Saccharomyces cerevisiae kinetochore protein (rDsn1p) induced apoptosis in Chinese hamster ovary cells

Dsn1p is a member of the MIND complex that forms part of the yeast kinetochore, which is essential for the proper chromosomal segregation during cell division. Its functionality is gene dosage dependent and it has characteristics of haploinsufficiency. Bioinformatics alignments predicted the existence of nuclear homologues in higher eukaryotic organisms. Literature on the possibility of Dsn1p being a functional homologue of these organisms is scarce. In this study we employed recombinant DNA expression technology to explore whether Dsn1p can function in a mammalian cell line, Chinese Hamster Ovary (CHO). Expression of rDsn1p in CHO cells induced cytopathic effects including changes in cellular morphology and cell size. Inhibition of cell growth was observed at the beginning the fourth post-transfection week. The recombinant CHO cell culture showed cytotoxic effects following the accumulation of the Dsn1p, resulting in apoptotic cell death; as evidenced by the presence of nuclear fragmentation and surface blebbing in the dying cells. This suggests that rDsn1p may interact with the counterpart/ligand of the nuclear homologue of this protein in CHO cells, resulting in nuclear anomalies and inhibition of cell growth, as observed in our previous study using yeast cells

Compatibility of the Omnican (R) Pen Needles with Insulin Pens, Humapen (R) and Novopen (R)

This study was carried out to assess accuracy of Omnican (R) insulin pen needles (29G and 30G) by measuring the weight of insulin delivered in each of 10 depressions of the plunger comparing these using other pen-injectors (Humapen (R) and Novopen (R) for each gauge. We found that the needle-to-needle variation was not statistically significant when the needles were used to dispense insulin using either of the insulin pens (Humapen (R) and Novopen (R). HumaPen (R) insulin pen was found to deliver the insulin closer to set target volume using either gauge (29G and 30G) of the Omnican (R) needles in some of the insulin ranges used in this study

Effects of supplementation with tocotrienol-rich fraction on immune response to tetanus toxoid immunization in normal healthy volunteers

Background/Objectives: Vitamin E is an essential fat-soluble vitamin that has been shown to induce favorable effects on animal and human immune systems. The objective of this study was to assess the effects of tocotrienol-rich fraction (TRF) supplementation on immune response following tetanus toxoid (TT) vaccine challenge in healthy female volunteers. Subjects/Methods: In this double-blinded, placebo-controlled clinical trial, participants were randomly assigned to receive either placebo (control group) or 400 mg of TRF (study group) supplementation daily. Over the 2-month period of the study, volunteers were asked to attend three clinical sessions (that is, on days 0, 28 and 56) and blood samples were obtained from the volunteers during the follow-up. On day 28, all volunteers were also vaccinated with the TT vaccine (20 Lf) intramuscularly. Results: The results from the clinical trial showed that TRF supplementation significantly increased the total vitamin E level in the plasma of the TRF-supplemented volunteers compared with the placebo group, indicating overall compliance. Volunteers supplemented with TRF showed a significantly (P0.05) enhanced production of interferon-γ and interleukin (IL)-4 by the mitogen or TT-stimulated leukocytes compared with the control group. Volunteers from the TRF group produced significantly (P < 0.05) lower amounts of IL-6 compared with the placebo group. Anti-TT IgG production was also significantly (P < 0.05) augmented in the TRF-supplemented group compared with the placebo group. Conclusions: We conclude that TRF has immunostimulatory effects and potential clinical benefits to enhance immune response to vaccines

Tocotrienols are good adjuvants for developing cancer vaccines

<p>Abstract</p> <p>Background</p> <p>Dendritic cells (DCs) have the potential for cancer immunotherapy due to their ability to process and present antigens to T-cells and also in stimulating immune responses. However, DC-based vaccines have only exhibited minimal effectiveness against established tumours in mice and humans. The use of appropriate adjuvant enhances the efficacy of DC based cancer vaccines in treating tumours.</p> <p>Methods</p> <p>In this study we have used tocotrienol-rich fraction (TRF), a non-toxic natural compound, as an adjuvant to enhance the effectiveness of DC vaccines in treating mouse mammary cancers. In the mouse model, six-week-old female BALB/c mice were injected subcutaneously with DC and supplemented with oral TRF daily (DC+TRF) and DC pulsed with tumour lysate from 4T1 cells (DC+TL). Experimental mice were also injected with DC pulsed with tumour lysate and supplemented daily with oral TRF (DC+TL+TRF) while two groups of animal which were supplemented daily with carrier oil (control) and with TRF (TRF). After three times vaccination, mice were inoculated with 4T1 cells in the mammary breast pad to induce tumour.</p> <p>Results</p> <p>Our study showed that TRF in combination with DC pulsed with tumour lysate (DC+TL+TRF) injected subcutaneously significantly inhibited the growth of 4T1 mammary tumour cells as compared to control group. Analysis of cytokines production from murine splenocytes showed significant increased productions of IFN-γ and IL-12 in experimental mice (DC+TL+TRF) compared to control, mice injected with DC without TRF, mice injected with DC pulsed with tumour lysate and mice supplemented with TRF alone. Higher numbers of cytotoxic T cells (CD8) and natural killer cells (NK) were observed in the peripheral blood of TRF adjuvanted DC pulsed tumour lysate mice.</p> <p>Conclusion</p> <p>Our study show that TRF has the potential to be an adjuvant to augment DC based immunotherapy.</p

Bisphenol A Exposure at Puberty Disrupts Expression of the Oestrogen Receptor-Alpha in the Hypothalamus of Male and Female Mice

ABSTRACT Bisphenol A is a xenoestrogen that interacts with both of the oestrogen receptors, oestrogen receptor alpha (ERα) and oestrogen receptor beta (ERβ). At environmentally low doses, bisphenol A has been reported to influence behaviour and affect development of the brain and reproductive tissues in rodents. In the present study mice were treated daily with a low (50 µg/kg) oral dose of bisphenol A starting on postnatal day (PND) 32 until sacrifice (PND50 or PND100), and at autopsy brain, testis, and uterus of all animals were harvested for analysis. Immunohistochemistry was used to evaluate the changes in the number of cells expressing ERα in the arcuate nucleus and ventromedial nucleus of the hypothalamus. Histological evaluations were performed on the testis and uterus, while analysis of estradiol and testosterone was carried out on mouse serum. At PND50, bisphenol A exposure increased ERα expressing neurons in the arcuate nucleus and ventromedial nucleus of the hypothalamus of both male and female mice (p&lt;0.05), while at PND100, bisphenol A exposure only increased ERα expressing neurons in the ventromedial nucleus of the hypothalamus of female mice (p&lt;0.05). No adverse effects were observed in the testis of exposed males, except for consistent thickening of the basement membrane. In the females, mild simple hyperplasia of the endometrial glands was observed with no differences between BPA50 and BPA100 groups. Male mice had decreased testosterone and elevated estradiol serum concentrations at PND50 while females were observed to have increased serum estradiol at PND100. Our results show that bisphenol A can modulate oestrogen receptors in the hypothalamus and indicate a comparable effect of exposure in male and female mice during adolescence but a differential effect during adulthood

Gamma-tocotrienol modifies methylation of HOXA10, IRF4 and RORα genes in CD4⁺ T-lymphocytes:Evidence from a syngeneic mouse model of breast cancer

DNA methylation plays a crucial role in polarising naïve lymphocytes towards their various sub-populations to fight against many immune challenges including establishment of tumour. Gamma-tocotrienol (γT3) is a natural form of vitamin E, reported to possess anticancer and immunomodulatory effects. This study reports the anticancer effects of γT3 through modulation of DNA methylation in several genes in CD4(+) T-lymphocytes using a syngeneic mouse model of breast cancer. Female BALB/c mice were fed with γT3 or vehicle (soy oil) for two-weeks via oral gavage before they were inoculated with 4T1 mouse mammary cancer cells. Supplementation continued until the mice were sacrificed. At autopsy, blood was collected via cardiac puncture and CD4(+) T-cells were isolated for DNA extraction. The DNA was analysed using the EpiTech Methyl II mouse T-helper cell differentiation PCR array. γT3 supplementation reduced tumour growth in the tumour-induced animals and modulated host immune system by inducing changes in DNA methylation patterns of the HOXA10, IRF4 and RORα genes, which are involved in differentiation and clonal expansion of CD4(+) T-cells. Results suggest that γT3 may enhance cell-mediated immune response in mice with breast cancer by inducing changes in DNA methylation pattern

Refining ostrich oil and its stabilization with curcumin

Ostrich Oil is particularly known for its cosmetic and therapeutic use. Being rich in polyunsaturated fatty acids, it is prone to oxidation causing undesirable changes in peroxide value, color, flavour and odor, which deteriorates its nutritional value. The objective of the study is to establish an efficient separation method to refine ostrich oil and stabilize it with curcumin or the extract of Curcuma longa. The reduction in peroxide value, color intensity, moisture content and odor were used as indicators of the purity of the oil. The modified refining process developed saw a reduction of 75% free fatty acid, 90% peroxide value, 54% moisture content, 60% in color intensity and odor. Curcumin, a natural antioxidant, prevented lipid peroxidation by 56% while vitamin E by only 8%, when tested at the same concentration (0.04%). Curcumin proved to be an effective stabilizer in a 6-month accelerated stability test. This study presents a method to refine ostrich oil efficiently and the ability of a natural antioxidant, curcumin, to stabilize the refined ostrich oil.This simple and straight forward method to refine and stabilize the oil can be adapted to be used for any ratite oils. The use of a natural alternative, curcumin as the anti-oxidant is not only safe but provides further added health benefits