast and Curcumin counteracted the Indomethacin induced gastropathy by a combination of its effect on inhibition of neutrophil adherence and through decrease in related production of free radicals that disrupts integrity of stomach mucosa. ConclusionsExperimental data clearly demonstrated the role of LTs was indomethacin induced gastric ulcers. However, inhibition of ulcerogenic events by Montelukast and Curcumin is suggestive of an important balance between COX and 5-LOX products.

Effect of anisodamine on lung edema after ischemia-reperfusion in rabbits

Objective To investigate the protective efficacy of anisodamine against lung edema after ischemia-reperfusion in rabbits,and to explore the possible mechanism.Methods In vivo lung ischemia-reperfusion model of rabbit was developed. Twenty-four Chinese rabbits of either gender,aged 10-12 weeks,weighing 2200-2600 g,were divided into 3 groups randomly:anisodamine group,control group and sham group,with 8 rabbits in each group.In anisodamine group,the left pulmonary hilum was occluded for 1 hour and then released for 3 hours,and anisodamine(3 mg/kg) was given by intravenously injection before occlusion. In control group,only ischemia-reperfusion was performed.In sham group,left lung received no ischemia-reperfusion.Left lung tissue in each group was harvested for histopathological analysis,lung water content determination and myeloperoxidase(MPO) level detection.Also,2%Evens blue(1.5 ml/kg) was injected into femoral venous to measure lung vascular permeability.Results After ischemia-reperfusion,the lung tissue water content,MPO level and Evens blue level in control group were(6.6±0.5) g/g dry tissue, (1.16±0.14) U/g protein and(173±16) ug/g wet tissue respectively,significantly higher than those in sham group[(4.3±0.4) g/g dry tissue,(0.53±0.09) U/g protein and(103±11)μg/g wet tissue,P0.05].These indices in anisodamine group were(5.6±0.4) g/g dry tissue,(0.82±0.11)U/g protein and(124±18)μg/g wet tissue,obviously lower than those in control group(P0.05).Left lung displayed prominent edema,septal swelling,hemorrhage and neutrophil infiltration in control group,while which was significantly improved in anisodamine group.Conclusion Anisodamine can significantly prevent lung edema after ischemia-reperfusion in rabbits. The mechanism may be ascribed to inhibition of neutrophil infiltration and decrease in vascular permeability.

Ghrelin Prevents Disruption of the Blood–Brain Barrier after Traumatic Brain Injury

Significant effort has been focused on reducing neuronal damage from post-traumatic brain injury (TBI) inflammation and blood-brain barrier (BBB)-mediated edema. The orexigenic hormone ghrelin decreases inflammation in sepsis models, and has recently been shown to be neuroprotective following subarachnoid hemorrhage. We hypothesized that ghrelin modulates cerebral vascular permeability and mediates BBB breakdown following TBI. Using a weight-drop model, TBI was created in three groups of mice: sham, TBI, and TBI/ghrelin. The BBB was investigated by examining its permeability to FITC-dextran and through quantification of perivascualar aquaporin-4 (AQP-4). Finally, we immunoblotted for serum S100B as a marker of brain injury. Compared to sham, TBI caused significant histologic neuronal degeneration, increases in vascular permeability, perivascular expression of AQP-4, and serum levels of S100B. Treatment with ghrelin mitigated these effects; after TBI, ghrelin-treated mice had vascular permeability and perivascular AQP-4 and S100B levels that were similar to sham. Our data suggest that ghrelin prevents BBB disruption after TBI. This is evident by a decrease in vascular permeability that is linked to a decrease in AQP-4. This decrease in vascular permeability may diminish post-TBI brain tissue damage was evident by decreased S100B.