Effect Of Combined Treatment Research Articles

The Anti-proliferative Effect of Caffeic Acid and Dactolisib on Human Cervical Carcinoma HeLa Cell Line

Cervical carcinoma is a common gynecological cancer with high mortality rate among women worldwide. Caffeic acid exerts an antiproliferative effect against cervical carcinoma. Dactolisib is a dual PI3K and mTOR inhibitor that has a therapeutic potential for cervical carcinoma. This study aimed to reveal the anti-proliferative effect of combination treatment of caffeic acid and Dactolisib on cervical carcinoma HeLa cell line. Cytotoxicity of caffeic acid and Dactolisib on HeLa cell line was assessed by MTS assay. Colony formation of HeLa cells treated with caffeic acid and Dactolisib was determined by staining colonies with crystal violet and visualizing under light microscope. Dactolisib decreased cell proliferation of HeLa cells in time and dose dependent manner. 5 μM caffeic acid did not show any significant change in cell viability of HeLa cells. Combination treatment of 5 μM caffeic acid and 0.5 μM Dactolisib showed decrease in cell viability of HeLa cells when compared to Dactolisib treated cells. Combination of caffeic acid and Dactolisib decreased colony diameter of HeLa cells significantly when compared to control group. Caffeic acid and Dactolisib shows anti-proliferative effect on human cervical carcinoma HeLa cell line, so further studies should be performed to reveal the mechanism of action.

The effectiveness of combined laser treatment technology in patients with glaucoma

Purpose: to study the effectiveness of combined laser treatment in patients with the advanced primary open-angle glaucoma (POAG).Material and methods: the present study included 63 patients with POAG. They were divided into the studied group (32 eyes) and control group (31 eye). Patients age ranged from 65 to 78 years (mean 70.68 ± 4.39 years in the studied group and 69.48 ± 3.82 in the control group). Intraocular pressure (IOP) in all patients was compensated with antihypertensive drops, and in average was 20.36 ± 1.27 mm Hg in the studied group and 20.02 ± 1.43 mm Hg in the control group. The number of antihypertensive drugs used before surgery ranged from 2 to 3 (average 2.40 ± 0.49 in the studied group and 2.51 ± 0.50 in the control group). YAG-LAT was performed with Lightlas YAG Nd-YAG laser (Lightmed, USA). Microimpulse transcleral cyclophotocoagulation (mTS-CPC) was performed with Cyclo G6 Glaucoma Laser System (IRIDEX, USA). In the studied group, all patients had YAG-LAT with mTS-CPC; in the control group – mTS-CPC. Patients were examined on days 1 and 7; and then in 1, 3 and 6 months after the treatment. The obtained results were summarized in a table and were statistically processed.Results: in the studied group, on day 1 after the intervention IOP was (mm Hg): 19.36 ± 1.36, in the control group – 19.45 ± 1.15; on day 7: studied group – 20.59 ± 1.5, control group – 19.9 ± 2.68; in one month: studied group – 13.95 ± 1.67, control group – 15.8 ± 1.5; in three months: studied group – 15.04 ± 1.13, control group – 16.48 ± 1.12; in 6 months: studied group – 15.27 ± 1.77, control group – 16.23 ± 1.41.Conclusion: the combined laser technology which includes YAG-laser activation of the trabecula and mTS-CPC is an effective and safe method for lowering intraocular pressure in patients with glaucoma

Abstract 704: Targeting hypoxic survival pathways to overcome radiotherapy-related treatment resistance

Abstract Tumor cells undergo biological adaptations in order to survive stress conditions, such as hypoxia and nutrient deprivation. These biological adaptations include the activation of hypoxic survival pathways that play an important role in acquiring radioresistance which becomes most challenging with treatment regimens using high doses of radiation/fraction (e.g., hypofractionated radiotherapy). Therefore, inhibiting key regulators within these pathways and thereby exploiting these survival mechanisms, gives rise to new potential drug targets for combined treatment modalities. This novel strategy investigated in our laboratory relies on the concept of biological cooperation: while ionizing radiation kills primarily well-oxygenated cells, such inhibitors of survival kinases will drive quiescent tumor cells under hypoxia and/or nutrient deprivation either directly into cell death or into re-entry into a proliferative and thereby more ionizing radiation-sensitive state. One of the survival kinases we are investigating in combination with ionizing radiation (IR) is DYRK1B (dual-specificity tyrosine regulated kinase 1B). We demonstrated that the expression of DYRK1B was upregulated under serum-starvation and hypoxia, but not in response to IR. The small molecule DYRK1B inhibitor AZ191 and shRNA-mediated DYRK1B knockdown significantly reduced proliferative activity and clonogenicity of SW620 tumor cells alone and in combination with IR under serum-starved conditions, which correlated with increased ROS levels and DNA damage. Furthermore, AZ191 successfully targeted the hypoxic core of tumor spheroids while IR preferentially targeted normoxic cells in the rim of the spheroids. A combined treatment effect was also observed in patient-derived colorectal carcinoma organoids but not in healthy tissue organoids. This data shows that inhibition of DYRK1B in quiescent tumor cells could drive tumor cells on their own into crisis or into a more radiosensitive cell cycle phase and thus supports our strategy of biological cooperation. In the continuation of this project, we have performed a drug-screen with approx. 3’000 clinically relevant compounds in colorectal tumor cells lines under normoxic and hypoxic conditions in order to explore additional hypoxia-mediated survival pathways contributing to radioresistance. Thereby we aim to identify novel combined treatment modalities to overcome radiotherapy-induced hypoxia and hypoxia-related resistance mechanisms. Citation Format: Claire Beckers, Lazaros Vasilikos, Alba Sanchez-Fernandez, Lorena Moor, Martin Pruschy. Targeting hypoxic survival pathways to overcome radiotherapy-related treatment resistance [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 704.

Abstract 6490: Pt1a and sorafenib combination treatment exerts synergistic anti-hepatocellular carcinoma effect through RNA m6A methylation modification

Abstract Hepatocellular carcinoma (HCC) is the most common type of liver cancer and has a poor prognosis. The survival benefits of sorafenib in patients with advanced HCC are modest owing to acquired resistance. Accumulating evidence has demonstrated that cancer stemness is an important cause of drug resistance and the root cause of tumor recurrence. [PtII(C^N^N)(NHC2Bu)]PF6 (Pt1a) is a newly developed platinum-based drug containing N-heterocyclic carbene with proven anticancer properties against a range of tumor types. However, its role in targeting cancer stemness and its ability to sensitize cancer cells to overcome molecular-targeted therapy drug resistance in HCC remains elusive. Herein, we demonstrate the synergistic inhibitory effect of sorafenib and Pt1a combination treatment on HCC through RNA m6A methylation modification. In cultured HCC cells and patient-derived organoids, the drug combination synergistically reduced the cell viability and promoted cell apoptosis compared to single-drug treatment. Combination treatment also attenuated self-renewal, as evidenced by a marked decrease in tumor-initiating cancer cell frequency, as measured by limiting dilution spheroid formation assay. These findings were further validated in HCC patient-derived xenograft mouse models and a mouse HCC model induced by hydrodynamic tail vein delivery of NRAS+AKT coactivation plasmids. Notably, the combination treatment with sorafenib and Pt1a, but not a single treatment, led to a significant reduction in tumor burden. The combination treatment was also effective in decreasing tumor-initiating cell frequency, as measured by subjecting the harvested tumor cells for limiting dilution spheroid formation assay ex vivo. Mechanistically, integrative RNA-sequencing and thermal proteome profiling analyses both indicated the enrichment of m6A-related pathways in combination drug treatment. An increased global m6A level upon combination treatment was validated using m6A dot blotting, confirming that the synergistic effect may be mediated by the elevated global m6A level in HCC cells. Collectively, our study demonstrated that Pt1a and sorafenib combination treatment synergistically inhibited the viability and stemness of HCC cells by increasing global RNA m6A level. These findings supported the potential of Pt1a as a therapeutic option for HCC and overcoming sorafenib drug resistance. Ongoing studies are focused on identifying m6A regulators and downstream targets involved in combination treatment. Citation Format: Yinjia Huang, Yalu Cui, Ianto Bosheng Huang, Yanyan Wang, Yang Xuan, Pui Ki Wan, Chun Nam Lok, Chi Ming Che, Stephanie Ma. Pt1a and sorafenib combination treatment exerts synergistic anti-hepatocellular carcinoma effect through RNA m6A methylation modification [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 6490.

Abstract 7196: Evaluation of nab-sirolimus in combination with fulvestrant or PI3K pathway inhibitors to overcome resistance in breast cancer cell lines

Abstract Background: Endocrine therapy (ET) is the standard of care for hormone receptor positive (HR+) breast cancer (BC), but ET resistance develops in most patients (pts). One resistance mechanism involves cross talk between the PI3K-AKT-mTOR and estrogen receptor (ER) signaling pathways, resulting in increased ER signaling and tumor proliferation. This is mitigated by mTOR inhibition as demonstrated by improved outcomes with exemestane plus the mTOR inhibitor (mTORi) everolimus in pts with HR+ advanced BC. While PI3K inhibitors (PI3Ki [e.g. alpelisib]) are efficacious in pts with HR+, PIK3CA-mutated BC, resistance can occur due to acquired genomic alterations in the PI3K-AKT-mTOR pathway. Combination therapy that targets a critical downstream node, mTOR, could delay or overcome acquired resistance due to activation of upstream components of the PI3K-AKT-mTOR pathway. nab-Sirolimus is an intravenously administered, albumin-bound nanoparticle form of the mTORi sirolimus, with significantly greater tumor accumulation, mTOR target suppression, and improved antitumor effects than oral mTORis in nonclinical tumor models. We evaluated the antiproliferative and cytotoxic effects of nab-sirolimus in combination with ET or PI3Kis in BC cell lines. Methods: HR+, PI3K-mutated BC cell lines (MCF7 or MDA-MB-361) were incubated for 5 days with increasing concentrations of nab-sirolimus and the selective ER degrader fulvestrant. MDA-MB-361 cells or HR-negative (HR−), PI3K-mutated BC cells, MDA-MD-453, were incubated for 5 days with increasing concentrations of nab-sirolimus and PI3K-AKT-mTOR pathway inhibitors gedatolisib or alpelisib. Antiproliferative/cytotoxic effects of single-agent and combination treatment were assessed via an automated trypan blue exclusion assay. Results: Addition of 20 or 80 nM nab-sirolimus to fulvestrant (12.5-500 nM) markedly decreased cell viability and nearly doubled cell death in MCF7 and MDA-MB-361 cells. Antiproliferative effects of low-dose gedatolisib (2.5-10 nM) were enhanced by 61-71% when nab-sirolimus (20 or 80 nM) was added to MDA-MB-453 cells. While low doses (2.5, 5, and 10 nM) of gedatolisib alone led to limited cytotoxicity (4.2-9.6%), up to 48.9% of cell death occurred when 20 or 80 nM nab-sirolimus was added. Results were similar with alpelisib (1 uM) plus nab-sirolimus (20 or 80 nM). Western blot showed increased p4EBP1 in response to fulvestrant or PI3Ki treatment, which was reversed with the addition of nab-sirolimus. Conclusions: nab-Sirolimus enhanced the cytotoxic effects of fulvestrant in HR+ BC cells and of PI3Kis in HR+ and HR− BC cells. Addition of nab-sirolimus to ET or other PI3K-AKT-mTOR pathway inhibitors may overcome mechanisms of resistance; further exploration is warranted to elucidate clinical relevance. Citation Format: Khine Nyein Myint, Sean Wallace, Shihe Hou, Maria Zalath, Brian McMorran, Andrew Kwon, Igor Vivanco. Evaluation of nab-sirolimus in combination with fulvestrant or PI3K pathway inhibitors to overcome resistance in breast cancer cell lines [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 7196.

Abstract 2821: Development of therapeutic monoclonal antibodies against DKK1 peptide-HLA-A2 complex to treat human cancers

Abstract Immunotherapy is a promising option for cancer treatment. Targeted immunotherapy with monoclonal antibodies (mAbs) is an effective and safe method for the treatment of malignancies. In recent years, efforts have been made to identify potential therapeutic mAbs by defining alternative or novel target antigens. We reported that Dickkopf-1 (DKK1) is a good target for immunotherapy of human cancers based on its wide expression in different cancers but not in normal tissues. As DKK1 is a secreted protein, mAbs directly binding to DKK1 may have limited effects on cancer cells in vivo. Our previous studies demonstrated that DKK1 peptide (such as P20 and P66v, which bind with HLA-A*0201 (HLA-A2) molecule) specific cytotoxic T cells specifically kill myeloma and other cancer cells that express DKK1 and HLA-A2, but not HLA-A2+ normal cells, indicating that DKK1+ tumor cells naturally express these peptides, in the context of HLA-A2 molecules, on their surface. To develop cancer therapeutic antibodies, DKK1 peptide P20-HLA-A2 monomer was synthesized and used to immunize mice. Hybridomas secreting mAbs recognizing cell surface-expressed DKKl P20-HLA-A2 complex (DKK1-A2) were obtained and analyzed. In this study, we generated novel (DKK1-A2) mAbs recognizing DKK1 (P20) peptide in the context of human HLA-A*0201 (HLA-A2) molecules (DKK1-A2 complexes) that are naturally expressed by HLA-A2+DKK1+ cancer cells. These mAbs bound to HLA-A2+DKK1+ hematologic (myeloma, lymphoma, and leukemia) and solid (breast, pancreatic, and prostate) cancer cells and directly induced apoptosis in the cancer cells by activating the caspase-9 cascade. DKK1-A2 mAbs also effectively lysed cancer cells in vitro by mediating CDC and ADCC activities. Furthermore, DKK1-A2 mAbs were therapeutic against established hematologic and solid cancers in their xenograft mouse models. To enhance the therapeutic efficacy of DKK1-A2 mAbs, we investigated the effect of combination treatment of DKK1-A2 mAbs with lenalidomide. Our results showed that there is an additive therapeutic effect between DKK1-A2 mAbs and lenalidomide in vivo, underscoring a potential clinical development strategy for combining DKK1-A2 mAb with lenalidomide to treat cancer patients. As DKK1 is not detected in most human tissues, DKK1-A2 mAbs did not bind to or kill HLA-A2+ blood cells in vitro nor caused tissue damage in vivo in tumor-free or tumor-bearing HLA-A2-transgenic mice. Thus, our study suggests that DKK1-A2 mAbs may be a promising therapeutic agent to treat human cancers and warrants further preclinic studies to determine their translatability to clinic. Citation Format: Jianfei Qian, Wang Qiang, Liuling Xiao, Wei Xiong, Miao Xian, Chuanchao Zhang, Yabo Li, Qing Yi. Development of therapeutic monoclonal antibodies against DKK1 peptide-HLA-A2 complex to treat human cancers [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 2821.

Abstract 4748: Aronia berry overcomes gemcitabine resistance by regulating the MYD88/NF-kB/P-glycoprotein pathway in pancreatic ductal adenocarcinoma

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is lethal, which is partly due to the limited efficacy of Gemcitabine (Gem)-based chemotherapy, resulting from the development of acquired resistance to this regimen. Aronia berry extract (ABE) is a natural source of phenolic compounds known for its anti-cancer properties. Previous research has highlighted the ability of ABE to help overcome chemoresistance to conventional chemotherapies in various cancers. In this study, we interrogated the ability of ABE to overcome Gem-resistance (Gem-R) in PDAC and identify specific pathway(s) responsible for this anti-cancer efficacy. Methods: We performed a series of in-vitro experiments in Gem-R cells to evaluate the synergistic effect of combined treatment with Gem and ABE. Furthermore, we conducted a genomewide transcriptomic analysis to identify the critical regulatory pathways associated with chemoresistance and potential therapeutic targets of ABE in Gem-R PDAC cells. The cell culture findings were validated in patient-derived 3D organoids. Results: The combined treatment with ABE and Gem in Gem-R cells exhibited significant synergistic anticancer effects on cell viability, proliferation, migration, and invasion. The transcriptomic analysis revealed that the NF-κB signaling pathway was associated with Gem-R (p<0.05), with a significant upregulation of MYD88. Additionally, MYD88 was significantly associated with overall survival in PDAC patients in the TCGA cohort (HR=1.58, P=0.045). The MYD88/NF-κB signaling is associated with chemoresistance as it can upregulate the efflux transporters like P-glycoprotein (P-gp). Our results confirmed that combined treatment inhibited the NF-κB signaling pathway by suppressing MYD88 and downregulating P-gp expression to overcome Gem-R. Finally, the combined treatment demonstrated superior anti-cancer activity in 3D organoids by reducing both their number and size (P<0.05). Conclusion: We provide novel insights into the potential of ABE in overcoming Gem-R in PDAC cells by targeting the MYD88/NF-κb/P-gp axis, which offers a safe and inexpensive approach for improving therapeutic outcomes in this fatal malignancy. Citation Format: Yuan Li, Caiming Xu, Haiyong Han, Ajay Goel. Aronia berry overcomes gemcitabine resistance by regulating the MYD88/NF-kB/P-glycoprotein pathway in pancreatic ductal adenocarcinoma [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 4748.

Abstract 4044: Potentiated antitumor activity of poly-γ-glutamic acid combined with anti PD-L1 blockade on murine models of colorectal cancer

Abstract Objective: Immune checkpoint inhibitors(ICIs) have been shown to be key role in cancer therapy. Especially, antibodies targeting programmed cell death ligand1(PD-L1) promoted T cells to activate immune response against cancer cells. However, the current ICIs still have insignificant to motivate effective antitumor immunity by blocking PD-1/PD-L1 pathway and the regulatory network of PD-L1 by cytokines in the tumor microenvironment remains to be uncovered. Previously, we have reported the anti-cancer effects of combination treatment with a natural product, poly-γ-glutamic acid(γ-PGA), and PD-1 blockade. Now, we show that combination therapy with anti PD-L1 and γ-PGA can overcome the limits of ICIs and enhance anticancer effect in murine colorectal cancer. Design: In vivo efficacy studies were performed by intraperitoneal injection of antibody targeting PD-L1 and oral administration of γ-PGA in C57BL/6 model transplanted with mouse tumor cell lines. Systemic changes in immunophenotype was measured by tumor volume size and body weight. The splenocyte was isolated from mice and the populations of myeloid-driven suppressor cells(MDSCs) and CD4/CD8 T cells were analyzed by FACS. Also, the significant cytokines activated by T- cell, IFN-γ and IL-2, were measured by ELISA. Results and Conclusions: In murine model of colorectal cancer, γ-PGA and anti PD-L1 blockade combination therapy significantly improved anti-tumor activity compared with single agents. In addition, tumoral infiltration of CD8+ T cells and IFN-γ productions were increased by γ-PGA, and its antitumoral activity was attenuated by antibody-mediated depletion of these cells, indicating that g-PGA suppressed tumor growth in a CD8 T cell-dependent manner. Therefore, our results suggest that γ-PGA promote ICI-mediated antitumor responses by the transformation of "cold tumors" into “hot tumors” Citation Format: Jae-Pyung Jang, Chan-Hee Yoo, Kyung-Soo Hahm, Do Young Lee, Young-Chul Park, Jae Chul Choi. Potentiated antitumor activity of poly-γ-glutamic acid combined with anti PD-L1 blockade on murine models of colorectal cancer [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 4044.

Abstract 4593: A synergy between MDM2 and AURKB pathways in driving cell cycle and cell survival in lung cancer cells

Abstract Lung cancer is the leading cause of cancer-related deaths worldwide and non-small cell lung cancer (NSCLC) is the most prevalent type among lung cancers, contributing to the high mortality seen with this deadly disease. Despite aggressive treatments, the 5-year survival rate for lung cancer is only 23.7% in the United States. MDM2 and AURKB are two cancer-driving genes that are highly expressed in a broad range of cancer types including lung cancers. Furthermore, MDM2 overexpression can positively impact AURKB expression and contribute to oncogenesis and cancer progression. Therefore, blocking MDM2 with specific inhibitors has become a promising strategy for the restoration of p53 function that could be used for the treatment of different types of cancers including lung cancers. In this regard, both MDM2 and AURKB inhibitors have shown promising antitumor activity against NSCLC. Therefore, the main objective of this study was to understand the molecular mechanisms linking MDM2 and ARKB using RG7388, CM272, BMS-582949 (p38 inhibitor), and Ralimetinib (p38 inhibitor) in A549 and H460 lung cancer cells. We analyzed the individual or combination treatment effects of these drugs on epigenetic modifications, cell cycle arrest, and cell death mechanisms, using the in vitro experimental model. When RG7388 was used individually, or in combination with BMS-582949 or Ralimetinib, a significant increase in the cytotoxic effect was observed in A549 and H460 cells. Induction of cell cycle arrest by RG7388 in cancer cells was evidenced by elevated p21 expression levels. In addition, RG7388 and combination treatments in NSCLC cells showed significant upregulation of p53, p27, Bax, cleaved PARP, phospho-FOXO3a and down-regulation in DNMT3a, DNMT1, FOXM1, AURKB, CDC25, and CDK4 levels. It appears that during treatments with a combination of MDM2 with DNMT or p38 inhibitor, two pathways might be blocked simultaneously, which could lead to increased cancer cell death. RG7388 appears to induce cell death in a p21-dependent manner and CM272, BMS-582949, and Ralimetinib seem to induce cell death via a p21-independent manner by inhibiting a downstream pathway. However, further studies are needed to fully understand the intracellular link between MDM2 and AURKB that is disrupted by the anti-cancer agents used in our study. (This project was supported by the PFRDG Grant of Nova Southeastern University and by the Generous support from the Royal Dames of Cancer Research Ft. Lauderdale, FL) Citation Format: Umamaheswari Natarajan, Appu Rathinavelu. A synergy between MDM2 and AURKB pathways in driving cell cycle and cell survival in lung cancer cells [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 4593.

Individual and combined treatments of 2,4-epibrassinolide (EBR) and calcium chloride (CaCl2) maintain the postharvest quality of baby mustard

Baby mustard is a characteristic vegetable of the Southwest China, and its quality, including appearance quality and nutritional quality, deteriorated during the postharvest senescence process. This study investigated the effects of individual and combined treatments with 2,4-epibrassinolide (EBR) and calcium chloride (CaCl2) on the visual quality and health-promoting compounds of baby mustard stored at room temperature. In terms of appearance, individual treatments with EBR and CaCl2 were more effective than the control and combined treatment in sensory evaluation and weight loss inhibition. All three exogenous treatments maintained the content of pigments, especially individual treatments. The total chlorophyll content was 1.6-, 1.6-, and 1.1-fold higher in EBR-, CaCl2-, and EBR + CaCl2-treated samples than in control samples at 4 days, respectively. In terms of nutritional quality, all three exogenous treatments delayed the decrease in glucosinolates and soluble protein content, with EBR treatment being the most effective in preserving the content of indolic glucosinolates. Additionally, three exogenous treatments effectively maintained antioxidant levels, thereby reducing hydrogen peroxide accumulation. Combined treatment inhibited brassinosteroid signaling, revealing a potential antagonistic relationship between EBR and CaCl2. The results suggest that the three exogenous treatments delayed senescence and preserved the postharvest quality of baby mustard. EBR treatment was the most effective in preserving pigments. This experiment provides potential technical and theoretical basis for the postharvest preservation of baby mustard.

Modulation of TNF-α, interleukin-6, and interleukin-10 by nebivolol-valsartan and nebivolol-lisinopril polytherapy in SHR rats.

Antihypertensive drug therapies have demonstrated their capacity to modulate the inflammatory processes associated with hypertension, leading to improvements in disease progression. Given the prevalent use of polytherapy in treating most hypertensive patients, comprehending the time-dependent effects of combination treatments on inflammation becomes imperative. In this study, spontaneously hypertensive rats (SHR) were divided into seven groups (n = 6): (i) SHR + vehicle, (ii) SHR + nebivolol, (iii) SHR + valsartan, (iv) SHR + lisinopril, (v) SHR + nebivolol-valsartan, (vi) SHR + nebivolol-lisinopril, and (vii) WKY + vehicle. Blood pressure was measured using the tail-cuff method. Temporal alterations in inflammatory cytokines TNF-α, IL-6, and IL-10 were assessed in serum through ELISA and mRNA expression in aortic tissue via qPCR after 1, 2, and 4 weeks of treatment with nebivolol, lisinopril, valsartan, and their respective combinations. Histological alterations in the aorta were assessed. The findings indicated that combined treatments reduced systolic and diastolic blood pressure in SHR. The nebivolol and lisinopril combination demonstrated a significant decrease in IL-6 serum and mRNA expression at both 1 week and 4 weeks into the treatment. Additionally, TNF-α mRNA expression also showed a reduction with this combination at the same time points. Particularly, nebivolol-valsartan significantly decreased TNF-α serum and mRNA expression after one and four weeks of treatment. Furthermore, an elevation in serum IL-10 levels was observed with both combination treatments starting from the second week onwards. This study provides compelling evidence that concurrent administration of nebivolol with lisinopril or valsartan exerts time-dependent effects, reducing proinflammatory cytokines TNF-α and IL-6 while modifying IL-10 levels in an experimental hypertensive model.

Induction of Exocytosis Rescues Lysosomal GM2 Accumulation in Tay-Sachs Disease

Introduction. The Tay-Sachs disease is a progressive neurodegenerative disorder that is caused by a genetic mutation in the HEXA gene coding the lysosomal α-subunit of β-hexosaminidase A. Currently, there is no effective treatment for Tay-Sachs. Induction of exocytosis as a potential treatment approach is suggested to restore lysosomal enlargement in several lysosomal storage diseases. Here, we aimed to test the therapeutic potential of two small molecules, δ-tocopherol and hydroxypropyl-β-cyclodextrin, in fibroblast and neuroglia cells derived from Hexa-/-Neu3-/- mice and Tay-Sachs patients. Method. The effect of two small molecules on lysosomal enlargement and GM2 accumulation in lysosomes was examined by LysoTracker staining and immunocytochemical colocalization analysis for GM2 and LAMP1. qRT-PCR and fluorometric enzyme assay were also used to investigate the effect of combined treatment on the level of neuraminidase 1, a negative regulator of exocytosis. Results. Single treatment with δ-tocopherol (5-40 μM) and hydroxypropyl-β-cyclodextrin (10-50 μM) for 48 hours led to significant induction of lysosomal exocytosis. We demonstrated that the combined treatment with δ-tocopherol (10 μM) and hydroxypropyl-β-cyclodextrin (25 μM) resulted in a significant reduction of lysosomal GM2 and downregulation of lysosomal Neu1 expression. Conclusion. In this study, we demonstrated that inducing exocytosis by δ-tocopherol and hydroxypropyl-β-cyclodextrin might have therapeutic potential to reduce GM2 storage and pathology in Tay-Sachs cells.

Synergistic effect of mesoporous silica nanocarrier-assisted photodynamic therapy and anticancer agent activity on lung cancer cells

Investigating combined treatment methodologies is crucial for addressing the complex nature of cancer. As an emerging strategy, nano-biotechnology encourages the design of unique nanocarriers possessing simultaneous therapeutic application properties. This study aims to explore the combined effects of photodynamic and anticancer treatments using a multifunctional nanocarrier system co-administering the photosensitizer IR780 and the anticancer agent curcumin (Cur) on lung cancer cells. Nanocarriers were prepared by encapsulation IR780 and Cur inside polyethylene glycol-capped mesoporous silica nanoparticles (Cur&IR780@MSN). Various concentrations of nanocarriers were evaluated on A549 cells following 5 min NIR laser light (continuous wave, 785 nm, 500 mW/cm2) irradiation. The internalization of nanocarriers was observed through the fluorescence of Cur. Changes in cell viability were determined using the MTT assay and AO/PI staining. A scratch assay analysis was also performed to examine the impact of combined treatments on cell migration. Characterization of the nanocarriers revealed adequate hydrophobic drug loading, temperature-inhibited feature, enhanced reactive oxygen species generation, a pH-dependent curcumin release profile, and high biocompatibility. Cur&IR780@MSN, which enabled the observation of synergistic treatment efficacy, successfully reduced cell viability by up to 78%. In contrast, monotherapies with curcumin-loaded nanocarriers (Cur@MSN) and IR780-loaded nanocarriers (IR780@MSN) resulted in a 38% and 56% decrease in cell viability, respectively. The constructed Cur&IR780@MSN nanocarrier has demonstrated remarkable performance in the application of combination therapies for lung cancer cells. These nanocarriers have the potential to inspire future studies in tumor treatment methods.

An Investigation into the Effect of Length Scale of Reinforcement on the Cryogenic Response of a Mg/2wt.%CeO2 Composite

The present study attempted for the first time an investigation on the effect of deep cryogenic treatment in liquid nitrogen (LN) on magnesium–cerium oxide (Mg/2wt.%CeO2) composites containing equal amounts of different length scales (micron and nanosize) cerium oxide (CeO2) particles. The disintegrated melt deposition method was used to synthesize Mg-2CeO2 micro- and nanocomposites, followed by hot extrusion as the secondary processing. Further liquid nitrogen treatment was performed at a cryogenic temperature of −196 °C. The combined effects of cryogenic treatment and reinforcement length scale on physical, mechanical, and thermal behaviors were studied. The results indicate that LN-treated micro- and nanocomposite samples exhibit, in common, a reduction in porosity, similar grain size, and a limited effect on the original texture of the matrix. However, microhardness, 0.2% Compressive Yield Strength (CYS), failure strain, and energy absorbed increased for both micro- and nanocomposite samples. Overall, results clearly indicate the capability of deep cryogenic treatment with LN to positively diversify the properties of both micro- and nanocomposite samples.

Alteration of growth, phenology, and yield of lily flowers through the synergetic effect of light spectra and endophytic bacterial priming

The production of ornamental flowers in controlled environment agriculture (CEA) has gained significant attention due to its potential for efficient cultivation. This study aimed to evaluate the influence of light quality (red, blue, red+blue, and white) after priming bulbs with Bacillus subtilis (Bs) on lily plant growth, phenology, yield, ornamental attributes, and pigmentation. Both light quality and Bs treatment significantly affected vegetative and generative organ development and overall plant growth. Bulbs primed with Bs and grown under blue light exhibited the earliest budding and flowering phases, along with an extended flowering period. The highest flower yield and productivity were achieved in Bs-primed bulbs under blue light. Regarding qualities, the best scores were obtained for Bs-primed bulbs grown under blue or red+blue light, compared to control plants grown under red light. The combined effects of Bs treatment and light quality also impacted photosynthetic pigment content in lily leaves. Throughout the growing season, the highest levels of photosynthetic pigments were detected under white light. Although the ratio between Chl a with other photosynthetic pigments were decreased during the flowering period, Bs treatment positively influenced those rations during the flowering phase when plants grown under monochromatic lights (red and blue). In summary, to achieve high-quality lily flowers in CEAs, the most promising approach involves utilizing a red+blue light spectrum in combination with Bs priming. This research highlights the importance of optimizing growing conditions and understanding the interplay between light quality and biological treatments for enhancing ornamental flower production in CEA systems.

Cardioprotective effect of antioxidant combination therapy: A highlight on MitoQ plus alpha-lipoic acid beneficial impact on myocardial ischemia-reperfusion injury in aged rats

Objective(s): Considering the poor prognosis of ischemic heart disease and the diminished effectiveness of cardioprotective interventions in the elderly, it becomes necessary to investigate the interaction of aging with protection during myocardial ischemia/reperfusion injury (IRI). This study was conducted to assess the impact of mitoquinone (MitoQ) and alpha-lipoic acid (ALA) preconditioning on cardioprotection following IRI in aged rats. MethodsFifty aged male Wistar rats (22–24 months old) were divided into five groups including Sham, IR, and treatment groups receiving ALA and/or MitoQ. Treatment groups were received 100 mg/kg/day ALA by oral gavage and/or 10 mg/kg/day MitoQ by intraperitoneal injection for 14 consecutive days. An in vivo model of myocardial IRI was established through ligation of coronary artery for 30 min and it's reopening for 24 h. The left ventricles were removed at the end of reperfusion to assess oxidative stress indicators, mitochondrial function, and expression of mitochondrial dynamic genes. Myocardial infarct size (IS), hemodynamic parameters, and serum lactate dehydrogenase (LDH) level were also measured. ResultsCombination of MitoQ and ALA reduced oxidative stress, LDH level, and IS in aged hearts subjected to IRI. It also enhanced mitochondrial function and upregulated Mfn1, Mfn2, and Foxo1 and downregulated Drp1 and Fis1 gene expression. Co-administration of MitoQ and ALA partially restored IRI-induced hemodynamic changes to normal state. In all measured parameters, the effect of combined treatment was greater than monotherapies. ConclusionThe combination therapy of MitoQ and ALA demonstrated considerable therapeutic potential in protecting the aging heart against IRI by improving oxidative stress, mitochondrial function, and dynamics in aged rats.

Combined effects of high-intensity ultrasound treatment and hydrogen peroxide addition on the thermal stabilities of myofibrillar protein emulsions at low ionic strengths

In this study, the effects of high-intensity ultrasound (HIU) treatment combined with hydrogen peroxide (H2O2) addition on the thermal stability of myofibrillar protein (MP)–stabilized emulsions in low-salt conditions were investigated. Results showed that compared to using either HIU or H2O2 treatment alone, HIU treatment combined with H2O2 was most effective in enhancing the physical stability of emulsions. Moreover, the emulsion stabilized by MPs co-treated with HIU and H2O2 exhibited the most uniform distribution, highest absolute zeta potential, and optimal rheological properties upon heating. This combination effect during heating was caused by the inhibition of disulfide bond cross-linking of myosin heads by H2O2 and the dissociation of filamentous myosin structures using the HIU treatment. In addition, the results of oxidative stability analysis indicated that the addition of H2O2 increased the content of oxidation products; however, the overall influence on the oxidative stability of emulsions was not significant. In conclusion, the combination of HIU and H2O2 treatment is a promising approach to suppress heat-induced MP aggregation and improve the thermal stability of corresponding emulsions.

Can the combination of antiplatelet or alteplase thrombolytic therapy with argatroban benefit patients suffering from acute stroke? a systematic review, meta-analysis, and meta-regression.

The effectiveness of administering argatroban as a treatment approach following antiplatelet therapy or alteplase thrombolytic therapy in patients with acute stroke is presently uncertain. However, it is important to highlight the potential benefits of combining this medication with known thrombolytics or antiplatelet therapy. One notable advantage of argatroban is its short half-life, which helps minimize excessive anticoagulation and risk of bleeding complications in inadvertent cases of hemorrhagic stroke. By conducting a meticulous review and meta-analysis, we aim to further explore the common use of argatroban and examine the plausible advantages of combining this medication with established thrombolytic and antiplatelet therapies. In this study, we performed a rigorous and methodical search for both randomized controlled trials and retrospective analyses. Our main objective was to analyze the impact of argatroban on the occurrence of hemorrhagic events and the mRS scores of 0-2. We utilized a meta-analysis to assess the relative risk (RR) associated with using argatroban versus not using it. In this study, we analyzed data from 11 different studies, encompassing a total of 8,635 patients. Out of these patients, 3999(46.3%) received argatroban treatment while the remaining 4636(53.7%)did not. The primary outcome of 90-day functional independence (modified Rankin scale (mRS) score≤2) showed that the risk ratio (RR) for patients using argatroban after alteplase thrombolytic therapy compared to those not using argatroban was(RR, 1.00 ([95% CI, 0.92-1.09]; P = 0.97), indicating no statistical significance. However, for patients using argatroban after antiplatelet therapy, was (RR,1.09 [95% CI, 1.04-1.14]; P = 0.0001), which was statistically significant. In terms of hemorrhagic events, the RR for patients using argatroban compared to those not using argatroban was (RR,1.08 [95% CI, 0.88-1.33]; P = 0.46), indicating no statistical significance. The results of this study suggest that further research into combination therapy with argatroban and antiplatelet agents may be warranted, however more rigorous RCTs are needed to definitively evaluate the effects of combination treatment.

The effects of E-beam irradiation and microwave combination treatment on the morphology, structure and physicochemical properties of red adzuki bean starch

This study investigated the effects of microwave (MW, 300W, 900W), electron beam irradiation (EBI, 8, 24 KGy), and their combination (MW-EBI, EBI-MW) on the morphology, structure and physicochemical properties of red adzuki bean starch. MW or EBI treatments had little effect on starch morphology, but high-frequency MW and high doses of EBI combined modification resulted in starch granule cracking. Both EBI and MW treatments retained the C-type crystalline structure of the starch but reduced the relative crystallinity, Mw, and short-range ordering. Besides, starch's short chain (A, B1) content, pasting temperature, solubility, and RDS content were significantly increased. In contrast, long chain (B2, B3), SDS + RS content, ΔH, SP, and RDS content were decreased considerably after MW and EBI treatments. Furthermore, the effect of MW and EBI combined modification on starch was higher than the single modification, and the MW-EBI modification was more effective than EBI-MW. Therefore, this study provides a green and efficient method to modify starch, which helps to broaden the scope of starch application.

An injectable thermosensitive hyaluronic acid/pluronic F-127 hydrogel for deep penetration and combination therapy of frozen shoulder

Frozen shoulder (FS) is a common and progressive shoulder disorder that causes glenohumeral joint stiffness, characterized by inflammation and fibrosis. The treatment options are quite limited, and the therapeutic response is hindered by the fibrous membrane formed by excessive collagen and the rapid removal by synovial fluid. To address these challenges, we designed a hyaluronic acid/Pluronic F-127 (HP)-based injectable thermosensitive hydrogel as a drug carrier loaded with dexamethasone and collagenase (HPDC). We screened for an optimal HP hydrogel that can sustain drug release for approximately 10 days both in vitro and in vivo. In the meanwhile, we found that HP hydrogel could inhibit the proliferation and diminish the adhesion capacity of rat synovial cells induced by transforming growth factor-β1. Furthermore, using an established immobilization rat model of FS, intra-articular injection of HPDC significantly improved joint range of motion compared to medication alone. Relying on sustained drug release, the accumulated collagen fibers were degraded by collagenase to promote the deep delivery of dexamethasone. These findings showed a positive combined treatment effect of HPDC, providing a novel idea for the comprehensive treatment of FS.