Hypoxia is one of the fundamental biological phenomena that are intricately associated with the development and aggressiveness of a variety of solid tumors. Hypoxia-inducible factors (HIF) function as a master transcription factor, which regulates hypoxia responsive genes and have been recognized to play critical roles in tumor invasion, metastasis, and chemo-radiation resistance, and contributes to increased cell proliferation, survival, angiogenesis and metastasis. Therefore, tumor hypoxia with deregulated expression of HIF and its biological consequence lead to poor prognosis of patients diagnosed with solid tumors, resulting in higher mortality, suggesting that understanding of the molecular relationship of hypoxia with other cellular features of tumor aggressiveness would be invaluable for developing newer targeted therapy for solid tumors. It has been well recognized that cancer stem cells (CSCs) and epithelial-to-mesenchymal transition (EMT) phenotypic cells are associated with therapeutic resistance and contributes to aggressive tumor growth, invasion, metastasis and believed to be the cause of tumor recurrence. Interestingly, hypoxia and HIF signaling pathway is known to play an important role in the regulation and sustenance of CSCs and EMT phenotype. However, the molecular relationship between HIF signaling pathway with the biology of CSCs and EMT remains unclear although NF-κB, PI3K/Akt/mTOR, Notch, Wnt/β-catenin, and Hedgehog signaling pathways have been recognized as important regulators of CSCs and EMT. In this article, we will discuss the state of our knowledge on the role of HIF-hypoxia signaling pathway and its kinship with CSCs and EMT within the tumor microenvironment. We will also discuss the potential role of hypoxia-induced microRNAs (miRNAs) in tumor development and aggressiveness, and finally discuss the potential effects of nutraceuticals on the biology of CSCs and EMT in the context of tumor hypoxia.

PURPOSE: Multidrug resistance (MDR) has become an obstacle for chemotherapy of cancer. p53 is reported to participate in the regulation of MDR, but the association between p53 status and MDR are complicated and conditional. It has been verified that apoptosis is not the only mechanism for MDR regulation by p53, the roles of autophagy in MDR is less studied. PATIENTS AND METHODS: Human ovarian carcinoma cell lines SKOV3 and multidrug resistant phenotype SKVCR cells were used and wild-type p53 (wt p53) and mutant 175H constructs were introduced into cells to establish cell models with different p53 status by gene engineering, the sensitivity to vincristine (VCR), cisplatin (DDP), pirarubicin (THP) and etoposide (VP-16) were detected by MTT assay, Western blot and quantitative real-time PCR were used to detect the expression of protein and mRNA, especially, monodansylcadaverine (MDC) staining was used for autophagy rate, Hoechst 33342/propidium iodide (PI) were used to assess apoptosis and necrosis. RESULTS: SKVCR cells induced by VCR shown overexpression of P-glycoprotein (P-gp) and MDR, and also displayed an enhanced autophagy compared with parental SKOV3. Wt p53 and 175H has no influence on drug sensitivity in SKOV3, while both sensitized SKVCR cells to VCR, THP and VP-16, especially 175H. The introduction of wt p53-induced apoptosis only, while 175H trigged autophagic cell death, necrosis and apoptosis so as to reverse the MDR. CONCLUSION: The enhancement of autophagy in MDR cells allows to survive during chemotherapy stress, autophagy plays important role in wt p53 and mutant p53-immediated MDR. The different influence of p53 status on drug sensitivity hint the individual treatment strategies based on p53 status in patients.

Objective: To evaluate right ventricular (RV) regional volume and systolic function in patients with pulmonary arterial hypertension (PAH) using real time three-dimensional echocardiography (RT3DE), and to explore the relationship between parameters measured by RT3DE and right heart catheterization (RHC). Methods: RT3DE images were acquired from 24 patients with PAH and 27 normal controls for evaluation and analysis to obtain RV regional end-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF) in three compartments (inflow, body, and outflow). Conventional echocardiographic parameters were calculated and recorded. RHC was performed in 17 patients to obtain pulmonary artery systolic pressure (PASP) and pulmonary vascular resistance (PVR). Results: RV regional EDV and ESV were significantly higher while regional EF was significantly lower in the PAH patients when compared with controls (P < 0.001). In the PAH group, EDV was similar in the inflow and body compartment, both higher than that in the outflow compartment (P < 0.05); EF was the highest in the inflow compartment and the lowest in the body compartment (P < 0.05). RV regional EF in the inflow compartment and global EF were negatively correlated with PASP (r =-0.766,-0.816, P < 0.001) and PVR (r =-0.529,-0.656, P < 0.05). Conclusions: In patients with PAH, RV regional volume was enlarged and systolic function was impaired with distinct characteristics; regional EF in the inflow compartment and global EF were inversely correlated with PASP and PVR. Evaluation of RV regional systolic function using RT3DE may play a potential role in the noninvasive assessment of the severity of PAH.(Echocardiography, 2012;**:1-7).

With the goal of establishing experimental protocols for cloning sika deer, various conditions for in vitro maturation (IVM) and artificial activation of sika deer oocytes were examined. In vitro maturation was evaluated in seven different culture media. The highest rate of oocyte maturation was 75.4% in 10 μg/ml follicle-stimulating hormone (FSH), 1 μg/ml LH, 0.2 mm cysteamine and 50 ng/ml epidermal growth factor (EGF) after 24 h of IVM. The efficiency after 24 h of IVM did not differ significantly (p > 0.05) from that observed after 20 h. Cysteamine (0.2 mm) significantly increased the maturation rates after 20 h (from 59.1% to 67.2%, p < 0.05) and after 24 h (from 63.2% to 71.6%, p < 0.05) of IVM. The IVM rates of oocytes collected during the oestrous season (75.4%) and the anoestrous season (23.3%) were significantly different at 24 h. The 20 μg/ml FSH, 2 μg/ml LH, 0.4 mm cysteamine and 100 ng/ml EGF significantly increased the maturation rates (from 23.3% to 54.2%, p < 0.01) at 24 h during the anoestrous season. For the activation experiments, the most effective method was chemical activation [ionomycin + 6-dimethylaminopurine (6-DMAP)], which promoted the development of sika deer oocytes to the blastocyst stage (32.4%). Our results indicate that in vitro matured sika deer oocytes are good candidates for parthenogenetic activation and that chemical treatment is needed for relatively efficient activation of the oocytes. These optimized conditions for IVM and parthenogenetic activation may be useful for efforts to restore populations of the endangered sika deer using the somatic cell nuclear transfer technique.

The differentiation of stomata provides a convenient model for studying pattern formation in plant tissues. Stomata formation is induced by a set of basic helix loop helix transcription factors (bHLH TF) and inhibited by a signal transduction pathway initiated by TOO MANY MOUTHS (TMM) and ERECTA family (ERf) receptors. The formation of a proper stomata pattern is also dependent upon the restriction of symplastic movement of bHLH TF into neighboring cells, especially in the backgrounds where function of the TMM/ERf signaling pathway is compromised. Here we describe a novel mutant of KOBITO1. The kob1-3 mutation leads to formation of stomata clusters in the erl1 erl2 background but not in the wild type. Cell-to-cell mobility assays demonstrated an increase in intercellular protein trafficking in kob1-3 including increased diffusion of SPEECHLESS , suggesting that the formation of stomata clusters is due to escape of cell fate specifying factors from stomatal lineage cells. While plasmodesmatal permeability is increased in kob1-3, we did not detect drastic changes in callose accumulation at the neck regions of the plasmodesmata. Previously, KOBITO1 has been proposed to function in cellulose biosynthesis. Our data demonstrate that disruption of cellulose biosynthesis in the erl1 erl2 background does not lead to formation of stomata clusters, indicating that cellulose biosynthesis is not a major determining factor for regulating plasmodesmatal permeability. Analysis of KOBITO1 structure suggests that it is a glycosyltransferase-like protein. KOBITO1 might be involved in a carbohydrate metabolic pathway that is essential for both cellulose biosynthesis and regulation of plasmodesmatal permeability.

In the past few years, there have been significant advances in the research on cancer stem cells (CSCs). The emerging evidences have demonstrated that CSCs and epithelial-mesenchymal transition (EMT)-type cells, which share molecular characteristics with CSCs, play critical roles in drug resistance, invasion, and metastasis. Pancreatic cancer (PC) has a high mortality due to both intrinsic (de novo) and extrinsic (acquired) drug resistance, leading to increased invasive and metastatic potential of PC cells. Therefore, targeting pancreatic CSCs and EMT-type cells could be a novel therapeutic strategy for the treatment of PC. In this article, we will review the current state of our knowledge on the role of pancreatic CSCs and EMT-type cells, and summarize the novel therapeutic strategies that could target pancreatic CSCs and EMT-type cells, leading to the reversal of EMT phenotype, the induction of drug sensitivity, and the inhibition of invasion and metastasis of PC, which is expected to yield better treatment outcome.

The emergence of castrate-resistant prostate cancer (CRPC) contributes to the high mortality of patients diagnosed with prostate cancer (PCa), which in part could be attributed to the existence and the emergence of cancer stem cells (CSCs). Recent studies have shown that deregulated expression of microRNAs (miRNAs) contributes to the initiation and progression of PCa. Among several known miRNAs, let-7 family appears to play a key role in the recurrence and progression of PCa by regulating CSCs; however, the mechanism by which let-7 family contributes to PCa aggressiveness is unclear. Enhancer of Zeste homolog 2 (EZH2), a putative target of let-7 family, was demonstrated to control stem cell function. In this study, we found loss of let-7 family with corresponding over-expression of EZH2 in human PCa tissue specimens, especially in higher Gleason grade tumors. Overexpression of let-7 by transfection of let-7 precursors decreased EZH2 expression and repressed clonogenic ability and sphere-forming capacity of PCa cells, which was consistent with inhibition of EZH2 3'UTR luciferase activity. We also found that the treatment of PCa cells with BR-DIM (formulated DIM: 3,3'-diindolylmethane by Bio Response, Boulder, CO, abbreviated as BR-DIM) up-regulated let-7 and down-regulated EZH2 expression, consistent with inhibition of self-renewal and clonogenic capacity. Moreover, BR-DIM intervention in our on-going phase II clinical trial in patients prior to radical prostatectomy showed upregulation of let-7 consistent with down-regulation of EZH2 expression in PCa tissue specimens after BR-DIM intervention. These results suggest that the loss of let-7 mediated increased expression of EZH2 contributes to PCa aggressiveness, which could be attenuated by BR-DIM treatment, and thus BR-DIM is likely to have clinical impact.

Sleep deprivation (SD) has been shown to affect selective attention but it is not known how two of its component processes: target enhancement and distractor suppression, are affected. To investigate, young volunteers either attended to houses or were obliged to ignore them (when attending to faces) while viewing superimposed face-house pictures. MR signal enhancement and suppression in the parahippocampal place area (PPA) were determined relative to a passive view control condition. Sleep deprivation was associated with lower PPA activation across conditions. Critically SD specifically impaired distractor suppression in selective attention, leaving target enhancement relatively preserved. These findings parallel some observations in cognitive aging. Additionally, following SD, attended houses were not significantly better recognized than ignored houses in a post-experiment test of recognition memory contrasting with the finding of superior recognition of attended houses in the well-rested state. These results provide evidence for co-encoding of distracting information with targets into memory when one is sleep deprived.

The model catalysts of ZrO₂-supported Au nanoparticles have been prepared by deposition of Au atoms onto the surfaces of thin ZrO₂ films with different morphologies. The adsorption and thermal stability of Au nanoparticles on thin ZrO₂ films have been investigated using synchrotron radiation photoemission spectroscopy (SRPES) and X-ray photoelectron spectroscopy (XPS). The thin ZrO₂ films were prepared by two different methods, giving rise to different morphologies. The first method was utilizing the wet chemical impregnation to synthesize the thin ZrO₂ film through the procedure of first spin-coating a zirconium ethoxide (Zr(OC₂H₅)₄) precursor onto a SiO₂/Si(100) substrate at room temperature followed by calcination at 773 K for 12 h. Scanning electron microscopy (SEM) investigations indicate that highly porous "sponge-like nanostructures" were obtained in this case. The second method was epitaxial growth of a ZrO₂(111) film through vacuum evaporation of Zr metal onto Pt(111) in 1×10^-6 Torr of oxygen at 550 K followed by annealing at 1000 K. The structural analysis with low energy electron diffraction (LEED) of this film exhibits well long-range ordering. It has been found that Au forms smaller particles on the porous ZrO₂ film as compared to those on the ordered ZrO₂(111) film at a given coverage. Thermal annealing experiments demonstrate that Au particles are more thermally stable on the porous ZrO₂ surface than on the ZrO₂(111) surface, although on both surfaces Au particles experience significant sintering at elevated temperatures. In addition, by annealing the surfaces to 1100 K, Au particles desorb completely from ZrO₂(111) but not from porous ZrO₂. The enhanced thermal stability for Au on porous ZrO₂ can be attributed to the stronger interaction of the adsorbed Au with defects and the hindered migration or coalescence resulting from the porous structures.

Prostate cancer (PCa) bone metastases have long been believed to be osteoblastic because of bone remodeling leading to the formation of new bone. However, recent studies have shown increased osteolytic activity in the beginning stages of PCa bone metastases, suggesting that targeting both osteolytic and osteoblastic mediators would likely inhibit bone remodeling and PCa bone metastasis. In this study, we found that PCa cells could stimulate differentiation of osteoclasts and osteoblasts through the up-regulation of RANKL, RUNX2 and osteopontin, promoting bone remodeling. Interestingly, we found that formulated isoflavone and 3,3'-diindolylmethane (BR-DIM) were able to inhibit the differentiation of osteoclasts and osteoblasts through the inhibition of cell signal transduction in RANKL, osteoblastic, and PCa cell signaling. Moreover, we found that isoflavone and BR-DIM down-regulated the expression of miR-92a, which is known to be associated with RANKL signaling, EMT and cancer progression. By pathway and network analysis, we also observed the regulatory effects of isoflavone and BR-DIM on multiple signaling pathways such as AR/PSA, NKX3-1/Akt/p27, MITF, etc. Therefore, isoflavone and BR-DIM with their multi-targeted effects could be useful for the prevention of PCa progression, especially by attenuating bone metastasis mechanisms.