Palm oil γ-tocotrienol and α-tocopherol act as potent inducers in the immune response of mouse splenocytes in vitro / Narimah Abdul Hamid Hasani, Khalid bin Abdul Kadir and Wan Zurinah Wan Ngah

Vitamin E may have anti carcinogenesis effect in human and animal models via the mechanism of cell cycle arrest and enhancement of immune system. The cell-mediated immune inducing properties of palm oil vitamin E, for example, γ-tocotrienol and α-tocopherol, were investigated by measuring the mitogenesis response of splenocytes, extracted from normal male Mus musculus to splenic T-lymphocytes mitogens, phytohemagglutinin (PHA; 0.25 μg/mL) and concanavalin A (Con A; 1.0 μg/mL); and B-lymphocytes mitogen i.e., lipopolysaccharide (LPS; 1.0 μg/mL). Both γ-tocotrienol and α-tocopherol enhanced the cell proliferation of mitogen untreated splenocytes as determined by 5-Bromo-2'-deoxy-uridine (BrdU) detection method. Both compounds also enhanced the T-lymphocytes response to PHA and Con A, as well as B-lymphocytes responses to LPS at all concentration used (0-300 μM). γ-Tocotrienol was observed to affect cell proliferation more than α-tocopherol. The uptake of γ-tocotrienol and α-tocopherol into the splenocytes was determined by high performance liquid chromatography (HPLC). γ-Tocotrienol was absorbed into the cells at markedly higher levels than α-tocopherol with the ratio of 4.8 : 1 (p<0.01, n=4) at 300 μM of treatment. This may be the reason of the higher proliferation affect of γ-tocotrienol as compared to α-tocopherol. In conclusion, we are of the opinion that palm oil γ-tocotrienol and α-tocopherol are able to synergistically influence splenocytes proliferation thus enhancing the cellmediated immune system

Aloe emodin induces apoptosis in ER+-breast cancer cells; MCF-7 through IGF-1R signalling pathway

Two-third of breast cancer patients expressed estrogen receptors (ER)s and received endocrine treatment with established anti-estrogens such as tamoxifen. But the action and acquired resistance during treatment are largely unknown. In contrary, phytochemicals are more selective and less cytotoxic to normal cells. Accordingly, we found aloe emodin, an anthraquinone to inhibit the proliferation of ER+-breast cancer cells, MCF-7 with IC50 of 80 μM, but not affecting control breast cells, MCF-10A. Tamoxifen was non-selective to both cells with IC50 of 27 and 38 μM, respectively. Thus, we aimed to investigate the anti-proliferative mechanism of aloe emodin on MCF-7 and its underlying signalling compared to tamoxifen. Cells were treated separately with aloe emodin and tamoxifen at respective IC50 for 72 h. Apoptosis was determined using Annexin V-FITC/PI staining. The expression of insulin-like growth factor-1 receptor (IGF-1R), insulin-like growth factor binding protein (IGFBP)-2 and B-raf gene was investigated using QuantiGene 2.0 Plex assay. Paired-student t-test and ANOVA test were used to compare between untreated and treated cells on the measured parameters. Each treatment was conducted in triplicate and repeated three times. Significance was set at p<0.05. The presences of early and late apoptosis in MCF-7 were seen in both treatments. All target genes were down regulated. The anti-proliferation effect of aloe emodin on MCF-7 is similar with tamoxifen which mediates inhibition of IGF-1R signalling pathway. This suggests aloe emodin as a potential anti-cancer agent to be used in combined anti-estrogen therapy to enhance its efficacy in ER+-breast cancer treatment

Effects of bisphenol a on neonatal cardiomyocytes beating rate and morphology

Bisphenol A (BPA) has been utilised excessively at a global capacity of 2.9 billion kg/year. It is widely used in manufacturing polycarbonate polymers and epoxy resins. Hence, humans are potentially exposed to this chemical substance in their daily life. As a typical endocrine disruptor, BPA exhibits detectable hormone-like properties. Many studies have been linking BPA exposure in humans with the risk of developing cardiovascular disease, however the direct exposure of BPA on cardiomyocytes beating rates and morphology have not been entirely explored. Therefore, in this study, we aimed to investigate the effects of BPA on cells structure and function of neonatal rat cardiomyocytes culture. Cardiomyocytes were isolated from 0 to 2 days old newborn rats and treated with 0.001 to 100 µM concentration of BPA. All cardiomyocytes were subjected to immunostaining, beating frequency assessment assay, MTS assay and Scanning Electron microscopy (SEM). In immunostaining, cardiomyocytes showed positive staining for F-actin. This staining allows identification of the cells thus differentiate cardiomyocytes from other cell types. Significance effects of BPA on cardiomyocytes were observed in MTS assay (p<0.05) and beating rates (p<0.01). Significant reduction (48%-64%, ± 1.5280) was observed in beating rate of cardiomyocytes exposed to 0.1 to 100 µM of BPA. Meanwhile in MTS assay, significant reduction (54%, 0.067 ± 0.0026) in cell viability was observed in cells exposed to 0.1 µM of BPA only. Interestingly, under SEM, cardiomyocytes showed altered cell surface homogeneity after BPA exposure. Exposure of 0.1 to 100 µM BPA lead to flatten of cardiomyocytes cell surface and blurring of the cell borders. This study offers an in vitro evidence of BPA effects on cardiomyocytes morphology and beating rates, thus suggest the potential adverse effect of BPA exposure. However, further investigation would be required to understand how BPA effects normal cells morphology and beating rates of heart cells

Development of custom lead shield and strainer for targeted irradiation for mice in the gamma cell chamber

We presented a development of a custom lead shield and mouse strainer for targeted irradiation from the gamma-cell chamber. This study was divided into two parts i.e., to (i) fabricate the shield and strainer from a lead (Pb) and (ii) optimize the irradiation to the mice-bearing tumour model with 2 and 8 Gy absorbed doses. The lead shielding was fabricated into a cuboid shape with a canal on the top and a hole on the vertical side for the beam path. Respective deliveries doses of 28 and 75 Gy from gamma-cell were used to achieve 2 and 8 Gy absorbed doses at the tumour sites

The impacts of intrauterine Bisphenol A exposure on pregnancy and expression of miRNAs related to heart development and diseases in animal model

This study aimed to examine the impact of BPA exposure on pregnancy and foetuses on cardiac tissues and the expression of cardiac microRNAs (miRNAs) related to heart development and diseases. Pregnancy is known to be the “critical windows” in determining the offspring physical and cells development in their life after birth. The increment of the risk of cardiovascular disease (CVD) in a later stage of life has been reported by few studies demonstrated from prenatal exposure of BPA. BPA has been shown to alter miRNAs expression profiles for organ development, regeneration and metabolic functions. These alterations have been associated with the risk of CVDs. However, the associations between pregnancy outcomes and miRNAs expression in cardiac of mother- and foetuses-exposed to BPA are still not entirely explored. In BPA-exposed pregnant rat groups, a significant weight gained was observed in comparison to control (p < 0.05). Interestingly, significant changes in systolic and diastolic blood pressure between the first and third trimester of BPA-exposed pregnant rats were also observed (p < 0.05). In BPA-exposed pregnant rats, miR-499-5p was significantly altered in the heart (p < 0.01). Meanwhile, altered miR-17-5p, -208-3p, and -210-3p expressions were observed in all heart of the foetuses from BPA-exposed pregnant rats (p < 0.05). In H&E staining, BPA-exposed foetal hearts showed a sign of fibrosis while BPA-exposed pregnant rats showed muscle remnant. Masson trichrome staining further confirmed the presence of fibrosis observed in BPA-exposed foetal heart and reduced expression of cardiac troponin I (cTnI) was also observed in BPA-exposed foetal heart. In summary, altered cardiac miRNAs with histological changes were observed in both mother- and foetus-exposed BPA These findings put forward the importance of future work to further understand how prenatal BPA exposure affect foetuses in their later stage of life