Apigenin-7-O-glucoside versus apigenin

Bioactive potential of apigenin derivative apigenin-7-O-glucoside related to its antifungal activity on Candida spp. and cytotoxic effect on colon cancer cells was studied and compared with bioactive potential of apigenin. Antifungal activity was tested on 14 different isolates of Candida spp. using membrane permeability assay, measuring inhibition of reactive oxidative species and inhibition of CYP51 C. albicans enzyme. Cytotoxic potential of apigenin-7-O-glucoside was tested on colon cancer HCT116 cells by measuring cell viability, apoptosis rate and apoptosis- and colon cancer-related gene expression. Obtained results indicated considerable antifungal activity of apigenin-7-O-glucoside towards all Candida isolates. Breakdown of C. albicans plasma membrane was achieved upon treatment with apigenin-7-O-glucoside for shorter period of time then with apigenin. Reduction of intra- and extracellular reactive oxidative species was achieved with minimum inhibitory concentrations of both compounds, suggesting that reactive oxidative species inhibition could be a mechanism of antifungal action. None of the compounds exhibited binding affinity to C. albicans CYP51 protein. Besides, apigenin-7-O-glucoside was more effective compared to apigenin in reduction of cell’s viability and induction of cell death of HCT116 cells. Treatment with both compounds resulted in chromatin condensation, apoptotic bodies formation and apoptotic genes expression in HCT116 cells, but the apigenin-7-O-glucoside required a lower concentration to achieve the same effect. Compounds apigenin-7-O-glucoside and apigenin displayed prominent antifungal potential and cytotoxic effect on HCT116 cells. However, our results showed that apigenin-7-O-glucoside has more potent activity compared to apigenin in all assays that we used

The Role of SOX Transcription Factors in Ageing and Age-Related Diseases

The quest for eternal youth and immortality is as old as humankind. Ageing is an inevitable physiological process accompanied by many functional declines that are driving factors for age-related diseases. Stem cell exhaustion is one of the major hallmarks of ageing. The SOX transcription factors play well-known roles in self-renewal and differentiation of both embryonic and adult stem cells. As a consequence of ageing, the repertoire of adult stem cells present in various organs steadily declines, and their dysfunction/death could lead to reduced regenerative potential and development of age-related diseases. Thus, restoring the function of aged stem cells, inducing their regenerative potential, and slowing down the ageing process are critical for improving the health span and, consequently, the lifespan of humans. Reprograming factors, including SOX family members, emerge as crucial players in rejuvenation. This review focuses on the roles of SOX transcription factors in stem cell exhaustion and age-related diseases, including neurodegenerative diseases, visual deterioration, chronic obstructive pulmonary disease, osteoporosis, and age-related cancers. A better understanding of the molecular mechanisms of ageing and the roles of SOX transcription factors in this process could open new avenues for developing novel strategies that will delay ageing and prevent age-related diseases

Hippocampal asymmetry in expression of catecholamine synthesizing enzyme and transporters in socially isolated rats

OBJECTIVES: Right-left asymmetry of human brain function has been known for a century. Brain asymmetry and lateralization has been observed at the neurochemical level. At the neurochemical level, it is important to further correlate changes in monoaminergic activity with the synthesis and reuptake of these monoamines. The aim of the present study was to analyze the effect of social isolation on catecholamine stores as well as on the regulation of catecholamine synthesis and uptake in the right and left hippocampus. METHODS: We examined changes in protein levels of dopamine-beta-hydroxylase (DBH), norepinephrine transporter (NET) and vesicular monoamine transporter 2 (VMAT2) in the right and left hippocampus of socially isolated adult male rats during 12 weeks by Western blot analysis. RESULTS: Chronic isolation stress reduced norepinephrine content in the right hippocampus. No changes were observed in protein levels of DBH and NET in the right hippocampus, whereas expression of this norepinephrine synthetizing enzyme and transporter were elevated in the left hippocampus. On the other hand, chronic isolation stress caused reduction of VMAT2 protein in the right hippocampus. CONCLUSION: Our results reveale not only the lateralization of stress regulatory system but they also show that long-term isolation stress produces right-left asymmetry of the hippocampus norepinephrine, different regulation of the catecholamines synthesis and reuptake

Hippocampal asymmetry in expression of catecholamine synthesizing enzyme and transporters in socially isolated rats

OBJECTIVES: Right-left asymmetry of human brain function has been known for a century. Brain asymmetry and lateralization has been observed at the neurochemical level. At the neurochemical level, it is important to further correlate changes in monoaminergic activity with the synthesis and reuptake of these monoamines. The aim of the present study was to analyze the effect of social isolation on catecholamine stores as well as on the regulation of catecholamine synthesis and uptake in the right and left hippocampus. METHODS: We examined changes in protein levels of dopamine-beta-hydroxylase (DBH), norepinephrine transporter (NET) and vesicular monoamine transporter 2 (VMAT2) in the right and left hippocampus of socially isolated adult male rats during 12 weeks by Western blot analysis. RESULTS: Chronic isolation stress reduced norepinephrine content in the right hippocampus. No changes were observed in protein levels of DBH and NET in the right hippocampus, whereas expression of this norepinephrine synthetizing enzyme and transporter were elevated in the left hippocampus. On the other hand, chronic isolation stress caused reduction of VMAT2 protein in the right hippocampus. CONCLUSION: Our results reveale not only the lateralization of stress regulatory system but they also show that long-term isolation stress produces right-left asymmetry of the hippocampus norepinephrine, different regulation of the catecholamines synthesis and reuptake

Expression Analysis of SOX14 during Retinoic Acid Induced Neural Differentiation of Embryonal Carcinoma Cells and Assessment of the Effect of Its Ectopic Expression on SOXB Members in HeLa Cells

<div><p>SOX14 is a member of the SOXB2 subgroup of transcription factors implicated in neural development. Although the first <i>SOX14</i> gene in vertebrates was cloned and characterized more than a decade ago and its expression profile during development was revealed in various animal model systems, the role of this gene during neural development is largely unknown. In the present study we analyzed the expression of SOX14 in human NT2/D1 and mouse P19 pluripotent embryonal carcinoma cells. We demonstrated that it is expressed in both cell lines and upregulated during retinoic acid induced neural differentiation. We showed that SOX14 was expressed in both neuronal and non-neuronal differentiated derivatives, as revealed by immunocytochemistry. Since it was previously proposed that increased SOXB2 proteins level interfere with the activity of SOXB1 counteracting partners, we compared expression patterns of SOXB members during retinoic acid induction of embryonal carcinoma cells. We revealed that upregulation of SOX14 expression is accompanied by alterations in the expression patterns of SOXB1 members. In order to analyze the potential cross-talk between them, we generated SOX14 expression construct. The ectopic expression of <i>SOX14</i> was demonstrated at the mRNA level in NT2/D1, P19 and HeLa cells, while an increased level of SOX14 protein was detected in HeLa cells only. By transient transfection experiments in HeLa cells we showed for the first time that ectopic expression of SOX14 repressed SOX1 expression, whereas no significant effect on SOX2, SOX3 and SOX21 was observed. Data presented here provide an insight into SOX14 expression during <i>in vitro</i> neural differentiation of embryonal carcinoma cells and demonstrate the effect of its ectopic expression on protein levels of SOXB members in HeLa cells. Obtained results contribute to better understanding the role of one of the most conserved SOX proteins.</p></div

SOX14 expression analysis during RA induced neural differentiation of NT2/D1 and P19 cells.

<p><b>A</b>: Western blot analysis of SOX14 expression in undifferentiated NT2/D1 cells treated with RA for 1, 2, 3 and 4 weeks. <b>B</b>: Comparison of SOX14 protein level between undifferentiated NT2/D1, cells differentiated for 4 weeks (NT2 4W) and a population of neurons (NT2-N). <b>C</b>: qRT-PCR of SOX14 mRNA isolated from NT2/D1, NT2 4W and NT2-N cells. The relative quantities of SOX14 mRNA were calculated as a percentage of the quantity in undifferentiated NT2/D1 cells, which was set as 1. Data are presented as the means ± SD of two independent NT2/D1 differentiation experiments. <b>D</b>: Western blot analysis of SOX14 expression in undifferentiated P19 and cells during RA-induced differentiation, including embryoid bodies (P19 EB) and a differentiated neuronal population (P19-N). Progression of neural differentiation was examined by expression analysis of β-III Tubulin and GFAP, as markers of differentiated neurons and astroglial cells, while GAPDH was used as the loading control.</p

Immunocytochemical detection of MAP2, GFAP and SOX14 in undifferentiated P19 and differentiated P19-N cells.

<p>Panel I: Immunocytochemical detection of MAP2 and GFAP-positive cells in P19-N. Panel II: Immunocytochemical detection of MAP2 and SOX14-positive cells in P19 and P19-N. The P19-N population consists of a large number of MAP2 terminally differentiated neurons (C and D), and a few GFAP-positive astroglial cells (B and D). Specific SOX14 immunoreactivity/punctated nuclear signal was detected in a majority of cells in differentiated P19-N cultures (I, K and M), and at basal level in P19 cells (F). DIC transmitted light images show morphology of SOX14+ cells in P19-N population (G and H). Yellow arrowhead in G-I marks flat cells with large nuclei which show strong SOX14 immunoreactivity. SOX14 is expressed in MAP2-positive neurons (K, arrows in L and M) and in non-neuronal cells (K, arrowheads in L and M). Boxed regions in J and K are enlarged in the same figures. Cell nuclei were counterstained with DAPI (A, D, E, H, J and L). Scale bars: A–K 50 μm, L and M 20 μm.</p

Ectopic expression of human SOX14 in NT2/D1, P19 and HeLa cells.

<p><b>A, C, E</b>: Semi-quantitative RT-PCR on mRNA obtained from NT2/D1, P19 and HeLa cells, respectively, transiently transfected with pcDNA3.1/SOX14 expression construct. <b>B, D and F</b>: Western blot analyses on whole cell lysates obtained from NT2/D1, P19 and HeLa cells, respectively, transiently transfected with pcDNA3.1/SOX14 expression construct. SOX14 mRNA and protein levels were analyzed 24 h, 48 h, and 72 h after transfection. Transfection with pcDNA3.1 vector (designated as C) was used as a control for transfection. Negative PCR control is designated as N. GAPDH, α-Tubulin or <i>actin</i> were used as loading controls for Western blot and RT-PCR analyses.</p

Expression of SOXB members during RA induced neural differentiation of EC cells.

<p><b>A, C</b>: Western blot analysis of SOX14, SNAP25 and OCT4 expression in undifferentiated NT2/D1 and cells at final phase of RA induction (NT2 4W) and undifferentiated P19 and P19 cells at final phase of RA induction (P19 EB), respectively. <b>B</b>: Western blot analysis of SOX1, SOX2, SOX3 and SOX21 expression in NT2/D1 and cells treated with RA for 1, 2, 3 and 4 weeks. <b>D</b>: Western blot analysis of SOX1, SOX2, SOX3 and SOX21 expression in P19 and cells at final phase of RA induction (P19 EB). Protein extract from HeLa cells transiently transfected with SOX21 expression construct was used as a positive control for SOX21 expression. GAPDH was used as a loading control. Quantitative data of relative SOX14, SOX1, SOX2 and SOX3 protein levels during RA induction of NT2/D1 cells (<b>E</b>) and P19 (<b>F</b>) are summarized by the histogram. The quantities were calculated as a percentage of the quantity in control, untreated NT2/D1/P19 cells which were set as 100%. Data are presented as the means±SD of at least two independent differentiation experiments.</p