Photodynamic therapy (PDT) utilizes reactive oxygen species (ROS) for eradication of cancer cells. Its effectiveness is governed by the oxygen content, which is scarce in the hypoxic tumor microenvironment. We report herein two zinc(II) phthalocyanines substituted with two or four nitric oxide (NO)-releasing moieties, namely ZnPc-2NO and ZnPc-4NO , which can suppress the mitochondrial respiration, thereby sparing more intracellular oxygen for PDT. Using HT29 human colorectal adenocarcinoma cells and A549 human lung carcinoma cells, we have demonstrated that both conjugates release NO upon interaction with the intracellular glutathione, which can reduce the cellular oxygen consumption rate and adenosine triphosphate generation and alter the mitochondrial membrane potential. They can also relieve the hypoxic status of cancer cells and decrease the expression of hypoxia-inducible factor protein HIF-1α. Upon light irradiation, both conjugates can generate ROS and induce cytotoxicity even under a hypoxic condition, overcoming the oxygen-dependent nature of PDT. Interestingly, the photodynamic action of ZnPc-2NO elicits the release of damage-associated molecular patterns, inducing the maturation of dendritic cells and triggering an antitumor immune response. The immunogenic cell death caused by this oxygen-economized PDT has been demonstrated through a series of in vitro and in vivo experiments.

Introduction: Vernonia amygdalina Delile (VAD), also known as bitter leaf, is widely utilized in traditional medicine for the treatment of various ailments, including cancer. The presence of bioactive compounds in VAD is believed to be responsible for its characteristic bitterness. In Ghana, it is a common practice to mitigate the bitterness of VAD by combining it with Citrus aurantifolia (Christm.) Swingle (lime) juice extracts, although this method lacks scientific evidence and documentation. Therefore, the antioxidant and anticancer activities of VAD and lime juice extracts (V5) and their combined effects were evaluated in vitro. Method: The antioxidant activity and cytotoxic effects of VAD extracts were determined against Jurkat, MCF-7, HepG2, and PNT2 cells using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay to quantify antioxidant activity and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to assess cytotoxicity. The statistical analysis of the data was conducted using Microsoft Excel and GraphPad Prism 8.0. Linear regression was employed to determine the correlation between the concentration and the percentage of antioxidant activity, while p values were calculated using Student's t-test. Results: The laboratory analysis focused on the extracts V1, V2, V3, V4, and V5. Briefly, V1 and V2 contained equal amounts of saponins and terpenoids. Among these, V2 exhibited the highest free radical scavenging activity, as indicated by an EC50 value of 2.14 ± 0.06 mg/mL. V2 also demonstrated cytotoxicity against the MCF-7, HepG2, Jurkat, and PNT2 cell lines. On the other hand, V3 and V4 did not show any cytotoxic effects across all tested cell lines. In contrast, V5 was toxic to HepG2 and MCF-7 cells but had no cytotoxic effect on Jurkat cell lines. V2 exhibited dose-dependent cytotoxicity (0-1000 μg/mL), with the strongest inhibition observed against Jurkat cells (IC50 value = 96.341 μg/mL) and a selective index of 3.567. The difference in activity between the extracts from different parts of the plant and the extract combined with lime juice was significant (p < 0.05), indicating a synergistic effect of the phytochemicals in both VAD and lime juice. Conclusion: V2 and V5 demonstrated a remarkable antioxidant property, and they are effective in inhibiting cancer cell lines, respectively.

Plasmacytoid dendritic cells are a rare subset of dendritic cells that exhibit antiviral functions in response to toll-like receptor 7/8 stimulations. Alternative toll-like receptors such as TLR4 have been known to be active in plasmacytoid dendritic cells for immune regulatory functions. However, it is unclear whether these toll-like receptors differentially activate plasmacytoid dendritic cells as compared with canonical toll-like receptor 7/8 stimulation. Here, we assessed alternative plasmacytoid dendritic cell activation states mediated by toll-like receptors other than endosomal toll-like receptors via the RNA sequencing approach. We found that toll-like receptor 4 stimulation induced a high degree of similarity in gene expression pattern to toll-like receptor 7/8 stimulation in plasmacytoid dendritic cells. Despite high resemblance to toll-like receptor 7/8, we discovered unique genes that were activated under toll-like receptor 4 activation only, as well as genes that were induced at a higher magnitude in comparison to toll-like receptor 7/8 activation. In comparison between toll-like receptor 4-activated plasmacytoid dendritic cells and conventional dendritic cells, we revealed that plasmacytoid dendritic cells and conventional dendritic cells expressed distinct gene sets, whereby conventional dendritic cells mostly favored antigen presentation functions for adaptive immune response regulation while plasmacytoid dendritic cells leaned toward immune response against infectious diseases. Last, we determined that toll-like receptor 4 activation sensitized plasmacytoid dendritic cells against SARS-CoV-2 (COVID-19) single-stranded RNA by enhancing antiviral-related responses and type I interferon production. These findings provided greater insights into the toll-like receptor 4 activation state in plasmacytoid dendritic cells, which can be beneficial for alternative therapeutic interventions involving plasmacytoid dendritic cells for various diseases.

Phytochemical investigation of the rhyzomes of Rumex abyssinicus (Polygonaceae) afforded six anthraquinones viz chrysophanol (1), physcion (2), emodin (3), mixture of physcion-8-O-beta,D-glucopyranoside (4) and chrypsophanol-8-O-beta,D-glucopyranoside (5), and emodin-8-O-beta,D-glucopyranoside (6). All the compounds were characterised and identified by comparison of their MS and NMR data with available literature data. The isolated compounds were evaluated for their antileishmanial activity. Emodin (3) was the most active compounds with IC50 13.82 and 0.26 mu g/mL against Leishmania donovani amastigotes and promastigotes, respectively. Emodin-8-O-beta,D-glucopyranoside (6) also showed a moderate activity with IC50 27.53 and 37.08 mu g/mL. This is the first report of antileishmanial compounds from R. abyssinicus and the antileishmanial activities of compounds 2, 4, 5 and 6 are here reported for the first time.

The aim of this study was to investigate antioxidant and anticancer potentials of natural cocoa extracts. Antioxidant properties of the extracts were evaluated by DPPH assay, total phenolic content by Folin-Ciocalteau and reduced glutathione content by O-phthalaldehyde assays. Antiproliferative activity was also evaluated on leukaemia and prostate cancer cell lines using MTT assay. The hydroethanolic extracts of cocoa roots and beans and aqueous cocoa leaves showed high total antioxidant activities with the root extract having the strongest scavenging effect on DPPH and the highest total phenolics content. The hydroethanolic cocoa bean crude extract showed the highest anticancer activity against LNCaP cells, whiles the ethyl acetate fraction of hydroethanolic cocoa leaf exhibited the highest anticancer against PC3. Jurkat and PC3 cells had similar susceptibilities to the natural cocoa extracts. Thus, cocoa has antioxidant and anticancer activity, and the specific bioactive compounds responsible for these activities need to be elucidated in future research.

Immune modulation is a hallmark of cancer. Cancer–immune interaction shapes the course of disease progression at every step of tumorigenesis, including metastasis, of which circulating tumor cells (CTCs) are regarded as an indicator. These CTCs are a heterogeneous population of tumor cells that have disseminated from the tumor into circulation. They have been increasingly studied in recent years due to their importance in diagnosis, prognosis, and monitoring of treatment response. Ample evidence demonstrates that CTCs interact with immune cells in circulation, where they must evade immune surveillance or modulate immune response. The interaction between CTCs and the immune system is emerging as a critical point by which CTCs facilitate metastatic progression. Understanding the complex crosstalk between the two may provide a basis for devising new diagnostic and treatment strategies. In this review, we will discuss the current understanding of CTCs and the complex immune-CTC interactions. We also present novel options in clinical interventions, targeting the immune-CTC interfaces, and provide some suggestions on future research directions.

Malaria affects about half of the world's population. The sub-Saharan African region is the most affected. Plant natural products have been a major source of antimalarial drugs; the first (quinine) and present (artemisinin) antimalarials are of natural product origin. Some secondary metabolites demonstrate adjuvant antioxidant effects and selective activity. The focus of this study was to investigate the anti-plasmodial activity, cytotoxicities and antioxidant properties of eight (8) Ghanaian medicinal plants. The anti-plasmodial activity was determined using the SYBR green assay and the tetrazolium-based colorimetric assay (MTT) was employed to assess cytotoxicity of extracts to human RBCs and HL-60 cells. Antioxidant potential of plant extracts was evaluated using Folin-Ciocalteu and superoxide dismutase assays. Phytochemical contstituents of the plant extracts were also assessed. All the extracts demonstrated anti-plasmodial activities at concentrations <50 μg/ml. Parkia clappertoniana and Terminalia ivorensis elicited the strongest anti-plasmodial activities with 50% inhibitory concentrations (IC50) of 1.13 μg/ml and 0.95 μg/ml, respectively. This is the first report on anti-plasmodial activities of Baphia nitida, Tabernaemontana crassa and Treculia Africana. T. Africana showed moderate anti-plasmodial activity with IC50 value of 6.62 µg/mL. Extracts of P. clappertoniana, T. Africana and T. ivorensis (0.4 mg/mL) showed >50% antioxidant effect (SOD). The extracts were not cytotoxicity towards RBCs at the concentration tested (200 μg/ml) but were weakly cytotoxic to HL-60 cell. Selectivity indices of most of the extracts were greater than 10. Our results suggest that most of the plant extracts have strong anti-plasmodial activity and antioxidant activity which warrants further investigations.

Most of the current cancer chemotherapeutics are associated with harsh and undesirable side effects, including toxicity and chemoresistance, driving the need for safer and more effective alternatives. In this study, the antiproliferative activities of the methanolic extract of Tetrapleura tetraptera fruits and nine different fractions (C1–C9) from the column chromatographic separation of the extract against leukemia (Jurkat) and human breast cancer (MCF-7) cell lines were investigated using a tetrazolium-based colorimetric assay. Phytochemical screening of the extract and fractions found alkaloids, carbohydrates, flavonoids, glycosides, phenols, saponins, steroids, tannins, and terpenoids in the methanolic extract. Most of the fractions exhibited antiproliferative activity (>100 μg/mL) with the Jurkat cells being more susceptible than the MCF-7 cells. Four of the collected fractions C4, C3, C5, and C2 had good selective indices in decreasing order of activity, in the case of Jurkat cells. Liquid chromatography-mass spectrometry analysis of all samples (except for C4 and C9) revealed that C1, C2, C3, and C5 each had a single component. More importantly, fractions C2, C3, and C5, which were selective to Jurkat cells, also had the same retention time of 1.846 min. Fractions C6 and C8 had two components, with C7 having four components. This study serves as a basis for further work to isolate and characterize potential anticancer agents from the fractions of extracts of T. tetraptera fruits.

Immunological detection methods play an important role in identifying specific biomarkers for disease diagnosis, prognosis, and treatment in precision medicine. With the recent attention shifted towards personalized treatments, immunological detection methods provide a platform for acquiring precise and accurate information using immunoassays. These assays operate on the principle of antigen–antibody interaction to identify disease-specific biomarkers in clinical application. In immunoassays, the antibody–antigen complex is detected through end-reaction outputs that can be analyzed both qualitatively and quantitatively. This book chapter discusses common immunoassays employed in clinical and research settings, emphasizing specificity and accuracy of each method. The chapter will also highlight some recent advancements in immunoassay technology.

Aim: To investigate the antileishmanial potency of Sargassum vulgare C. Agardh-derived products and the in silico inhibition of trypanothione reductase by fucosterol. Materials & methods: Sargassum vulgare crude extract and its derived fractions, subfractions and fucosterol were screened against Leishmania major and Leishmania donovani using the MTS and trypanothione reductase colorimetric assays. Macrophages viability was evaluated using the resazurin assay. The inhibition of trypanothione reductase by fucosterol was predicted in silico. Results: The crude extract, fractions 2, 4 and 7, subfractions 8.2 and 8.3 and fucosterol-exhibited antileishmanial activity on promastigote (IC50 = 18.99–156.02 μg/ml), while fraction 1, subfraction 8.2 and fucosterol were active on L. major and L. donovani amastigote (IC50 = 18.47–65.34 μg/ml) with low cytotoxicity. Interestingly, fucosterol showed the best activity against both parasites (IC50 = 18.47–58.21 μg/ml). Strong binding affinities were recorded between fucosterol and Leishmania spp. trypanothione reductases. Conclusion: Fucosterol, which was abundant in S. vulgare, might be responsible for the antileishmanial activity.

Immunological detection methods play an important role in identifying specific biomarkers for disease diagnosis, prognosis, and treatment in precision medicine. With the recent attention shifted towards personalized treatments, immunological detection methods provide a platform for acquiring precise and accurate information using immunoassays. These assays operate on the principle of antigen–antibody interaction to identify disease-specific biomarkers in clinical application. In immunoassays, the antibody–antigen complex is detected through end-reaction outputs that can be analyzed both qualitatively and quantitatively. This book chapter discusses common immunoassays employed in clinical and research settings, emphasizing specificity and accuracy of each method. The chapter will also highlight some recent advancements in immunoassay technology.

Background: Globally, interest in herbal medicines is increasing. In Ghana, most herbalist use herbal medicines for treatment of various ailments including prostate cancer, although no empirical evidence on their efficacies exists. Objective: The aim of the presentstudy was to test for antioxidant and anti-prostate cancer activities of Moringa oleifera, Phyllanthus amarusand Carica papaya. Methods: Plants parts used were air-dried, ground and sequentially extracted using solvents with increasing order of polarity (petroleum ether, dichloromethane, ethyl acetate, ethanol and aqueous). The 2, 2-diphenyl-1-picrylhydrazyl assay, Folin Ciocalteu method and tetrazolium-based calorimetric assay were used to determine total antioxidant capacities, total phenolic content of extracts and cytotoxicities of the extracts against LNCaP and PC3 prostate cancer cells, respectively. Results: Ethanolic extract of P. amaruspossessed the highest phenolic content while its aqueous extract showed the strongest antioxidant activity (EC50=19.32±1.13 μg/mL). Aqueous extract of C. papayaexhibited anti-prostate cancer activity with good selectivity towards PC3 cells [IC50= 45.68 ±1.16 μg/mL,selectivity index (SI) =18], whereas dichloromethane extract of P. amarusshowed the strongest anticancer activity against LNCaP cells (IC50= 43.97±1.14 μg/mL). Conclusion: These findings lend pharmacological credence to the anecdotal evidence of the anti-prostate cancer property of the plants. Further studies must be performed to identify the active principles in the bioactive plant components.

One new limonoid, trigilgianin (1), one new phenyl alkene, epoxy gilgialkene (2), together with five known compounds: scopoletin (3), sitosteryl-6'-O-undecanoate-β-D-glucoside (4), sitosteryl-O-β-D-glucopyranoside (5), cinchonain A (6) and cinchonain B (7) were isolated from the stem bark of Trichilia gilgiana Harms. (Meliaceae). All compounds were isolated for the first time from this species. The structures were elucidated on the basis of spectral studies and by comparison of these data with those from the literature. Compounds 1, 2, 3, 6 and 7 were tested for in vitro antileishmanial activity against visceral leishmaniasis parasite Leishmania donovani and cytotoxicity against macrophage RAW 264.7 cell line. Compounds 1 and 3 showed the highest antileishmanial activity (IC50 values of 6.044 and 6.804 µg/mL, respectively) with low cytotoxicity (CC50 values of >200 and 47.47 µg/mL, respectively), while compound 2 was moderately active on L. donovani promastigotes (IC50 56.81 µg/mL).

This study sought to evaluate the heavy metal content of the raw powder and extract of the plant, the phytochemical constituents, antioxidant effect by the use of DPPH assay, the total phenolic content using Folin Ciocalteu assay and the cytotoxic effect using the MTT Assay of Ficus pumila ethanolic extract, methanolic and hydro fractions on liver cancer cells (HepG2). Leukemic cells (Jurkat) and normal liver cells (Chang). FTIR and Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the functional groups and major constituents of the most active fraction of F. pumila. Alkaloids, terpenoids, flavonoids, cardiac glycoside, saponins, and tannins were present in the ethanolic extract of F. pumila. The heavy metal analysis revealed the presence of Iron in both the raw powder (1.97 +/- 0.11 mg/l) and extract (0.92 +/- 0.02 mg/L). Zinc was also detected in both the raw powder (1.19 +/- 0.00 mg/l) and extract (0.6595 +/- 0.02 mg/l). The results from the FTIR revealed the presence of alkvnes, alkyl halides, aromatics and aliphatic amines common to all fractions and compounds such as Phenol, 2,4-bis (1,1-Dimethylethyl) and Dodecane, 2,6, 10-trimethyl were detected in the samples by GC-MS. The DPPH assay also showed that all the fractions scavenged DPPH free radical in a dose-dependent manner as compared to the positive control (Ascorbic acid) and positively correlated to the phenolic contents. The MTT assay revealed that methanolic fraction was selective towards the Jurkat cell lines (Selectivity Index = 2.822). This increases the prospects that this plant contains compound(s) which could serve as leads for novel anticancer drugs.

Background There is considerable evidence that many people take dietary supplements including those of herbal origin as an alternative therapy to improve their health. One such supplement, with an amalgam of constituents, is CellGevity®. However, the effect of this dietary supplement on drug-metabolizing enzymes is poorly understood, as it has not been studied extensively. Therefore, we investigated the effect of CellGevity dietary supplement on selected rat liver microsomal cytochrome P450 (CYP) enzymes, the most common drug-metabolizing enzymes. We also determined the total antioxidant potential of this dietary supplement in vitro. Methods To determine the antioxidant potential of CellGevity dietary supplement, 2,2-diphenyl-2-picryl-hydrazyl (DPPH), total phenolic, and flavonoid assays were used after initial preparation of a solution form of the supplement (low dose, LD; 4 mg/kg and high dose, HD; 8 mg/kg). Rats received oral administration of these doses of the supplement for 7 days, after which the effect of the supplement on selected liver CYP enzymes was assessed using probe substrates and spectroscopic and high-performance liquid chromatographic methods. Rats which received daily administration of 80 mg/kg of phenobarbitone and distilled water served as positive and negative controls, respectively. Results The IC50 value of the supplement 0.34 ± 0.07 mg/ml compared to 0.076 ± 0.03 mg/ml of the BHT (positive control). The total phenolic content of the supplement at a concentration of 2.5 mg/ml was 34.97 g gallic acid equivalent (GAE)/100 g while its total flavonoid content at a concentration of 2.5 mg/ml was 6 g quercetin equivalent (QE)/100 g. The supplement significantly inhibited rat CYP2B1/2B2 (LDT 92.4%; HDT 100%), CYP3A4 (LDT 81.2%; HDT 71.7%), and CYP2C9 (LDT 21.7%; HDT 28.5%) while it had no significant inhibitory effect on CYPs 1A1/1A2, CYP1A2, and CYP2D6. Conclusion CellGevity dietary supplement possesses moderate antioxidant activity in vitro and has an inhibitory effect on selected rat liver CYP enzymes, suggesting its potential interaction with drugs metabolized by CYP enzymes.

This study is aimed at isolating colorectal cancer stem-like cells in vitro using a neurosphere assay method employed in isolating gliobastoma multiforme tumor cells. This was followed with confirmation of the isolated cells by flow cytometry, pluripotent genes expression and in vivo tumorigenicity assay. Using this culture assay, stem-like and non-stem-like CRC cells were isolated and expanded in vitro from purchased Balb/c mice induced with CT26 colorectal cancer (CRC) cell line. The procedure includes an initial mechanical dissociation and chemical digestion of tumor tissue and subsequently plating the resulting single cell suspension in serum-free medium (SFM) or serum-containing medium (SCM). This selectively permits growth of cancer stem-like cells in SFM and eliminates non-stem-like cancer cells through the process of anoikis or apoptosis. CRC stem cells derived cultures proliferated as non-adherent spheres in vitro in different shapes and sizes. These cells expressed cell surface markers previously reported for tumor stem cells, including CD44, CD133, CD166 and CD26 and formed tumors when implanted in severe combined immunodeficient mice in a concentration dependent manner. Importantly, the stem-like cells had self-renewal properties with significantly higher expression of the pluripotent stem cell genes NANOG, OCT4, and SOX2 compared to the adherent non-stem cells. Collectively, the results of this study indicate that SFM is a defined culture medium that enriches for CRC stem-like cells and represents a suitable in vitro model for the study of CRC stem-like cells. This finding may be useful in developing therapeutic strategies aimed at eradicating the tumorigenic subpopulation within colorectal cancer.

Background: Weanimix is an important food for children in Ghana.Mothers are trained to prepare homemade weanimix from beans, groundnuts and maize for their infants.Groundnuts and maize are prone to aflatoxin contamination while fumonisin contaminates maize.Aflatoxin, is produced by the Asperguillus fungi while fumonisin, is produced by Fusarium fungi.These mycotoxins occur in tropical areas worldwide due to favorable climate for their growth.Objective: The objective of the study was to determine the levels of aflatoxin and fumonisin in homemade weanimix in the Ejura-Sekyedumase district in the Ashanti Region of Ghana.Methods: Thirty six homemade weanimix samples (50g each) were collected from households.Aflatoxin and fumonisin were measured using a fluorometric procedure described by the Association of Official Analytical Chemist (AOAC official method 993.31,V1 series 4).Results: Aflatoxin and fumonisin were detected in all 36 samples, range 7.9-500ppb.Fumonisin levels range: 0.74-11.0ppm).Thirty (83.3%) of the thirty six samples were over the action limit of 20ppb for aflatoxin with an overall mean of 145.2 ppb whiles 58.3% of the samples had fumonisins above the action limit of 4 ppm with an overall mean of 4.7 ppm .Conclusion: There were significant aflatoxin and fumonisin contamination of homemade weanimix.Children fed on this nutritional food were being exposed to unacceptable levels of aflatoxin and fumonisin.Therefore there is a critical need to educate mothers on the dangers of mycotoxin exposure and to develop strategies to eliminate exposure of children fed homemade weanimix to aflatoxin and fumonisin.

Aflatoxin B1 is a persistent public health issue in Ghana. Assessment of AFB1 intervention efficacy is currently dependent on long-term biomarkers. This study was designed to determine whether daily AFM1 biomarker levels could be utilized as an early detection method for intervention efficacy. Participants were treated with a refined calcium montmorillonite clay (UPSN) or a placebo (calcium carbonate) in a crossover study. Urine samples were assessed for AFM1 levels daily. UPSN treatment reduced AFM1 biomarkers by 55% compared to the placebo. This is the first study to show that daily urinary AFM1 levels can be used as a biomarker of internal aflatoxin B1 exposure in short-term intervention trials to determine efficacy.