Significant Reduction In Lesion Size Research Articles

Analysis of middle meningeal artery embolization for the treatment of chronic, acute on chronic, and subacute subdural hematomas.

Chronic subdural hematoma (cSDH) is a common sequela of traumatic brain injury. Middle meningeal artery embolization (MMAE) has shown promising results as an emerging minimally invasive alternative treatment. The purpose of this study is to examine the safety and efficacy of MMAE performed in patients with cSDH, acute-on-chronic, and subacute SDH with a traumatic etiology. This retrospective study included cases performed at a Level II Trauma Center between January 2019 and December 2020 for MMAE of cSDHs. Data collected included patient demographic characteristics and comorbidities, SDH characteristics, complications, and efficacy outcomes. The lesion measurements were collected before the procedure, 4-6 weeks and 3-6 months post-procedure. In our patient population, 78% (39) either had lesions improve or completely resolved. The sample included 50 patients with a mean age of 74 years old. Statistically significant reductions in lesion size were found from pre- to post-procedure in the left lesions, right lesions, and midline shifts. The left lesions decreased from 13.88 ± 5.70 mm to 3.19 ± 4.89 mm at 3-6 months with P < 0.001. The right lesions decreased from 13.74 ± 5.28 mm to 4.93 ± 7.46 mm at 3-6 months with P = 0.02. Midline shifts decreased from 3.78 ± 3.98 mm to 0.48 ± 1.31 mm at 3-6 months with P = 0.02. No complications were experienced for bleeding, hematoma, worsening SDH, pseudoaneurysm, or stroke. Our pilot study from a single center utilizing MMAE demonstrates that MMAE is successful without increasing treatment-related complications not only for cSDH but also in acute-on-cSDH and SDH with a subacute component.

Mammalian target of rapamycin inhibitors: A new-possible approach for in-utero medication therapy.

The mammalian/mechanistic target of rapamycin (mTOR) is a protein kinase that plays a crucial role in regulating cellular growth, metabolism, and survival. Although there is no absolute contraindication for the use of mTOR inhibitors during pregnancy, the specific fetal effects remain unknown. Available data from the past 2decades have examined the use of mTOR inhibitors during pregnancy in patients with solid organ transplantation, showing no clear link to fetal complications or structural abnormalities. Recently, a handful of case reports and series have described transplacental therapy of mTOR inhibitors to control symptomatic and complicated pathologies in the fetus. The effect of these agents includes a significant reduction in lesion size in the fetus and a reduced need for mechanical ventilation in neonates. In this context, we delve into the potential of mTOR inhibitors as in-utero therapy for fetal abnormalities, with a primary focus on lymphatic malformation (LM) and cardiac rhabdomyoma (CR). While preliminary reports underscore the efficacy of mTOR inhibitors for the treatment of fetal CR and fetal brain lesions associated with tuberous sclerosis complex, chylothorax, and LMs, additional investigation and clinical trials are essential to comprehensively assess the safety and efficacy of these medications.

Amphotericin B is usually underdosed in the treatment of experimental cutaneous leishmaniasis

Introduction: Amphotericin B is an effective drug for the treatment of the different clinical forms of leishmaniasis. However, there are reports of its ineffectiveness in animals experimentally infected with Leishmania spp. That is why, the objective of the present work was to evaluate the balance of activity-toxicity at amphotericin B doses over 1 mg/kg, so that its use as a positive control antileishmanial drug were adequate.&#x0D; Method: BALB/c mice were experimentally infected with L. amazonensis and treated with amphotericin B by intraperitoneal route at doses from 5 mg/kg to 12.5 mg/kg, beginning 21 days after infection. The size of the lesions and the body weight of the mice were measured for eleven weeks after the commencement of treatment. The number of parasites was also determined three days after the end of treatment.&#x0D; Results: Amphotericin B at 5 mg/kg retarded lesions growth but neither reduced lesion size nor the parasite load at lesion site. Doses of 7.5 mg/kg to 10 mg/kg, every 48 h for 14 days (7 doses) caused a significant reduction of lesion size and parasite load without evident loss of body weight and without signs of toxicity. Amphotericin B at 12.5 mg/kg was more effective but produced unacceptable toxicity.&#x0D; Conclusions: The results support the use of amphotericin B as a positive control drug in BALB/c mice experimentally infected with L. amazonensis at doses of 7.5 mg/kg to 10 mg/kg to achieve an effect comparable to that observed in clinical practice.

Human pluripotent stem cell-derived ectomesenchymal stromal cells promote more robust functional recovery than umbilical cord-derived mesenchymal stromal cells after hypoxic-ischaemic brain damage.

Aims: Hypoxic-ischaemic encephalopathy (HIE) is one of the most serious complications in neonates and infants. Mesenchymal stromal cell (MSC)-based therapy is emerging as a promising treatment avenue for HIE. However, despite its enormous potential, the clinical application of MSCs is limited by cell heterogeneity, low isolation efficiency and unpredictable effectiveness. In this study, we examined the therapeutic effects and underlying mechanisms of human pluripotent stem cell-derived ectomesenchymal stromal cells (hPSC-EMSCs) in a rat model of HIE.Methods: hPSC-EMSCs were induced from either human embryonic stem cells or induced pluripotent stem cells. Stem cells or the conditioned medium (CM) derived from stem cells were delivered intracranially or intranasally to neonatal rats with HIE. Human umbilical cord-derived MSCs (hUC-MSCs) were used as the therapeutic comparison control and phosphate-buffered saline (PBS) was used as a negative control. Lesion size, apoptosis, neurogenesis, astrogliosis and microgliosis were evaluated. The rotarod test and Morris water maze were used to determine brain functional recovery. The PC-12 cell line, rat primary cortical neurons and neural progenitor cells were used to evaluate neurite outgrowth and the neuroprotective and neurogenesis effects of hPSC-EMSCs/hUC-MSCs. RNA-seq and enzyme-linked immunosorbent assays were used to determine the secretory factors that were differentially expressed between hPSC-EMSCs and hUC-MSCs. The activation and suppression of extracellular signal-regulated kinase (ERK) and cAMP response element-binding protein (CREB) were characterised using western blotting and immunofluorescent staining.Results: hPSC-EMSCs showed a higher neuroprotective potential than hUC-MSCs, as demonstrated by a more significant reduction in lesion size and apoptosis in the rat brain following hypoxia-ischaemia (HI). Compared with PBS treatment, hPSC-EMSCs promoted endogenous neurogenesis and alleviated astrogliosis and microgliosis. hPSC-EMSCs were more effective than hUC-MSCs. hPSC-EMSCs achieved a greater recovery of brain function than hUC-MSCs and PBS in rats with HIE. CM derived from hPSC-EMSCs had neuroprotective and neurorestorative effects in vitro through anti-apoptotic and neurite outgrowth- and neurogenesis-promoting effects. Direct comparisons between hPSC-EMSCs and hUC-MSCs revealed the significant enrichment of a group of secretory factors in hPSC-EMSCs, including nerve growth factor (NGF), platelet-derived growth factor-AA and transforming growth factor-β2, which are involved in neurogenesis, synaptic transmission and neurotransmitter transport, respectively. Mechanistically, the CM derived from hPSC-EMSCs was found to potentiate NGF-induced neurite outgrowth and the neuronal differentiation of NPCs via the ERK/CREB pathway. Suppression of ERK or CREB abolished CM-potentiated neuritogenesis and neuronal differentiation. Finally, intranasal delivery of the CM derived from hPSC-EMSCs significantly reduced brain lesion size, promoted endogenous neurogenesis, mitigated inflammatory responses and improved functional recovery in rats with HIE.Conclusion: hPSC-EMSCs promote functional recovery after HI through multifaceted neuromodulatory activities via paracrine/trophic mechanisms. We propose the use of hPSC-EMSCs for the treatment of HIE, as they offer an excellent unlimited cellular source of MSCs.

PFKFB3 gene deletion in endothelial cells inhibits intraplaque angiogenesis and lesion formation in a murine model of venous bypass grafting

Vein grafting is a frequently used surgical intervention for cardiac revascularization. However, vein grafts display regions with intraplaque (IP) angiogenesis, which promotes atherogenesis and formation of unstable plaques. Graft neovessels are mainly composed of endothelial cells (ECs) that largely depend on glycolysis for migration and proliferation. In the present study, we aimed to investigate whether loss of the glycolytic flux enzyme phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3) in ECs inhibits IP angiogenesis and as such prevents unstable plaque formation. To this end, apolipoprotein E deficient (ApoE−/−) mice were backcrossed to a previously generated PFKFB3fl/fl Cdh5iCre mouse strain. Animals were injected with either corn oil (ApoE−/−PFKFB3fl/fl) or tamoxifen (ApoE−/−PFKFB3ECKO), and were fed a western-type diet for 4 weeks prior to vein grafting. Hereafter, mice received a western diet for an additional 28 days and were then sacrificed for graft assessment. Size and thickness of vein graft lesions decreased by 35 and 32%, respectively, in ApoE−/−PFKFB3ECKO mice compared to controls, while stenosis diminished by 23%. Moreover, vein graft lesions in ApoE−/−PFKFB3ECKO mice showed a significant reduction in macrophage infiltration (29%), number of neovessels (62%), and hemorrhages (86%). EC-specific PFKFB3 deletion did not show obvious adverse effects or changes in general metabolism. Interestingly, RT-PCR showed an increased M2 macrophage signature in vein grafts from ApoE−/−PFKFB3ECKO mice. Altogether, EC-specific PFKFB3 gene deletion leads to a significant reduction in lesion size, IP angiogenesis, and hemorrhagic complications in vein grafts. This study demonstrates that inhibition of endothelial glycolysis is a promising therapeutic strategy to slow down plaque progression.

Transcriptome analysis of scions grafted to potato rootstock for improving late blight resistance

BackgroundLate blight seriously threatens potato cultivation worldwide. The severe and widespread damage caused by the fungal pathogen can lead to drastic decreases in potato yield. Although grafting technology has been widely used to improve crop resistance, the effects of grafting on potato late blight resistance as well as the associated molecular mechanisms remain unclear. Therefore, we performed RNA transcriptome sequencing analysis and the late blight resistance testing of the scion when the potato late blight-resistant variety Qingshu 9 and the susceptible variety Favorita were used as the rootstock and scion, respectively, and vice versa. The objective of this study was to evaluate the influence of the rootstock on scion disease resistance and to clarify the related molecular mechanisms.ResultsA Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that the expression levels of genes related to plant–pathogen interactions, plant mitogen-activated protein kinase (MAPK) signaling pathways, and plant hormone signal transduction pathways were significantly up-regulated in the scion when Qingshu 9 was used as the rootstock. Some of these genes encoded calcium-dependent protein kinases (CDPKs), chitin elicitor receptor kinases (CERKs), LRR receptor serine/threonine protein kinases (LRR-LRKs), NPR family proteins in the salicylic acid synthesis pathway, and MAPKs which were potato late blight response proteins. When Favorita was used as the rootstock, only a few genes of late blight response genes were upregulated in the scion of Qingshu 9. Grafted plants using resistant variety as rootstocks inoculated with P. infestans spores showed significant reductions in lesion size while no significant difference in lesion size was observed when susceptible variety was used as the rootstock. We also showed that this induction of disease resistance in scions, especially scions derived from susceptible potato varieties was mediated by the up-regulation of expression of genes involved in plant disease resistance in scions.ConclusionsOur results showed that potato grafting using late blight resistant varieties as rootstocks could render or enhance resistance to late blight in scions derived from susceptible varieties via up-regulating the expression of disease resistant genes in scions. The results provide the basis for exploring the molecular mechanism underlying the effects of rootstocks on scion disease resistance.

Efficacy of histamine H1 receptor antagonists azelastine and fexofenadine against cutaneous Leishmania majorinfection.

Current drug therapies for cutaneous leishmaniasis are often difficult to administer and treatment failure is an increasingly common occurrence. The efficacy of anti-leishmanial therapy relies on a combination of anti-parasite activity of drugs and the patient’s immune response. Previous studies have reported in vitro antimicrobial activity of histamine 1-receptor antagonists (H1RAs) against different pathogens. We used an ex vivo explant culture of lymph nodes from mice infected with Leishmania major to screen H1RAs compounds. Azelastine (AZ) and Fexofenadine (FX) showed remarkable ex vivo efficacy (EC50 = 0.05 and 1.50 μM respectively) and low in vitro cytotoxicity yielding a high therapeutic index. AZ significantly decreased the expression of H1R and the proinflammatory cytokine IL-1ẞ in the ex vivo system, which were shown to be augmented by histamine addition. The anti-leishmanial efficacy of AZ was enhanced in the presence of T cells from infected mice suggesting an immune-modulatory mechanism of parasite suppression. L. major infected BALB/c mice treated per os with FX or intralesionally with AZ showed a significant reduction of lesion size (FX = 69%; AZ = 52%). Furthermore, there was significant parasite suppression in the lesion (FX = 82%; AZ = 87%) and lymph nodes (FX = 81%; AZ = 36%) with no observable side effects. AZ and FX and potentially other H1RAs are good candidates for assessing efficacy in larger studies as monotherapies or in combination with current anti-leishmanial drugs to treat cutaneous leishmaniasis.

Inhibition of choroidal neovascularization by systemic delivery of gold nanoparticles

Choroidal neovascularization (CNV) is the abnormal growth of blood vessels that sprout from the choroid vasculature and grow beneath and into the retina. The newly formed blood vessels in CNV often leak blood and fluid which deteriorates vision over time, eventually leading to blindness. In the present study, we examined the efficacy of intravenously injected gold nanoparticles in the laser-induced CNV animal model. Using optical coherence tomography (OCT) and fluorescein angiography, we evaluated CNV lesions longitudinally, over a period of 21 days, with and without nanoparticle treatment. Intravenously injected low concentration of bare gold nanoparticles showed significant anti-angiogenic properties by suppressing CNV development and progression. The treatment group showed significantly decreased fluorescein leakage at the CNV site compared to vehicle injected control mice. OCT assisted CNV volume measurement at all time points showed a significant reduction in lesion size in the treatment group compared with controls.

Topical Recombinant Human Nerve Growth Factor (Cenegermin) for Neurotrophic Keratopathy: A Multicenter Randomized Vehicle-Controlled Pivotal Trial

To evaluate the efficacy and safety of topical cenegermin (recombinant human nerve growth factor) in patients with neurotrophic keratopathy. Multicenter, randomized, double-masked, vehicle-controlled trial. Patients with neurotrophic persistent epithelial defect with or without stromal thinning. The NGF0214 trial, conducted among 11 sites in the United States, randomized 48 patients 1:1 to cenegermin 20 μg/ml or vehicle eye drops, 6 drops daily for 8 weeks of masked treatment. Follow-up was 24 weeks. Safety was assessed in all patients who received study drug. Efficacy was assessed by intention to treat. The primary end point was healing of the neurotrophic lesion (persistent epithelial defect or corneal ulcer) after 8 weeks of masked treatment. Masked central readers measured neurotrophic lesions in randomized clinical pictures, then assessed healing status conventionally (<0.5 mm of fluorescein staining in the greatest dimension of the lesion area) and conservatively (0-mm lesion staining and no other residual staining). Secondary variables included corneal healing at 4 weeks of masked treatment (key secondary end point), overall changes in lesion size, rates of disease progression, and changes in visual acuity and corneal sensitivity from baseline to week8. Conventional assessment of corneal healing showed statistically significant differences at week 8: compared to 7 of 24 vehicle-treated patients (29.2%), 16 of 23 cenegermin-treated patients (69.6%) achieved less than 0.5 mm of lesion staining (+40.4%; 95% confidence interval [CI], 14.2%-66.6%; P= 0.006). Conservative assessment of corneal healing also reached statistical significance at week 8: compared to 4 of 24 vehicle-treated patients (16.7%), 15 of 23 cenegermin-treated patients (65.2%) achieved 0 mm of lesion staining and no other residual staining (+48.6%; 95% CI, 24.0%-73.1%; P < 0.001). Moreover, the conservative measure of corneal healing showed statistical significance at week 4 (key secondary end point). Compared to vehicle, cenegermin-treated patients showed statistically significant reductions in lesion size and disease progression rates during masked treatment. Cenegermin was well tolerated; adverse effects were mostly local, mild, and transient. Cenegermin treatment showed higher rates of corneal healing than vehicle in neurotrophic keratopathy associated with nonhealing corneal defects.

The combination therapy of meglumine antimoniate and oxiranes (epoxy-α-lapachone and epoxymethyl-lawsone) enhance the leishmanicidal effect in mice infected by Leishmania (Leishmania) amazonensis

Current treatment of cutaneous leishmaniasis includes pentavalent antimonials as first-line drugs, but this therapy has shown severe adverse effects. An alternative to minimize this issue is based on combination therapy scheme with other drugs. In this study we analyzed the potential of the association of meglumine antimoniate (MA) with the oxiranes epoxy-α-lapachone (LAP) or epoxymethyl-lawsone (LAW). Results demonstrated that association between these drugs enhanced leishmanicidal activity on Leishmania (Leishmania) amazonensis infection. The compounds were tested in monotherapy or in combinations (3:1; 1:1 and 1:3) and reduced intracellular parasite numbers, measured by the endocytic index, in all tested conditions. The most effective combination regimens were MA/LAP or MA/LAW in 3:1 ratio, which achieved a reduction of 98.3% and 93.6% in the endocytic index, respectively. BALB/c mice challenged with L. (L.) amazonensis showed significant reduction in lesion size and parasite load in both footpad and lymph nodes, after four weeks of treatment. Although, MA, LAP or LAW monotherapy were able to control the evolution of lesions when compared to untreated animals (30%, 40% and 40% of reduction, respectively), the combination of MA/LAP and LAW in 3:1 ratio showed better results reducing 61.7 and 54.4%, respectively. The results indicate that the association of meglumine antimoniate to oxiranes lead to an increment in the antileishmanial activity and represent a promising approach for the cutaneous leishmaniasis treatment.

MARK4 (Microtubule Affinity-Regulating Kinase 4)-Dependent Inflammasome Activation Promotes Atherosclerosis-Brief Report.

MARK4 (microtubule affinity-regulating kinase 4) regulates NLRP3 (nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain containing 3) inflammasome activation. The aim of the study is to examine the role of MARK4 in hematopoietic cells during atherosclerosis. We show increased MARK4 expression in human atherosclerotic lesions compared with adjacent areas. MARK4 is coexpressed with NLRP3, and they colocalize in areas enriched in CD68-positive but α-SMA (α-smooth muscle actin)-negative cells. Expression of MARK4 and NLRP3 in the atherosclerotic lesions is associated with the production of active IL (interleukin)-1β and IL-18. To directly assess the role of hematopoietic MARK4 in NLRP3 inflammasome activation and atherosclerotic plaque formation, Ldlr (low-density lipoprotein receptor)-deficient mice were lethally irradiated and reconstituted with either wild-type or Mark4-deficient bone marrow cells, and were subsequently fed a high-fat diet and cholesterol diet for 9 weeks. Mark4 deficiency in bone marrow cells led to a significant reduction of lesion size, together with decreased circulating levels of IL-18 and IFN-γ (interferon-γ). Furthermore, Mark4 deficiency in primary murine bone marrow-derived macrophages prevented cholesterol crystal-induced NLRP3 inflammasome activation, as revealed by reduced caspase-1 activity together with reduced production of IL-1β and IL-18. MARK4-dependent NLRP3 inflammasome activation in the hematopoietic cells regulates the development of atherosclerosis.

Synergy Between Peroxisome Proliferator-Activated Receptor γ Agonist and Radiation Therapy in Cancer

Approximately 60% of newly diagnosed cancer patients receive radiotherapy to accomplish neoadjuvant, definitive, adjuvant or palliative purpose as required. However, radiotherapy may trigger host-mediated proangiogenic and proinflammatory effects that lead to tumor recurrence. Angiogenesis and inflammation are crucial processes, through which tumor microenvironment (TME) influences tumor growth, invasiveness and metastasis. Peroxisome proliferator-activated receptor γ (PPARγ) agonist rosiglitazone has anti-inflammatory, antiangiogenic and antineoplastic activities from preclinical data. We hypothesize that rosiglitazone and radiation therapy synergistically amplify the antitumor effect, which is based on the evidence that rosiglitazone remodels tumor vasculature and limits tumor-associated macrophages (TAMs) infiltration. The aim of this study is to evaluate the synergistic effect of PPARγ agonist rosiglitazone combined with radiation therapy. Each female Balb/c mouse was inoculated with 5X105 murine colon carcinoma cell line CT26 or breast cancer 4T1. When tumor size approximately reached 250-300 mm3, 12Gy in a fraction as hypofractionated irradiation therapy (HFRT) or 12Gy in 4 daily fractions of 3Gy as conventional fractionated irradiation therapy (CFRT) was delivered to the tumor, meanwhile administration of 100mg/kg rosiglitazone was started daily for 2 weeks. Tumors were harvested according to the schedule. Flow cytometry, ELISA, immunofluorescence, western blot, and so forth were performed to test the synergistic mechanisms. We presented that rosiglitazone has antiangiogenic effect by inhibiting VEGF-VEGFR2 signaling pathways. Immunofluorescence work identified that rosiglitazone can remodel tumor vasculature contributing to decrease of microvessel density, promotion of pericyte coverage, reduction of TAMs infiltration, and even further accompanied improvement of intratumoral hypoxia and vessel perfusion compared with the control (P<0.05). 4T1 or CT26 tumor-bearing mice treated with rosiglitazone in combination with HFRT or CFRT showed a significant reduction in lesion size and lung metastases compared with the control (P<0.001), rosiglitazone(P<0.001) or radiotherapy group (P<0.05). Analysis of ELISA, immunofluorescence and flow cytometry showed that tumor-derived chemokine CCL2 in reaction to HFRT facilitates the recruitment of migrating CD11b+ myeloid monocytes and TAMs into irradiated sites that initiates vasculogenesis and allows tumor recurrence after HFRT. We found that rosiglitazone decreased CCL2 and suppressed CD11b+ myeloid monocytes and TAMs infiltrations after HFRT, which delayed tumor recurrence. Rosiglitazone may elicit normalization of tumor vasculature and limit TAMs infiltration, and thereby delay tumor progression with a fresh perspective. Rosiglitazone, as an adjuvant therapy to HFRT, inhibits CCL2 mediating tumor recurrence.

Synergy between peroxisome proliferator-activated receptor γ agonist and radiotherapy in cancer.

Angiogenesis and inflammation are crucial processes through which the tumor microenvironment (TME) influences tumor progression. In this study, we showed that peroxisome proliferator‐activated receptor γ (PPARγ) is not only expressed in CT26 and 4T1 tumor cell lines but also in cells of TME, including endothelial cells and tumor‐associated macrophages (TAM). In addition, we showed that rosiglitazone may induce tumor vessel normalization and reduce TAM infiltration. Additionally, 4T1 and CT26 tumor‐bearing mice treated with rosiglitazone in combination with radiotherapy showed a significant reduction in lesion size and lung metastasis. We reported that a single dose of 12 Gy irradiation strongly inhibits local tumor angiogenesis. Secretion of C‐C motif chemokine ligand 2 (CCL2) in response to local irradiation facilitates the recruitment of migrating CD11b+ myeloid monocytes and TAM to irradiated sites that initiate vasculogenesis and enable tumor recurrence after radiotherapy. We found that rosiglitazone partially decreases CCL2 secretion by tumor cells and reduces the infiltration of CD11b+ myeloid monocytes and TAM to irradiated tumors, thereby delaying tumor regrowth after radiotherapy. Therefore, combination of the PPARγ agonist rosiglitazone with radiotherapy enhances the effectiveness of radiotherapy to improve local tumor control, decrease distant metastasis risks and delay tumor recurrence.

Efficacy of Paromomycin-Chloroquine Combination Therapy in Experimental Cutaneous Leishmaniasis.

ABSTRACTThe 4-aminoquinoline chloroquine (CQ) is clinically used in combination with doxycycline to cure chronic Q fever, as it enhances the activity of the antibiotic against the causative bacterium Coxiella burnetii residing within macrophage phagolysosomes. As there is a similar cellular host-pathogen biology for Leishmania parasites, this study aimed to determine whether such an approach could also be the basis for a new, improved treatment for cutaneous leishmaniasis (CL). We have evaluated the in vitro and in vivo activities of combinations of CQ with the standard drugs paromomycin (PM), miltefosine, and amphotericin B against Leishmania major and Leishmania mexicana. In 72-h intracellular antileishmanial assays, outcomes were variable for different drugs. Significantly, the addition of 10 μM CQ to PM reduced 50% effective concentrations (EC50s) by over 5-fold against L. major and against normally insensitive L. mexicana parasites. In murine models of L. major and L. mexicana CL, daily coadministration of 50 mg/kg of body weight PM and 25 mg/kg CQ for 10 days resulted in a significant reduction in lesion size but not in parasite load compared to those for mice given the same doses of PM alone. Overall, our data indicate that PM-CQ combination therapy is unlikely to be a potential candidate for further preclinical development.

Abstract 162: A prodrug of green tea polyphenol (-)-epigallocatechin-3-gallate served as a potentially novel angiogenesis inhibitor in endometrioid endometrial cancer

Abstract Background and aim: Anti-angiogenesis is a promising treatment in endometrioid endometrial cancer (EEC) while the present anti-angiogenic drugs still have limitations such as drug tolerance, toxicity profile and substantial costs. (-)-Epigallocatechin-3-gallate (EGCG), the major polyphenol of green tea, is associated with anti-cancer benefits, but it is unstable and has poor bioavailability. These limitations can be solved through peracetate ester of EGCG as a prodrug (ProEGCG), while little is known about its anti-angiogenesis effect in cancer. The objective is to study the effect and underling mechanism of ProEGCG on EEC angiogenesis. Methods and Results: Tumor xenografts were established by subcutaneous injection of a human EEC cell line (RL95-2) into nude mice, and ProEGCG or control was orally administered every day. 35 days later, mice treated with ProEGCG showed significant reduction in lesion size and MVD without major toxic effects compared to control. To find out how ProEGCG inhibits tumor progress and angiogenesis, microarray analysis of excised lesions was conducted and indicated that HIF-1α, VEGFA and CXCR4 were downregulated by ProEGCG. Compared to control, immunohistochemistry showed that ProEGCG reduced expression of HIF-1α and VEGFA in RL95-2 cells and decreased CXCR4 expression in tumor-associated macrophages (TAMs) and endothelial cells (ECs). To further study the role of ProEGCG on EEC cells, qRT-PCR and Western blotting proved that hypoxia (1% O2) upregulated the level of HIF-1α and VEGFA in EEC cell lines, which was reversed by ProEGCG through inhibiting hypoxia-induced ROS formation. It is known that TAMs play an important role in tumor angiogenesis, and double immunofluorescence of F4/80 and CD206 antibodies in xenografts was applied and found that TAMs infiltration was reduced in ProEGCG group with decreased VEGFA in TAMs. Moreover, proliferation and migration of ECs are critical for tumor angiogenesis. To study the effect of ProEGCG on ECs, tube formation and invasion assays were conducted. The addition of ProEGCG on ECs showed less capillary tube-like structure formation and less invaded ECs by inhibition of CXCR4 expression on ECs. However, it was still unclear if CXCL12, the ligand of CXCR4, was affected. Immunostaining for CXCL12 in tumor lesions indicated that ProEGCG decreased CXCL12 expression in cancer and stromal cells, which was further confirmed in primary human endometrial stromal cells by addition of ProEGCG. Conclusion: Our study was the first to reveal the mechanism of anti-angiogenesis of ProEGCG, which decreased HIF1α/VEGFA expression in tumor cells, inhibited recruitment and differentiation of TAMs and tube formation of endothelial cells via CXCL12/CXCR4 reduction. These findings provided that ProEGCG would be an effective, safe and economic anti-angiogenic drug for EEC. Citation Format: Jianzhang Wang, Chi Wai Man, Xueying Yang, Joseph Kwong, Q. Ping Dou, Tak Hang Chan, Chi Chiu Wang. A prodrug of green tea polyphenol (-)-epigallocatechin-3-gallate served as a potentially novel angiogenesis inhibitor in endometrioid endometrial cancer [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 162. doi:10.1158/1538-7445.AM2017-162

The role of bone marrow edema on osteochondral lesions of the talus

BackgroundTo assess the functional and radiological outcomes after arthroscopic talus autologous matrix-induced chondrogenesis (AT-AMIC®) in 2 groups: patients with and without bone marrow edema (BME). MethodsThirty-seven patients of which 24 without edema (GNE) and 13 with edema (GE) were evaluated. All patients were treated with AT-AMIC® repair for symptomatic osteochondral talar lesion. Clinical and radiological parameters were evaluated with VAS score for pain, AOFAS and SF-12 at T0 (preoperatively), T1 (6 months), T2 (12 months), T3 (24 months) and MRI and CT-scan at T0, T1, T2 and T3. ResultsNo patients were lost to the final follow-up. In both groups we found a significant difference for clinical and radiological parameters with ANOVA for repeated measures through four time points (p<0.001). In GNE, AOFAS improved significantly at each follow-up (p<0.05); while CT and MRI showed a significant reduction in lesion size between T1 and T2 and T2 and T3 (p<0.05). In GE, AOFAS improved significantly between T0 and T1 and T2 and T3 (p<0.05); lesion size, measured with CT, decreased between T1 and T2 (p<0.05), while with MRI the lesion showed a reduction at each follow-up (p<0.05). Lesion size was significantly higher both in MRI and CT in GE compared to GNE (p<0.05). In GNE no patients presented edema at T3, while in GE only 23.08% of the patients presented edema at T3. ConclusionsThe study revealed that osteochondral lesions of the talus were characterized by bigger size both in MRI and CT in patients with edema. We conclude that AT-AMIC® can be considered a safe and reliable procedure that allows effective healing, regardless of edema and more than half of patients did not present edema six months after surgery.

Interplay between HSP90 and Nrf2 pathways in diabetes-associated atherosclerosis

IntroductionOxidative stress and inflammation are determinant processes in the development of diabetic vascular complications. Heat shock protein 90 (HSP90) overexpression in atherosclerotic plaques plays a role in sustaining inflammatory mechanisms, and its specific inhibition prevents atherosclerosis. The present work investigates, in a mouse model of diabetes-driven atherosclerosis, whether atheroprotection by pharmacological HSP90 inhibition is accomplished by bolstering antioxidant defense mechanisms headed by nuclear factor erythroid-derived 2-like 2 (Nrf2). MethodsStreptozotocin-induced diabetic apolipoprotein E-deficient mice were randomized to receive vehicle or HSP90 inhibitor (17-dimethylaminoethylamino-17-demethoxygeldanamycin, 4mg/kg) for 10 weeks. Aortic root sections were analyzed for plaque size and composition, transcription factor activity, and expression of inflammatory and antioxidant markers. In vitro studies were performed in murine macrophages cultured under hyperglycemic conditions. ResultsTreatment with HSP90 inhibitor promoted the activation of Nrf2 in the aortic tissue of diabetic mice (predominantly localized in macrophages and smooth muscle cells) and also in cultured cells. Nrf2 induction was associated with a concomitant inhibition of nuclear factor-κB (NF-κB) in atherosclerotic plaques, thus resulting in a significant reduction in lesion size and inflammatory component (leukocytes and cytokines). Furthermore, atheroprotection by HSP90 inhibition was linked to the induction of cytoprotective HSP70, antioxidant enzymes (heme oxygenase-1, superoxide dismutase and catalase) and autophagy machinery (LC3 and p62/SQSTM1) in aortic tissue. ConclusionHSP90 inhibition protects from atherosclerosis in experimental diabetes through the induction of Nrf2-dependent cytoprotective mechanisms, reinforcing its therapeutic potential.

Interplay between HSP90 and Nrf2 pathways in diabetes-associated atherosclerosis

IntroductionOxidative stress and inflammation are determinant processes in the development of diabetic vascular complications. Heat shock protein 90 (HSP90) overexpression in atherosclerotic plaques plays a role in sustaining inflammatory mechanisms, and its specific inhibition prevents atherosclerosis. The present work investigates, in a mouse model of diabetes-driven atherosclerosis, whether atheroprotection by pharmacological HSP90 inhibition is accomplished by bolstering antioxidant defense mechanisms headed by nuclear factor erythroid-derived 2-like 2 (Nrf2). MethodsStreptozotocin-induced diabetic apolipoprotein E-deficient mice were randomized to receive vehicle or HSP90 inhibitor (17-dimethylaminoethylamino-17-demethoxygeldanamycin, 4mg/kg) for 10 weeks. Aortic root sections were analyzed for plaque size and composition, transcription factor activity, and expression of inflammatory and antioxidant markers. In vitro studies were performed in murine macrophages cultured under hyperglycemic conditions. ResultsTreatment with HSP90 inhibitor promoted the activation of Nrf2 in the aortic tissue of diabetic mice (predominantly localized in macrophages and smooth muscle cells) and also in cultured cells. Nrf2 induction was associated with a concomitant inhibition of nuclear factor-κB (NF-κB) in atherosclerotic plaques, thus resulting in a significant reduction in lesion size and inflammatory component (leukocytes and cytokines). Furthermore, atheroprotection by HSP90 inhibition was linked to the induction of cytoprotective HSP70, antioxidant enzymes (heme oxygenase-1, superoxide dismutase and catalase) and autophagy machinery (LC3 and p62/SQSTM1) in aortic tissue. ConclusionHSP90 inhibition protects from atherosclerosis in experimental diabetes through the induction of Nrf2-dependent cytoprotective mechanisms, reinforcing its therapeutic potential.

Exacerbated Leishmaniasis Caused by a Viral Endosymbiont can be Prevented by Immunization with Its Viral Capsid.

Recent studies have shown that a cytoplasmic virus called Leishmaniavirus (LRV) is present in some Leishmania species and acts as a potent innate immunogen, aggravating lesional inflammation and development in mice. In humans, the presence of LRV in Leishmania guyanensis and in L. braziliensis was significantly correlated with poor treatment response and symptomatic relapse. So far, no clinical effort has used LRV for prophylactic purposes. In this context, we designed an original vaccine strategy that targeted LRV nested in Leishmania parasites to prevent virus-related complications. To this end, C57BL/6 mice were immunized with a recombinant LRV1 Leishmania guyanensis viral capsid polypeptide formulated with a T helper 1-polarizing adjuvant. LRV1-vaccinated mice had significant reduction in lesion size and parasite load when subsequently challenged with LRV1+ Leishmania guyanensis parasites. The protection conferred by this immunization could be reproduced in naïve mice via T-cell transfer from vaccinated mice but not by serum transfer. The induction of LRV1 specific T cells secreting IFN-γ was confirmed in vaccinated mice and provided strong evidence that LRV1-specific protection arose via a cell mediated immune response against the LRV1 capsid. Our studies suggest that immunization with LRV1 capsid could be of a preventive benefit in mitigating the elevated pathology associated with LRV1 bearing Leishmania infections and possibly avoiding symptomatic relapses after an initial treatment. This novel anti-endosymbiotic vaccine strategy could be exploited to control other infectious diseases, as similar viral infections are largely prevalent across pathogenic pathogens and could consequently open new vaccine opportunities.

Nimotuzumab in Management of Brain Stem Glioma: A Case Report

Brainstem glioma is rare tumour in adults accounting for 1% - 2% of intracranial gliomas. In this case study, a 28-year-old female diagnosed with BG, and lesions were observed in the pons and medulla region of the brain stem. She was initially treated with radiotherapy (54 Gy in 30 fractions for 6 weeks) but no change in her clinical condition and size of tumor was observed. Temozolomide (250 mg/daily for 5 days) was prescribed as first line chemotherapy. After completion of three cycles of Temozolomide, patient presented with diplopia and MRI showed increase in the size of lesions. After unsatisfactory response to radiation and chemotherapy, the patient was treated with Nimotuzumab therapy. MRI scan demonstrated the reduction of lesion size after 8 cycles of Nimotuzumab (200 mg/week). This treatment continued for another 8 cycles and the MRI scan of patients showed a significant reduction in lesion size. Nimotuzumab was found to be an effective and safe treatment option for brainstem glioma patient who was resistant to radiotherapy and chemotherapy.