Complete Stasis Research Articles

A Single-Center Experience with a Shear-Thinning Conformable Embolic

The purpose of this study was to evaluate the technical success, effectiveness, and safety of transarterial embolization for acute bleeding management with a shear-thinning conformable embolic. This single-center retrospective study evaluated outcomes after embolization using Obsidio conformable embolic (OCE). Technical success was defined as performing transarterial embolization within the target vessel to complete stasis of antegrade flow. Treatment effectiveness was defined as cessation of bleeding for patients. Eleven patients underwent 11 embolization procedures. A total of 16 arteries were embolized. Indications for embolization were spontaneous tumor bleeding (6/11), hematuria (2/11), active duodenal bleeding (1/11), portal hypertensive bleeding (1/11), and rectus sheath hematoma (1/11). The technical success rate was 100%. The median vessel diameter was 2 mm (range, 1–3 mm). There were no adverse events or off-target embolization. OCE demonstrated technical success and treatment effectiveness with a short-term safety profile for transarterial embolization interventions.

Inhibiting PRMT5 induces DNA damage and increases anti-proliferative activity of Niraparib, a PARP inhibitor, in models of breast and ovarian cancer

BackgroundInhibitors of Poly (ADP-Ribose) Polymerases (PARP) provide clinical benefit to patients with breast and ovarian cancers, by compromising the DNA repair activity of cancer cells. Although these agents extend progression-free survival in many patients, responses can be short lived with many patients ultimately progressing. Identification of combination partners that increase dependence of cancer cells to the DNA repair activity of PARPs may represent a strategy to increase the utility of PARP inhibitors. Protein arginine methyltransferase 5 (PRMT5) regulates DNA damage response pathways through splicing and protein modification, and inhibitors of PRMT5 have recently entered clinical trials.MethodsThe effect of PRMT5 inhibition on the levels of DNA damage and repair markers including γH2AX, RAD51, and 53BP1 was determined using high content immunofluorescent imaging. The anti-proliferative activity of the combination of PRMT5 and PARP inhibitors was evaluated using in vitro models of breast and ovarian cancers using both cell lines and ex vivo patient derived xenografts. Finally, the combinations of PRMT5 and PARP inhibitors were evaluated in cell line xenograft models in vivo.ResultsInhibition of PRMT5 by GSK3326595 led to increased levels of markers of DNA damage. The addition of GSK3326595 to the PARP inhibitor, niraparib, resulted in increased growth inhibition of breast and ovarian cancer cell lines and patient derived spheroids. In vivo, the combination improved the partial effects on tumor growth inhibition achieved by either single agent, producing complete tumor stasis and regression.ConclusionThese data demonstrate that inhibition of PRMT5 induced signatures of DNA damage in models of breast and ovarian cancer. Furthermore, combination with the PARP inhibitor, Niraparib, resulted in increased anti-tumor activity in vitro and in vivo. Overall, these data suggest inhibition of PRMT5 as a mechanism to broaden and enhance the clinical application of PARP inhibitors.

Abstract 243: Local Periadventitial Delivery Of Dexamethasone Improves Murine Deep Vein Thrombosis Resolution

Background: Post deep vein thrombosis (DVT), up to 30-50% of patients will develop post-thrombotic syndrome (PTS), despite anticoagulation. Currently no preventive therapies, including catheter-based thrombectomy/thrombolysis (CDT), have shown to reduce PTS. Local steroid therapy adjunct to CDT is being tested clinically for DVT patients in the DEXTERITY trials, but minimal mechanistic data exists on DVT resolution. Here, we investigate effects of local dexamethasone (DEX) therapy on murine DVT resolution. Method: C57/BL6 male mice (n=146) underwent inferior vena cava (IVC) complete ligation to develop stasis DVT on day 0 (D0) followed by mechanical de-ligation on D2 to spur restoration of blood flow (RBF). On D2, mice were injected with PBS/DEX (0.1, 0.2, or 0.5 mg/mouse, 60μl volume, across four zones) in the periadventitial tissue of thrombosed IVC, with or without de-ligation (Fig. A-B). Mice underwent D4&D8 serial ultrasound imaging of IVC blood flow to assess RBF-defined as IVC mean flow velocity > 0 mL/sec. D8 thrombus burden was measured followed by histological/picrosirius red assessment of vein wall fibrosis. Results: N=6-11 mice per group underwent IVC DVT creation. Low-dose (0.1 mg) but not higher doses of DEX reduced thrombus burden (p=0.02) and weight (p=0.01) on D8 (Fig. C-D). Low-dose DEX increased D4 RBF rates and mean flow velocity (MFV) (Fig. E-F). Complete IVC stasis without de-ligation but with DEX non-significantly reduced thrombus burden and weight (Fig. G-H). Low-dose DEX further decreased vein wall fibrosis (Fig. I; p=0.001). D8 thrombi exhibited reduced macrophage content with DEX (Fig. J; n=15-18 sections/group, p=0.007). Conclusion: Following stasis DVT induction, locally delivered low-dose DEX improves DVT resolution by reducing thrombus burden, vein wall injury, macrophage content, and increases RBF rate. These data provide a scientific foundation for ongoing DEXTERITY trail testing adjunct DEX for DVT patients undergoing CDT.

Comparison of Fibered versus Nonfibered Coils for Venous Embolization in an Ovine Model

PurposeTo compare nylon fibered (F) with nonfibered (NF) coils for embolization in an ovine venous model. Materials and MethodsFour- to 8-mm-diameter, 0.035-inch F and NF coils were deployed in 24 veins in 6 sheep. The number of coils, total length of the coils, and length of implanted coil pack required to achieve complete stasis were recorded, as were vessel diameter, radiation dose, ease of packing, damage to embolized vessel, and time to stasis. Venography at 1 and 3 months was used to assess the migration and durability of vessel occlusion. Veins were harvested at 3 months. ResultsF and NF coils were deployed in 24 veins, and stasis was achieved, without immediate coil migration or vessel damage. The mean numbers of F and NF coils per vein were 5 and 8.75, respectively (P = .007). The vessel diameter between the groups was not statistically different. The total coil length (F, 70 cm vs NF, 122.5 cm; P = .0007), coil pack length (F, 29.3 mm vs NF, 39.4 mm; P = .003), time to stasis (F, 5.3 minutes vs NF, 9.0 minutes; P = .008), and radiation dose (F, 25.3 mGy vs NF, 34.9 mGy; P = .037) were significantly different between the groups. Challenges with the animal model prevented conclusive long-term results. Migration occurred with 8 of 11 (72%) coil packs in the femoral veins and 0 of 13 (0%) coil packs in the internal iliac and deep femoral veins. Venography demonstrated that of 16 remaining coil packs, 11 were occluded at 1 month and 10 remained occluded at 3 months. ConclusionsFibers allow for significantly fewer coils to achieve immediate venous occlusion.

Photoacoustic imaging a PDT response marker for monitoring vasculature changes

Photoacoustic imaging (PAI) is an evolving modality for studying and imaging tumor hypoxia. The experimental results from animals treated with ALA for Pre-PDT, Post-PDT, 4hr PDT, 24 hr PDT show a short-term increase in the BV followed by a strong decrease; however, no complete stasis. An alternate method is created by looking at the spatial and frequency domain histogram to observe the tumor vasculature. The images are represented in frequency domains by application of two-dimensional Fast Fourier Transform (FFT) on spatial domain images. The variations in acoustic impedance after Photodynamic Therapy (PDT) are studied. The photoacoustic signals implemented on tumor models indicated PDT influenced variations in the hemoglobin levels against different time periods and studied their outcome on the tumor necrosis.

Pathophysiology of Venous Thromboembolism: Putative role of HDL

IntroductionIn 2019, cardiovascular disease (CVD) was the main reason of all global deaths (32%) and out of these deaths, 85% were due to heart attack and stroke alone. Venous thromboembolism (VTE), which is the third leading cause of CVD, has a very high incidence rate in Indian Population. Data suggest that thromboembolic disorders are due to multiple genetic and environmental variables and hence considered as a complex multifactorial trait. High Density Lipoprotein (HDL) has attracted a great deal of interest in recent years because of its implications in the prevention of various CVD. Various data suggests the relation between HDL and the expression of coagulation/fibrinolytic factors among the patients of CVD. Protein content of HDL is largely responsible for antioxidant mechanism by metabolizing lipid hydro peroxides and preventing their accumulation. Association of venous thrombosis and dyslipidemia has been investigated in several clinical studies, but results are inconsistent. Therefore, we sought to identify the role of HDL in pathophysiology of VTE in both human and animal studies.MethodologyDVT was detected in male patients using color Doppler followed by D‐Dimer and Prothrombin Fragment 1+2 (PF1+2) levels to confirm thrombus formation. HDL associated proteins, endothelial adhesion molecules, thrombotic molecules, inflammatory molecules and oxidized lipoprotein levels were measured. For animal study, a complete stasis animal model was standardized to generate thrombus formation in the SD rats. Thrombus length/weight was measured along with D‐Dimer & PF 1+2 levels. HDL was isolated from plasma and was used in kinetic reaction of LDL oxidation. Progression of LDL oxidation was measured through DCFH‐method. All experimental procedures were performed in accordance to ethical guidelines.Major FindingsA well written consent was taken from all the participants before conducting human study. High levels of D‐Dimer and PF 1+2 in DVT patients as well as IVC ligated group confirmed thrombosis. HDL associated molecules were found to be significantly lower, whereas adhesion molecules, inflammatory molecules, and thrombotic molecules were significantly higher in both DVT patients and IVC ligated group. Oxidized lipoproteins and the lipooxygenase enzymes responsible for lipoprotein oxidation were found significantly higher. Isolated HDL used in LDL oxidation kinetics reaction found to be non‐functional in preventing LDL oxidation. P‐value of <0.05 was used as significance for all the tests.Conclusion & DiscussionLow levels of HDL proteins along with high levels of adhesion molecules, thrombotic markers and inflammatory molecules indicate the suppression of HDL activity. High lipooxygenase enzymes and oxidized lipoprotein levels support the hypothesis of dysfunctional HDL. HDL isolated from the ligated group was unable to prevent the LDL oxidation which clarifies that HDL loses its functionality in case of VTE. Furthermore, finding of this study will also help in understanding the pathophysiology of VTE which might give an insight in prevention/early diagnosis of the disease.

CCS1477, a Novel p300/CBP Bromodomain Inhibitor, Enhances Efficacy of Azacitidine and Venetoclax in Pre-Clinical Models of Acute Myeloid Leukaemia and Lymphoma

E1A binding protein (p300) and CREB binding protein (CBP) are two closely related histone acetyl transferases that co-activate key oncogenes such as MYC and IRF4 which are relevant in a number of haematological malignancies. Here we describe the effects of CCS1477, a new orally available inhibitor of the p300/CBP bromodomains, given alone or in combination with azactidine or venetoclax in pre-clinical models of AML and B-cell lymphoma.As a monotherapy, daily oral dosing with CCS1477 (5, 10 and 20mg/kg) caused a significant dose-dependent reduction in tumour growth in a MOLM-16 xenograft model of AML, with regressions observed at the highest and well tolerated dose of 20mg/kg/qd. The inhibition of tumour growth during drug treatment was accompanied by a significant reduction in tumour expression of MYC by qPCR. In murine retroviral transduction and transplantation models of human AML initiated by either MLL-AF10 or MLL-AF9, daily monotherapy by oral gavage with CCS1477 at 30mg/kg for 42 days, caused a highly significant prolongation of survival. All seven control treated mice died of AML within 160 days whereas only one of seven CCS1477 treated mice had died in each model by 300 days.In the MOLM-16 xenograft model, a sub-maximal dose of CCS1477 (10mg/kg) demonstrated superior tumour growth inhibition by comparison with azacitidine (0.5mg/kg i.p) and with significant combination benefit when these two agents were combined. In this same model venetoclax (50 or 100 mg/kg/qd oral) had no effect on tumour growth, indicating that MOLM-16 tumours are intrinsically resistant. CCS1477 dosed daily at 10mg/kg restored sensitivity to venetoclax as evidenced by significantly greater tumour growth inhibition when compared with CCS1477 given alone. In a venetoclax-sensitive xenograft model of AML (MV-4-11), CCS1477 (10mg/kg) and venetoclax (100mg/kg) resulted in a similar and partial reduction in tumour growth over a 21 day dosing period. When venetoclax and CCS1477 were combined there was a significant combination benefit compared to either drug given alone. The DOHH-2 xenograft model of B-cell lymphoma was also sensitive to venetoclax (50mg/kg) and with a similar impact on tumour growth compared with CCS1477 (10mg/kg). In this model the combination of CCS1477 and venetoclax was more profound and caused complete tumour stasis during the dosing period.These data support the clinical testing of CCS1477 in combination with azacitidine and/or venetoclax in AML and B-cell lymphomas. CCS1477 is currently in Phase I/II clinical trials in AML, Non-Hodgkin lymphoma (including B-cell lymphoma) and multiple myeloma. (NCT04068597). DisclosuresBrooks: CellCentric Ltd: Current Employment, Current holder of individual stocks in a privately-held company. Knurowski: CellCentric Ltd: Current Employment, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Hughes: CellCentric Ltd: Consultancy, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Clegg: CellCentric Ltd: Current Employment, Current holder of individual stocks in a privately-held company. West: CellCentric Ltd: Current Employment, Current holder of individual stocks in a privately-held company. Pegg: CellCentric Ltd: Current Employment, Current holder of individual stocks in a privately-held company. Somervaille: Novartis: Consultancy, Honoraria.

Protective Effects of Kininogen-1 Gene Deficiency in Mouse Models of Venous Thrombosis

Background- High molecular weight Kininogen (HK) is a nonenzymatic co-factor of the contact activation system. HK binds prekallikrein (PK) and FXI to surfaces in proximity to FXII, amplifying PK activation by FXIIa and the reciprocal activation of FXII by activated PK (PKa), as well as FXI activation of FXIIa. PKa cleavage of HK also liberates bradykinin-a proinflammatory and vasoactive nanopeptide. The aim of this study was to define the pro-thrombotic role of kininogen in venous thrombosis (VT) and to use in vivo serial analysis of thrombus development to understand the recruitment and retention of platelets in the growing thrombus in the absence and presence of kininogen.Methods- The development of VT in mice deficient in kininogen (mKng1-/-) was compared to that in their wild-type littermates. A femoral-saphenous stasis VT model was prepared by ligating both saphenous and femoral veins. Next VT formation, growth, and dissolution (n=3 for each group) was monitored using intravital microscopy (IVM) via a multichannel epifluorescence microscope (Nikon Eclipse 90i). To induce stasis VT, FITC-dextran (10 mg/kg, ex/em 488/520 nm) was injected retro-orbitally, and then continuous light irradiation (20x objective, 475nm/35nm) of the saphenous vein was applied for 5 minutes. FITC-dextran fluorescence angiography monitored thrombus formation and dissolution. Immediately after VT formation, platelet accumulation at the thrombus site was monitored in the Cy5 channel (630/38 nm) via injection of a GPIbβ antibody conjugated with Dylight-649 (150nmol/kg), over time. All images were identically windowed in each channel, and thrombus area was measured using NIH ImageJ software. To corroborate IVM studies, we also evaluated a complete stasis model of inferior vena cava (IVC) ligation (n=7-8 per group). Thrombi were harvested after 48 hours and thrombus weight and length were measured to estimate thrombus mass. FXI circulates in blood as a homodimer along with HK. We determined the effect of kininogen deficiency on FXI activity. FXI activity assay used a combination of inhibitors, serially, to monitor the cleavage of substrate specific to activated FXI and release of chromogen, as a function of FXI activity. Finally, to determine the effects of Kng1 deficiency on bleeding, tail vein bleeding times were also determined (n=8 per group).Results- In femoral-saphenous stasis VT, thrombus developed in both groups immediately following FITC-channel light irradiation. However, thrombus size was smaller in Kng1-/- as compared to WT (Figure 1). Results from serial IVM of VT indicated faster thrombus dissolution in the Kng1-/- group. Lower platelet signals, as shown at 2 and 6 hours in the Kng1-/- mice may be consistent with this hypothesis. Thrombus area analysis suggested decreased thrombus formation in the Kng1-/- animals, and temporal analysis indicated faster dissolution by 6 hours (Figure 2). IVC ligation results corroborated the findings of femoral-saphenous DVT model, demonstrating that thrombus weight was significantly lower in Kng1-/- mice as compared to WT (p<0.001, Figure 3). FXI activity was also decreased in the Kng1-/- group (p<0.10). Tail vein bleeding times, however, showed no increased bleeding in Kng1-/- mice.Conclusion- These initial results suggest a pro-thrombotic role of kininogen and a protective role of kininogen deficiency in two murine venous thrombosis models, without incurring a bleeding penalty. Thrombus dissolution was faster and platelet accumulation was inhibited in Kng1-/- mice. These findings suggest that targeting kininogen may provide a new approach to prevent and treat venous thrombosis. [Display omitted] DisclosuresMcCrae: Dova, Novartis, Rigel, and Sanofi Genzyme: Consultancy; Sanofi, Novartis, Alexion, and Johnson & Johnson: Consultancy, Honoraria. Jaffer: Mercator, Inc.: Other: Sponsred research.

S2623 An Unexpected Cause of Melena: A Case Report on Hepatic Pseudoaneurysm Rupture

INTRODUCTION: Hepatic Pseudoaneurysms (HAP) are rare, yet life- threatening vascular disorders due to rupture with the right hepatic artery most commonly being involved. Many of these cases present as obscure gastrointestinal bleeding (GIB), often leading to a delay in diagnosis and treatment. We present a 56-year-old female with a history of metastatic adenocarcinoma who was evaluated in the ICU for hemorrhagic shock due to an upper GIB. CASE DESCRIPTION/METHODS: Patient is a 56-year-old female with a past medical history of metastatic adenocarcinoma of the gallbladder with a surgical history of Roux-en-Y, hepaticojejunostomy, cholecystectomy and biliary stent placement who presented with complaints of epigastric pain and fever of two days duration. The patient was admitted to the ICU for septic shock secondary to cholangitis and gram-negative bacteremia. The patient's hospital course was additionally complicated by hemorrhagic shock secondary to presumed upper GIB due to melena with her receiving multiple transfusions and an emergent upper endoscopy (EGD). The EGD demonstrated multiple clots throughout the stomach and the duodenal bulb filled with blood without an identifying source. Interventional radiology was consulted at which time an abdominal angiogram was performed identifying a pseudoaneurysm of the right hepatic artery at the site of a tumor in the common bile duct. The vessel was successfully coiled and embolized with complete stasis of bleeding. DISCUSSION: HAP is a serious pulsatile hematoma caused by extravasation of blood through disruption of the arterial wall into the surrounding tissue, often leading to haemobilia. HAP is often secondary to trauma, most commonly iatrogenic in nature from procedures. HAP often leads to obscure gastrointestinal bleeding, making the diagnosis of this condition difficult and leading to numerous unnecessary procedures. While there have been numerous diagnostic modalities that have been found to be helpful, the gold standard in diagnosing HAP is conventional angiography. The treatment of choice for pseudoaneurysms had always been surgery, however endovascular treatment with embolization has become more popular based on the high success rate with fewer complications. Although most of these patients remain asymptomatic, the diagnosis of this condition remains clinically significant due to the high-risk of rupture, regardless of size. Therefore, early diagnosis and prompt treatment leads to the best result in these patients.Figure 1.: CT Abdomen/Pelvis.Figure 2.: Example of EGD Demonstrating Haemobilia.Figure 3.: Abdominal Angiogram.

Abstract B09: Selective depletion of YAP1 with next-generation (constrained ethyl-cEt) antisense oligonucleotides results in tumor regression in mouse models of HCC with YAP1 activation

Abstract The Hippo pathway plays important roles in tissue homeostasis. When dysregulated in adult tissues, however, it contributes to the development of a variety of cancers including hepatocellular carcinoma (HCC). YAP1 is the final regulator of the Hippo pathway and is associated with poor prognosis of HCC patients, and more recently has been suggested to be involved in tumor immune evasion. Here, we employed next-generation constrained ethyl (cEt) antisense oligonucleotides (ASOs) to selectively target YAP1 in the genetically engineered mouse (GEM) model of HCC (Salvador KO mice) as well as in a spontaneous carcinogen-induced model of HCC (diethylnitrosamine, DEN-induced HCC). In YAP1-activated Salvador KO mice with established HCC tumors, systemic treatment with YAP1 ASO at 25 mg/kg resulted in a marked reduction in YAP1 protein level in tumor (&amp;gt;90%) and regression of existing tumors, in contrast to the tumors in vehicle or control ASO-treated animals that significantly increased in size. In the DEN-induced HCC model, YAP1 was strongly activated in tumors as demonstrated by nuclear localization of YAP1 protein and the gene signature we previously identified (SOH). YAP1 ASOs produced either complete tumor stasis when DEN tumors were grown in immune-deficient mice, or complete regressions when tumors were grown in immune-competent animals. In accordance with this, YAP1 ASO treatment resulted in significant infiltration of T cells and myeloid lineage cells in the tumors in immune-competent mice. Importantly, when tumor-bearing animals were taken off the treatment after tumor regression was observed with YAP1 ASO, the antitumor response was sustained for more than 1 year, with 4 out of 16 animals remaining tumor-free in YAP1 ASO group. Furthermore, YAP1 ASO in combination with anti-PD1 antibody led to enhanced antitumor activity in the same tumor model. Collectively, these results suggest a potential therapeutic use of YAP1 ASO in the treatment of HCC with YAP1 activation as a single agent or in combination with immuno-oncology drugs. Broad screens have identified the human YAP1 ASO clinical development compound YAP1RX. Citation Format: Youngsoo Kim, Joanna Schmidt, Minji Jo, Jing Yi, Xiaolin Luo, Rachel Fleming, Sun Young Yim, Ju-Seog Lee, Brett P. Mornia, Randy L. Johnson, Gordon Mills, A. Robert MacLeod. Selective depletion of YAP1 with next-generation (constrained ethyl-cEt) antisense oligonucleotides results in tumor regression in mouse models of HCC with YAP1 activation [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr B09.

Minimally Invasive Repair of Ascending Aortic Pseudoaneurysms: An Alternative to Open Surgical Repair in High-Risk Patients

Development of a pseudoaneurysm of the ascending aorta is an uncommon complication of aortic surgery. Several nonsurgical techniques are available for treatment of ascending aortic pseudoaneurysms (AAPs). This report outlines a single-center retrospective experience with 14 nonsurgical procedures for treatment of AAPs in 10 patients. Modified stent grafts, septal defect occlusion devices, coil embolics, and liquid embolics were deployed by transthoracic and endovascular approaches. Complete stasis of the AAP was achieved in 7 of 10 patients (70%). Mean postprocedural recoveries occurred within 3.5 days. Nonsurgical techniques for repair of AAPs offer a comparatively safe and effective alternative to open surgical repair.

How do small birds evolve in response to climate change? Data from the long-term record at La Brea tar pits.

Biology textbooks describe the small changes in the beaks of the Galápagos finches as exemplars of how birds evolve in response to environmental changes. However, recent studies of the abundant fossil birds at Rancho La Brea find no evidence of evolutionary responses to the dramatic climate changes of the glacial-interglacial cycle over the past 35000 years: none of the large birds exhibit any change in body size or limb proportions, even during the last glacial maximum approximately 18000-20000 years ago, when the southern California chaparral was replaced by snowy coniferous forests. However, these are all large birds with large ranges and broad habitat preferences, capable of living in many different environments. Perhaps the smaller birds at La Brea, which have smaller home ranges and narrower habitats, might respond to climate more like Galápagos finches. The only 3 common small birds at La Brea are the western meadowlark, the yellow-billed magpie and the raven. In this study, we demonstrate that these birds also show complete stasis over the last glacial-interglacial cycle, with no statistically significant changes between dated pits. Recent research suggests that the small-scale changes over short timescales seen in the Galápagos finches are merely fluctuations around a stable morphology, and rarely lead to long-term accumulation of changes or speciation. Instead, the prevalence of stasis supports the view that long-term directional changes in morphology are quite rare. While directional changes in morphology occur frequently over short (<1ka) timescales, in the long term such changes only rarely remain stable for long enough to appear in the fossil record.

The efficacy of clove and thyme against experimentally induced Candidiasis in broilers

Objectives: The present study was designed to investigate the efficacy of clove powder and thyme oil against experimentally-induced candidiasis in broiler chicken. Design: Randomized controlled experimental study. Animals: One hundred and twenty five chicken of one month old were used in this study Procedures: Chicken were randomly allocated into 5 equal groups of 25 bird each. The first group was used as a negative control which received no treatment. The second group was experimentally infected with candida (C. albicans), and served as a positive control. Meanwhile, the third, fourth and fifth groups were also experimentally infected with candida, in addition to receiving clove powder (2 %), thyme oil (200 ml/kg) in diet and fluconazole (5 mg/kg) respectively, for 10 days after the appearance of the symptoms of candida. The symptoms included delayed gastrointestinal transit times, static crop or sour crop, anorexia and regurgitation. The normal thin wall of the crop become thickened with a complete crop stasis. Results: Administration of clove powder and thyme oil improved the symptoms induced by C. albicans and increased PCV %, ALT and AST level, with a decrease in erythrocyte count and total leukocyte count. Clove powder increased creatinine levels, while thyme oil decreased them. The tested plants induced marked histopathological alteration in crop, lung, intestine and gizzard of treated birds. Conclusion and clinical relevance: Clove powder and thyme oil can be used as an alternative medicine for the treatment of candidiasis in chicken.

High Density Lipoprotein Dysfunctionality in Venous Thromboembolism

Venous thromboembolism (VTE) is complex multi‐factorial cardiovascular disease that encompasses deep vein thrombosis (DVT) and pulmonary embolism (PE) and has a high incidence in several populations across the world with unknown pathophysiological mechanism driven by various environmental, acquired and inherited risk factors. DVT, a condition where a blood clot forms inside deeply situated veins in the body (usually in the lower limbs) is very common among Indian population. Lipoproteins modulate the expression and function of coagulation/fibrinolytic factors in patients of various cardiovascular diseases. HDL associated proteins have been proven to show antioxidant, anti‐inflammatory and anti‐thrombotic properties. Thus, the present study aims to examine the functionality of HDL in VTE using an animal model and also validating the same in DVT patients.MethodologyA complete stasis model was used for inducing thrombus in Sprague Dawley rats by ligating their Inferior Vena Cava (IVC). Animal experiments were divided into two groups: Ligated group (with IVC ligation) and SHAM Control group (without ligation). D‐Dimer and Prothrombin Fragment (PF) 1+2 levels were checked to confirm thrombus formation. Further, HDL‐associated proteins and other molecules were quantified using their respective ELISA kits. Similar tests were done in a cohort of human patients diagnosed with DVT (confirmed by a color Doppler test) and the results were compared with healthy individuals. All experiments were done in accordance with their respective ethical guidelines.ResultsD‐Dimer, PF 1+2 and inflammatory molecules were significantly higher in the ligated group, confirming the formation of thrombus. HDL associated proteins (APO‐A1, PON‐1, PAF‐AH &amp; S‐1‐P) were significantly lower whereas oxidized lipoproteins (Ox‐HDL and Ox‐LDL) and lipooxygenase enzymes (15/12‐LOX) were found to be higher in the ligated group as compared to the SHAM control group. DVT patients showed a similar trend of D‐Dimer and PF 1+2 as in animals, when compared to healthy individuals. Lipid profile (Total cholesterol, Triglycerides, LDL and HDL) of patients did not show any significant changes albeit levels of HDL associated proteins were lower than those in healthy volunteers, wherein oxidized lipoproteins were significantly higher. A significance level (p‐value) of &lt;0.05 was used for all the tests.Conclusion and DiscussionPositive D‐Dimer test, high PF 1+2 and inflammatory molecules in DVT patients &amp; in the ligated group of animals, confirm the biochemical changes that occur during thrombosis. Low levels of HDL associated proteins and high levels of lipooxygenase enzymes, responsible for lipoprotein oxidation (proven by high levels of oxidised lipoproteins) support the dysfunctional HDL hypothesis in case of VTE. These findings give more insight in understanding the pathophysiology of VTE and might help in disease prevention/diagnosis/treatment.Support or Funding InformationDefence Research and Development Organisation

Left Gastric Artery Embolization for Weight Loss

Abstract Purpose To conduct a prospective clinical trial to evaluate the feasibility, safety and efficacy of left gastric artery embolization as a minimally-invasive way for the treatment of obesity. Materials &amp; Methods Patients were recruited with respect to detailed inclusion and exclusion criteria. A baseline clinical history taking and physical examination including anthropometric measurements was undertaken. A baseline EGD was performed to rule out any pre-existing pathology in the esophagus, stomach or duodenum. Baseline serum Ghrelin level was assessed. Embolization was performed using non-spherical polyvinyl alcohol particles sized 355–500 µm (Bearing nsPVA®, Merit Medical) until complete stasis of flow within the LGA and its branches. Patients were followed up weekly for 1 month then monthly for 6 months by anthropometric measurements by the clinical nutritionists from the Physical Medicine department. At 1 month, EGD was repeated to assess for any complications or adverse effects. Another blood sample was withdrawn to assess the change in serum Ghrelin level. Results Left gastric artery embolization was performed on six obese to morbidly obese patients (four women and two men; average age, 44 years; range, 24–62 years), with a mean weight of 113.67 kg (range 86–133 kg), a mean BMI of 44.45 kg/m2 (range 33.6–51.2 kg/m2) Technical success rate was 100%. There was one minor adverse event in the form of a small asymptomatic superficial gastritis in the fundus/lesser curvature. There were no major adverse events. Mean excess weight loss of 5.58% and 2.91% was noted at 1 month and at 6 months, respectively. Mean change in serum ghrelin was −35.57% at 1 month. Conclusion LGA embolization is technically feasible and safe. There is considerable drop in serum ghrelin level at 1 month. However, the weight loss appears to be transient. Future studies can opt for smaller-sized particles and probably embolization of the right gastro-epiploic artery together with the left gastric artery. Abbreviations BMI = Body Mass Index, EGD = Esophagogastroduodenoscopy, LGA = Left Gastric Artery, NSAID = Non-steroidal anti-inflammatory drugs, PPI = Proton pump inhibitors, WHO = World Health Organization.

Acute experimental venous thrombosis impairs venous relaxation but not contraction

ObjectiveVenous thrombosis (VT) damages the vein wall, both physically by prolonged distension and from inflammation. These factors contribute to post-thrombotic syndrome (PTS). Interleukin (IL)-6 might play a role in experimental PTS and vein wall responses. Previous assessments of post-thrombotic vein wall injury used static measures such as histologic examination and immunologic assays. The purpose of the present study was to use myography to quantify the changes in contraction and relaxation of murine vessels exposed to an acute VT. MethodsWild-type (WT) C57BL/6 mice were used to determine the baseline vein wall passive tension on a DMT 610m myograph (DMT-USA, Inc., Ann Arbor, Mich), including dosing concentrations of phenylephrine (Phe) and acetylcholine (Ach). WT and IL-6−/− mice underwent VT using inferior vena cava (IVC) ligation (complete stasis) and stenosis (partial stasis), with no-surgery mice used as controls. The mice were harvested at 2 days (2D) and analyzed using a myograph. The vessels were stimulated with Phe and Ach to stimulate a contraction and relaxation response. The endothelial responses to VT were quantified by CD31 immunohistochemistry, Greiss assay, polymerase chain reaction, and Evans blue assay. ResultsOptimal passive tension was determined to be 2 mN, with an optimal concentration of Phe and Ach of 7E-3M and 1E-5M, respectively. No significant differences were found in the contractions when exposed to Phe between the WT control, WT 2D ligation, and WT 2D stenosis IVC segments and the IL-6−/− mice with and without thrombus (P > .05 for all). When treated with Ach, significantly more relaxation was found in the nonthrombosed control IVC segments than in those IVC segments that had had a 2D thrombus from either ligation- or stenosis-derived thrombotic mechanisms in both WT and IL-6−/− mice. CD31 staining showed ∼20% less luminal endothelium after stasis thrombosis (P ≤ .01) but no loss in the controls (P > .05). Evans blue staining showed a trend toward increased leakiness in post-thrombotic vein walls. No significant difference in the endothelial gene markers or nitric oxide production was found. ConclusionsCompared with the controls, acute thrombosis in the total or partial stasis models did not impair IVC contractile responses, suggesting no effect on the medial vascular smooth muscle response. The relaxation response was significantly reduced in the post-thrombotic groups, likely from direct endothelial injury. These findings suggest, at acute points, that VT impairs the endothelial function of a vein wall while retaining the vascular smooth muscle cell function and might be a mechanism that promotes PTS.

Impaired Inferior Vena Cava Relaxation but not Contraction in Two Models of Murine Acute Deep Venous Thrombosis

Venous thrombosis (VT) damages the vein wall, both physically with prolonged distention and from inflammation. These factors underlie post-thrombotic syndrome (PTS). Interleukin 6 (IL-6) may play a role in experimental PTS and vein wall responses. Prior assessment of post-thrombotic vein wall injury has used static measures such as histology and immunologic assays. The purpose of this study was to use myography to quantify the changes in contraction and relaxation of murine vessels exposed to an acute thrombus. Wild-type (WT) C57BL/6 mice were used to determine baseline vein wall passive tension on a DMT 610M myograph (DMT, Ann Arbor, Mich), including dosing concentrations of phenylephrine (Phe) and acetylcholine (Ach). WT and IL-6 knockout mice underwent VT using inferior vena cava (IVC) ligation (complete stasis) and stenosis (partial stasis), and no-surgery mice were used as controls. Mice were harvested at 2 days (2D) and analyzed using a myograph. Vessels were stimulated with Phe and Ach to stimulate a contraction and relaxation response. Immunohistochemistry for CD31 was used to identify endothelial cells on WT no-surgery and WT 2D ligation vessels. Optimal passive tension was determined to be 2 mN, with the optimal concentrations of Phe and Ach being 7E-3M and 1E-5M, respectively. There was no significant difference in contraction on exposure to Phe between the WT no-thrombus mice, WT 2D ligation, and WT 2D stenosis mice and the IL-6 knockout mice with or without thrombus (all P > .05). When mice were treated with Ach, there was significantly more relaxation in the nonthrombosed control IVC segments than in those mice IVC segments that had a 2D thrombus of either ligation- or stenosis-derived mechanisms (Fig). The CD31 staining confirmed the presence of endothelium in the WT 2D IVC thrombus mice, ruling out tissue lost during harvesting. Compared with controls, acute thrombosis with total or partial stasis did not impair IVC contractile responses, suggesting no effect on medial vascular smooth muscle response. The relaxation response, however, was significantly reduced in the post-thrombotic groups. These findings suggest that at acute time points, VT impairs the endothelial function of a vein wall while retaining the vascular smooth muscle cell function and may be a mechanism that promotes PTS.

3:09 PM Abstract No. 132 Complete stasis embolization may convey survival benefit in hepatocellular carcinoma patients treated with doxorubicin eluting bead transcatheter arterial chemoembolization

3:09 PM Abstract No. 132 Complete stasis embolization may convey survival benefit in hepatocellular carcinoma patients treated with doxorubicin eluting bead transcatheter arterial chemoembolization

Abstract B140: PT2977, a novel HIF-2a antagonist, has potent antitumor activity and remodels the immunosuppressive tumor microenvironment in clear cell renal cell cancer

Abstract Hypoxia-inducible factor 2a (HIF-2a), a transcription factor, has been established as an oncogenic driver in clear cell renal cell cancer (ccRCC). The first HIF-2a antagonist being evaluated in clinical development, PT2385, has demonstrated clinical activity in ccRCC patients who had previously been treated with multiple lines of therapy. There is continuing effort to characterize additional HIF-2a antagonists possessing attributes that may contribute to enhanced clinical activity. PT2977 is a novel HIF-2a antagonist with improved potency in preclinical tumor models compared to PT2385. This improvement arises from enhanced biochemical and cellular potency, an improvement in plasma protein binding, and diminished metabolic clearance in vivo relative to PT2385. PT2977 exhibits favorable metabolic stability and pharmacokinetic characteristics when dosed orally in multiple preclinical species. Allometric scaling of the preclinical data predicts PT2977 to be suitable for oral once-daily dosing in humans. PT2977 inhibits expression of HIF-2a target genes in tumor cells and induces complete stasis or regression in ccRCC xenografts. A strong pharmacokinetics/pharmacodynamics correlation is observed in tumors from xenograft models treated with PT2977. Gene expression analyses of ccRCC xenografts treated with PT2977 reveal extensive modulation of genes in the tumor cells as well as in immune cells. Immune phenotyping of tumors treated with PT2977 confirms that HIF-2a antagonism results in a reduction in the number of immunosuppressive myeloid-derived cells, including neutrophils and macrophages. Treatment with PT2977 also results in an influx of mature dendritic cells. These observations are consistent with HIF-2a exerting an immunosuppressive effect on the tumor microenvironment, in addition to driving angiogenesis and the proliferation and viability of tumor cells. With its favorable preclinical profile, PT2977 is well positioned to further reveal the broader therapeutic potential of HIF-2a antagonism for the treatment of cancer, as a single agent or in combination with other immune-modulating agents. Citation Format: Tai W. Wong, Rajeev Shrimali, Cristina Contreras, Tzuling Cheng, Robert M. Czerwinski, Daryl D. Dixon, Xinlin Du, Craig Fett, Jessica Goree, Jonas A. Grina, Guangzhou Han, Heli Huang, Jim Rizzi, Stephen T. Schlachter, Bin Wang, Keshi Wang, Paul M. Wehn, Shanhai Xie, Rui Xu, Hanbiao Yang, John A. Josey, Eli M. Wallace. PT2977, a novel HIF-2a antagonist, has potent antitumor activity and remodels the immunosuppressive tumor microenvironment in clear cell renal cell cancer [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr B140.

Duodenal Ulceration as a Complication of Yttrium-90 Microsphere Radioembolization of Intrahepatic Cholangiocarcinoma

Yttrium-90 (Y-90) microsphere radioembolization (RE) can be an effective regional treatment for primary hepatobiliary malignancies. RE with Y-90 has been described as safe for patients with intrahepatic cholangiocarcinoma (ICC)1. However, gastrointestinal ulceration is a rarely reported complication of Y-90 microsphere RE2. Given the increasing incidence of primary hepatobiliary tumors and an expected rise in use of Y-90 RE, we present a case of Y-90 RE-associated duodenal ulceration. A 76-year-old woman presented to the hospital with lower abdominal pain and was evaluated with a CT scan which revealed an ill-defined liver mass adjacent to the gallbladder which was diagnosed as stage IV intrahepatic cholangiocarcinoma involving segment 4 of the liver. She was treated with Y-90 RE with the intent of tumor down-sizing prior to resection. As part of the treatment application, the gastroduodenal artery was densely packed with coils with complete stasis of flow achieved. Through the medial branch of the left hepatic artery, the Y-90 spheres were super-selectively deployed through a micro-catheter. Two months after the radioembolization, she experienced mid-epigastric abdominal pain, not responsive to proton pump inhibitor. Upper endoscopy revealed two non-bleeding superficial duodenal ulcers with no stigmata of bleeding in the duodenal bulb. Pathology from biopsies of the ulcer bed revealed the presence of Y-90 beads in the lamina propria, consistent with radiation-induced duodenitis. She is currently being treated with high dose proton pump inhibitor (PPI) therapy by mouth twice daily, with good response. This case highlights a potential complication associated with radioactive microsphere embolization. Given the increasing use of this treatment modality, gastroenterologists should recognize this entity, and its delayed occurrence in particular. New-onset, or change in abdominal discomfort or pain should prompt endoscopic evaluation in patients with prior Y-90 RE. We suggest high dose PPI therapy as initial management, but the optimal treatment strategy to treat gastrointestinal ulceration as a complication of RE should be further investigated.