Application of Soil Bacteria as Bioinoculants to Promote Growth of Cowpea (Vigna unguiculata)

This work aimed to evaluate the capacity of soil bacteria as bioinoculants (biofertilizers) to promote cowpea (Vigna unguiculata) growth. Three pure bacterial cultures namely Acinetobacter pittii PT1.3.4 (AP), Achromobacter sp.C2.23 (AS), and Achromobacter xylosoxidans N3.4 (AX) were used as bioinoculants to enhance germination and development of cowpea seeds. Pre-decide formulations of single or mixed cultures were prepared, soaked with cowpea seeds, and cultivated on agar in a growth chamber for 7 days at 25°C. Shoot and root length were measured and percentage germination was determined. Similarly, bacterial formulations were prepared in talcum powder and were used as bioinoculants to adhere to cowpea seeds. The inoculated seeds were cultivated in pots for 28 days for the shoot and root length, fresh and dry weight, and percentage germination. Among the tested various formulations, treatment has A. pittii (AP) displayed the highest shoot length (14.67 cm) and fresh weight (0.58 g/plant) of cowpea under laboratory conditions after seven days of inoculation. Similarly, cowpea plants treated with A. pittii (AP) also have the tallest shoots (14.25 cm) under natural conditions after 7 days of inoculation, while the highest root length (10.5 cm) and fresh weight (1.57 g/plant) were recorded from the treatment of Achromobacter sp. (AS). Further, the results of the study also revealed that soil bacteria can survive for one month in talcum powder at 4°C and room temperature storage. These bioinoculants can be used for agricultural application by local farmers to mitigate the cost of chemicals that cause environmental concerns to promote sustainable agriculture in Thailand

Cytotoxicity and antiproliferative activity of essential oils from lemon, wild orange and petitgrain against MCF-7, HepG2 and HeLa cancer cells

The purpose of this study was to determine the chemical composition and biological properties of the citrus essential oils (EOs) derived from orange rinds (peels) of lemon (Citrus limon), wild orange (Citrus sinensis) from Brazil extracted by the cold pressed/expressed method, and leaves and twigs of petitgrain (Citrus aurantium) from Paraguay extracted by steam distillation. These food grade EOs were evaluated for their cytotoxic activity in breast, liver, and cervical cancer cells (MCF-7, HepG2 and HeLa) via MTT assay, antiproliferative activity via colony formation assay, and antimigratory activity via wound healing assay, and apoptosis via DNA fragmentation and morphology assessment. The major compounds found in lemon EO were D-limonene (66.75%), beta-pinene (12.82%), and gamma-terpinene (11.57%), totaling over 90% of the identified compounds. For wild orange, the only predominant compound was limonene (96.60%), and the rest, found in minor amounts, included alpha-pinene, bicyclohexane, beta-pinene, beta-myrcene, 3-carene, and o-cymene. For petitgrain EO, linalyl isobutyrate (51.76%) and linalool (26.86%) were mainly detected. Based on the MTT assay, petitgrain EO was the most effective against MCF-7, HepG2 and HeLa. However, wild orange EO was the most antiproliferative and antimigratory against all three cells using the anticolony formation assay and wound healing assay, respectively. The results showed that cell death is associated with the apoptotic process, with morphological hallmarks of apoptosis including membrane blebbing and DNA fragmentation. These findings imply that the three citrus EOs might be used as active components in functional food products for chemopreventive benefits

Antioxidant properties and cytotoxic effects of Alternanthera sissoo and Alternanthera bettzickiana extracts against cancer cells

This work aimed to evaluate the antioxidant capacities and cytotoxic attributes of Alternanthera sissoo (AS) and Alternanthera bettzickiana Green (AB) which are commonly grown in Thailand, yet not extensively-studied. Dried leaves were extracted by maceration using ethanol for 48 h and ethanolic extract solution was used. Cytotoxic, anti-proliferative, anti-migratory capacities and apoptosis-related gene expressions of AS and AB extracts on MCF-7, HepG2, and HeLa cell lines were investigated using 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetra zolium bromide (MTT) assay, anti-colony formation and wound healing test, respectively. The antioxidant activities by DPPH assay and FRAP assay of AS (130.62±1.61 µg TE/g extract and 24.76 ± 0.54 mg Fe(II)/g extract, respectively) were significantly higher than those of AB (104.34±3.68 µg TE/g extract and 23.13± 0.90 mg Fe(II)/g extract, respectively). Likewise, total phenolic content and the total flavonoid content of AS (7.43±0.39 mg GAE/g and 184.22±3.20 mg RE/g extract, respectively) were significantly higher than AB (6.10±0.29 mg GAE/g extract and 168.07±7.90 mg RE/g extract, respectively). HPLC analysis showed the predominance of myricetin, rutin and ferulic acid. For MCF-7, HepG2 and HeLa, the lowest IC50 values of 48.53 µg/mL, 69.94 µg/mL and 43.98 µg/mL, respectively were found in AS extract at 72 h exposure using MTT assay. Apoptotic bodies, gene expression and protein expressions related to apoptosis were more pronounced in AS treatment. Both AS and AB can be served as local foods with chemopreventive effects against breast, liver and cervical cancers. This work provided a foundation for future pharmacological research on Alternanthera plant extracts

Anticancer effects of Rhinacanthus nasutus and Acanthus ebracteatus extracts against human cervical cancer cells

Cervical cancer is second only to breast cancer in terms of incidence; however, it is the most lethal form of cancer among Thai women due to the asymptomatic nature of its early stages. This work aimed to examine cytotoxic and antiproliferative capacities of Rhinacanthus nasutus (RN) and Acanthus ebracteatus (AE) extracts against human cervical cancer cells (HeLa). Plant leaves were used for ethyl acetate extraction. The antioxidant assays, HPLC analysis, a cytotoxic MTT assay, a clonogenic assay and real-time PCR were conducted. Both RN and AE displayed similar DPPH scavenging activity (3.97 and 4.05 mg TE/g DW) and ferric reducing antioxidant power (4.79 and 4.35 mg Fe2+/g DW). However, AE was richer in total phenolic content than RN (13.30 and 10.84 mg GAE/g DW, respectively). Rutin, catechin, chlorogenic acid, and cinnamic acid were found in AE, whilst only cinnamic acid with much higher content was found in RN. Higher cytotoxicity of 91.73% against HeLa cells was found in RN (IC50 value of 62.06 µg/mL). RN showed higher antiproliferative effect (IC50 of 25.24 µg/mL) than AE (34.35 µg/mL). Genes (Bcl-2, Bax, MMP-2, MMP-9, caspase-3, p21, and cyclin D1) and proteins (cytochrome c, caspase-3 and p21) linked to apoptosis and migration were substantially more affected by RN. To conclude, both RN and AE hold promise as anticancer herbal plants against human cervical cancer; however, RN was more cytotoxic and antiproliferative in HeLa cells. RN offers a better alternative herbal medicine or complementary remedy to the standard drug for human cervical cancer treatment

Cytotoxic effects of Saccharomyces cerevisiae TC6 and Lactobacillus brevis TBRC 3003 isolated from Thai fermented foods

Purpose: To determine the cytotoxic effect, anti-colony formation effect and antimigratory effect of Saccharomyces cerevisiae TC6 isolated from Thai water kefir, and Lactobacillus brevis TBRC 3003 isolated from picked cabbage. Methods: Crude microbial extracts were obtained from whole cultures (cells and broths) using ethyl acetate as extracting solvent, and the dried extracts were redissolved in ethanol before use. Cytotoxic, antiproliferative and antimigratory effects of the two microbial extracts on MCF-7, HepG2, and HeLa were tested using 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetra zolium bromide (MTT), clonogenic formation and wound healing assays. Results: Lb. brevis TBRC 3003 showed the highest cytotoxicity toward HepG2 cells (IC50 of 669.72 µg/mL), while S. cerevisiae TC6 showed the highest cytotoxicity against MCF-7 (IC50 of 691.49 µg/mL) and HeLa (IC50 of 379.16 µg/mL) based on MTT assay. Anti-colony formation test showed that S. cerevisiae TC6 was most the most effective in inhibiting colony formation of HepG2 (IC50 of 311.12 µg/mL) and HeLa (IC50 of 494.64 µg/mL), while Lb. brevis TBRC 3003 was the most potent in inhibiting colony formation of MCF-7 (IC50 of 267.88 µg/mL). Conclusion: Both microbes can potentially be implemented in functional foods as bio-therapeutics with chemopreventive properties against breast, liver and cervical cancers

Cytotoxic and antiproliferative effects of Streblus asper from northeastern Thailand on A549 lung cancer cells

Thailand's second-leading cause of death is lung cancer. Thai indigenous herbal plants are sought after as an alternative treatment against lung cancer. This work aimed to examine cytotoxic, antiproliferative and antimigratory capacities of different parts of Streblus asper (SA) from northeastern Thailand on A549 lung cancer cells. Plant leaves, twigs, bark and wood were used for ethanolic extraction by maceration. The highest cytotoxicity of 85.46% was found in twig extract (IC50 57.46 µg/mL) assessed by an MTT assay. The lowest IC50 (18.42 µg/mL) was also found in twig extract using a clonogenic assay indicating its most antiproliferative activity in a long-term therapy. In addition, all SA extracts displayed antimigratory activity against A549 cells in a dose-dependent fashion, especially twig extract. Apoptotic characteristics were noticeable in SA extract treated cells. The maximum DPPH-scavenging activity, FRAP value, total phenolic and flavonoid content were found in twig extract. GC-MS analysis revealed that twig extract contained four prominent components namely ethyl-α-D-glucopyranoside, hexadecanoic acid, ethyl ester, lupeol and γ-sitosterol. Real-time PCR results showed that genes (Bcl-2, Bax, p21, and cytochrome c) linked to apoptosis were significantly affected by all SA extracts. The various SA components' ethanolic extracts exhibited moderate-to-high cytotoxic action towards A549 cells. This work will significantly advance the utilization of the plant as an alternative source of medicine for rural Thais, and it paves the way for future research to determine the active compound(s) and anticipate new drug candidates

Formation of Sulforaphane and Iberin Products from Thai Cabbage Fermented by Myrosinase-Positive Bacteria

Myrosinase-positive bacteria from local fermented foods and beverages in Thailand with the capacity to metabolize glucosinolate and produce isothiocyanates (ITCs) were isolated and used as selected strains for Thai cabbage fermentation. Enterobacter xiangfangensis 4A-2A3.1 (EX) from fermented fish and Enterococcus casseliflavus SB2X2 (EC) from fermented cabbage were the two highest ITC producers among seventeen strains identified by 16S rRNA technique. EC and EX were used to ferment Thai cabbage (Brassica oleracea L. var. capitata) containing glucoiberin, glucoraphanin and 4-hydroxyglucobrassicin at 430.5, 615.1 and 108.5 µmol/100 g DW, respectively for 3 days at 25 °C. Different amounts of iberin nitrile, iberin, sulforaphane and indole 3-acetonitrile were produced by spontaneous, EX- and EC-induced cabbage fermentations, and significantly higher ITCs were detected (p < 0.01) with increased antioxidant activities. Iberin and sulforaphane production in EX-induced treatment peaked on day 2 at 117.4 and 294.1 µmol/100 g DW, respectively, significantly higher than iberin at 51.7 µmol/100 g DW but not significantly higher than sulforaphane at 242.6 µmol/100 g DW in EC-induced treatment at day 2. Maximum health-promoting benefits from this functional food can be obtained from consumption of a liquid portion of the fermented cabbage with higher ITC level along with a solid portion

Antioxidant activity of mustard green and Thai rat-tailed radish grown from cold plasma treated seeds and their anticancer efficacy against A549 lung cancer cells

Lung cancer is Thailand's second-highest cause of mortality. Mustard green (MG) and rat-tailed radish (RTR) 7-day-old microgreens were previously shown to exhibit cytotoxicity against MCF-7 and HepG7 cells. However, both plants have yet to be tested on A549 lung cancer cells. This study evaluated the antioxidant activity of MG and RTR plants grown from cold plasma treated seeds at different early growth periods and their anticancer activities against A549 lung cancer cells. Plant seeds primed with cold plasma at 21 kV (for MG) and 19 kV (for RTR) for 5 min were grown on vermiculite for 14, 21 and 28 days. Results showed that RTR-P and MG-P (plasma-treated seeds) showed significantly higher ferric reducing antioxidant power (FRAP) and total antioxidant phosphomolybdate activity than RTR-C and MG-C (control seeds). Highest cytotoxicity (Emax) of 95.41% against A549 cells was found in MG-P of 14 days at 72 h exposure with IC50 value of 67.11 µg/mL. Lower IC50 of 30.93 µg/mL was found in RTR-P of 14 days at 72 h exposure and Emax of 93.38%. MG-P and RTR-P had significantly more pronounced effects on apoptosis and migration-related gene expressions (Bax, Bcl-2, caspase-3, p21, MMP-9 and cyclin D1) and also protein expressions (caspase-3, cytochrome c and p21). The RTR-P extract was more cytotoxic and antiproliferative than MG-P in human lung cancer cells. Cold plasma played a key role in enhancing cytotoxicity in these two plants with improved chemopreventive benefits for consumers