Nutrient-deprived Conditions Research Articles

Influence of nutrient deprivations on lipid accumulation in a dominant indigenous microalga Chlorella sp., BUM11008: Evaluation for biodiesel production

Abstract Microalgae are a potential source of biodiesel. The urgent need for an alternative and sustainable energy has created renewed interest to analyze the microalgae for biodiesel production. In this study, a dominant indigenous freshwater unicellular microalgal strain Chlorella sp., BUM11008 , was examined for its efficiency towards biodiesel production. The organism was evaluated for ability to yield high of biomass and lipid productivity under normal and various nutrient-deprived conditions (nitrogen, phosphate-potassium, iron, and all three combined). Under normal conditions, after 20 days of cultivation in Chu10 medium, the organism yielded a biomass of 2.58 ± 0.07 g/L, with lipid content of 312.16 ± 2.38 mg/g. In a two-phase culturing system upon nutrition deprivation, the organism was able to respond with different levels of lipid accumulation. Among the various post-harvest treatments, nitrogen deprivation yielded the highest lipid productivity of 53.96 ± 0.63 mg/L d, followed by the combined deprivation condition (49.16 ± 1.36 mg/L d). FAME profiles of the isolate were found to meet the requirements of international standards for biodiesel. The study leads to the conclusion that the two-phase culturing system with nitrogen starvation as post-harvest treatment would be suitable for gaining maximum biomass productivity, and lipid content of high quality fatty acids. Thus, it is proposed that Chlorella sp., BUM11008 , would be a promising candidate for sustainable biodiesel production.

Mitophagy in neurodegeneration and aging.

Macroautophagy is a cellular catabolic process that involves the sequestration of cytoplasmic constituents into double-membrane vesicles known as autophagosomes, which subsequently fuse with lysosomes, where they deliver their cargo for degradation. The main physiological role of autophagy is to recycle intracellular components, under conditions of nutrient deprivation, so as to supply cells with vital materials and energy. Selective autophagy also takes place in nutrient-rich conditions to rid the cell of damaged organelles or protein aggregates that would otherwise compromise cell viability. Mitophagy is a selective type of autophagy, whereby damaged or superfluous mitochondria are eliminated to maintain proper mitochondrial numbers and quality control. While mitophagy shares key regulatory factors with the general macroautophagy pathway, it also involves distinct steps, specific for mitochondrial elimination. Recent findings indicate that parkin and the phosphatase and tensin homolog-induced putative kinase protein 1 (PINK1), which have been implicated in the pathogenesis of neurodegenerative diseases such as Parkinson’s disease, also regulate mitophagy and function to maintain mitochondrial homeostasis. Here, we survey the molecular mechanisms that govern the process of mitophagy and discuss its involvement in the onset and progression of neurodegenerative diseases during aging.

The Role of Autophagy in Clinical Practice

Resisting cell death is one of the six hallmarks of cancer. Autophagy is a highly adaptable metabolic process that plays an important role in stressful conditions, such as nutrient deprivation and hypoxia. In these conditions, it is becoming evident that autophagy protects cells, by providing an alternative energy source and by eliminating dysfunctional organelles or proteins. In tumourigenesis, autophagy plays a dual role, which may be related to the different stages in cancer development. The autophagy-mediated removal of damaged proteins and organelles may prevent cancer initiation by limiting tissue inflammation. In contrast, autophagy has been shown to allow established tumours to survive in nutrient-deprived or hypoxic conditions during cancer progression. Key regulators of the autophagy pathway are modulated or aberrantly expressed in cancer and modulating autophagy is an attractive concept for cancer therapy. The difficulties, however, lie in the complexity of the crosstalk between apoptosis and autophagy and the lack of robust tissue biomarkers and in vivo assessment of autophagic flux. Currently there are 19 clinical trials in both solid and haematogenous cancers investigating the efficacy and toxicity of adding an autophagy inhibitor to standard treatment. Hydroxychloroquine, a drug routinely used in the treatment of malaria and autoimmune disorders, is the most common autophagy inhibitor under investigation due to its more favourable toxicity profile. This overview summarises the role of autophagy in cancer initiation, progression and resistance to treatment and thereby the therapeutic benefit that may be gained by modulating its effects.

The RpoS-Mediated General Stress Response inEscherichia coli

Under conditions of nutrient deprivation or stress, or as cells enter stationary phase, Escherichia coli and related bacteria increase the accumulation of RpoS, a specialized sigma factor. RpoS-dependent gene expression leads to general stress resistance of cells. During rapid growth, RpoS translation is inhibited and any RpoS protein that is synthesized is rapidly degraded. The complex transition from exponential growth to stationary phase has been partially dissected by analyzing the induction of RpoS after specific stress treatments. Different stress conditions lead to induction of specific sRNAs that stimulate RpoS translation or to induction of small-protein antiadaptors that stabilize the protein. Recent progress has led to a better, but still far from complete, understanding of how stresses lead to RpoS induction and what RpoS-dependent genes help the cell deal with the stress.

Comparative spatiotemporal analysis of root aerenchyma formation processes in maize due to sulphate, nitrate or phosphate deprivation

Nitrate (N), phosphate (P) or sulphate (S) deprivation causes aerenchyma formation in maize (Zea mays L.) nodal roots. The exact mechanisms that trigger the formation of aerenchyma under these circumstances are unclear. We have compared aerenchyma distribution across the nodal roots of first whorl (just emerging in 10-day-old seedlings), which were subject to S, N or P deprivation over a period of 10days in connection with oxygen consumption, ATP concentration, cellulase and polygalacturonase activity in the whole root. The effect of deprivation on aerenchyma formation was examined using light and electron microscopy, along with in situ detection of calcium and of reactive oxygen species (ROS) by fluorescence microscopy. Aerenchyma was not found in the root base regardless of the deprivation. Programmed cell death (PCD) was observed near the root tip, either within the first two days (-N) or a few days later (-S, -P) of the treatment. Roots at day6 under all three nutrient-deprived conditions showed signs of PCD 1cm behind the cap, whereas only N-deprived root cells 0.5cm behind the cap showed severe ultrastructural alterations, due to advanced PCD. The lower ATP concentration and the higher oxygen consumptions observed at day2 in N-, P- and S-deprived roots compared to the control indicated that PCD may be triggered by perturbations in energy status of the root. The peaks of cellulase activity located between days3 (-N) and 6 (-P), along with the respective alterations in polygalacturonase activity, indicated a coordination which preceded aerenchyma formation. ROS and calcium seemed to contribute to PCD initiation, with ROS possessing dual roles as signals and eliminators. All the examined parameters presented both common features and characteristic variations among the deprivations.

Pancreatic anticancer activity of a novel geranylgeranylated coumarin derivative

A series of hydroxycoumarin derivatives has been synthesized and evaluated against human pancreatic PANC-1 cancer cells under nutrient-deprived conditions. Several compounds exhibited 100% preferential cytotoxicity at low micromolar concentrations under nutrition starvation, and showed no cytotoxicity under nutrient-rich conditions. In this study, a novel geranylgeranylated ether coumarin derivative 9 was found to exhibit the highest cytotoxic activity of 6.25 μM within 24h. The preferential anti-tumor activity exhibited by compound 9 against PANC-1 under low oxygen and nutrient environment illustrates its great potential as a promising lead structure for the development of novel agents to combat pancreatic cancer.

Eukaryotic elongation factor‐2 (eEF2): its regulation and peptide chain elongation

Regulation at the level of translation in eukaryotes is feasible because of the longer lifetime of eukaryotic mRNAs in the cell. The elongation stage of mRNA translation requires a substantial amount of energy and also eukaryotic elongation factors (eEFs). The important component of eEFs, i.e. eEF2 promotes the GTP-dependent translocation of the nascent protein chain from the A-site to the P-site of the ribosome. Mostly the eEF2 is regulated by phosphorylation and dephosphorylation by a specific kinase known as eEF2 kinase, which itself is up-regulated by various mechanisms in the eukaryotic cell. The activity of this kinase is dependent on calcium ions and calmodulin. Recently it has been shown that the activity of eEF2 kinase is regulated by MAP kinase signalling and mTOR signalling pathway. There are also various stimuli that control the peptide chain elongation in eukaryotic cell; some stimuli inhibit and some activate eEF2. These reports provide the mechanisms by which cells likely serve to slow down protein synthesis and conserve energy under nutrient deprived conditions via regulation of eEF2. The regulation via eEF2 has also been seen in mammary tissue of lactating cows, suggesting that eEF2 may be a limiting factor in milk protein synthesis. Regulation at this level provides the molecular understanding about the control of protein translocation reactions in eukaryotes, which is critical for numerous biological phenomenons. Further the elongation factors could be potential targets for regulation of protein synthesis like milk protein synthesis and hence probably its foreseeable application to synthetic biology.

Activation of autophagy in mesenchymal stem cells provides tumor stromal support

Recent studies have implicated multipotential mesenchymal stem cells (MSCs) as an aid to breast cancer cell proliferation and metastasis, partly as a result of the MSCs secretome. As the tumor gets beyond 2 mm in diameter, the stromal cells could undergo starvation due to the lack of sufficient nutrients in solid tumor microenvironment. In this study, we investigated the survival mechanisms used by stressed stromal cells in breast cancers. We used serum-deprived mesenchymal stem cells (SD-MSCs) and MCF-7 breast cancer cells as model system with a hypothesis that stromal cells in the nutrient-deprived core utilize survival mechanisms for supporting surrounding cells. We tested this hypothesis using in vivo tumor xenografts in immunodeficient mice, which indicated that SD-MSCs supported MCF-7 tumor growth by protection from apoptosis. Histochemical assays showed that SD-MSCs-injected tumors exhibited higher cellularity, decreased apoptosis and decreased differentiation. Beclin-1 staining indicated autophagic areas surrounded by actively proliferating cells. Furthermore, in vitro studies demonstrate that SD-MSCs survive using autophagy and secrete paracrine factors that support tumor cells following nutrient/serum deprivation. Western blot and immunocytochemistry analysis of SD-MSCs demonstrated upregulation and perinuclear relocation of autophagy key regulators such as beclin-1, ATG10, ATG12, MAP-LC3 and lysosomes. Electron microscopic analysis detected a time-dependent increase in autophagosome formation and HDAC6 activity assays indicated the upregulation of autophagy. Taken together, these data suggest that under nutrient-deprived conditions that can occur in solid tumors, stromal cells utilize autophagy for survival and also secrete anti-apoptotic factors that can facilitate solid tumor survival and growth.

Tetrahydroxanthones by Sequential Pd-Catalyzed C−O and C−C Bond Construction and Use in the Identification of the “Antiausterity” Pharmacophore of the Kigamicins

Readily available C-acylated cycloalkanones undergo efficient Pd catalyzed ring closure/cross-coupling providing 7-substituted tetrahydroxanthones in a single operation. One of the synthesized derivatives (depicted) is shown to selectively kill pancreatic cancer (PANC-1) cells under conditions of nutrient deprivation indicating that the tetrahydroxanthone is responsible, in part, for the "antiausterity" effects of the naturally occurring kigamicins.

MCL-1 is a stress sensor that regulates autophagy in a developmentally regulated manner

Apoptosis has an important role during development to regulate cell number. In differentiated cells, however, activation of autophagy has a critical role by enabling cells to remain functional following stress. In this study, we show that the antiapoptotic BCL-2 homologue MCL-1 has a key role in controlling both processes in a developmentally regulated manner. Specifically, MCL-1 degradation is an early event not only following induction of apoptosis, but also under nutrient deprivation conditions where MCL-1 levels regulate activation of autophagy. Furthermore, deletion of MCL-1 in cortical neurons of transgenic mice activates a robust autophagic response. This autophagic response can, however, be converted to apoptosis by either reducing the levels of the autophagy regulator Beclin-1, or by a concomitant activation of BAX. Our results define a pathway whereby MCL-1 has a key role in determining cell fate, by coordinately regulating apoptosis and autophagy.

COMP-Ang1, a Variant of Angiopoietin 1, Inhibits Serum-Deprived Apoptosis of Mesenchymal Cells via PI3K/Akt and Mitogen-Activated Protein Kinase Pathways

Background/Aims: Cartilage oligomeric matrix protein (COMP)-angiopoietin 1 (Ang1) is a soluble and stable form of Ang1 which plays important roles in vessel formation and the survival of endothelial cells, neurons and cardiomyocytes. However, the effects of COMP-Ang1 on the survival of mesenchymal cells are unknown. Mesenchymal cells have been transplanted with some scaffolds for bone tissue regeneration, but they occasionally underwent cell death due to a lack of nutrient supply. This study examined the effects of COMP-Ang1 on the survival of mesenchymal cells under nutrient-deprived conditions. Methods: Primary and C3H10T1/2 mesenchymal cells were cultured under serum deprivation with or without COMP-Ang1. The effects of COMP-Ang1 on mesenchymal cell survival and its molecular mechanism were determined using a viability test, RT-PCR, Western blotting and fluorescence-activated cell sorting analysis. Results and Conclusion: COMP-Ang1 inhibited the nutrient-deprived apoptotic cell death of mesenchymal cells through the Akt, p38 and extracellular-signal-regulated kinase (ERK) pathways. In addition, COMP-Ang1 reversed the nutrient-deprived suppression of cyclin D1 mRNA expression. These results suggest that COMP-Ang1 has a protective role in the survival of nutrient-deprived mesenchymal cells. The use of COMP-Ang1 with some scaffolds might be useful for bone tissue engineering.

Failure and exploitation of autophagy in human pathologies—cellular integrity between inflammation, infection, and cell survival

Autophagy is a cellular disposal system directing cargoes into lysosomes, where the substrates, ranging from proteins to pathogens, are finally subjected to proteolytic cleavage. It is well-known as a “provider” of anabolic building blocks under conditions of nutrient deprivation, but it is also evident that autophagy has implications far beyond a simple adaption to starvation in mammalian cells and organisms. This special review issue aims to summarize current knowledge regarding the involvement of autophagy in human diseases and to encourage future research in reliably detecting autophagy and manipulating autophagosome biogenesis for clinical applications. Macroautophagy represents the most extensively studied form of autophagy and involves the formation of a double membrane compartment known as the autophagosome. Within these introductory notes, we will only use the term “autophagy,” which for all topics discussed includes macroautophagy. The reviews presented in this special issue provide a sophisticated discussion on the possible involvement of other forms of autophagy in unique settings. Chloe Burman and Nicholas T. Ktistakis describe the concepts and key players of autophagosome formation, thus introducing the diversity of cellular processes that are controlled or influenced by autophagy. Furthermore, the authors demonstrate the remarkable progress made in understanding the regulation of autophagy by delineating how different cellular states like low energy levels, growth factor signaling, and starvation, can lead to increased or decreased rates of autophagosome formation. Mouse models provide crucial insights into the function of autophagy in vivo; however, the generation of genetically modified mouse strains is timeand labor-intensive, explaining why only a small portion of the mouse genome has been targeted to date. Additionally, mice lacking core autophagy genes are embryonic or neonatal lethal, thus making conditional gene targeting necessary. Further, reconstitution experiments allowing mutational analyses in vivo are not readily feasible in mice. Therefore, other model organisms are an attractive option to examine autophagy functions and pathways in vivo. One example is Drosophila, which is suitable for genetic loss-of-function induced by RNAi, as well as expression reconstitution by cDNAs immune to RNAi. This allows for the validation of specific versus off-target effects and a replacement of the wild-type protein with well-defined mutant forms [1 and references therein]. Jonathan Zirin and Norbert Perrimon give a comprehensive overview of autophagy genes and pathways that are conserved in Drosophila. Given the amount of ready-to-use tools available for Drosophila genetics, it is somewhat surprising that these resources are not more frequently and rigorously applied towards the study of autophagy functions in vivo. While some aspects of autophagy cannot be addressed, particularly the function of autophagy in adaptive immunity, Drosophila serves as a valuable model organism to study the removal of protein aggregates, autophagosome–pathogen interactions, the role of autophagy in metabolism, and other processes that control cellular logistics. To date, the autophagy pathway has been linked to a variety of human diseases. A role in cancer, especially in P. Kuballa : R. J. Xavier Center for Computational and Integrative Biology and Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

P53-dependent regulation of autophagy protein LC3 supports cancer cell survival under prolonged starvation

The p53 tumor suppressor is mutated in a high percentage of human tumors. However, many other tumors retain wild-type (wt) p53 expression, raising the intriguing possibility that they actually benefit from it. Recent studies imply a role for p53 in regulation of autophagy, a catabolic pathway by which eukaryotic cells degrade and recycle macromolecules and organelles, particularly under conditions of nutrient deprivation. Here, we show that, in many cell types, p53 confers increased survival in the face of chronic starvation. We implicate regulation of autophagy in this effect. In HCT116 human colorectal cancer cells exposed to prolonged nutrient deprivation, the endogenous wt p53 posttranscriptionally down-regulates LC3, a pivotal component of the autophagic machinery. This enables reduced, yet sustainable autophagic flux. Loss of p53 impairs autophagic flux and causes excessive LC3 accumulation upon starvation, culminating in apoptosis. Thus, p53 increases cell fitness by maintaining better autophagic homeostasis, adjusting the rate of autophagy to changing circumstances. We propose that some cancer cells retain wt p53 to benefit from the resultant increased fitness under limited nutrient supply.

The Human Papillomavirus Type 16 E6 Oncoprotein Activates mTORC1 Signaling and Increases Protein Synthesis

The mammalian target of rapamycin (mTOR) kinase acts as a cellular rheostat that integrates signals from a variety of cellular signal transduction pathways that sense growth factor and nutrient availability as well as intracellular energy status. It was previously reported that the human papillomavirus type 16 (HPV16) E6 oncoprotein may activate the S6 protein kinase (S6K) through binding and E6AP-mediated degradation of the mTOR inhibitor tuberous sclerosis complex 2 (TSC2) (Z. Lu, X. Hu, Y. Li, L. Zheng, Y. Zhou, H. Jiang, T. Ning, Z. Basang, C. Zhang, and Y. Ke, J. Biol. Chem. 279:35664-35670, 2004; L. Zheng, H. Ding, Z. Lu, Y. Li, Y. Pan, T. Ning, and Y. Ke, Genes Cells 13:285-294, 2008). Our results confirmed that HPV16 E6 expression causes an increase in mTORC1 activity through enhanced phosphorylation of mTOR and activation of downstream signaling pathways S6K and eukaryotic initiation factor binding protein 1 (4E-BP1). However, we did not detect a decrease in TSC2 levels in HPV16 E6-expressing cells. We discovered, however, that HPV16 E6 expression causes AKT activation through the upstream kinases PDK1 and mTORC2 under conditions of nutrient deprivation. We show that HPV16 E6 expression causes an increase in protein synthesis by enhancing translation initiation complex assembly at the 5' mRNA cap and an increase in cap-dependent translation. The increase in cap-dependent translation likely results from HPV16 E6-induced AKT/mTORC1 activation, as the assembly of the translation initiation complex and cap-dependent translation are rapamycin sensitive. Lastly, coexpression of the HPV16 E6 and E7 oncoproteins does not affect HPV16 E6-induced activation of mTORC1 and cap-dependent translation. HPV16 E6-mediated activation of mTORC1 signaling and cap-dependent translation may be a mechanism to promote viral replication under conditions of limited nutrient supply in differentiated, HPV oncoprotein-expressing proliferating cells.

Possibilities of Gutta-Percha–centered Infection in Endodontically Treated Teeth: An In Vitro Study

Introduction Implanted biomaterials could provide surface for bacterial adherence and formation of biofilm, eventually leading to biomaterial-centered infections (BCIs). In this in vitro study, we examined the biofilm-forming capacity of Enterococcus faecalis on gutta-percha (GP) points under different nutrient status and surface conditioning with saliva and serum. Methods GP points (n = 420) coated with different root canal sealers (Sealapex or Roth) were conditioned with saliva or serum for 2-, 4-, and 12-week intervals and subsequently were incubated with E. faecalis for 2 weeks under nutrient-rich and nutrient-deprived environments. The biofilm-forming capacity and the nature of biofilm formed on GP were assessed by using a viable cell assay and scanning electron microscopy (SEM). Results E. faecalis produced biofilms on GP points (with and without root canal sealers) under both nutrient-rich and nutrient-deprived environments after conditioning with saliva or serum. SEM reflected that the biofilms formed under nutrient-rich conditions were regular, whereas the biofilms formed under nutrient-deprived conditions were irregular and scanty. Under nutrient-deprived conditions, longer conditioning periods in saliva or serum were required for bacterial adherence. Conclusions Under the conditions of this study, saliva and serum conditioning of GP points plays an important role in the biofilm formation under tough environmental conditions.

Immunogenic Potential of Enterococcus faecalis Biofilm under Simulated Growth Conditions

IntroductionThe aim of this study was to evaluate the effect of growth modes (planktonic and biofilms) of Enterococcus faecalis on the intracellular survival and ability to produce tumor necrosis factor α (TNF-α) and interleukin (IL)-6 after interacting with monocytes/in vitro–differentiated macrophages. MethodsIn vitro biofilms of three E. faecalis strains (ATCC-29212, OG1RF, and FA2-2) were grown on dentin under simulated nutrient-rich and nutrient-deprived conditions. Biofilm-derived E. faecalis cells were incubated with monocytes/in vitro–differentiated human macrophages. A fluorometric assay was used to quantify the surface adherent bacteria, whereas an antibiotic protection assay was used to quantify the internalized E. faecalis cells 6 to 48 hours after interaction with macrophages. TNF-α and IL-6 produced during this interaction were quantified by an enzyme-linked immunosorbent assay. ResultsThe surface adherence and intracellular survival of E. faecalis within macrophages was significantly higher in biofilm bacteria when compared with planktonic cells (p ≤ 0.05). E. faecalis strains survived within the macrophages for up to 48 hours and produced higher levels of IL-6 with planktonic cells when compared with biofilm cells. Except in FA2-2, the planktonic cells of other strains did not show a significant increase in TNF-α compared with the biofilm cells (p > 0.05). ConclusionBiofilm cells of E. faecalis interacting with macrophages showed higher potential for surface adherence, intracellular survival and produced lesser amounts of IL-6 and TNF-α when compared to planktonic cells. Further experiments are required to understand the clinical implication of the intracellular survival of E. faecalis biofilm.

Identification of microRNAs and their targets in switchgrass, a model biofuel plant species

In recent years, several plant species such as switchgrass, Miscanthus and Brachypodium have been recognized as potential model plant species for cellulosic bioenergy production. Of these, switchgrass has attracted much attention in the United States and worldwide because it can grow well on marginal lands and tolerate frequent drought spells. However, little is known about the basic biology of the traits that control these important characteristics in switchgrass. Genome-encoded approximately 21-24nt microRNAs (miRNAs) have emerged as critical regulators of gene expression important for normal growth and development and adaptation to abiotic stress, including nutrient-deprived conditions. To understand miRNA-guided post-transcriptional gene regulatory networks in this plant species, we sought to identify miRNAs in switchgrass. Using computational and experimental approaches, we identified approximately 20 conserved miRNA families. Temporal expression analysis indicated that some miRNAs have distinct tissue-specific expression, although most are ubiquitously expressed. Unlike in Arabidopsis and other plants, miR395 and miR399 were detected in plants grown on optimal levels of sulfate or phosphate in switchgrass, and were only slightly altered when exposed to sulfate or phosphate deficit conditions. Thirty-seven genes were predicted as targets for miRNAs, and 4 target mRNAs (Squamosa promoter binding-like factor, apetala 2-like, NAC domain containing transcription factor and HD-Zip homologs) were validated by 5'-RACE assays. These findings provide a snapshot of the miRNA component and possible targets in switchgrass.

Mitochondrial bioenergetic adaptations of breast cancer cells to aglycemia and hypoxia

Breast cancer cells can survive and proliferate under harsh conditions of nutrient deprivation, including limited oxygen and glucose availability. We hypothesized that such environments trigger metabolic adaptations of mitochondria, which promote tumor progression. Here, we mimicked aglycemia and hypoxia in vitro and compared the mitochondrial and cellular bioenergetic adaptations of human breast cancer (HTB-126) and non-cancer (HTB-125) cells that originate from breast tissue. Using high-resolution respirometry and western blot analyses, we demonstrated that 4 days of glucose deprivation elevated oxidative phosphorylation five-fold, increased the spread of the mitochondrial network without changing its shape, and decreased the apparent affinity of oxygen in cancer cells (increase in C ( 50 )), whereas it remained unchanged in control cells. The substrate control ratios also remained constant following adaptation. We also observed the Crabtree effect, specifically in HTB-126 cells. Likewise, sustained hypoxia (1% oxygen during 6 days) improved cell respiration in non-cancer cells grown in glucose or glucose-deprived medium (+ 32% and +38%, respectively). Conversely, under these conditions of limited oxygen or a combination of oxygen and glucose deprivation for 6 days, routine respiration was strongly reduced in cancer cells (-36% in glucose medium, -24% in glucose-deprived medium). The data demonstrate that cancer cells behave differently than normal cells when adapting their bioenergetics to microenvironmental conditions. The differences in hypoxia and aglycemia tolerance between breast cancer cells and non-cancer cells may be important when optimizing strategies for the treatment of breast cancer.

The Effect of Autophagy to Cell Death in Nutrient-Deprived H460 Cells

Background: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. Methods: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. Results: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. Conclusion: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.

Imaging tumour physiology and vasculature to predict and assess response to heat

The vascular supply of tumours and the tumour microenvironment both play an important role when tumours are treated with hyperthermia. Blood flow is one of the major vehicles by which heat is dissipated thus the vascular supply will influence the ability to heat the tumour. It also influences the type of microenvironment that exists within tumours, and it is now well-established that cells existing in areas of oxygen deficiency, nutrient deprivation and acidic conditions are more sensitive to the effect of hyperthermia. The vascular supply and microenvironment are also affected by hyperthermia. In general, mild heat temperatures transiently improve blood flow and oxygenation, while higher hyperthermia temperatures cause vascular collapse and so increase the adverse microenvironmental conditions. Being able to image these vascular and microenvironmental parameters both before and after heating will help in our ability to predict and assess response. Here we review the various techniques that can be applied to supply this information, especially using non-invasive imaging approaches.