GC-ToF-MS Profiling and In Vitro Inhibitory Effects of Selected South African Plants against Important Mycotoxigenic Phytopathogens.

The harmful effects following the ingestion of mycotoxin-contaminated food include the induction of cancers, mutagenicity, immune suppression, and toxicities that target organs of the digestive, cardiovascular, and central nervous systems. Synthetic fungicides are generally associated with a high toxic residue in food and the development of excessive fungal resistance. This study aimed to determine the antifungal activities against mycotoxigenic fungi of selected South African plant leaves and potentially develop plant-derived bio-fungicides, and, furthermore, to explore the in vitro antioxidant activity and the phytochemical spectra of the compounds of the selected medicinal plant extracts. The extracts were tested for antifungal activity against phytopathogenic strains using a microdilution broth assay. Bauhinia galpinii extracts exhibited the lowest minimum inhibitory concentration (MIC) against C. cladospoides and P. haloterans at 24 h incubation periods. C. caffrum had good antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) with 50% inhibitory concentration (IC50) values of 0.013 mg/mL while B. galpini had IC50 values of 0.053 against free radicals of 2,2'-azinobis (3-ethylbenzthiazoline-6-suphonic acid (ABTS). The antimycotoxigenic and antioxidant activity exerted by both B. galpinii and C. caffrum may well be attributed to high TPC. In the GC-ToF-MS analysis, all the selected medicinal plants exhibited the presence of Hexadecanoic acid at varying % areas, while both B. galpinii and C. caffum exhibited the presence of lupeol at % area 2.99 and 3.96, respectively. The compounds identified, particularly the ones with higher % area, may well explain the biological activity observed. Although the selected medicinal plants exhibited a notable biological activity, there is a need to explore the safety profiles of these plants, both in vitro and in vivo.

Microbiomics in Collusion with the Nervous System in Carcinogenesis: Diagnosis, Pathogenesis and Treatment.

The influence of the naturally occurring population of microbes on various human diseases has been a topic of much recent interest. Not surprisingly, continuously growing attention is devoted to the existence of a gut brain axis, where the microbiota present in the gut can affect the nervous system through the release of metabolites, stimulation of the immune system, changing the permeability of the blood-brain barrier or activating the vagus nerves. Many of the methods that stimulate the nervous system can also lead to the development of cancer by manipulating pathways associated with the hallmarks of cancer. Moreover, neurogenesis or the creation of new nervous tissue, is associated with the development and progression of cancer in a similar manner as the blood and lymphatic systems. Finally, microbes can secrete neurotransmitters, which can stimulate cancer growth and development. In this review we discuss the latest evidence that support the importance of microbiota and peripheral nerves in cancer development and dissemination.

Apoptosis in Cancer Cells Is Induced by Alternative Splicing of hnRNPA2/B1 Through Splicing of Bcl-x, a Mechanism that Can Be Stimulated by an Extract of the South African Medicinal Plant, Cotyledon orbiculata.

Alternative splicing is deregulated in cancer and alternatively spliced products can be linked to cancer hallmarks. Targeting alternative splicing could offer novel effective cancer treatments. We investigated the effects of the crude extract of a South African medicinal plant, Cotyledon orbiculata, on cell survival of colon (HCT116) and esophageal (OE33 and KYSE70) cancer cell lines. Using RNASeq, we discovered that the extract interfered with mRNA regulatory pathways. The extract caused hnRNPA2B1 to splice from the hnRNPB1 to the hnRNPA2 isoform, resulting in a switch in the BCL2L1 gene from Bcl-xL to Bcl-xS causing activation of caspase-3-cleavage and apoptosis. Similar splicing effects were induced by the known anti-cancer splicing modulator pladienolide B. Knockdown of hnRNPB1 using siRNA resulted in decreased cell viability and increased caspase-3-cleavage, and over-expression of hnRNPB1 prevented the effect of C. orbiculata extract on apoptosis and cell survival. The effect of the hnRNPA2/B1 splicing switch by the C. orbiculata extract increased hnRNPA2B1 binding to Bcl-xl/s, BCL2, MDM2, cMYC, CD44, CDK6, and cJUN mRNA. These findings suggest that apoptosis in HCT116, OE33, and KYSE cancer cells is controlled by switched splicing of hnRNPA2B1 and BCL2L1, providing evidence that hnRNPB1 regulates apoptosis. Inhibiting this splicing could have therapeutic potential for colon and esophageal cancers. Targeting hnRNPA2B1 splicing in colon cancer regulates splicing of BCL2L1 to induce apoptosis. This approach could be a useful therapeutic strategy to induce apoptosis and restrain cancer cell proliferation and tumor progression. Here, we found that the extract of Cotyledon orbiculata, a South African medicinal plant, had an anti-proliferative effect in cancer cells, mediated by apoptosis induced by alternative splicing of hnRNPA2B1 and BCL2L1.

In vitro antibacterial and cytotoxic activity of leaf extracts of Centella asiatica (L.) Urb, Warburgia salutaris (Bertol. F.) Chiov and Curtisia dentata (Burm. F.) C.A.Sm - medicinal plants used in South Africa.

BACKGROUND: Compounds having both anticancer and antimicrobial activity have promising therapeutic potential due to their selective cytotoxicity and their potential to reduce the occurrence of bacterial and fungal infections in immune-compromised cancer patients. In our quest to find new antimicrobial agents with potent anticancer activity, the biological potential of leaves from the three medicinal plants Centella asiatica, Warburgia salutaris and Curtisia dentata as used by Zulu traditional healers for the treatment of cancer is investigated. METHODS: Extracts were assayed for antibacterial activity using the agar well diffusion and micro plate dilution assay. In addition, minimum bactericidal concentrations (MBC), lactate dehydrogenase (LDH) release assay and rhodamine 6G intake assay were used to ascertain the antibacterial activity. The cytotoxic effects of the plant extracts were determined using tetrazolium-based colorimetric (MTT) cell proliferation assay against MCF-7, human colorectal carcinoma cells (Caco-2), A549 and HeLa cancerous cell lines. RESULTS: The acetone extracts from Waburgia salutaris revealed noteworthy anti-proliferative effect yielding IC50 value of 34.15 µg/ml against MCF-7 cell line, while acetone extract from Curtisia dentata significantly (P ≤ 0.05) revealed promising IC50 values of 41.55, 45.13, 57.35 and 43.24 µg/ml against A549, HeLa, CaCo-2 and MCF-7 cell lines. The extracts further revealed a broad-spectrum antibacterial activity against bacterial strains used in the study. An acetone extract from W. salutaris revealed the highest zone of inhibition and the lowest minimum inhibitory concentration (MIC) of 21.0 mm and 0.16 mg/ml respectively against Staphylococcus aureus. Methanol extract from W. salutaris and ethyl acetate extract from C. dentata revealed 53% inhibition of R6G inside the cell against Staphylococcus aureus and Escherichia coli respectively in a cytosolic lactate dehydrogenase assay, suggesting that the mode of action of such extracts may be through efflux pump. CONCLUSIONS: Overall, the extracts had good antibacterial activity and anti-proliferative effects against selected cancerous cell lines. Given the good antibacterial activity of the extracts the plants may act as an immune booster and prevent infection in immunosuppressed cancer patients. This is further supported by the plants' anti-proliferative potential, bacteriostatic, bactericidal properties and also their ability to block bacterial efflux pump systems.

Ethnobotanical knowledge of the lay people of Blouberg area (Pedi tribe), Limpopo Province, South Africa.

BACKGROUND: Limpopo province, South Africa, has a rich plant diversity and is referred to as one of the hotspots areas within the country. The aim of the current work was to identify and document medicinal plant species used by the indigenous Pedi people of Blouberg area, Limpopo Province, South Africa. METHODS: A total of 40 informants which includes both traditional healers and medicinal plant sellers were randomly selected and asked about the plant species used in treatment of variety of infections using a structured questionnaire. Follow-up visits and various field walks were also used to identify and document various plant species used in Traditional medicine (TM). The interviews were carried out from April 2008 to June 2016 using indigenous language (Sehananwa). RESULTS: A total of 82 medicinal plants species belonging to 42 families have been collected, identified and documented. About 46.34% of the plant species were herbs, followed by trees (25.61%), shrubs (20.73%) and climbers (7.32%). The most used plant parts are roots and rhizomes (58.58%). Peltophorum africanum Sond revealed frequency index of greater than 70 and is used in combination with other plants species to treat various pathogenic infections. Most of the plant species reported are used in the treatment of sexually transmitted infections (24), management of HIV-AIDS (15) and stomach ache (14). Our informants indicated that the use of plant medicines in combinations is also applied to cure pathogenic infections. CONCLUSION: The current study demonstrate that the indigenous people of Blouberg area, Limpopo Province harbours an important information about the vegetation around them. The plant species are used in the treatment of various pathogenic infections, offers fruits as additional source of food and form integral part of other medicinal products that may in turn produce income.

Isolation and characterization of the compounds responsible for the antimutagenic activity of Combretum microphyllum (Combretaceae) leaf extracts.

BACKGROUND: Mutations play a major role in the pathogenesis and development of several chronic degenerative diseases including cancer. It follows, therefore that antimutagenic compound may inhibit the pathological process resulting from exposure to mutagens. Investigation of the antimutagenic potential of traditional medicinal plants and compounds isolated from plant extracts provides one of the tools that can be used to identify compounds with potential cancer chemopreventive properties. The aim of this study was to isolate and characterise the compounds responsible for the antimutagenic activity of Combretum microphyllum. METHODS: The methanol leaf extract of C. microphyllum was evaluated for antimutagenicity in the Ames/microsome assay using Salmonella typhimurium TA98. TA100 and TA102. Solvent-solvent fractionation was used to partition the extracts and by using bioassay-guided fractionation, three compounds were isolated. The antimutagenic activity of the three compounds were determined in the Ames test using Salmonella typhimurium TA98, TA100 and TA102. The antioxidant activity of the three compounds were determined by the quantitative 2,2-diphenyl-1-picrylhydrazyl (DPPH)-free radical scavenging method. The cytotoxicity was determined in the MTT assay using human hepatocytes. RESULTS: A bioassay-guided fractionation of the crude extracts for antimutagenic activity led to the isolation of three compounds; n-tetracosanol, eicosanoic acid and arjunolic acid. Arjunolic acid was the most active in all three tested strains with a antimutagenicity of 42 ± 9.6%, 36 ± 1.5% and 44 ± 0.18% in S. typhimurium TA98, TA100 and TA102 respectively at the highest concentration (500 µg/ml) tested, followed by eicosanoic acid and n-tetracosanol. The antioxidant activity of the compounds were determined using the quantitative 2,2 diphenyl-1-picryhydrazyl (DPPH)-free radical scavenging method. Only arjunolic acid had pronounced antioxidant activity (measured as DPPH-free scavenging activity) with an EC50 value of 0.51 µg/ml. The cytotoxicity of the isolated compounds were determined in the MTT assay using human hepatocytes. The compounds had low cytotoxicity at the highest concentration tested with LC50 values >200 µg/ml for n-tetracosanol and eicosanoic acid and 106.39 µg/ml for arjunolic acid. CONCLUSIONS: Based on findings from this study, compounds in leaf extracts of C. microphyllum protected against 4-NQO and MMC induced mutations as evident in the Ames test. The antimutagenic activity of arjunolic acid may, at least in part, be attributed to its antioxidant activity resulting in the detoxification of reactive oxygen species produced during mutagenesis.

The correlation between antimutagenic activity and total phenolic content of extracts of 31 plant species with high antioxidant activity.

BACKGROUND: Antimutagenic activity of plant extracts is important in the discovery of new, effective cancer preventing agents. There is increasing evidence that cancer and other mutation-related diseases can be prevented by intake of DNA protective agents. The identification of antimutagenic agents present in plants presents an effective strategy to inhibit pathogenic processes resulting from exposure to mutagenic and/or carcinogenic substances present in the environment. There are no reports on the antimutagenic activities of the plant species investigated in this study. Many mutations related to oxidative stress and DNA damage by reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been identified in numerous human syndromes. Oxidative DNA damage plays a significant role in mutagenesis, cancer, aging and other human pathologies. Since oxidative DNA damage plays a role in the pathogenesis of several chronic degenerative diseases, the decrease of the oxidative stress could be the best possible strategy for prevention of these diseases. Antioxidant compounds can play a preventative role against mutation-related diseases, and thus have potential antimutagenic effects. METHODS: The number of antioxidant compounds present in methanol leaf extracts of 120 plant species was determined using a combination of Thin Layer Chromatography (TLC) and spraying with 2, 2-diphenyl-1-picrylhydrazyl (DPPH). The 31 most promising extracts were selected for further assays. The quantitative antioxidant activity was determined using DPPH free radical scavenging spectrophotometric assay. Total phenolic contents were determined using the Folin-Ciocalteu colorimetric assay. The mutagenicity of 31 selected extracts was determined in the Ames test using Salmonella typhimurium strains TA98 and TA100. The antimutagenicity of the plant extracts against 4-nitroquinoline 1-oxide (4-NQO) was also determined using the Ames test. RESULTS: Of the 120 plant extracts assayed qualitatively, 117 had some antioxidant activity. The selected 31 extracts contained well defined antioxidant compounds. These species had good DPPH free radical antioxidant activity with EC50 values ranging from 1.20 to 19.06 µg/ml. Some of the plant extracts had higher antioxidant activity than L-ascorbic acid (vitamin C). The total phenolic contents ranged from 5.17 to 18.65 mg GAE (gallic acid equivalent)/g plant extract). The total phenolic content of the plant extracts correlated well with the respective antioxidant activity of the plant extracts. No plant extract with good antioxidant activity had mutagenic activity. Several extracts had antimutagenic activity. The percentage inhibition of 4-NQO ranged from 0.8 to 77% in Salmonella typhimurium TA98 and from 0.8 to 99% in strain TA100. There was a direct correlation between the presence of antioxidant activity and antimutagenic activity of the plant extracts. Although no plant extract had mutagenic activity on its own, some of the plant extracts enhanced the mutagenicity of 4-NQO, a phenomenon referred to as comutagenicity. CONCLUSIONS: Some of the plant extracts investigated in this study had potential antimutagenic activities. The antimutagenic activities may be associated with the presence of antioxidant polyphenols in the extracts. From the results plant extracts were identified that were not mutagenic, not cytotoxic and that may be antimutagenic in the Ames test. For most plant extracts, at the highest concentration used (5 mg/ml), the level of antimutagenicity was below the recommended 45% to conclude whether plants have good antimutagenic activity. However, in most screening studies for antimutagenesis, a 20% decrease in the number of revertants must be obtained in order to score the extract as active. Psoralea pinnata L. had the highest percentage antimutagenicity recorded in this study (76.67 and 99.83% in S. typhimurium TA98 and TA100 respectively) at assayed concentration of 5 mg/ml. The results indicate that investigating antioxidant activity and the number of antioxidant compounds in plant extracts could be a viable option in searching for antimutagenic compounds in plants.

Synthesis and In Vitro Cytotoxic Properties of Polycarbo-Substituted 4-(Arylamino)quinazolines.

Herein, we describe the synthesis of novel unsymmetrical polycarbo-substituted 4-anilinoquinazolines derived from the 2-aryl-6-bromo-8-iodoquinazolines via one-pot three-step reaction sequences involving initial amination and subsequent double cross-coupling (bis-Suzuki, Sonogashira/Stille or Sonogashira/Suzuki-Miyaura) reactions with different cross coupling partners for the two carbon-carbon bond formation steps. The 4-anilinoquinazolines were evaluated for potential cytotoxicity against three cancer cell lines, namely, human breast adenocarcinoma (MCF-7) cells, human cervical cancer (HeLa) and human lung cancer (A549) cells. The most active compounds, 2b, 2c, 3c, 4a, 4c and 5a, were found to be more selective against the MCF-7 and HeLa cell lines than the human lung carcinoma (A549) cells. We selected compounds 2c, 3c and 7a as representatives for further evaluation for potential to induce apoptosis and/or necrotic properties in the three cancer cell lines. Compound 2c induced apoptosis of MCF-7 cells through cell membrane alteration. Treatment of Hela and A549 cell lines with compounds 3c and 7a, respectively, led to caspase-3 activation in both cell lines. Compound 3c, on the other hand, caused more necrosis than apoptosis induction in the membrane alteration assay.

Novel Polycarbo-Substituted Imidazo[1,2-c]quinazolines: Synthesis and Cytotoxicity Study.

Amination of the 2-aryl-6-bromo-4-chloro-8-iodoquinazolines with 2-aminoethanol followed by acid-promoted cyclodehydration of the incipient 2-((6,8-dihalo-2-phenylquinazolin-4-yl)amino)ethanols afforded the corresponding novel 5-aryl-9-bromo-7-iodo-2,3-dihydro-2H-imidazo[1,2-c]quinazolines. The latter were, in turn, subjected to sequential (Sonogashira and Suzuki-Miyaura) and one-pot two-step (Sonogashira/Stille) cross-coupling reactions to afford diversely functionalized polycarbo-substituted 2H-imidazo[1,2-c]quinazolines. The imidazoquinazolines were screened for in vitro cytotoxicity against human breast adenocarcinoma (MCF-7) cells and human cervical cancer (HeLa) cells.

Synthesis, Biological Evaluation and Molecular Docking Studies of 6-Aryl-2-Styrylquinazolin-4(3H)-Ones.

Suzuki-Miyaura cross-coupling of 6-bromo-2-styrylquinazolin-4(3H)-ones with arylboronic acids afforded a series of novel 6-aryl-2-styrylquinazolin-4(3H)-ones. These compounds were evaluated for potential anticancer properties against the human renal (TK-10), melanoma (UACC-62) and breast cancer (MCF-7) cell lines. Their antimicrobial properties were also evaluated against six Gram-positive and four Gram-negative bacteria, as well as two strains of fungi. Molecular docking studies (in silico) were conducted on compounds 5a, b, d and 6a, b, d-f to recognize the hypothetical binding motif of the title compounds within the active site of the dihydrofolate reductase and thymidylate synthase enzymes.

Novel polycarbo-substituted alkyl (thieno[3,2-c]quinoline)-2-carboxylates: synthesis and cytotoxicity studies.

Direct one-pot base-promoted conjugate addition-elimination of 6,8-dibromo-4-chloroquinoline-3-carbaldehyde with methyl mercaptoacetate and subsequent cyclization afforded methyl [(6,8-dibromothieno[3,2-c]quinoline)]-2-carboxylate. The latter undergoes Suzuki-Miyaura cross-coupling with arylboronic acids to yield exclusively the corresponding alkyl [(6,8-diarylthieno[3,2-c]quinoline)]-2-carboxylates,. The cytotoxicity of the prepared compounds was evaluated against the human breast cancer cell line MCF-7 using the MTT assay. The effects of compounds 2, 3c and 4d on cell kinetics were further determined using the xCELLigence Real Time Cell Analysis (RTCA) system. In both the MTT assay and Real Time Cell Analysis, the compounds inhibited cancer cell growth in a dose- and time-dependent manner. Furthermore, on the basis of the calculated LC50 values, the compounds compared favourably with nocodazole, a well-established anticancer drug.