Increase In Mean Survival Time Research Articles

Abstract 945: In vivo anti-tumor activity of onvansertib, a PLK1 inhibitor, combined with gemcitabine or carboplatin in platinum- resistant ovarian carcinoma patient-derived xenograft models

Abstract Background: The standard treatment for high grade ovarian carcinoma (HGOC) is cytoreductive surgery followed by a platinum based therapy. Despite an initial high rate of complete remission, most of the patients relapse with a platinum resistant disease. Patients with resistant tumors have few options, including monotherapy with gemcitabine, and new effective therapeutic alternatives are needed. The Polo-like kinase 1 (PLK1) is a master regulator of mitosis and recent evidences suggest its role in interfering with several DNA repair mechanisms. We investigated the effect of onvansertib (ONVA), a highly selective ATP-competitor PLK1 inhibitor, in combination with carboplatin (CARBO) and gemcitabine (GEM) in platinum-resistant HGOC patient-derived xenografts (PDXs). Methods: The combination of ONVA with either CARBO or GEM was investigated in two platinum-resistant HGOC PDXs, one intrinsically resistant to platinum (MNHOC#315), the other made resistant after repeated in vivo treatments with cisplatin (MNHOC#266R). Tumors were subcutaneously (s.c.; MNHOC#315) or orthotopically (i.p.; MNHOC#266R) transplanted in nude mice, mice were treated for 4 weeks with vehicle, single agents or drug combinations and followed for tumor growth (s.c. model) and survival. Results: CARBO- and GEM-ONVA combinations were well tolerated, with a maximum body weight loss of 6.6%. Both combinations showed strong anti-tumor activities in the two tested models. In MNHOC#315 PDX, CARBO- and GEM-ONVA combinations caused a tumor stabilization that extended beyond the dosing period and resulted in a higher tumor growth inhibition as compared to vehicle and single agents treated mice. In i.p. MNHOC#266R PDX, single agents were inactive, while a statistically significant increase in survival was observed in CARBO/ONVA and in GEM/ONVA treated mice as compared to vehicle treated mice, with a 6.6- and 7-fold increase in mean survival time, respectively. Pharmacodynamic studies are ongoing to decipher the mechanisms underlying the synergic effects of both combinations. Conclusions: The combinations of ONVA with either CARBO or GEM showed potent anti-tumor activities in two platinum-resistant HGOC PDXs. Additional studies in other HGOC platinum-resistant models, both in vivo and in vitro, as well pharmacodynamic studies are ongoing to validate these results and to get insights in the mechanisms underlying their efficacy. Citation Format: Michela Chiappa, Federica Guffanti, Ilaria Mengoli, Maya Ridinger, Giovanna Damia. In vivo anti-tumor activity of onvansertib, a PLK1 inhibitor, combined with gemcitabine or carboplatin in platinum- resistant ovarian carcinoma patient-derived xenograft models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 945.

Mitochondrial-mediated apoptosis as a therapeutic target for FNC (2′-deoxy-2′-b-fluoro-4′-azidocytidine)-induced inhibition of Dalton’s lymphoma growth and proliferation

PurposeT-cell lymphomas, refer to a diverse set of lymphomas that originate from T-cells, a type of white blood cell, with limited treatment options. This investigation aimed to assess the efficacy and mechanism of a novel fluorinated nucleoside analogue (FNA), 2′-deoxy-2′-β-fluoro-4′-azidocytidine (FNC), against T-cell lymphoma using Dalton’s lymphoma (DL)-bearing mice as a model.MethodsBalb/c mice transplanted with the DL tumor model received FNC treatment to study therapeutic efficacy against T-cell lymphoma. Behavioral monitoring, physiological measurements, and various analyses were conducted to evaluate treatment effects for mechanistic investigations.ResultsThe results of study indicated that FNC prevented DL-altered behavior parameters, weight gain and alteration in organ structure, hematological parameters, and liver enzyme levels. Moreover, FNC treatment restored organ structures, attenuated angiogenesis, reduced DL cell viability and proliferation through apoptosis. The mechanism investigation revealed FNC diminished MMP levels, induced apoptosis through ROS induction, and activated mitochondrial-mediated pathways leading to increase in mean survival time of DL mice. These findings suggest that FNC has potential therapeutic effects in mitigating DL-induced adverse effects.ConclusionFNC represents an efficient and targeted treatment strategy against T-cell lymphoma. FNC’s proficient ability to induce apoptosis through ROS generation and MMP reduction makes it a promising candidate for developing newer and more effective anticancer therapies. Continued research could unveil FNC’s potential role in designing a better therapeutic approach against NHL.

Abstract B043: Dual epigenetic therapy combined with anti-PD1 rescues HMA-induced suppressive myeloid phenotype and reduces tumor growth in a PDAC model

Abstract INTRODUCTION. We previously showed, using the PDAC mouse model KPC (KrasLSL G12D/+; p53 r172H/+; Pdx1-Cre), that sequential treatment with the DNA hypomethylating drug decitabine (5aza-dC; DAC) followed by anti-PD-1 led to increased tumor necrosis, slower tumor growth, increased tumor-infiltrating CD8+ and CD4+ T-cells (TILs), and significantly increased mean survival time. However, despite these positive outcomes, the KPC tumors eventually escaped from treatment. We identified a unique myeloid population as a specific subtype of M2-polarized putatively immunosuppressive Chil3+ macrophages (TAMs). Here we further characterize the recruitment mechansism of these cells and show that HDAC inhibition can further improve the efficacy of DAC+PD-1 treatment. METHODS. Primary macrophages were isolated from bone marrow (BMDM) or the peritoneum of 6-8 weeks mice and RAW264.7 cells were also used and exposed to conditioned media from primary KPC cell lines after HMA treatment (KPC-SN-DAC). Gene set enrichemnt analysis was done on RNASeq data from myeloid cells and validated by RT-qPCR. We used cytokine arrays to identify secreted cytokines from KPC cells. KPC cells were orthtopically injected in C57BL/6 mice, and tumor growth was followed by untrasound. Treatments were started when tumor reaced a size of 1 mm3. RESULTS. Expression profiling reveled that exposure of the myeloid cells to KPC-SN-DAC resulted in an HMA-specific M2-like gene signature. Notably DAC alone had minimal effect on myeloid cells polarization. To assess which factors might trigger this polarization we employed cytokine arrays and identified a number of candidate cytokines induced or secreted from KPC cells specifically following HMA treatment. In order to revert this unfavorable cytokine profile, we have evaluated the combination of DAC with four different HDAC inhibitors with reported immunomodulatory effects: Romidepsin (ROMI), Domatinostat (DOMA), Pracinostat (PRAC) and Entinostat (ENT). In vitro synergy studies through cell viability assays showed that ROMI, PRAC and DOMA synergized at low doses with DAC. Pofiling of supernatants from KPC cells treated with a combination of DAC+HDACis showed that, while ROMI had limited effect, PRAC and DOMA reverted DAC-induced cytokine secretion. Lastly, we tested in vivo the combination of DAC+ROMI/DOMI/PRAC+PD1 using mice orthotopically injected with KPC cells. The combination DAC+PRAC+PD1 showed reduced tumor volume compared to DAC+PD1. Moreover, PRAC+PD1 significantly improved overall survival. Interestingly, HDAC inhibition rescued the increased Chil3 expression in DAC+PD1 treated tumors. CONCLUSIONS. Our results show that paracrine rather than direct effect of hypomethylating therapy is responsible for suppressive myeloid polarization and characterize a specific effect of hypomethylating therapy on immature myeloid cells. Moreover, we have identified HDAC inhibition as a viable strategy to prevent unfavorable myeloid polarization and improve the therapeutic efficacy of the DAC+PD1 paradigm. Citation Format: Arturo Orlacchio, Stephen Muzyka, Catherine Do, Diane M. Simeone, Tamas Gonda. Dual epigenetic therapy combined with anti-PD1 rescues HMA-induced suppressive myeloid phenotype and reduces tumor growth in a PDAC model [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B043.

Exploring the efficacy of ethnomedicinal plants of Himalayan region against the malaria parasite

Ethnopharmacological relevancePlasmodium falciparum multi-drug resistant (MDR) strains are a great challenge to global health care. This predicament implies the urgent need to discover novel antimalarial drugs candidate from alternative natural sources. The Himalaya constitute a rich repository of medicinal plants which have been used traditionally in the folklore medicine since ages and having no scientific evidence for their activity. Crambe kotschyana Boiss. and Eremurus himalaicus Baker are used for their antipyretic and hepatoprotective properties in Kinnaur district of Himachal Pradesh, India. Aim of the studyThis study would investigate the antiplasmodial efficacy of C. kotschyana and E. himalaicus extracts, their fractions and active components using in vitro, in vivo and in silico approaches to provide a scientific insight into their activity. MethodsThe methanol extracts of C. kotschyana (CKME) and E. himalaicus (EHME) were prepared by maceration followed by fractionation using ethyl acetate. The isolation of flavonoid glycosides isorhamnetin-3, 7-di-O-glucoside from C. kotschyana and luteolin-6-C-glucoside (isoorientin) from E. himalaicus was carried out by antiplasmodial activity-guided isolation. In vitro antimalarial activity was assessed by WHO method while in vitro cytotoxicity was ascertained employing the MTT assay. Molecular docking and molecular dynamics simulation were performed using the Glide module of Schrödinger Software and Gromacs-2022 software package respectively. In vivo curative activity was assessed by Ryley and Peters method. ResultsThe methanol extracts of both the plants illustrated the best antiplasmodial activity followed by the ethyl acetate fractions. Iso-orientin (IC50 6.49 μg/ml) and Isorhamnetin-3,7-di-O-glucoside (IC50 9.22 μg/ml) illustrated considerable in vitro activity even against P. falciparum resistant strain. Extracts/fractions as well as the isolated compounds were found to be non-toxic with CC50 > 640 μg/ml. Molecular docking studies were performed with these 2 O-glucosides against four malaria targets to understand the binding pose of these molecules and the results suggested that these molecules have selectivity for lactate dehydrogenase enzyme. CKME and EHME exhibited curative activity in vivo along with increase in Mean Survival Time of mice. ConclusionThe research delineated the scientific evidence that both the therapeutic herbs possessed antimalarial activity and notably, bioactive compounds responsible to exhibit the antimalarial activity have been isolated, identified and characterized. Further studies are underway to assess the antiplasmodial efficacy of isolated compounds alone and in combination with standard antimalarials.

In vitro and in vivo anticancer activity of mebendazole in colon cancer: a promising drug repositioning.

Colon cancer is one of the most common cancers and one of the main causes of death worldwide. Therefore, new treatment methods with better efficiency and fewer risks are very necessary. Mebendazole (MBZ), a drug commonly used for helminthic infections, has recently received attention as a suitable candidate for the treatment of various cancers. This study aimed to investigate, in vitro and in vivo, anticancer activity and selectivity Index of MBZ on colon cancer. HT-29 (human colorectal adenocarcinoma) and MCF-10 (non-tumorigenic epithelial) cell lines were treated with MBZ and Doxorubicin (DOX; positive control drug). IC50 values were estimated using methyl thiazole diphenyl-tetrazolium bromide (MTT) assay. We employed flow cytometry using annexin V-FITC and propidium iodide dyes. For the animal study, colon cancer was subcutaneously induced by CT26 cells (mouse colon cancer) in Bulb/C mice. The mice were treated with 0.05 of LD50, intraperitoneal, every other day for 35 days. Finally, the survival rate, tumor volume, and tumor weight were calculated. Our results demonstrated that IC50 values after 72 h for HT29 and MCF-10 cell lines were 0.29 ± 0.04 µM and 0.80 ± 0.02 µM, respectively. MBZ was more selective than DOX in inhibiting the proliferation of cancer cells compared to normal cells (2. 75 vs. 2.45). Annexin V/PI staining demonstrated that MBZ treatment at IC50 concentrations induced (78 ± 12%) apoptosis in the HT29 cancer cell line after 48 h (P ≤ 0.0001). Also, in mice bearing colon cancer, MBZ significantly reduced the tumor volume (1177 ± 1109 mm3; P ≤ 0.001) and tumor weight (2.30 ± 1.97 g; P ≤ 0.0001) compared to the negative control group (weight 12.45 ± 2.0 g; volume 7346 ± 1077). Also, MBZ increases mean survival time (MST) and increase life span (ILS) percentage in the animal study (51.2 ± 37% vs 93%, respectively). This study suggests that mebendazole strongly and selectively inhibits proliferation and induces apoptosis in colon cancer cells. It may be, accordingly, a promising drug for clinical research and application.

Antitumor Activity of Ferulic Acid Against Ehrlich Solid Carcinoma in Rats via Affecting Hypoxia, Oxidative Stress and Cell Proliferation.

Background Ferulic acid is a natural compound commonly found in fruits and vegetables like tomatoes, sweet corn, rice bran, and dong quai. It has various beneficial effects on the body, such as anti-inflammatory, anti-apoptotic, hepatoprotective, cardioprotective, and neuroprotective properties. Aims We conducted a study to investigate the antitumor activity of ferulic acid against Ehrlich solid carcinoma (ESC), specifically by affecting hypoxia-inducible factor (HIF)-1α and its subsequent effects on other factors like nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), cellular Myc (cMyc), cyclin D1, mammalian target of rapamycin (mTOR), and signal transducer and activator of transcription 3 (STAT3). Materials and methods The study involved implanting rats with ESC cells and administering 50 mg/kg of ferulic acid orally daily for eight days. Sections of the muscles with ESC were stained with toluidine blue or immunostained with anti-HIF-1α antibodies. The tumor samples were used to evaluate the expression of HIF-1α, Nrf2, HO-1, cMyc, cyclin D1, mTOR, and STAT3. Results Ferulic acid increased mean survival time, reduced tumor volume and weight, and improved the appearance of the tumor tissue. Furthermore, ferulic acid significantly elevated the expression of Nrf2 and HO-1, while reducing the expression of HIF-1α, Nrf2, HO-1, cMyc, cyclin D1, mTOR, and STAT3. Conclusions Ferulic acid can reduce tumor size and weight while improving the structure of muscle cells, suggesting it may have antineoplastic activity against ESC. Further investigation revealed that ferulic acid downregulatesHIF-1α, increasing the expression of antioxidant proteins Nrf2 and HO-1. Additionally, ferulic acid decreases the expression of proliferation markers cMyc and cyclin D1 and downregulates cellular regulators mTOR and STAT3.

Design, synthesis and evaluation of novel pyrrole-hydroxybutenolide hybrids as promising antiplasmodial and anti-inflammatory agents

In the pursuance of novel scaffolds with promising antiplasmodial and anti-inflammatory activity, a series of twenty-one compounds embraced with most promising penta-substituted pyrrole and biodynamic hydroxybutenolide in single skeleton was designed and synthesized. These pyrrole-hydroxybutenolide hybrids were evaluated against Plasmodium falciparum parasite. Four hybrids 5b, 5d, 5t and 5u exhibited good activity with IC50 of 0.60, 0.88, 0.97 and 0.96 μM for chloroquine sensitive (Pf3D7) strain and 3.92, 4.31, 4.21 and 1.67 μM for chloroquine resistant (PfK1) strain, respectively. In vivo efficacy of 5b, 5d, 5t and 5u was studied against the P. yoelii nigeriensis N67 (a chloroquine-resistant) parasite in Swiss mice at a dose of 100 mg/kg/day for 4 days via oral route. 5u was found to show maximum 100% parasite inhibition with considerably increased mean survival time. Simultaneously, the series of compounds was screened for anti-inflammatory potential. In preliminary assays, nine compounds showed more than 85% inhibition in hu-TNFα cytokine levels in LPS stimulated THP-1 monocytes and seven compounds showed more than 40% decrease in fold induction in reporter gene activity analyzed via Luciferase assay. 5p and 5t were found to be most promising amongst the series, thus were taken up for further in vivo studies. Wherein, mice pre-treated with them showed a dose dependent inhibition in carrageenan induced paw swelling. Moreover, the results of in vitro and in vivo pharmacokinetic parameters indicated that the synthesized pyrrole-hydroxybutenolide conjugates abide by the required criteria for the development of orally active drug and thus this scaffold can be used as pharmacologically active framework that should be considered for the development of potential antiplasmodial and anti-inflammatory agents.

Enhanced antitumor effect of curcumin loaded solid lipid nanoparticles in Dalton’s ascites lymphoma mice

Curcumin is widely known for its antibacterial, antioxidant and anti inflammatory effects and has been reported to possess anticancerous activity as well. However, its medical application is limited because of poor bioavailability and rapid metabolism. In this study, we encapsulated curcumin in solid lipid nanoparticles and studied its anticancerous effect in Dalton’s Ascites Lymphoma (DAL) mice model. The physicochemical characteristics of curcumin solid lipid nanoparticles (CUR-SLN) were assessed and the anticancer efficacy was determined by in vivo studies. The curcumin solid lipid nanoparticles were synthesized by solvent emulsification evaporation method with particle size less than 100 nm. Antitumor effect of nanocurcumin (50 mg/kg) and curcumin (100 mg/kg) was evaluated in Dalton’s Ascites Lymphoma bearing mice. Pathological and immunohistochemical parameters were studied. Mean survival time and percentage increase in lifespan were assessed. Nanocurcumin group showed more significant influence in reducing tumor volume and weight, inducing apoptosis, reducing angiogenesis and invasion restoring antioxidant parameters and increased mean survival time. Curcumin and nanocurcumin inhibited the activation of nuclear factor-kappa B (Nf-kB), and thereby proved the pathway by which it induced anti-angiogenic and anti-invasive property.

Antiplasmodial Activity of 80% Methanolic Extract and Solvent Fractions of Stem Bark of Acacia tortilis in Swiss Albino Mice.

Malarial infection has significant negative impact on the health of the world population. It is treated by modern and traditional medicines. Among traditional medicinal plants, Acacia tortilis is used by different communities as antimalarial agent. Therefore, the objective of this study is to validate antimalarial activity of the stem bark of Acacia tortilis in mice. To evaluate antimalarial activity of the plant, 4-day suppressive, curative, and prophylactic antimalarial test models were used. Parasitemia, packed cell volume (PCV), survival time, rectal temperature, and body weight were used to evaluate the effect of the plant extracts. Data were analyzed using SPSS version 26 followed by Tukey's post hoc multiple comparison test. The crude extract and dichloromethane fraction significantly suppressed the level of parasitemia (p < 0.001) and increased mean survival time (p < 0.01) at all tested doses. Similarly, significant effects were observed in mean survival time, % change of PCV, weight, and temperature in both curative and prophylactic antimalarial test models. The methanolic extract and solvent fractions of the stem bark of Acacia tortilis has shown antimalarial activity, and the finding supports the traditional use and the in vitro studies. Thus, this study can be used as an initiation for researchers to find the most active phytochemical entity and to conduct additional safety and efficacy tests.

Evaluating the Safety and Quality of Life of Colorectal Cancer Patients Treated by Autologous Immune Enhancement Therapy (AIET) in Vinmec International Hospitals.

(1) Colorectal cancer (CRC) is an increasingly prevalent disease with a high mortality rate in recent years. Immune cell-based therapies have received massive attention among scientists, as they have been proven effective as low-toxicity treatments. This study evaluated the safety and effectiveness of autologous immune enhancement therapy (AIET) for CRC. (2) An open-label, single-group study, including twelve patients diagnosed with stages III and IV CRC, was conducted from January 2016 to December 2021. Twelve CRC patients received one to seven infusions of natural killer (NK)-cell and cytotoxic T-lymphocyte (CTL). Multivariate modelling was used to identify factors associated with health-related quality-of-life (HRQoL) scores. (3) After 20–21 days of culture, the NK cells increased 3535-fold, accounting for 85% of the cultured cell population. Likewise, CTLs accounted for 62.4% of the cultured cell population, which was a 1220-fold increase. Furthermore, the QoL improved with increased EORTC QLQ-C30 scores, decreased symptom severity, and reduced impairment in daily living caused by these symptoms (MDASI-GI report). Finally, a 14.3 ± 14.1-month increase in mean survival time was observed at study completion. (4) AIET demonstrated safety and improved survival time and HRQoL for CRC patients in Vietnam.

Abstract 1295: Small molecule reactivators of Y220C mutant p53 modulate tumor infiltrating leukocytes and synergize with immune checkpoint inhibitors

Abstract Half of human cancers possess mutations in the TP53 gene, with most clustering as hotspots in the DNA binding domain. A tyrosine to cysteine substitution at amino acid 220 of the p53 protein (Y220C) is one such hotspot mutation (~1% of all solid tumors). Small molecules that reactivate Y220C mutant p53 activity to be like wild type have been developed at PMV Pharmaceuticals and the lead compound PC14586 is in Phase I clinical development. The molecules were designed to bind tightly to a crevice within the Y220C p53 mutant protein and stabilize it to a wild type conformation allowing reactivation of p53 transcriptional activity and expression of target proteins (e.g. p21, MDM2), resulting in tumor regression in Y220C p53 mutant xenograft models grown in immunocompromised mice. p53 has been documented to contribute to immune responses by activating regulators of immune signaling pathways and to result in T-cell exclusion in tumor models. To investigate the role of p53 and the immune system in tumorigenesis, a Y220C human p53 knock-in (HUPKI) mouse was generated. Mice that are homozygous mutant for the Y220C humanized alleles succumbed to lymphomas and sarcomas within 6 months and cell lines generated from these tumors were shown to be sensitive in an in vitro proliferation assay to Y220C p53 reactivators (IC50 ~ 192-722 nM) and used to generate Y220C mutant syngeneic mouse models. Administration of Y220C p53 reactivators in Y220C p53 syngeneic mouse models resulted in a dose responsive anti-tumor effect, with maximally efficacious doses resulting in durable cures in nearly all animals, more than observed in immunocompromised mouse models. Addition of checkpoint agents with sub-efficacious administration of Y220C reactivating compounds resulted in synergistic increases in mean survival time of mice, longer than with either single agent. Immunophenotyping analysis of HUPKI Y220C mutant tumors exposed to p53 reactivator compounds showed modulation of the tumor immune environment. Changes include dose responsive increases in T-cells (CD4+, CD8+), T-regulatory cells, natural killer T cells and dose responsive decreases in macrophages (M2) and g-MDSC cells. The synergy observed with an anti-PD-1 combination appears driven by an increase in CD8+ T cells, given a decrease in efficacy in mice depleted of CD8+ T cells, following exposure to anti-CD8. Further investigation into the role of Y220C p53 reactivation in immune modulation showed decreased cytokines and chemokines involved in inflammation (i.e. CXCL10, Ccl2) and an increase in a Tumor Inflammation Signature (nanoString) in a dose and time dependent manner. Taken together, these data suggest a role for p53 signaling in encouraging an immunologically hot tumor microenvironment and provide support for testing small molecule reactivators of mutant p53 in combination with immune checkpoint inhibitors. Citation Format: Anna M. Puzio-Kuter, Chris Mulligan, Brandon Russo, Amy Wiebesiek, Lizhong Xu, Hong Yang, Binh Vu, Melissa Dumble. Small molecule reactivators of Y220C mutant p53 modulate tumor infiltrating leukocytes and synergize with immune checkpoint inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1295.

Survival in patients with surgically treated dural based metastases: A single-center retrospective study

PurposeDural-based metastases (DBM) are present in 4% of all patients diagnosed with metastatic cancer. DBM most commonly arise from primary cancers of the lung, breast, and prostate. While diagnostic challenges are relatively well studied, the epidemiology, common intracranial locations, and therapeutic outcomes of DBM remain poorly understood. Our current study aims to define the epidemiological characteristics of patients presenting with DBM, the most common locations of metastasis within the dura, and clinical outcomes after surgery and radiation therapy. MethodsPatients presenting to the Montreal Neurological Institute for intracranial metastasis between the years 2010 and 2016 were reviewed, and patients with DBM were identified based on radiological characteristics. Data were extracted from patient charts and MRIs. ResultsA total of 46 patients met inclusion criteria and were included in the study. The mean age at DBM diagnosis was 57 years old. Lung cancer was the most common origin (57%), while breast cancer was the second most common (22%). DBM were more likely to present in patients with active systemic disease (63%) and were found within 1 cm of a major venous sinus in 52% of cases. 87% of patients underwent gross total resection of their tumors. In these patients, adjuvant SRS was associated with a 2-fold increase in mean survival time (p = 0.013). ConclusionAlthough DBM are a marker of advanced systemic cancer, surgical resection with adjuvant radiotherapy may play a role in control of intracranial disease and overall patient survival.

Prognostic Significance of GPR55 mRNA Expression in Colon Cancer.

G protein-coupled receptor 55 (GPR55) probably plays a role in innate immunity and tumor immunosurveillance through its effect on immune cells, such as T cells and NK cells. In this study, the prognostic value of GPR55 in colon cancer (CC) was investigated. mRNA expression levels of GPR55 were determined in 382 regional lymph nodes of 121 CC patients with 12 years observation time after curative surgery. The same clinical material had previously been analyzed for expression levels of CEA, CXCL16, CXCL17, GPR35 V2/3 and LGR5 mRNAs. Clinical cutoffs of 0.1365 copies/18S rRNA unit for GPR55 and 0.1481 for the GPR55/CEA ratio were applied to differentiate between the high- and low-GPR55 expression groups. Kaplan–Meier survival analysis and Cox regression risk analysis were used to determine prognostic value. Improved discrimination between the two groups was achieved by combining GPR55 with CEA, CXCL16 or CXCL17 compared with GPR55 alone. The best result was obtained using the GPR55/CEA ratio, with an increased mean survival time of 14 and 33 months at 5 and 12 years observation time, respectively (p = 0.0003 and p = 0.003) for the high-GPR55/CEA group. The explanation for the observed improvement is most likely that GPR55 is a marker for T cells and B cells in lymph nodes, whereas CEA, CXCL16 and CXCL17, are markers for tumor cells of epithelial origin.

Pithecellobium dulce induces apoptosis and reduce tumor burden in experimental animals via regulating pro-inflammatory cytokines and anti-apoptotic gene expression

The present study demonstrates the efficacy of fruit extract of Pithecellobium dulce (FPD) against Dalton's lymphoma ascites (DLA) cell lines in vitro and in vivo (DLA induced ascitic and solid tumor). Administration of FPD induced apoptosis in DLA cells via p53 regulation both in vitro and in vivo. Cell viability was quantified by MTT assay. Apoptotic cells were determined by qualitative (staining methods) and quantitative analysis (Annexin-propidium iodide based flow cytometry). Expression of pro-apoptotic markers (Caspase 3, Caspase 9, and Bax) were markedly elevated, while expression of anti-apoptotic proteins (Bcl 2 and Bcl XL) were downregulated in tumor cells. FPD administration effectively reduced tumor burden, increased mean survival time via modulating NF-kB, and reduced the level of proinflammatory cytokines (IL-6, IL-1β, GM-CSF and TNF-α). Phytochemical screening of FPD by GC/MS analysis divulged the presence of several novel bioactive chemical constituents. Further, bioactive components identified from extract were evaluated for drug-like properties by Lipinski rule of five and properties. Naringenin, nootkatone, and gallic acid showed good drug-like properties and good pharmacokinetic profiles compared to other bioactive constituents in the extract.

Rutin ameliorates malaria pathogenesis by modulating inflammatory mechanism: an in vitro and in vivo study.

Rutin (3, 3', 4' 5 and 7-pentahydroxyflavone-3-rhamnoglucoside) is a flavonoid glycoside, found in many edible plants such as buckwheat and berries. Severe malaria is an inflammatory response triggered by oxidative stress that results in multi-organ pathologies and a high mortality rate in children and pregnant women worldwide. Rutin is recommended as a food supplement for the treatment of various diseases due to its anti-oxidative and anti-inflammatory properties, which prompted us to investigate its ameliorative effects in severe malaria pathogenesis against oxidative stress and inflammatory response using in vitro and in vivo bioassays. Rutin was examined in this work for its anti-plasmodial activity against chloroquine-sensitive and resistant Plasmodium falciparum strains, as well as its anti-oxidative and anti-inflammatory activity against LPS-stimulated macrophage cells. The in vitro data were subsequently verified in mice fed orally with rutin alone or in combination with chloroquine in Plasmodium berghei-induced malaria pathogenesis. The anti-plasmodial and anti-inflammatory properties of rutin were demonstrated in in vitro results. Apart from its anti-inflammatory and anti-oxidant effects in malaria pathogenesis, in vivo efficacy studies indicated that oral treatment with rutin reduced parasitaemia, increased mean survival time, and restored haemoglobin and glucose levels in mice at lower dose. Interestingly, both rutin and chloroquine demonstrated synergy in in vitro and in vivo experiments. The findings of the present study thus highlighted the suitability of rutin for further study in the management of drug resistant malaria in combination with standard anti-malarial drugs.

Evaluation of Antimalarial Activity of 80% Methanolic Root Extract of Dorstenia barnimiana Against Plasmodium berghei-Infected Mice.

BackgroundDue to resistance of parasites, costs, and safety issues with currently available drugs, there is a need to discover new antimalarials. Medicinal plants are one of the most important sources of new drugs. The purpose of this study was to evaluate the antimalarial activity of a methanolic root extract of Dorstenia barnimiana in Swiss albino mice infected with Plasmodium berghei.MethodsFour-day suppressive, curative, and prophylactic tests were performed on mice infected with P. berghei to evaluate the antimalarial activity of a methanolic root extract of the plant. Parasitemia suppression, survival time, body-weight change, rectal temperature change, and packed-cell volume were used to evaluate the activity of the extract. Data were analyzed using one-way ANOVA followed by Tukey’s post hoc test for comparisons between and within groups, with P<0.05 considered statistically significant.ResultsThe D. barnimiana root extract showed significant (P<0.01) parasitemia-suppressive activities in all models compared to the negative control. The extract increased mean survival time and prevented weight loss, reduction in temperature, and anemia significantly in all tested doses in 4-day suppressive and curative tests.ConclusionBased on these findings, D. barnimiana root has promising antimalarial activity and can considered a potential source to develop new agents.

Vaccination with Prion Peptide-Displaying Polyomavirus-Like Particles Prolongs Incubation Time in Scrapie-Infected Mice.

Prion diseases like scrapie in sheep, bovine spongiform encephalopathy (BSE) in cattle or Creutzfeldt–Jakob disease (CJD) in humans are fatal neurodegenerative diseases characterized by the conformational conversion of the normal, mainly α-helical cellular prion protein (PrPC) into the abnormal β-sheet rich infectious isoform PrPSc. Various therapeutic or prophylactic approaches have been conducted, but no approved therapeutic treatment is available so far. Immunisation against prions is hampered by the self-tolerance to PrPC in mammalian species. One strategy to avoid this tolerance is presenting PrP variants in virus-like particles (VLPs). Therefore, we vaccinated C57/BL6 mice with nine prion peptide variants presented by hamster polyomavirus capsid protein VP1/VP2-derived VLPs. Mice were subsequently challenged intraperitoneally with the murine RML prion strain. Importantly, one group exhibited significantly increased mean survival time of 240 days post-inoculation compared with 202 days of the control group. These data show that immunisation with VLPs presenting PrP peptides may represent a promising strategy for an effective vaccination against transmissible spongiform encephalitis agents.

Antimalarial Activity of Methanolic Extracts and Solvent Fractions of Combretum molle Leaves in Plasmodium berghei Infected Mice.

BackgroundMalaria is an infectious disease-causing mortality and morbidity in children and adults. Our study planned to measure the antimalarial activity of Combretum molle extract in vivo assays against Plasmodium berghei in Swiss albino mice.MethodsPlasmodium berghei was inoculated to healthy mice and methanolic crude extract and solvent fractions of C. molle at 100, 200, and 400mg/kg/day were administered. Percent parasitemia inhibition, percent change in weight, temperature, packed cell volume, and mean survival time were determined. Data were analyzed using one-way ANOVA followed by the post hoc Tukey HSD test with SPSS software version 24.0 and P < 0.05 considered as statistically significant.ResultsChemosuppresive effect exerted by the methanolic crude extract ranged between 27 and 68%, chloroform fraction 52.93–67.73%, hexane fraction 23.61–44.97%, and aqueous fraction 29.48–51.13%. The curative effect of the crude extract was within the range of 25–49%, chloroform fraction 42.78–69.22%, and prophylactic effect of the crude extract was within the range of 51–76.2%% and chloroform fraction 46.57–71.96%. The utmost effect in all tests on chemosuppresive, curative, prophylactic, prevention of weight loss, temperature and packed cell volume, and an increase in mean survival time was observed at higher doses of the crude extract.ConclusionFrom this study, it will be concluded that crude extract of C. molle leaves has been shown promising antimalarial activity. The findings of this study may support the normal use of leaf as a part of the plant for malaria treatment.

4-chloro eugenol interacts synergistically with artesunate against drug resistant P. falciparum inducing oxidative stress

4-chloro eugenol (4CE), a semisynthetic analog of phytomolecule eugenol exhibited potent antiplasmodial activity with IC50 in the range of 1.5–5 μM against sensitive as well as drug resistant strain of P. falciparum. This analog also showed synergy with a clinically used antimalarial drug artesunate and was able to curtail the IC50 of artesunate up to 4–5 folds. Although, 4CE did not show any effect on heme polymerization, the most common drug target in the malaria parasite, it could increase the level of reactive oxygen species (ROS) and reactive nitrogen species (RNS) alone as well as in combination with artesunate. Further, 4CE induced oxidative stress was observed to affect the macromolecules in terms of DNA damage, protein carbonylation and lipid peroxidation. At the physiological level, cellular organelles like mitochondria and endoplasmic reticulum were observed to be get affected by 4CE in terms of membrane depolarization and calcium ion leakage respectively. These observations could be validated by expression analysis of oxidative stress responsive genes and proteins. Further, in in vivo assay, 4CE showed significant chemo-suppression of parasitemia as well as an increase in mean survival time in the murine malaria model. Interestingly, in combination with artesunate, 4CE showed higher chemo-suppression as well as enhanced mean survival time at a much lower concentrations of both the partners as compared to an individual dose of artesunate and 4CE. A combination of 4CE and artesunate was also observed to attenuate cerebral malaria pathogenesis.

Antiplasmodial Assessment of Artemether- Lumefantrine/Ivermectin on Mice Infected with Plasmodium berghei

Substantial progress made in the fight against malaria has been undermined by challenges including drug resistance instigating the search for new antimalarial drugs or the repurposing of already approved drugs. This study evaluated the antiplasmodial activity of ivermectin (IM) in combination with artemether/lumefantrine (A/L) in a mice model infected with Plasmodium berghei. Parasitized (Plasmodium berghei) adult mice weighing 23-28g were used. The mice were grouped and treated orally with 1M (0.17 mg/kg), A/L (2.3/13.7mg/kg) and A/L/IM daily in curative, suppressive and prophylactic tests. The negative control (NC) was treated orally with normal saline (0.2ml) whereas the positive control was treated orally with chloroquine (CQ) (10mg/kg). After drug treatment, blood samples were collected and evaluated for percentage parasitemia levels, percentage parasitemia inhibition, lipid and hematologic parameters. Mean survival time was also evaluated. The 4-day curative, suppressive and prophylactic test showed significant decreases in percentage parasitemia levels at IM (0.17 mg/kg) (p<0.01), A/L (2.3/13.7 mg/kg) (p<0.001) and A/L/IM (p<0.0001) when compared to negative control. IM (0.69 mg/kg), A/L (2.3/13.7mg/kg) and A/L/IM increased mean survival time significantly at p<0.05, p<0.01 and p<0.001 respectively when compared to negative control. Red blood cells, packed cell volume, hemoglobin, high density lipoprotein cholesterol levels were increased significantly whereas total cholesterol, white blood cell, low density lipoprotein cholesterol and triglyceride levels were decreased significantly at IM (0.17 mg/kg) (p<0.05), A/L (2.3/13.7mg/kg) (p<0.01) and A/L/IM (p<0.001) when compared to negative control. This study suggests the use of A/L/IM as a viable malaria treatment.