Chimeric antigen receptor (CAR) T cell therapies targeting single antigens have performed poorly in clinical trials for solid tumors due to heterogenous expression of tumor-associated antigens (TAAs), limited T cell persistence, and T cell exhaustion. Here, we aimed to identify optimal CARs against glypican 2 (GPC2) or CD276 (B7-H3), which were highly but heterogeneously expressed in neuroblastoma (NB), a lethal extracranial solid tumor of childhood. First, we examined CAR T cell expansion in the presence of targets by digital droplet PCR. Next, using pooled competitive optimization of CAR by cellular indexing of transcriptomes and epitopes by sequencing (CITE-Seq), termed P-COCC, we simultaneously analyzed protein and transcriptome expression of CAR T cells to identify high-activity CARs. Finally, we performed cytotoxicity assays to identify the most effective CAR against each target and combined the CARs into a bicistronic “OR” CAR (BiCisCAR). BiCisCAR T cells effectively eliminated tumor cells expressing GPC2 or CD276. Furthermore, the BiCisCAR T cells demonstrated prolonged persistence and resistance to exhaustion when compared with CARs targeting a single antigen. This study illustrated that targeting multiple TAAs with BiCisCAR may overcome heterogenous expression of target antigens in solid tumors and identified a potent, clinically relevant CAR against NB. Moreover, our multimodal approach integrating competitive expansion, P-COCC, and cytotoxicity assays is an effective strategy to identify potent CARs among a pool of candidates.

The chemical composition and biological activity of three parts (rind, flesh and seeds) of pumpkin fruits (Cucurbita pepo L.) cultivated in Egypt were studied. Chemical analysis of fibre, protein, β-carotene, carbohydrates, minerals and fatty acids present in the rind, flesh, seeds and defatted seeds meal was conducted. Chemical, GC-MS and biological assays of organic extracts of the main fruit parts, rind and flesh established their unique constituents. Chromatographic purification of the extracts afforded triglyceride fatty acid mixture (1), tetrahydro-thiophene (2), linoleic acid (3), calotropoleanly ester (4), cholesterol (5) and 13(18)-oleanen-3-ol (6). GC-MS analysis of the extract's unpolar fraction revealed the existence of dodecane and tetradecane. Structures of the isolated compounds (1-6) were confirmed by NMR and EI-MS spectrometry. Antimicrobial, antiviral and antitumour activities of the fruit parts were discussed. The promising combined extract of rind and flesh was biologically studied for microbial and cytotoxic activities in comparison with the whole isolated components.

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This study attempts to elucidate the secondary metabolite profiles of Ficus lyrata leaves and fruits grown in Egypt. Non-targeted metabolite profiling via ultra performance liquid chromatography (UPLC)-qTOF-MS was used to identify various chemical classes in F. lyrata fruits and leaves (i.e. flavonoids, phenolic acids and fatty acids) analysed by chemometrics. A total of 72 metabolites were evaluated via a UPLC-qTOF-MS-based metabolomic study. Seventeen flavonoids were characterised and tentatively identified with the main constituents being catechins/procyanidins, O- and C-linked flavonoid glycosides. The major procyanidins were dimers and trimers comprising (epi)catechin and (epi)afzelechin units, whereas the predominant flavones were C-glycosides of luteolin and apigenin. Aside from these major flavonoid classes, a group of benzoic acids, caffeoylquinic acids, fatty acid and sphingolipids were also annotated. This study provides the most complete map for polyphenol distribution in F. lyrata leaves and fruits and the basis for future investigation of its fruits nutritional value or possible nutraceutical uses.

Background: The objective of this study was to determine the impacts arising from the substitution of prickly pear fruits (Opuntia ficus indica) peels at different levels (0, 5, 10, and 15%) with yellow corn grain in broiler diets and meat quality. In this study, 200 male chick of commercial Cobb breed were used to study the effect of partially replacing of yellow corn grain (YC) with prickly pear fruits peel powder (PPP). Chicks were divided into four treatments (50 birds each), each treatment contained 5 replicates of 10 birds each. The experimental treatments were the following: 1-control diet (R1). 2-diet containing 5% pp (R2). 3-diet containing 10% pp (R3), and 4diet containing 15% pp (R4). Feeding trails extending to 6 weeks. Results: Obtained results illustrated that feed intake, live body weight, feed conversion ratio of broiler fed diet containing PPP were better (P < 0.05) than that of control. Broilers fed diet containing 5, 10, and 15% PPP were heavier (P < 0.05) by 1.60, 3.68, and 5.78%, respectively, over those fed the control diet. Inoculation of diets containing PPP achieved high (P < 0.05) carcasses weight and dressing% compared with control. Significant differences (P < 0.05) were detected for serum total protein, and total globulin of blood serum broiler as feeding inclusion levels of tested diets. Broiler fed diets contained 15% achieved higher degrees of taste, color, odor (aroma), texture, and overall acceptability. Highest (P < 0.05) biological value, true-digestibility and net protein utilization of broiler meat-fed PPP than other groups of control which was expected, as casein is a protein source with optimal digestibility. Conclusions: It could be concluded that feeding commercial Cobb chicken on diets containing 15% prickly pear peel (Opuntia ficus indica) peel substituted with yellow corn grain, resulted in superior nutrition status and better daily gain, feed conversion ratio, economical efficiency, and better meat quality, as compared with other groups could be recommended.