Clinical response to chemotherapy in oesophageal adenocarcinoma patients is linked to defects in mitochondria

Chemotherapeutic drugs kill cancer cells, but it is unclear why this happens in responding patients but not in non-responders. Proteomic profiles of patients with oesophageal adenocarcinoma may be helpful in predicting response and selecting more effective treatment strategies. In this study, pretherapeutic oesophageal adenocarcinoma biopsies were analysed for proteomic changes associated with response to chemotherapy by MALDI imaging mass spectrometry. Resulting candidate proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and investigated for functional relevance in vitro. Clinical impact was validated in pretherapeutic biopsies from an independent patient cohort. Studies on the incidence of these defects in other solid tumours were included. We discovered that clinical response to cisplatin correlated with pre-existing defects in the mitochondrial respiratory chain complexes of cancer cells, caused by loss of specific cytochrome c oxidase (COX) subunits. Knockdown of a COX protein altered chemosensitivity in vitro, increasing the propensity of cancer cells to undergo cell death following cisplatin treatment. In an independent validation, patients with reduced COX protein expression prior to treatment exhibited favourable clinical outcomes to chemotherapy, whereas tumours with unchanged COX expression were chemoresistant. In conclusion, previously undiscovered pre-existing defects in mitochondrial respiratory complexes cause cancer cells to become chemosensitive: mitochondrial defects lower the cells' threshold for undergoing cell death in response to cisplatin. By contrast, cancer cells with intact mitochondrial respiratory complexes are chemoresistant and have a high threshold for cisplatin-induced cell death. This connection between mitochondrial respiration and chemosensitivity is relevant to anticancer therapeutics that target the mitochondrial electron transport chain.

Emission factors from road dust resuspension in a Mediterranean freeway

Particulate matter emissions from paved roads are currently one of the main challenges for a sustainable transport in Europe. Emissions are scarcely estimated due to the lack of knowledge about the resuspension process severely hampering a reliable simulation of PM and heavy metals concentrations in large cities and evaluation of population exposure. In this study the Emission Factors from road dust resuspension on a Mediterranean freeway were estimated per single vehicle category and PM component (OC, EC, mineral dust and metals) by means of the deployment of vertical profiles of passive samplers and terminal concentration estimate. The estimated PM10 emission factors varied from 12 to 47 mg VKT?1 (VKT: Vehicle Kilometer Traveled) with an average value of 22.7 ? 14.2 mg VKT?1. Emission Factors for heavy and light duty vehicles, passenger cars and motorbikes were estimated, based on average fleet composition and EPA ratios, in 187e733 mg VKT?1, 33e131 VKT?1, 9.4e36.9 VKT?1 and 0.8e3.3 VKT?1, respectively. These range of values are lower than previous estimates in Mediterranean urban roads, probably due to the lower dust reservoir on freeways. PM emitted material was dominated by mineral dust (9e10 mg VKT?1), but also OC and EC were found to be major components and approximately 14 e25% and 2e9% of average PM exhaust emissions from diesel passenger cars on highways respectively.

Real time passenger information systems and quality of bus services

One of the main problems in urban areas is the steady growth in car ownership and traffic levels. Therefore, the challenge of sustainability is focused on a shift of the demand for mobility from cars to collective means of transport. For this end, buses are a key element of the public transport systems. In this respect Real Time Passenger Information (RTPI) systems help citizens change their travel behaviour towards more sustainable transport modes. This paper provides an assessment methodology which evaluates how RTPI systems improve the quality of bus services in two European cities, Madrid and Bremerhaven. In the case of Madrid, bus punctuality has increased by 3%. Regarding the travellers perception, Madrid raised its quality of service by 6% while Bremerhaven increased by 13%. On the other hand, the users ́ perception of Public Transport (PT) image increased by 14%.

Quality of bus services performance: benefits of real time passenger information systems

One of the main problems in urban areas is the steady growth in car ownership and traffic levels. Therefore, the challenge of sustainability is focused on a shift of the demand for mobility from cars to collective means of transport. For this purpose, buses are a key element of the public transport systems. In this respect Real Time Passenger Information (RTPI) systems help people change their travel behaviour towards more sustainable transport modes. This paper provides an assessment methodology which evaluates how RTPI systems improve the quality of bus services performance in two European cities, Madrid and Bremerhaven. In the case of Madrid, bus punctuality has increased by 3%. Regarding the travellers perception, Madrid raised its quality of service by 6% while Bremerhaven increased by 13%. On the other hand, the users¿ perception of Public Transport (PT) image increased by 14%.

Plastid redox state and sugars: Interactive regulators of nuclear-encoded photosynthetic gene expression

Feedback regulation of photosynthesis by carbon metabolites has long been recognized, but the underlying cellular mechanisms that control this process remain unclear. By using an Arabidopsis cell culture, we show that a block in photosynthetic electron flux prevents the increase in transcript levels of chlorophyll a/b-binding protein and the small subunit of Rubisco that typically occurs when intracellular sugar levels are depleted. In contrast, the expression of the nitrate reductase gene, which is induced by sugars, is not affected. These findings were confirmed in planta by using Arabidopsis carrying the firefly luciferase reporter gene fused to the plastocyanin and chlorophyll a/b-binding protein 2 gene promoters. Transcription from both promoters increases on carbohydrate depletion. Blocking photosynthetic electron transport with 3-(3′, 4′-dichlorophenyl)-1,1′-dimethylurea prevents this increase in transcription. We conclude that plastid-derived redox signaling can override the sugar-regulated expression of nuclear-encoded photosynthetic genes. In the sugar-response mutant, sucrose uncoupled 6 (sun6), plastocyanin-firefly luciferase transcription actually increases in response to exogenous sucrose rather than decreasing as in the wild type. Interestingly, plastid-derived redox signals do not influence this defective pattern of sugar-regulated gene expression in the sun6 mutant. A model, which invokes a positive inducer originating from the photosynthetic electron transport chain, is proposed to explain the nature of the plastid-derived signal.

Nitric oxide partitioning into mitochondrial membranes and the control of respiration at cytochrome c oxidase

An emerging and important site of action for nitric oxide (NO) within cells is the mitochondrial inner membrane, where NO binds to and inhibits members of the electron transport chain, complex III and cytochrome c oxidase. Although it is known that inhibition of cytochrome c oxidase by NO is competitive with O2, the mechanisms that underlie this phenomenon remain unclear, and the impact of both NO and O2 partitioning into biological membranes has not been considered. These properties are particularly interesting because physiological O2 tensions can vary widely, with NO having a greater inhibitory effect at low O2 tensions (<20 μM). In this study, we present evidence for a consumption of NO in mitochondrial membranes in the absence of substrate, in a nonsaturable process that is O2 dependent. This consumption modulates inhibition of cytochrome c oxidase by NO and is enhanced by the addition of exogenous membranes. From these data, it is evident that the partition of NO into mitochondrial membranes has a major impact on the ability of NO to control mitochondrial respiration. The implications of this conclusion are discussed in the context of mitochondrial lipid:protein ratios and the importance of NO as a regulator of respiration in pathophysiology.

Differential Control of Xanthophylls and Light-Induced Stress Proteins, as Opposed to Light-Harvesting Chlorophyll a/b Proteins, during Photosynthetic Acclimation of Barley Leaves to Light Irradiance

Barley (Hordeum vulgare L.) plants were grown at different photon flux densities ranging from 100 to 1800 μmol m−2 s−1 in air and/or in atmospheres with reduced levels of O2 and CO2. Low O2 and CO2 partial pressures allowed plants to grow under high photosystem II (PSII) excitation pressure, estimated in vivo by chlorophyll fluorescence measurements, at moderate photon flux densities. The xanthophyll-cycle pigments, the early light-inducible proteins, and their mRNA accumulated with increasing PSII excitation pressure irrespective of the way high excitation pressure was obtained (high-light irradiance or decreased CO2 and O2 availability). These findings indicate that the reduction state of electron transport chain components could be involved in light sensing for the regulation of nuclear-encoded chloroplast gene expression. In contrast, no correlation was found between the reduction state of PSII and various indicators of the PSII light-harvesting system, such as the chlorophyll a-to-b ratio, the abundance of the major pigment-protein complex of PSII (LHCII), the mRNA level of LHCII, the light-saturation curve of O2 evolution, and the induced chlorophyll-fluorescence rise. We conclude that the chlorophyll antenna size of PSII is not governed by the redox state of PSII in higher plants and, consequently, regulation of early light-inducible protein synthesis is different from that of LHCII.

Bioenergetic scaling: metabolic design and body-size constraints in mammals.

The cytosolic phosphorylation ratio ([ATP]/[ADP][P(i)]) in the mammalian heart was found to be inversely related to body mass with an exponent of -0.30 (r = 0.999). This exponent is similar to -0.25 calculated for the mass-specific O2 consumption. The inverse of cytosolic free [ADP], the Gibbs energy of ATP hydrolysis (delta G'ATP), and the efficiency of ATP production (energy captured in forming 3 mol of ATP per cycle along the mitochondrial respiratory chain from NADH to 1/2 O2) were all found to scale with body mass with a negative exponent. On the basis of scaling of the phosphorylation ratio and free cytosolic [ADP], we propose that the myocardium and other tissues of small mammals represent a metabolic system with a higher driving potential (a higher delta G'ATP from the higher [ATP]/[ADP][P(i)]) and a higher kinetic gain [(delta V/Vmax)/delta [ADP]] where small changes in free [ADP] produce large changes in steady-state rates of O2 consumption. From the inverse relationship between mitochondrial efficiency and body size we calculate that tissues of small mammals are more efficient than those of large mammals in converting energy from the oxidation of foodstuffs to the bond energy of ATP. A higher efficiency also indicates that mitochondrial electron transport is not the major site for higher heat production in small mammals. We further propose that the lower limit of about 2 g for adult endotherm body size (bumblebee-bat, Estrucan shrew, and hummingbird) may be set by the thermodynamics of the electron transport chain. The upper limit for body size (100,000-kg adult blue whale) may relate to a minimum delta G'ATP of approximately 55 kJ/mol for a cytoplasmic phosphorylation ratio of 12,000 M-1.

Estrutura espacial urbana e mobilidade: o caso da Região Metropolitana de São Paulo

A metrópole de São Paulo é a maior e mais importante aglomeração urbana do Brasil e está entre as dez maiores áreas urbanas do mundo. No entanto, a forma como acessibilidade espacial ocorre gera um fardo para a população e para a atividade econômica. Este trabalho pretende contribuir para a discussão de como melhorar a acessibilidade na Região Metropolitana de São Paulo estudando as características e impactos de estruturas espaciais urbana, analisando criticamente a estrutura espacial da metrópole e proporcionando sugestões de melhorias a fim de proporcionar uma mobilidade mais sustentável. Os procedimentos metodológicos incluem uma revisão bibliográfica sobre o tema e uma caracterização da estrutura espacial da Região Metropolitana de São Paulo, considerando a alocação de população, alocação de empregos e os padrões de deslocamento para os modais individual, coletivo e não motorizado. Apresentamos um relato da evolução recente, com dados das pesquisas de origem e destino realizadas pelo Metrô em 1997 e 2007 e da pesquisa de mobilidade de 2012. Também realizamos uma caracterização mais aprofundada com os dados da pesquisa de 2007. As cidades se desenvolvem com base no trade-off entre proximidade e mobilidade: a fim de maximizar as possibilidades de interação, as pessoas e as empresas tendem a se localizar onde o deslocamento necessário para executar essas interações requer menos custos financeiros, perda de tempo e desconforto. Esse processo molda a alocação espacial de atividades, que define parcialmente os hábitos de transporte. A estrutura espacial urbana pode ser caracterizada por sua escala (padrões compacto ou disperso), arranjo de densidades (padrão disperso ou clusterizado) e arranjo de atividade (padrão monocêntrico ou policêntrico). Estruturas espaciais com padrão mais compacto apresentam menores distâncias de viagem, reduzindo o impacto ambiental das viagens e viabilizando o transporte não motorizado e coletivo, e levam a um uso mais eficiente da terra, menor custo de infraestrutura e maior equidade no acesso ao transporte. Já estruturas clusterizadas policêntricas são associadas com maior facilidade de acesso à terra. Existe um debate sobre a capacidade de estruturas policêntricas resultarem em uma aproximação generalizada de empregos e residências. A Região Metropolitana de São Paulo apresenta um padrão monocêntrico na escala metropolitana, com fortes movimentos pendulares da periferia para o centro expandido da iii capital. Durante o período de análise, foi observada uma realocação da população para áreas mais centrais da cidade e uma centralização dos empregos ainda mais forte, resultando no agravamento dos movimentos pendulares. Existe uma clara divisão modal por renda: as classes mais altas utilizam majoritariamente automóveis, enquanto as classes mais baixas utilizam majoritariamente transporte coletivo e não motorizado. Para o futuro, o novo plano diretor tem o mérito de caminhar na direção do desenvolvimento urbano orientado pelo transporte sustentável, porém os níveis de densidade máxima permitidos ainda são parecidos com o do plano anterior e a largura dos eixos de adensamento é restrita. Acreditamos ser vantajoso um aumento do adensamento em áreas próximas dos empregos; geração de polos de adensamento em áreas mais afastadas dos empregos, mas próximas das infraestruturas de transporte coletivo de alta velocidade, e desencorajamento do adensamento em áreas com baixa acessibilidade. Também é necessária uma gestão integrada dos transportes, provendo infraestrutura para viagens não motorizadas e viagens intermodais, e uma gestão dos impactos negativos do adensamento.

Mitochondrial impairments contribute to Spinocerebellar ataxia type 1 progression and can be ameliorated by the mitochondria-targeted antioxidant MitoQ

Spinocerebellar ataxia type 1 (SCA1), due to an unstable polyglutamine expansion within the ubiquitously expressed Ataxin-1 protein, leads to the premature degeneration of Purkinje cells (PCs), decreasing motor coordination and causing death within 10-15 years of diagnosis. Currently, there are no therapies available to slow down disease progression. As secondary cellular impairments contributing to SCA1 progression are poorly understood, here, we focused on identifying those processes by performing a PC specific proteome profiling of Sca1154Q/2Q mice at a symptomatic stage. Mass spectrometry analysis revealed prominent alterations in mitochondrial proteins. Immunohistochemical and serial block-face scanning electron microscopy analyses confirmed that PCs underwent age-dependent alterations in mitochondrial morphology. Moreover, colorimetric assays demonstrated impairment of the electron transport chain complexes (ETC) and decrease in ATPase activity. Subsequently, we examined whether the mitochondria-targeted antioxidant MitoQ could restore mitochondrial dysfunction and prevent SCA1-associated pathology in Sca1154Q/2Q mice. MitoQ treatment both presymptomatically and when symptoms were evident ameliorated mitochondrial morphology and restored the activities of the ETC complexes. Notably, MitoQ slowed down the appearance of SCA1-linked neuropathology such as lack of motor coordination as well as preventing oxidative stress-induced DNA / RNA damage and PC loss. Our work identifies a central role for mitochondria in PC degeneration in SCA1 and provides evidence for the supportive use of mitochondria-targeted therapeutics in slowing down disease progression.

Altered fungal sensitivity to a plant antimicrobial peptide through over-expression of yeast cDNAs

A yeast cDNA expression library was screened to identify genes and cellular processes that influence fungal sensitivity to a plant antimicrobial peptide. A plasmid-based, GAL1 promoter-driven yeast cDNA expression library was introduced into a yeast genotype susceptible to the antimicrobial peptide MiAMP1 purified from Macadamia integrifolia. Following a screen of 20,000 cDNAs, three yeast cDNAs were identified that reproducibly provided transformants with galactose-dependent resistance to MiAMP1. These cDNAs encoded a protein of unknown function, a component (VMA11) of the vacuolar H+-ATPase and a component (cytochrome c oxidase subunit VIa) of the mitochondrial electron transport chain, respectively. To identify genes that increased sensitivity to MiAMP1, the yeast cDNA expression library was introduced into a yeast mutant with increased resistance to MiAMP1. From 11,000 cDNAs screened, two cDNA clones corresponding to a ser/thr kinase and a ser/thr phosphatase reproducibly increased MiAMP1 susceptibility in the mutant in a galactose-dependent manner. Deletion mutants were available for three of the five genes identified but showed no change in their sensitivity to MiAMP1, indicating that these genes could not be detected by screening of yeast deletion mutant libraries. Yeast cDNA expression library screening therefore provides an alternative approach to gene deletion libraries to identify genes that can influence the sensitivity of fungi to plant antimicrobial peptides.

Antioxidants, reactive oxygen and nitrogen species, gene induction and mitochondrial function

Redox-sensitive cell signalling Thiol groups and the regulation of gene expression Redox-sensitive signal transduction pathways Protein kinases Protein phosphatases Lipids and phospholipases Antioxidant (electrophile) response element Intracellular calcium signalling Transcription factors NF-?B AP-1 p53 Cellular responses to oxidative stress Cellular responses to change in redox state Proliferation Cell death Immune cell function Reactive oxygen and nitrogen species – good or bad? Reactive oxygen species and cell death Reactive oxygen species and inflammation Are specific reactive oxygen species and antioxidants involved in modulating cellular responses? Specific effects of dietary antioxidants in cell regulation Carotenoids Vitamin E Flavonoids Inducers of phase II enzymes Disease states affected Oxidants, antioxidants and mitochondria Introduction Mitochondrial generation of reactive oxygen and nitrogen species Mitochondria and apoptosis Mitochondria and antioxidant defences Key role of mitochondrial GSH in the defence against oxidative damage Mitochondrial oxidative damage Direct oxidative damage to the mitochondrial electron transport chain Nitric oxide and damage to mitochondria Effects of nutrients on mitochondria Caloric restriction and antioxidants Lipids Antioxidants Techniques and approaches Mitochondrial techniques cDNA microarray approaches Proteomics approaches Transgenic mice as tools in antioxidant research Gene knockout and over expression Transgenic reporter mice Conclusions Future research needs

Mechanism of attenuation of muscle protein degradation induced by tumor necrosis factor-alpha and angiotensin II by beta-hydroxy-beta-methylbutyrate

Both tumor necrosis factor-alpha (TNF-alpha)/interferon-gamma (IFN-gamma) and angiotensin II (ANG II) induced an increase in total protein degradation in murine myotubes, which was completely attenuated by treatment with beta-hydroxy-beta-methylbutyrate (HMB; 50 microM). There was an increase in formation of reactive oxygen species (ROS) within 30 min, as well as an increase in the activity of both caspase-3 and -8, and both effects were attenuated by HMB. Moreover, inhibitors of caspase-3 and -8 completely attenuated both ROS formation and total protein degradation induced by TNF-alpha/IFN-gamma and ANG II. There was an increased autophosphorylation of double-stranded RNA-dependent protein kinase (PKR), which was attenuated by the specific caspase-3 and -8 inhibitors. Neither ROS formation or protein degradation occurred in myotubes expressing a catalytically inactive PKR variant, PKRDelta6, in response to TNF-alpha/IFN-gamma, compared with myotubes expressing wild-type PKR, although there was still activation of caspase-3 and -8. HMB also attenuated activation of PKR, suggesting that it was important in protein degradation. Formation of ROS was attenuated by rotenone, an inhibitor of the mitochondrial electron transport chain, nitro-l-arginine methyl ester, an inhibitor of nitric oxide synthase, and SB 203580, a specific inhibitor of p38 mitogen-activated protein kinase (p38 MAPK), which also attenuated total protein degradation. Activation of p38 MAPK by PKR provides the link to ROS formation. These results suggest that TNF-alpha/IFN-gamma and ANG II induce muscle protein degradation by a common signaling pathway, which is attenuated by HMB, and that this involves the initial activation of caspase-3 and -8, followed by autophosphorylation and activation of PKR, which then leads to increased ROS formation via activation of p38 MAPK. Increased ROS formation is known to induce protein degradation through the ubiquitin-proteasome pathway.

Social sustainability initiatives in cocoa supply chains in Africa:an analysis of sustainability reporting of world's major chocolatiers

This research paper focuses on the self-declared initiatives of the four largest chocolate companies to tackle social problems within the context of establishing a sustainable supply chain. After the literature review of sustainability, supply chain management, and cocoa farming, this paper gives an assessment of the extant practices of the chocolatiers and makes a comparative analysis based on Corporate Social Responsibility (CSR) and Sustainability Reports. This paper uses a case study approach based on secondary-data. A roadmap and benchmarking of social sustainability initiatives were conducted for the supply chain management activities of the world's four largest chocolatiers. This paper analyses the extant sustainability practices of the chocolatiers and offers a model framework for comparison of the measures taken. This paper is based on self-declared secondary data. There is a chance that some practices were not documented by the case companies; or that companies claim what they don't actually do. This paper provides a framework for agricultural businesses to compare their sustainability efforts and improve the performance of their supply chains. Originality and value of this research reside in terms of both literature and methodology. The framework for analysing the social sustainability aspects of agricultural supply chains is original and gives an up-to-date view of sustainability practices. The use of secondary data to compare self-declared initiatives is also a novel approach to business sustainability research.

From the Cover: Arsenite Uncouples Mitochondrial Respiration and Induces a Warburg-like Effect in Caenorhabditis elegans.

Millions of people worldwide are chronically exposed to arsenic through contaminated drinking water. Despite decades of research studying the carcinogenic potential of arsenic, the mechanisms by which arsenic causes cancer and other diseases remain poorly understood. Mitochondria appear to be an important target of arsenic toxicity. The trivalent arsenical, arsenite, can induce mitochondrial reactive oxygen species production, inhibit enzymes involved in energy metabolism, and induce aerobic glycolysis in vitro, suggesting that metabolic dysfunction may be important in arsenic-induced disease. Here, using the model organism Caenorhabditis elegans and a novel metabolic inhibition assay, we report an in vivo induction of aerobic glycolysis following arsenite exposure. Furthermore, arsenite exposure induced severe mitochondrial dysfunction, including altered pyruvate metabolism; reduced steady-state ATP levels, ATP-linked respiration and spare respiratory capacity; and increased proton leak. We also found evidence that induction of autophagy is an important protective response to arsenite exposure. Because these results demonstrate that mitochondria are an important in vivo target of arsenite toxicity, we hypothesized that deficiencies in mitochondrial electron transport chain genes, which cause mitochondrial disease in humans, would sensitize nematodes to arsenite. In agreement with this, nematodes deficient in electron transport chain complexes I, II, and III, but not ATP synthase, were sensitive to arsenite exposure, thus identifying a novel class of gene-environment interactions that warrant further investigation in the human populace.