Differences in hypericin synthesis between experimentally induced seedling shoot cultures of Hypericum hookerianum Wight & Arn.

Varghese, Reji Joseph,Sooriamuthu, Seeni 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.4

During seed germination trials of Hypericum hookerianum, seedlings of Lake View accession from Palni hills of Southern India segregated into green- (97.44 %) and red-pigmented (2.56 %) types. Seedlings cultured in Murashige and Skoog (1962) basal medium developed into fast growing green and slow growing red plant types in 6 weeks, the latter showing increased concentrations of total phenols, anthocyanins and flavonoids and 19-fold higher concentration of hypericin. Hypocotyls/cotyledons of red seedlings cultured using $2.325{\mu}M$ kinetin (KIN) produced hypericin-rich ($4.38{\pm}0.18$ mg/g DW), stunted (0.5-1.2 cm) shoots which ceased to grow in 8 weeks. Segments (4-6 mm) of these shoots sub-cultured in the dark for 4 weeks followed by 2-week light exposure and repeated subculture enabled mass multiplication of productive ($3.93{\pm}0.06$ mg hypericin/g DW) shoots. Green hypocotyls and cotyledons subjected to 4+2 weak dark-light treatment also produced $9.18{\pm}2.44$ and $4.25{\pm}0.96$ comparable hypericin-rich ($3.73{\pm}0.21$ mg/g DW) shoots. Red and green seedling explants cultured in basal medium in the dark produced $6.82{\pm}0.75$ cm etiolated shoots with reduced leaves which synthesized $2.27{\pm}0.15$ mg hypericin/g DW on illumination. Green cotyledons cultured in the dark using $2.45{\mu}M$ indole-3 butyric acid (IBA) formed calluses which on illumination formed $12.64{\pm}3.8$ productive ($3.86{\pm}0.31$ mg hypericin/g DW) 0.5- to 1.5-cmlong shoot clusters. Phenotypic segregation of seedlings, the ability of both red and green seedling explants to multiply in the dark and produce hypericin on illumination, and IBA-induced indirect shoots producing significant amounts of metabolite compared to wild plants ($0.35{\pm}0.09$ mg/g DW) and green shoot cultures ($0.91{\pm}0.03$ mg/g DW) are new to Hypericum.

Differences in hypericin synthesis between experimentally induced seedling shoot cultures of Hypericum hookerianum Wight & Arn.

Reji Joseph Varghese,Seeni Sooriamuthu 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.4

During seed germination trials of Hypericum hookerianum, seedlings of Lake View accession from Palni hills of Southern India segregated into green- (97.44 %) and red-pigmented (2.56 %) types. Seedlings cultured in Murashige and Skoog (1962) basal medium developed into fast growing green and slow growing red plant types in 6 weeks, the latter showing increased concentrations of total phenols, anthocyanins and flavonoids and 19-fold higher concentration of hypericin. Hypocotyls/cotyledons of red seedlings cultured using 2.325 lM kinetin (KIN) produced hypericin-rich (4.38 ± 0.18 mg/g DW), stunted (0.5–1.2 cm) shoots which ceased to grow in 8 weeks. Segments (4–6 mm) of these shoots sub-cultured in the dark for 4 weeks followed by 2-week light exposure and repeated subculture enabled mass multiplication of productive (3.93 ± 0.06 mg hypericin/g DW) shoots. Green hypocotyls and cotyledons subjected to 4 ? 2 weak dark–light treatment also produced 9.18 ± 2.44 and 4.25 ± 0.96 comparable hypericin-rich (3.73 ± 0.21 mg/g DW) shoots. Red and green seedling explants cultured in basal medium in the dark produced 6.82 ± 0.75 cm etiolated shoots with reduced leaves which synthesized 2.27 ± 0.15 mg hypericin/ g DW on illumination. Green cotyledons cultured in the dark using 2.45 lM indole-3 butyric acid (IBA) formed calluses which on illumination formed 12.64 ± 3.8 productive (3.86 ± 0.31 mg hypericin/g DW) 0.5- to 1.5-cmlong shoot clusters. Phenotypic segregation of seedlings, the ability of both red and green seedling explants to multiply in the dark and produce hypericin on illumination, and IBAinduced indirect shoots producing significant amounts of metabolite compared to wild plants (0.35 ± 0.09 mg/g DW) and green shoot cultures (0.91 ± 0.03 mg/g DW) are new to Hypericum.

Effects of Rapamycin on Cell Apoptosis in MCF-7 Human Breast Cancer Cells

Tengku Din, Tengku Ahmad Damitri Al-Astani,Seeni, Azman,Khairi, Wirdatul-Nur Mohd,Shamsuddin, Shaharum,Jaafar, Hasnan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.24

Background: Rapamycin is an effective anti-angiogenic drug. However, the mode of its action remains unclear. Therefore, in this study, we aimed to elucidate the antitumor mechanism of rapamycin, hypothetically via apoptotic promotion, using MCF-7 breast cancer cells. Materials and Methods: MCF-7 cells were plated at a density of $1{\times}10^5$ cells/well in 6-well plates. After 24h, cells were treated with a series of concentrations of rapamycin while only adding DMEM medium with PEG for the control regiment and grown at $37^{\circ}C$, 5% $CO_2$ and 95% air for 72h. Trypan blue was used to determine the cell viability and proliferation. Untreated and rapamycin-treated MCF-7 cells were also examined for morphological changes with an inverted-phase contrast microscope. Alteration in cell morphology was ascertained, along with a stage in the cell cycle and proliferation. In addition, cytotoxicity testing was performed using normal mouse breast mammary pads. Results: Our results clearly showed that rapamycin exhibited inhibitory activity on MCF-7 cell lines. The $IC_{50}$ value of rapamycin on the MCF-7 cells was determined as $0.4{\mu}g/ml$ (p<0.05). Direct observation by inverted microscopy demonstrated that the MCF-7 cells treated with rapamycin showed characteristic features of apoptosis including cell shrinkage, vascularization and autophagy. Cells underwent early apoptosis up to 24% after 72h. Analysis of the cell cycle showed an increase in the G0G1 phase cell population and a corresponding decrease in the S and G2M phase populations, from 81.5% to 91.3% and 17.3% to 7.9%, respectively. Conclusions: This study demonstrated that rapamycin may potentially act as an anti-cancer agent via the inhibition of growth with some morphological changes of the MCF-7 cancer cells, arrest cell cycle progression at G0/G1 phase and induction of apoptosis in late stage of apoptosis. Further studies are needed to further characterize the mode of action of rapamycin as an anti-cancer agent.

A review of the phytochemistry and medicinal activities of the popular African food additive: Parkia biglobosa seed

Wasiu Gbolahan Balogun,Ismail Abiola Adebayo,Usman Yusuf,Azman Seeni 경희대학교 융합한의과학연구소 2018 Oriental Pharmacy and Experimental Medicine Vol.18 No.4

Food additives have been used for many decades, mainly to enhance food aesthetics and taste. However, considerable research attention has been placed on identifying food additives with additional medicinal benefits. Despite on-going efforts to document food additives with medicinal benefits, including the Food and Drug Administration database, many of such products have not yet been covered. Expanding the coverage of food additives with medicinal benefits could help accelerate drug discovery efforts. Here, we provide an update of the medicinal properties of the fermented seeds of the African locust bean (Parkia biglobosa), an example of less-known food additives. This condiment is commonly used in West and Central Africa. In this review, the chemical composition of P. biglobosa seeds has been highlighted as a cheap, reliable source of vital minerals and vitamins. The medicinal activities of P. biglobosa seeds include antihypertensive, anti-diabetes, wound healing and antioxidant activities. For a natural product to be accepted as a potential drug or supplement for food, its safety is important, hence, the safety concerns about this seed have been highlighted. The seeds can be considered safe as there are no reported side effects of the food additive. The review also discussed the future direction of research on the medicinal and nutritional values of the seeds. Finally, the medicinal activities identified from the seeds could be further explored for drug discovery.

Apoptosis Induction in MV4-11 and K562 Human Leukemic Cells by Pereskia sacharosa (Cactaceae) Leaf Crude Extract

Asmaa, Mat Jusoh Siti,Al-Jamal, Hamid Ali Nagi,Ang, Cheng Yong,Asan, Jamaruddin Mat,Seeni, Azman,Johan, Muhammad Farid Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.1

Background: Pereskia sacharosa is a genus of cacti widely used in folk medicine for cancer-related treatment. Anti-proliferative effects have been studied in recent years against colon, breast, cervical and lung cancer cell lines, with promising results. We here extended study of anti-proliferative effects to a blood malignancy, leukemia. Materials and Methods: Two leukemic cell lines, MV4-11 (acute myeloid leukemia) and K562 (chronic myeloid leukemia), were studied. $IC_{50}$ concentrations were determined and apoptosis and cell cycle regulation were studied by flow cytometric analysis. The expression of apoptosis and cell-cycle related regulatory proteins was assessed by Western blotting. Results: P sacharosa inhibited growth of MV4-11 and K562 cells in a dose-dependent manner. The mode of cell death was via induction of intrinsic apoptotic pathways and cell cycle arrest. There was profound up-regulation of cytochrome c, caspases, p21 and p53 expression and repression of Akt and Bcl-2 expression in treated cells. Conclusions: These results suggest that P sacharosa induces leukemic cell death via apoptosis induction and changes in cell cycle checkpoint, thus deserves further study for anti-leukemic potential.

Lactobacillus Strains Alleviated Aging Symptoms and Aging-Induced Metabolic Disorders in Aged Rats

HOR YAN YAN,Cheong-Hwa Ooi,Boon-Yin Khoo,Sy-Bing Choi,Azman Seeni,Shaharum Shamsuddin,Chern-Ein Oon,Kee-Leong Ong,정우식,Min-Tze Liong 한국식품영양과학회 2019 Journal of medicinal food Vol.22 No.1

Aging is an inevitable and ubiquitous progress that affects all living organisms. A total of 18 strains of lactic acid bacteria (LAB) were evaluated on the activation of adenosine monophosphate-activated protein kinase (AMPK), an intracellular energy sensor mediating lifespan extension. The cell-free supernatant (CFS) of Lactobacillus fermentum DR9 (LF-DR9), Lactobacillus paracasei OFS 0291 (LP-0291), and Lactobacillus helveticus OFS 1515 (LH-1515) showed the highest activation of AMPK and was further evaluated. The phosphorylation of AMPK by these three LAB strains was more evident in U2OS and C2C12 cells, compared to the other cell lines and control (P < .05). Using premature senescent Sprague-Dawley rats induced by D-galactose (D-gal), the administration of LAB (10 log CFU/rat/day) for 12 weeks prevented the shortening of telomere length in D-gal-treated rats compared to the untreated control (P < .05). LF-DR9 lowered gene expression of p53, a known senescent biomarker, in gastrocnemius muscle and tibia compared to the control. The selected LAB strains also enhanced lipid, renal, and liver profile of rats, suggesting added potential of the strains in preventing aging-induced metabolic diseases. Strain LP-0291 and LH-1515 showed ability to adhere to mucin, no antibiotic resistance, tolerated and proliferated under gastric and intestinal simulated conditions, and inhibited the growth of pathogens Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis, comparable to commercial probiotic LF-DR9 and Lactobacillus sakei Probio 65. This study provided an insight into the potential of LAB for exhibiting antisenescence effects, with potentials as new medicinal foods for targeted antiaging therapies.