Increased Blood Perfusion Research Articles

The Role of Nitric Oxide during Sonoreperfusion of Microvascular Obstruction.

Rationale: Microembolization during PCI for acute myocardial infarction can cause microvascular obstruction (MVO). MVO severely limits the success of reperfusion therapies, is associated with additional myonecrosis, and is linked to worse prognosis, including death. We have shown, both in in vitro and in vivo models, that ultrasound (US) and microbubble (MB) therapy (termed “sonoreperfusion” or “SRP”) is a theranostic approach that relieves MVO and restores perfusion, but the underlying mechanisms remain to be established.Objective: In this study, we investigated the role of nitric oxide (NO) during SRP.Methods and results: We first demonstrated in plated cells that US-stimulated MB oscillations induced a 6-fold increase in endothelial nitric oxide synthase (eNOS) phosphorylation in vitro. We then monitored the kinetics of intramuscular NO and perfusion flow rate responses following 2-min of SRP therapy in the rat hindlimb muscle, with and without blockade of eNOS with LNAME. Following SRP, we found that starting at 6 minutes, intramuscular NO increased significantly over 30 min and was higher than baseline after 13 min. Concomitant contrast enhanced burst reperfusion imaging confirmed that there was a marked increase in perfusion flow rate at 6 and 10 min post SRP compared to baseline (>2.5 fold). The increases in intramuscular NO and perfusion rate were blunted with LNAME. Finally, we tested the hypothesis that NO plays a role in SRP by assessing reperfusion efficacy in a previously described rat hindlimb model of MVO during blockade of eNOS. After US treatment 1, microvascular blood volume was restored to baseline in the MB+US group, but remained low in the LNAME group. Perfusion rates increased in the MB+US group after US treatment 2 but not in the MB+US+LNAME group.Conclusions: These data strongly support that MB oscillations can activate the eNOS pathway leading to increased blood perfusion and that NO plays a significant role in SRP efficacy.

Triple-phase 99mTc-3P-RGD2 imaging of peripheral primitive neuroectodermal tumor in the hip muscle group with bone metastasis.

Peripheral primitive neuroectodermal tumors (pPNETs) are a group of aggressive neoplasms that are most commonly encountered in pediatric patients and may be located in the abdomen, pelvis, thoracopulmonary region and, rarely, in the head and neck region. pPNETs in adults are extremely rare. The present study reports a case of pPNET located in the hip muscles with bone metastasis. The patient was a 44-year-old woman who complained of progressive pain and swelling with a mass near the left hip. Computed tomography (CT) and enhanced CT revealed a soft tissue mass lesion in the hip muscle group measuring 4.3×4.3×4.4 cm. The lesion was ill-defined, heterogeneous, exhibiting mild post-contrast enhancement. There was a large number of bent neovessels and several branches from the left internal iliac artery and deep femoral artery on enhanced CT scan. Triple-phase dynamic imaging with integrin αvβ3-targeted 99mTc-3P-RGD2 as the radiotracer revealed increased blood perfusion and radiotracer aggregation in the large, ill-defined, heterogeneous, hypodense mass and adjacent bone. The patient was suspected of having pPNET with bone metastasis, which was confirmed by histological examination of a sample obtained by needle aspiration. Due to the high blood perfusion of primary pPNETs and high RGD uptake by the primary and metastatic lesions, chemoembolization and anti-angiogenic therapy were considered to be the optimal therapeutic choice. This also suggested that 177Lu-labeled RGD has great potential for the targeted treatment of pPNETs with multiple metastases.

Overexpression of Gremlin1 in Mesenchymal Stem Cells Improves Hindlimb Ischemia in Mice by Enhancing Cell Survival

Mesenchymal stem cells (MSCs) are a promising cell resource for the treatment of ischemic diseases, partially through paracrine effects. One of the major obstacles of MSC treatment is the poor survival rate and low efficiency of transplanted stem cells due to ischemic or inflammatory environments. Gremlin1 (GREM1), a regulator of growth, differentiation and development, has been identified as a novel proangiogenic factor. However, the role and mechanism of GREM1 in MSCs remains unclear. Therefore, we assessed the putative beneficial effects of GREM1 on MSC-based therapy for hindlimb ischemia. The lentiviral vector, EF1a-GREM1, was constructed using the Multisite Gateway System and used to transduce MSCs. In vitro studies demonstrated increased survival of GREM1-MSCs exposed to H2 O2 , which is consistent with the activation of caspase-3. Conditional medium from GREM1-MSCs (GREM1-MSC-CM) increased the anti-apoptotic effects of human umbilical vein endothelial cells (HUVECs), and this effect was attenuated by treatment with the PI3K/Akt pathway inhibitor LY294002. MSCs modified with GREM1 could significantly increase blood perfusion of the ischemic hindlimb in vivo in a mouse model, which was correlated to improved MSC survival. This study demonstrates that overexpression of GREM1 in MSCs have greater therapeutic effects against ischemia compared with wild-type MSCs by enhancing the survival of MSCs and ECs, which may provide new tools for studies investigating the treatment of ischemic diseases. J. Cell. Physiol. 232: 996-1007, 2017. © 2016 Wiley Periodicals, Inc.

Acute supplementation of L-arginine affects neither strength performance nor nitric oxide production

L-arginine is a semi-essential amino acid involved in nitric oxide production. As nitric oxide is an important vasodilator, L-arginine supplementation would increase blood perfusion and, subsequently, muscle performance during exercises. The aim of this study was to determine the acute effect of L-arginine supplementation on strength performance and nitric oxide levels in healthy trained individuals. In a double-blind, placebo-controlled, cross-over study, 12 men were randomly assigned to L-arginine or placebo supplementation. Subjects received 6 g of L-arginine or placebo 60 minutes before strength test (maximum number of repetitions, 3 sets at 70% of one repetition maximum on bench press and at 80% of one repetition maximum on knee extensions, 2 minutes of rest between sets and exercises). Blood samples were collected before supplementation and 6 min after exercise. Plasma nitrite levels did not significantly change after L-arginine or placebo supplementation and strength-training exercise (placebo, from 13.01±1.18 to 11.83±2.81 mM; L-arginine, from 10.95±4.09 to 11.99±2.5 mM). There was a significant reduction in the number of repetitions performed from set 1 to set 3 in each set of both bench press and knee extension, but no significant interactions were observed between placebo and L-arginine. These results do not support the use of L-arginine as an ergogenic aid for strength performance, at least in context of acute use immediately before resistance exercise performance.

Engineering a Dual-Layer Chitosan–Lactide Hydrogel To Create Endothelial Cell Aggregate-Induced Microvascular Networks In Vitro and Increase Blood Perfusion In Vivo

Here, we report the use of chemically cross-linked and photo-cross-linked hydrogels to engineer human umbilical vein endothelial cell (HUVEC) aggregate-induced microvascular networks to increase blood perfusion in vivo. First, we studied the effect of chemically cross-linked and photo-cross-linked chitosan-lactide hydrogels on stiffness, degradation rates, and HUVEC behaviors. The photo-cross-linked hydrogel was relatively stiff (E = ∼15 kPa) and possessed more compact networks, denser surface texture, and lower enzymatic degradation rates than the relatively soft, chemically cross-linked hydrogel (E = ∼2 kPa). While both hydrogels exhibited nontoxicity, the soft chemically cross-linked hydrogels expedited the formation of cell aggregates compared to the photo-cross-linked hydrogels. Cells on the less stiff, chemically cross-linked hydrogels expressed more matrix metalloproteinase (MMP) activity than the stiffer, photo-cross-linked hydrogel. This difference in MMP activity resulted in a more dramatic decrease in mechanical stiffness after 3 days of incubation for the chemically cross-linked hydrogel, as compared to the photo-cross-linked one. After determining the physical and biological properties of each hydrogel, we accordingly engineered a dual-layer hydrogel construct consisting of the relatively soft, chemically cross-linked hydrogel layer for HUVEC encapsulation, and the relatively stiff, acellular, photo-cross-linked hydrogel for retention of cell-laden microvasculature above. This dual-layer hydrogel construct enabled a lasting HUVEC aggregate-induced microvascular network due to the combination of stable substrate, enriched cell adhesion molecules, and extracellular matrix proteins. We tested the dual-layer hydrogel construct in a mouse model of hind-limb ischemia, where the HUVEC aggregate-induced microvascular networks significantly enhanced blood perfusion rate to ischemic legs and decreased tissue necrosis compared with both no treatment and nonaggregated HUVEC-loaded hydrogels within 2 weeks. This study suggests an effective means for regulating hydrogel properties to facilitate a stable, HUVEC aggregate-induced microvascular network for a variety of vascularized tissue applications.

Value of correlative biomarkers in understanding tumor biology

We appreciate the insightful review of our article Batchelor et al . “Improved tumor oxygenation and survival in glioblastoma patients who show increased blood perfusion after cediranib and chemoradiation” by Dr. Burt Nabors (1,2). In his commentary, he skillfully highlights the initial enthusiasm followed by more measured interest for anti-angiogenic agents in glioblastoma (GBM) (1). Based on the robust angiogenesis that characterizes GBM, there exists a strong biological rationale for targeting tumor blood vessels and, fundamentally, blood flow and nutrient delivery are essential to tumor survival. The therapeutic challenge with this class of agents has been identifying those patients likely to benefit and achieve a durable response both clinically and radiographically.

The therapeutic effect of curcumin in male albino rats and its putative mechanisms on cerebral microvascular flow

The present study aimed to investigate the therapeutic effect of curcumin on hypertension and its putative mechanisms in the cerebral microcirculation. The surgical preparation was made to generate a cranial window for observation of the capillary network in the cerebral cortex region. Digital image processing, intravital videomicroscopy, and laser Doppler flow meter were used in this investigation. The number of open capillaries, arterial blood pressure, red cell velocity, microvascular diameter, circulating endothelial cells, relative blood flow and frequency were determined. Control rats showed severe dysfunction in the microcirculation with increased blood pressure. In curcumin treated mice, the blood pressure significantly reduced compared to their respective controls. Curcumin significantly increased blood velocity and LDF flow in hypertensive and normotensive rats. Curcumin significantly altered the circulating endothelial cells and open capillaries number in the male albino rats. Our results suggested that the curcumin exerts its therapeutic effect in male albino rats by regulating vasomotion function, increasing blood perfusion, releasing the peripheral resistance and opening efficiently capillaries. Taking all these data together, it is concluded that the curcumin might be useful in the regulation of the cerebral microcirculatory function and hypertension.

Combinatorial Treatment with Apelin-13 Enhances the Therapeutic Efficacy of a Preconditioned Cell-Based Therapy for Peripheral Ischemia.

Hypoxic pretreatment of peripheral blood mononuclear cells (PBMNCs) enhances therapeutic angiogenesis in ischemic tissues after cell transplantation. However, newly formed vessels generated using this approach are immature and insufficient for promoting functional recovery from severe ischemia. In this study, we examined whether apelin-13, a regulator of vessel maturation, could be an effective promoter of therapeutic angiogenesis, following severe limb ischemia. Combinatorial treatment of hypoxic preconditioned PBMNCs with apelin-13 resulted in increased blood perfusion and vascular reactivity in ischemic mouse hindlimbs compared with a monotherapy comprising each factor. Apelin-13 upregulated expression of PDGF-BB and TGF-β1 in hypoxic PBMNCs, as well as that of PDGFR-β in vascular smooth muscle cells (VSMCs). Proliferation and migration of VSMCs treated with apelin-13 was accelerated in the presence of PDGF-BB. Interestingly, expression of an apelin receptor, APJ, in PBMNC was increased under hypoxia but not under normoxia. In addition, an in vitro angiogenesis assay using a co-culture model comprising mouse thoracic aorta, hypoxic PBMNCs, and apelin-13 demonstrated that combinatorial treatment recruited mural cells to sprouted vessel outgrowths from the aortic ring, thereby promoting neovessel maturation. Thus, combinatorial injection of hypoxic PBMNCs and apelin-13 could be an effective therapeutic strategy for patients with severe ischemic diseases.

A shape-controlled tunable microgel cell delivery platform for low-dose delivery of primed stem cells for in vivo therapeutic neovascularization

Event Abstract Back to Event A shape-controlled tunable microgel cell delivery platform for low-dose delivery of primed stem cells for in vivo therapeutic neovascularization Dilip Thomas1, 2, Grazia Marsico1, Arun Thirumaran2, Bartlomiej Lukasz3, Kerry Thompson4, Peter Dockery4, Brian Rodriguez3, Martina Marchetti-Deschmann5, Timothy O'Brien1, 2 and Abhay Pandit1 1 National University of Ireland Galway, CÚRAM, Ireland 2 National University of Ireland, REMEDI, Ireland 3 University College Dublin, Conway Institute, Ireland 4 National University of Ireland Galway, Anatomy, Ireland 5 Vienna University of Technology, Institute of Chemical Technologies and Analytics, Austria Introduction: Hydrogel-based cell delivery platforms fabricated from synthetic or natural polymers have evolved from a cytoprotective design to a functional biomaterial design, which not only offers better cell survival but also modulates cellular behaviour via intrinsic biophysical or biochemical cues. Tissue regenerative processes are driven by reparative mesenchymal stem cells (MSCs) under the influence of biochemical cues dictated by the microenvironment. Prolonged ischemia and lack of blood supply drive cellular apoptosis and tissue death. However, delivery of primed stem cells via a tunable microenvironment triggers an ‘angiocrine’ response, driving therapeutic angiogenesis[1]. Hence, in this study, it is hypothesised that delivery of primed human mesenchymal stem cells on a shape-controlled microgel platform at a low-cell dose promotes angiogenesis in a murine model of hind-limb ischemia (HLI). The specific objectives of the study were to investigate the cellular behaviour on tunable parameters such as matrix concentration, cross-linker concentration and cell densities and therapeutic efficacy of the optimized group in a hind-limb ischemia model. Materials and Methods: Collagen-based microgels of concentrations 1, 2 and 3mg/ml were fabricated by neutralizing type-I collagen and dispensing a mixture composed of cells and cross-linker 4S-Star-PEG Mw 10000 KDa (Jenkem Tech, U.S.A) on to a hydrophobic surface. hMSCs at an optimized cell density of 0.8x106 were embedded within the microgels[2]. Cell embedded microgels were tested for changes in morphology by confocal microscopy; protein and gene expression quantitation by multiplex ELISA and gene arrays; and changes in matrix stiffness with atomic force microscopy and confocal microscopy. In vivo experiments were performed in a Balb/c nude hind-limb ischemia mouse model. Animal groups (n=12/group) were PBS; microgels alone; microgels with 50,000 hMSCs; 50,000 and 1,000,000 cells alone. Laser-Doppler perfusion and clinical signs of disease severity were assessed along with molecular evaluation using; multiplex ELISA, gene expression arrays and MALDI-imaging mass spectrometry. Statistical analysis was performed using two-way ANOVA with p<0.05. Results and Discussion: The use of 4S-StarPEG combined with the high surface hydrophobicity allowed gelation after 40 minutes, conferring a spherical shape to the microgels. Over 80% viability and low apoptosis was observed using flow-cytometry. 2mg/ml microgels at 0.8x106 cell density showed up-regulation of paracrine factors involved in angiogenesis and distinguishing integrin expression profile (Figure1). AFM analysis revealed insignificant changes in matrix stiffness and unique cell morphology in 2mg/ml microgels compared to 1 and 3mg/ml microgels. Results from the in vivo hind-limb ischemia studies (Figure 2) showed increased blood perfusion, up-regulation of protein and gene markers related to angiogenesis, and changes in glycan environment between the treatment and control groups. Conclusion: Tunable shape-controlled collagen based microgel platform can be used to deliver hMSCs at a low cell dose for angiogenesis in vivo. This publication has emanated from research supported in part by a research grant from Science Foundation Ireland (SFI) and is co-funded under the European Regional Development Fund under Grant Number 13/RC/2073. Science Foundation Ireland (SFI) under grant no.09/SRC/B1794.

Neural Mechanism of Exercise: Neurovascular Responses to Exercise.

Physical exercise is responsible for different metabolic and hemodynamic changes, including increased cerebral blood flow and perfusion. It is known that running increases vascular endothelial growth factor expression in the brain, which is critical for the anti-depressive effects of adult neurogenesis induced by physical exercise. Both animal and human studies revealed that neurovascular responses to physical exercise are well correlated to adult hippocampal neurogenesis and cognition improvement. Yet it is unknown if the increased blood perfusion to hippocampus is affecting the adult neurogenesis. Manipulating systemic blood pressure, or stimulating the cerebral blood flow with alternative measures, might provide useful tools to understand how much neurovascular plasticity contributes to the brain cognition enhancement by physical exercise. In addition, it will be interesting to examine the responses of brain cells (including neuron, glia and endothelia cells) to increased shear stress and oxygen load, to investigate the underlying molecular mechanisms.

Early effects of low dose bevacizumab treatment assessed by magnetic resonance imaging.

BackgroundAntiangiogenic treatments have been shown to increase blood perfusion and oxygenation in some experimental tumors, and to reduce blood perfusion and induce hypoxia in others. The purpose of this preclinical study was to investigate the potential of dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) and diffusion weighted MRI (DW-MRI) in assessing early effects of low dose bevacizumab treatment, and to investigate intratumor heterogeneity in this effect.MethodsA-07 and R-18 human melanoma xenografts, showing high and low expression of VEGF-A, respectively, were used as tumor models. Untreated and bevacizumab-treated tumors were subjected to DCE-MRI and DW-MRI before treatment, and twice during a 7-days treatment period. Tumor images of Ktrans (the volume transfer constant of Gd-DOTA) and ve (the fractional distribution volume of Gd-DOTA) were produced by pharmacokinetic analysis of the DCE-MRI data, and tumor images of ADC (the apparent diffusion coefficient) were produced from DW-MRI data.ResultsUntreated A-07 tumors showed higher Ktrans, ve, and ADC values than untreated R-18 tumors. Untreated tumors showed radial heterogeneity in Ktrans, i.e., Ktrans was low in central tumor regions and increased gradually towards the tumor periphery. After the treatment, bevacizumab-treated A-07 tumors showed lower Ktrans values than untreated A-07 tumors. Peripherial tumor regions showed substantial reductions in Ktrans, whereas little or no effect was seen in central regions. Consequently, the treatment altered the radial heterogeneity in Ktrans. In R-18 tumors, significant changes in Ktrans were not observed. Treatment induced changes in tumor size, ve, and ADC were not seen in any of the tumor lines.ConclusionsEarly effects of low dose bevacizumab treatment may be highly heterogeneous within tumors and can be detected with DCE-MRI.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-015-1918-1) contains supplementary material, which is available to authorized users.

Mastication induces long-term increases in blood perfusion of the trigeminal principal nucleus

Understanding mechanisms for vessel tone regulation within the trigeminal nuclei is of great interest because some headache syndromes are due to dysregulation of such mechanisms. Previous experiments on animal models suggest that mastication may alter neuron metabolism and blood supply in these nuclei. To investigate this hypothesis in humans, arterial spin-labeling magnetic resonance imaging (MRI) was used to measure blood perfusion within the principal trigeminal nucleus (Vp) and in the dorsolateral-midbrain (DM, including the mesencephalic trigeminal nucleus) in healthy volunteers, before and immediately after a mastication exercise consisting of chewing a gum on one side of the mouth for 1h at 1bite/s. The side preference for masticating was evaluated with a chewing test and the volume of the masseter muscle was measured on T1-weighted MRI scans.The results demonstrated that the mastication exercise caused a perfusion increase within the Vp, but not in the DM. This change was correlated to the preference score for the side where the exercise took place. Moreover, the basal Vp perfusion was correlated to the masseter volume.These results indicate that the local vascular tone of the trigeminal nuclei can be constitutively altered by the chewing practice and by strong or sustained chewing.

Mesenchymal Stem Cells Enhance Angiogenesis and Follicle Survival in Human Cryopreserved Ovarian Cortex Transplantation.

Transplantation of cryopreserved ovarian tissue is a novel technique to restore endocrine function and fertility especially for cancer patients. However, the main obstacle of the technique is massive follicle loss as a result of ischemia in the process of transplantation. Mesenchymal stem cells (MSCs) have been acknowledged to play an important role in supporting angiogenesis and stabilizing long-lasting blood vessel networks through release of angiogenic factors and differentiation into pericytes and endothelial cells. This study is aimed to investigate whether MSCs could be applied to overcome the above obstacle to support the ovarian tissue survival in the transplantation. Here we show that human MSCs could enhance the expression level of VEGF, FGF2, and especially the level of angiogenin, significantly stimulate neovascularization, and increase blood perfusion of the grafts in the cryopreserved ovarian tissue transplantation. Further studies reveal that MSCs could notably reduce the apoptotic rates of primordial follicles and decrease follicle loss in the grafted ovarian tissues. In summary, our findings demonstrate a previously unrecognized function of MSCs in improving human ovarian tissue transplantation and provide a useful strategy to optimize fertility preservation and restoration.

Neutrophil elastase induces inflammation and pain in mouse knee joints via activation of proteinase-activated receptor-2.

Neutrophil elastase plays a crucial role in arthritis. Here, its potential in triggering joint inflammation and pain was assessed, and whether these effects were mediated by proteinase-activated receptor-2 (PAR2). Neutrophil elastase (5 μg) was injected into the knee joints of mice and changes in blood perfusion, leukocyte kinetics and paw withdrawal threshold were assessed. Similar experiments were performed in animals pretreated with the neutrophil elastase inhibitor sivelestat, the PAR2 antagonist GB83, the p44/42 MAPK inhibitor U0126 and in PAR2 receptor knockout (KO) mice. Neutrophil elastase activity was also evaluated in arthritic joints by fluorescent imaging and sivelestat was assessed for anti-inflammatory and analgesic properties. Intra-articular injection of neutrophil elastase caused an increase in blood perfusion, leukocyte kinetics and a decrease in paw withdrawal threshold. Sivelestat treatment suppressed this effect. The PAR2 antagonist GB83 reversed neutrophil elastase-induced synovitis and pain and these responses were also attenuated in PAR2 KO mice. The MAPK inhibitor U0126 also blocked neutrophil elastase-induced inflammation and pain. Active neutrophil elastase was increased in acutely inflamed knees as shown by an activatable fluorescent probe. Sivelestat appeared to reduce neutrophil elastase activity, but had only a moderate anti-inflammatory effect in this model. Neutrophil elastase induced acute inflammation and pain in knee joints of mice. These changes are PAR2-dependent and appear to involve activation of a p44/42 MAPK pathway. Blocking neutrophil elastase, PAR2 and p44/42 MAPK activity can reduce inflammation and pain, suggesting their utility as therapeutic targets.

Short-term follow-up of digital ulcers by laser speckle contrast analysis in systemic sclerosis patients

ObjectiveTo monitor in systemic sclerosis (SSc) the evolution of digital ulcer (DU) status by analysing blood perfusion (BP) using laser speckle contrast analysis (LASCA). MethodsHand BP was recorded by LASCA in twenty SSc patients with recent onset fingertip DUs before and after 10days of local/systemic treatment. Regions of interest (ROIs) to analyse BP were created al the level of ulcer, peri-ulcer, periungual and fingertip areas. Visual analogue pain scale (VAS) was also administered to patients before and after follow-up. ResultsA statistically significant increase of BP was observed from T0 to T1 in the ROIs created at the level of the ulcer area (p<0.0001), as well as a significant decrease of BP was observed in the peri-ulcer area (p<0.0001). A statistically significant decrease of both ulcer size (p<0.0001) and VAS (p=0.001) was observed, whereas no significant variation of both periungual and fingertip BP was detected. ConclusionsLASCA may safely monitor DU evolution in SSc patients, by evaluating its blood perfusion and area during standard treatment. This may be useful to monitor DU evolution during treatment in clinical trials.

Tadalafil for lower urinary tract symptoms secondary to benign prostatic hyperplasia: a review of clinical data in Asian men and an update on the mechanism of action.

Tadalafil, a phosphodiesterase type 5 (PDE5) inhibitor, is approved worldwide for the treatment of lower urinary tract symptoms secondary to benign prostatic hyperplasia (BPH-LUTS). The purpose of this narrative review is to summarize the clinical data on tadalafil 5 mg once-daily, primarily focusing on Asian men with BPH-LUTS, and to update the current understanding of the mechanism of action underlying PDE5 inhibition. Findings from studies have demonstrated that PDE5 is highly expressed in the lower urinary tract and supporting vasculature, and that PDE5 inhibition potentially decreases smooth muscle cell proliferation in the prostate, relaxes smooth muscle in the prostate, bladder neck and supporting vasculature, increases blood perfusion to the lower urinary tract, and modulates bladder afferent nerve activity. A total of 11 larger, 12-week, double-blind, randomized, placebo-controlled studies of tadalafil, including four Asian studies, have been conducted globally, enrolling >3000 men with BPH-LUTS. In addition, two long-term (42- and 52-week) studies enrolled 394 Japanese and 428 North American men, respectively, with BPH-LUTS. Overall, tadalafil 5 mg once-daily resulted in significant improvements in the change from baseline to endpoint in total International Prostate Symptom Scores (IPSS), IPSS storage and voiding subscores, and IPSS quality of life index compared with placebo. Tadalafil was well tolerated and had a favorable safety profile. These findings support tadalafil 5 mg once-daily for treating men, including Asian men, with BPH-LUTS.

Slit2 modifies VEGF-induced angiogenic responses in rabbit skeletal muscle via reduced eNOS activity

Slit2 is a possible modulator of VEGF-induced angiogenesis, but its effects have not been tested on large animal models. We studied the effect of Slit2 on therapeutic angiogenesis induced by VEGF receptor 2 (VEGFR2) ligands Vammin and VEGF-D(ΔNΔC) in vivo in rabbit skeletal muscles. The Slit2 target genes were also studied by RNA sequencing in endothelial cells. Adenoviral intramuscular gene transfers were performed into New Zealand White rabbit hindlimbs. Confocal and multiphoton microscopes were used for blood vessel imaging. Signaling experiments and gene expression analyses were performed to study mechanisms of Slit2 action. Slit2 decreased VEGFR2-mediated vascular permeability. Slit2 also reduced VEGFR2-mediated increase in blood perfusion and capillary enlargement, whereas sprouting of the capillaries was increased. Slit2 gene transfer alone did not have any effects on vascular functions or morphology. VEGFR2 activation was not affected by Slit2, but eNOS phosphorylation was diminished. The transcriptome profiling showed Slit2 down-regulating angiogenesis-related genes such as Nuclear receptor subfamily 4 group A member 1 (NR4A1) and Stanniocalcin-1 (STC-1) as well as genes related to endothelial cell migration and vascular permeability. Combining Slit2 with VEGFs adjusts VEGFR2-mediated angiogenic effects into a more physiological direction. This possibly allows the use of higher VEGF vector doses to achieve a more widespread vector and VEGF distribution in the target tissues leading to a better therapeutic outcome while reducing excess vascular permeability.

VEGF gene transfected umbilical cord mesenchymal stem cells transplantation improve the lower limb vascular lesions of diabetic rats

BackgroundPrevious studies had explored the therapeutic effect of hUC-MSCs transplantation on ischemia; in this study, we further assessed the effectiveness of VEGF over-expressed hUC-MSCs cells transplantation on vascular proliferation in lower limb ischemia model in type 2 diabetic rats. MethodshUC-MSCs cells were over-expressed with VEGF, and transplanted to lower limb ischemia rats model of type 2 diabetes. ResultsVEGF over-expression increased the hUC-MSCs cells proliferation activity and VEGF secretion. VEGF gene transfected hUC-MSCs transplantation increased VEGF expression at a high level throughout 4weeks in skeletal muscle tissues of rats. Importantly, the vascular proliferation and blood perfusion of VEGF over-expressed hUC-MSCs transplanted limb were significantly improved compared with those of control group. The expression levels of ERK, AKT, MMP2 and MMP9 in VEGF over-expressed hUC-MSCs transplantation group increased dramatically compared with control group, while TIMP1 and TIMP2 expression had no significant change. ConclusionVEGF over-expressed hUC-MSCs transplantation was more effective to stimulate angiogenesis and increase blood perfusion than the simply hUC-MSCs transplantation, as maybe a new choice to improve the lower limb vascular lesions of diabetics.

Abstract 639: Carnosine Supplementation Promotes Revascularization in Mouse model of Hindlimb Ischemia

Revascularization surgeries are important for treatment of critical limb ischemia (CLI) however, advanced CLI patients are poor candidates for these surgeries. Here, we proposed that carnosine, a dipeptide of β-alanine and histidine which is highly abundant in the skeletal muscle (2-20mM) and has the abilities to quench reactive aldehydes and chelate divalent metals, can promote revascularization. WTC57Bl/6 mice were pre-treated with carnosine (1g/L) in drinking water for 7 days, subjected to hindlimb ischemia surgery (HLI) by femoral artery ligation and allowed to recover for 14 days with carnosine supplementation. Doppler analysis showed that carnosine supplementation increased blood perfusion to 63±6% compared to 19±3% in controls. Post-surgery carnosine treatment also increased blood flow to 49.4± 6% compared to 28.2± 4% in controls following 21 days of recovery. Histological analysis of the ischemic limb showed that tissue injury was attenuated by carnosine supplementation. Mobilization of angiogenic endothelial progenitor cells (EPCs) (Flk+/Sca+) and the expression of vascular endothelial growth factor (VEGF) in the ischemic muscle was enhanced by carnosine treatment. These results were associated with an increase in ischemic muscle carnosine levels in the carnosine treated HLI mice. Furthermore, expression of the carnosine transporter PHT1 was increased in carnosine treated HLI mice compared to HLI mice. Concomitant with the increased VEGF expression in carnosine treated HLI mice, we observed that pre-treatment of murine myoblast C2C12 cells with octyl-D-carnosine followed by hypoxia increased hypoxia inducible factor 1-alpha nuclear translocation and VEGF expression and secretion. Our results demonstrate that carnosine treatment increases blood flow in the ischemic limb by increasing EPC mobilization and therefore can be a potential therapeutic intervention for CLI patients.

Therapy of thyroid nodules

Thyroid nodules are frequent in Germany. In about every fourth person thyroid nodules can be detected. Most of them are benign. Signs for malignancy are hypoechogenicity, microcalcifications, an unregular margin and increased blood perfusion. There is no strict indication for the treatment of benign nodules. In most cases iodine supplementation is sufficient. A combination therapy with levothyroxine and iodine is more efficient for the treatment of larger nodules. Subclinical hyperthyroidism caused by an adenoma does not necessarily need to be treated, whereas manifest hyperthyroidism needs to treated in most cases with antithyroid drug therapy. Radioiodine therapy is the classical indication for the treatment of unifocal autonomous adenomas. A largely increased thyroid gland with and without uni- / multifocal adenomas are often operated.