Mol Microbiol 2005, 55:998–1014.PubMedCrossRef 14. Hazan R, He J, Xiao G, Dekimpe V, Apidianakis Y, Lesic B, Astrakas C, AG-881 cost Deziel E, Lepine F, Rahme LG: Homeostatic interplay between bacterial cell-cell signaling and iron in virulence. PLoS Pathog 2010, 6:e1000810.PubMedCrossRef 15. Kesarwani M, Hazan R, He J, Que Y, Apidianakis

Y, Lesic B, Xiao G, Dekimpe V, Milot S, Deziel E, et al.: A quorum sensing regulated small volatile molecule reduces acute virulence and promotes chronic infection phenotypes. PLoS Pathog 2011, 7:e1002192.PubMedCrossRef 16. Deziel E, Lepine F, Milot S, He J, Mindrinos MN, Tompkins RG, Rahme LG: Analysis of Pseudomonas aeruginosa 4-hydroxy-2-alkylquinolines (HAQs) reveals a role EPZ015666 for 4-hydroxy-2-heptylquinoline

in cell-to-cell communication. Proc Natl Acad Sci USA 2004, 101:1339–1344.PubMedCrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions RH, YQ, and LGR designed the SGT method. RH and YQ and DM carried out the experiments. RH, YQ, and LGR wrote the manuscript. All authors read and approved the final manuscript.”
“Background Horizontal gene transfer (HGT) is recognized as the major force in bacterial SB525334 mouse genome evolution (for review see: [1]). It has contributed to the diversity of bacterial species and to the success of bacterial colonization of almost all the possible niches on earth. HGT events have Vildagliptin been detected in most bacteria for which genome sequences are available. Yet many questions remain about the dynamics of gene exchange and the mechanisms underlying these DNA transfers. Some bacterial species seem particularly well equipped for sharing DNA at high frequency (for review see: [2]). These bacteria present an abundance of different mobile genetic elements (MGE) and have other characteristics such as natural competence, efficient machinery for homologous recombination and numerous secretion systems that favor gene exchange. For other bacteria

with limited MGE repertoire and routes of DNA transfer, the means of genetic exchange are not so obvious. The class Mollicutes is a group of wall-less bacteria that colonize a variety of hosts, from plants to humans, and are characterized by a small genome with a low G+C content [3, 4]. Mollicutes are thought to have evolved from a common ancestor with Firmicutes through successive genome losses [5]. This drastic evolution resulted in some mollicutes, such as Mycoplasma genitalium, having a cell with a highly reduced genome that is considered the best representative of a natural minimal cell [6]. However, genome down-sizing was not the sole force operating during evolution because it has been shown that mollicutes were also able to exchange genetic material through HGT.

aureus has led to the search for alternative drug targets. Amongst them, proteins indispensable for cellular viability are optimal candidates. There are currently about 15 essential proteins from bacterial

genomes used as antibiotic targets encompassing a CH5183284 cell line restricted set of microbial processes, including DNA replication and repair, fatty acid and protein biosynthesis, and cell wall synthesis [5]. A large number of essential proteins remain to be investigated for novel antimicrobial development. In a genome-wide study in Bacillus subtilis the IPTG-inducible Pspac conditional expression system was used to determine gene essentiality [6]. A subset of 15 genes identified in this screening had no significant homology to any gene of known function, and included the well-conserved Era/Obg family

of GTP binding proteins [6]. The latter belongs to a diverse superfamily of the often referred to as low molecular weight GTPases, which act as molecular switches in the Proteasome function regulation of crucial cellular processes across all domains of life, including: intracellular and membrane signalling, vesicular transport, cell division, chromosome partitioning, protein targeting and ribosomal function [7]. Although very few of the bacterial low molecular weight GTPases have well characterised roles, there is increasing evidence that members of the Era/Obg family of GTPases are involved in ribosome function, assembly or stability. Work on Era, Obg, YjeQ/YloQ, YlqF, YphC, and YsxC in E. coli and B. subtilis has indicated associations of these proteins ITF2357 ic50 with ribosomal subunits and changes in ribosomal profiles [8–10]. Ribosome profiles, created by separation of ribosome constituents on a sucrose gradient, show a decrease in whole 70 S ribosomes with an concomitant increase in 30 S and 50 S ribosomal subunits after

depletion of the protein of interest [9, 11–15]. YsxC in B. subtilis (YihA in E. coli) is an ortholog of the Era/Obg family of GTP-binding protein much that has been reported to be essential in B. subtilis, E. coli, S. pneumoniae, H. influenzae, and M. genitalium [9, 16, 17]. We have previously solved the crystal structure of the B. subtilis YsxC in its open and closed conformations, proven its ability to complex with GDP and GTP, and shown the conformational changes occurring upon nucleotide binding and GTP hydrolysis [18]. A B. subtilis mutant with ysxC under the control of the regulatable Pspank promoter has revealed that depletion of the protein led to the accumulation of intermediate 50 S subunits (described as 44.5 S subunits) different from those seen upon depletion of similar GTPases YphC and YlqF [9]. However, as with YlqF and YphC depletion, intermediates lacked ribosomal proteins L16, L36 and possibly L27. Other putative ribosomal interacting partners of YsxC have been suggested by Wicker-Planquart and co-authors [10]. YsxC is likely to be essential across eubacteria. In this study we demonstrate that YsxC of S.

This leads to the necessity to focus on breast cancer follow-up procedures for the high relevance they have for both patients and professional personnel [6]. The primary aim of routine post-operative surveillance after early stage breast cancer

surgery, referred to as ‘follow-up’, is to enhance survival, psychosocial and physical well-being of patients. The effectiveness of different breast cancer follow-up procedures for early detection of metastatic disease is an old issue, starting in the 1980s [7–10]. In the 1990s, evidences from phase III randomized trials (RCTs) demonstrated that intensive follow-up procedures do not improve outcome or quality of life when compared to patients’ RGFP966 educations about symptoms referral and regular physical ARN-509 cost examinations [11–18]. Nowadays, there is a general agreement on the utility of yearly mammography for detecting local recurrences and/or second primary cancers while intensive follow-up practices by imaging techniques (i.e. chest radiograph, bone scan and liver sonography) are not recommended by current international guidelines [19, 20]. Nevertheless, the appropriateness of screening tests to be used as well as the frequency of follow-up procedures and the optimal follow-up duration

are still object of debate [21–24], which reflects in the wide use of intensive surveillance and in the long-term follow-up period in everyday clinical practice [6, 25–28]. Based on these premises, we conducted a systematic review of the surveillance procedures utilized in phase III RCTs of adjuvant treatments in early stage breast cancer in order to asses if a similar variance exists in the scientific world. Methods Literature Cisplatin cost search and eligibility criteria We searched PXD101 ic50 PubMed (PubMed, available at URL: http://​www.​ncbi.​nlm.​nih.​gov/​pubmed) from January 1, 2002

to December 31, 2012 for phase III RCTs of early breast cancer medical adjuvant therapies with disease free survival (DFS) as primary endpoint of the study [29]. We selected only full text publications (not abstracts), written in English-language. Trials on neoadjuvant therapies, neoadjuvant followed by adjuvant therapies, adjuvant bisphosphonates alone, non medical treatments, radiation therapies, adjuvant chemotherapy for loco-regional relapses and non-phase III trials were excluded. When multiple publications of the same RCT were identified, the first publication was selected. We used as keywords: breast cancer adjuvant therapy, clinical trial, phase III, phase 3 and randomized. Data extraction Information extracted from each trial included: date of beginning of patients enrollment, geographic location, number of participating countries, sponsorship by pharmaceutical companies, number of participating centers, number of enrolled patients, follow-up description (modalities, frequency and duration).

In this study, we successfully used Ad-CALR/MAGE-A3 to express CALR and MAGE-A3 proteins in the glioblastoma cell line U87. In both in vitro and in vivo experiments

we demonstrate that tumor growth and invasive abilities are reduced, while apoptosis is induced, in Ad-CALR/MAGE-A3-transfected Omipalisib solubility dmso U87 cells. In addition, molecular mechanisms underlying the antitumor effects of Ad-CALR/MAGE-A3 are partially revealed, which could serve as a rationale for gene therapy in the treatment of glioblastoma. Methods Cell lines and cell culture Cells of the human embryo kidney cell line 293-LP and human glioblastoma cell line U87 were grown in Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 10% fetal bovine serum. Human umbilical vein endothelial cells (HUVECs) were grown in Kaighn’s modification of Ham’s F-12 medium (F-12 K), with 0.1 mg/mL heparin, 0.03-0.05 mg/mL endothelial Compound C solubility dmso cell growth supplement, and 10% fetal bovine serum (FBS), in a humidified atmosphere containing 5% CO2 at 37°C. All cells were purchased from the Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences. All media and sera were purchased from Gibco. Adenoviral vector construction and transfection To create Ad-CALR, a fragment of CALR was excised using EcoRI/KpnI and cloned into a pShuttle- green fluorescent protein (GFP)- cytomegalovirus (CMV) plasmid

to produce the shuttle DOK2 vector. CALR was subsequently excised from the shuttle vector using I-CeuI and I-SceI

and ligated into the pAd vector for the recombinant generation of Ad-CALR. To create Ad-CALR/MAGE-A3, a fragment of CALR was excised using NheI/PmeI and cloned into a pShuttle-GFP-CMV plasmid; a fragment of MAGE-A3 was excised by BglII/XhoI and cloned into the pShuttle-(ΔGFP)-CALR plasmid. CALR/MAGE-A3 was subsequently excised from the shuttle vector using I-CeuI and I-SceI and ligated into the pAd vector for the recombinant generation of Ad-CALR/MAGE-A3. CRT0066101 research buy Ad-CALR and Ad-CALR/MAGE-A3 were further amplified in HEK293LP cells. Viral particles were purified using cesium chloride density gradient centrifugation. 293-LP cells in serum-free DMEM were transfected with Ad-GFP to identify the optimal conditions. U87 cells (2 × 106) were transfected with Ad-vector, Ad-CALR, and Ad-CALR/MAGE-A3 at 100 multiplicity of infection (MOI), (calculated as the number of plaque-forming units [PFU] per cell), in a humidified atmosphere containing 5% CO2 at 37°C. Transfection with a null plasmid served as a control. The cells were harvested 48 h after transfection for analyses. Reverse transcription-PCR and real-time quantitative RT-PCR (qRT-PCR) All PCR kits were purchased from Takara, Japan. Total RNA was isolated from cultured cells using an RNAiso Plus kit (1 mL per 5 × 106 cells). The concentration and purity of RNA were detected by an ultraviolet spectrometer.

The cultures were maintained in a humidified 5% CO2 environment at 37°C. The medium was changed twice a week and the cells were trypsinized and subcultivated once a week. Somatostatin and Octreotide (Sigma) were prepared as described previously [24]. The cells were treated with 1 nM somatostatin

and 1 nM Octreotide for INCB28060 cost different periods of time (0, 1 h, 12 h, 24 h, 72 h), as described by Brevini [25]. Controls were untreated cells. RNA extraction and RT-PCR XAF1 mRNA was detected using reverse transcription PCR (RT-PCR). Total cellular RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA), according to the manufactures’ instruction. cDNA was synthesized using random primers (N6) and M-MLV reverse transcriptase. PCR was performed by using

XAF1 -specific primers as follows: forward: 5′-ATG GAA GGA GAC TTC TCG GT-3′; reverse: 5′-TTG CTG AGC LY2874455 TGC ATG TCC AG-3′ and the conditions were: denaturation at 94°C for 5 min, followed by 34 cycles of 94°C 30 s, 60°C 30 s, 72°C 45 s, and then a final cycle of 10 min at 72°C. Amplification products (290 bps) were electrophoresed onto 1.5% agarose gels and visualized by 0.5% ethidium bromide staining. The results of electrophoresis were analyzed by the Gel Image System Fluor Chem TM 9900 (Alpha Innotech). Western blot analysis Cells were lysed in buffer containing 50 mM Tris-HCl (pH 7.5), 250 mM NaCl, 0.1% NP-40 and 5 mM EGTA, 50 mM sodium flu-oride, 60 mM β-glycerol-phosphate, 0.5 mM sodium-vanadate, 0.1 mM PMSF, 10 μg/ml oxyclozanide aprotinin and 10 μg/ml leupeptin. Protein concentration was determined using the BCA protein assay kit (Pierce Bio-technology, Inc., USA). Protein GF120918 purchase samples (40 μg) were subjected to a 10% SDS-PAGE and electrophoretically transferred to PVDF membranes (Bio-Rad, Hercules, CA, USA). The membranes were first incubated with 5% nonfat milk in Tris-buffered saline (TBS). After washing three times in 0.1% Tween 20-TBS (TBST), the membranes were incubated with primary antibody (goat anti-human XAF1, 1:600;

Santa Cruz Biotecnology) and β-actin (rabbit anti-actin antibody R-22, 1:1000; Santa Cruz Biotecnology) separately at 4°C overnight, followed with the corresponding secondary antibodies separately (1:2500) for 1.5 h at room temperature and the antibody-bound proteins were detected by the ECL system (Amersham Biosciences, Little Chalfont Buckinghamshire, UK). Results Expression of XAF1 mRNA and protein in prostate cell lines The expression of XAF1 was detected at mRNA and protein levels with RT-PCR and Western blot. As shown in Figure 1, RT-PCR using cDNA primers specific for a segment of the human XAF1 mRNA provided a product of the expected size in four prostate cell lines. It showed lower expression of XAF1 mRNA in prostate cancer cells LNCaP, DU145 and PC3 compared with that in RWPE-1 cells which displayed the strongest expression of XAF1 mRNA among all four cell lines.

The Y-axis shows percentage of CD3 CD19 double-negative lymphocytes out of total CD45 positive splenic lymphocytes. Columns SPI2pos and SPI2neg show average values for all mice clustered into two groups according to being infected with either any of the SPI-2 positive or the SPI-2 negative mutants. * – t-test different from SPI2neg at P < 0.01. Figure 5 Presence of NK cells in spleen of mice 5 days post infection with the wild type S . selleck compound Enteritidis (wt) or ΔSPI2 mutant averaged from the animal infections see more 2, 3 and 4. n.i. – non-infected mice. * – t-test different from the non-infected or ΔSPI2

mutant infected mice at P < 0.01. Next we investigated whether the depletion of NK cells was associated specifically with the presence of SPI-2 or whether it was rather a general indicator of S. Enteritidis virulence. In this experiment we infected mice with another two attenuated mutants with defects in lon or rfaL and monitored NK cells in the spleen of infected mice. As shown in Figure 6, the NK cells decreased

in the spleen only after the infection with the wild type S. Enteritidis, but not after the infection with the rfaL or lon mutants. Figure 6 Presence of NK cells in spleen of mice 5 days post infection with the wild type S . Enteritidis (wt) or attenuated ΔSPI2, rfaL or lon mutants as determined in the animal infection 2. n.i. – non-infected mice. The NK cells depletion was not specific for the ΔSPI2 mutant but was a general indicator of the mutant’s virulence or avirulence. Selleckchem Ilomastat Since there were only 3 animals per group and greater variation was observed among the mice infected with the wild type S. Enteritidis, none of the differences in this 17-DMAG (Alvespimycin) HCl experiment reached statistical significance. Although it was obvious that the NK cells

decreased after infection with the wild type strain or virulent mutants, the reason for the NK cell depletion was unknown. We considered two alternative hypotheses – either the NK cells migrated out of the spleen possibly to the intestinal tract or they died as a result of the extensive killing of S. Enteritidis-positive splenocytes. To test these hypotheses, we performed two additional experiments. In the first experiment we analysed cytokine signaling in the intestinal tract of the infected mice and in the second experiment we determined NK cell counts not only in the spleen but also in blood and the caecal lamina propria. These experiments were performed only with the wild type S. Enteritidis and ΔSPI2 mutant. When the cytokine signaling in the ceacal samples was determined, we did not find any differences in the expression of IFNγ, iNOS and IL-12p40. Numerically low, but statistically significant induction of TNFα was observed in caecum of mice infected with the wild type S. Enteritidis. Mice also responded to S. Enteritidis infection by the upregulation of IL-18 although this cytokine was significantly upregulated in mice infected both by the wild type S. Enteritidis and the ΔSPI2 mutant (Figure 7).

Early onset disease usually results from mother-to-child transmission and can be prevented through intrapartum chemoprophylaxis. PFT�� manufacturer The routine use of screening protocols and intrapartum chemoprophylaxis has led to decrease in the incidence of early onset disease, whereas the incidence of late onset

disease is not affected [1, 2]. Streptococcus agalactiae also causes a considerable burden of disease in adults, with case fatality rates approximating 15% in countries in North America, Asia and Europe [2–4]. The incidence of GBS disease in non-pregnant adults has increased in recent years [3–5]. In adults, S. agalactiae may cause meningitis or septicaemia as well as localized infections such as subcutaneous abscesses, urinary tract infection or arthritis [3]. The drivers behind emergence of S. agalactiae disease in adults are poorly understood. To study the epidemiology of S. agalactiae, numerous molecular methods have been used. This includes comparative typing methods, such as pulsed field gel electrophoresis (PFGE), which is suitable for outbreak investigations [6–8]. For population genetic analyses, highly standardized and portable typing methods are preferable, e.g. multilocus sequence typing (MLST), which targets the core genome, or

3-set genotyping, which targets the accessory genome content of S. agalactiae[9–11]. MLST is an important tool for molecular epidemiology because the MLST databases for individual Savolitinib pathogen Celecoxib species currently cover far more isolates than have

been characterized based on whole genome sequencing [12]. Similarly, isolates that have been characterized by 3-set genotyping still outnumber isolates that have been characterized by whole genome sequencing, thus providing a less detailed but broader frame of reference than Selleck AMN-107 offered by whole genome sequences. MLST is an unambiguous method based on sequencing of the internal portion of selected housekeeping genes [13]. It is used to define sequence types (STs), which may be associated with specific disease syndromes. For example, ST17 is more prevalent among isolates from invasive disease in infants than among carriage isolates from pregnant adults [1, 13]. Three-set genotyping encompasses molecular serotyping (MS) and profiling of surface protein genes and mobile genetic elements (MGE), and allows for further differentiation of isolates belonging to the same ST [11]. For example, ST283 isolates with molecular serotype III-4, C-α protein and C-α protein repeating units and the MGEs IS1381, ISSag1, and ISSag2 are associated with the emergence of GBS meningitis in adults in Southeast Asia [7, 8]. Invasive disease due to S. agalactiae is not limited to humans. Other species affected include terrestrial mammals such as cattle, dogs and cats [14, 15] and aquatic or semi-aquatic species such as sea mammals [16, 17], crocodiles [6], bullfrogs [18] and fish [16, 19]. Outbreaks of streptococcosis due to S.

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Int 2013, 90(1):10–16.PubMedCrossRef 4. Chiong E, Kesavan A, Mahendran R, Chan YH, Sng JH, Lim YK, Kamaraj R, Tan TM, Esuvaranathan K: NRAMP1 and hGPX1 gene polymorphism and response to bacillus Calmette-Guerin therapy for bladder cancer. Eur Urol 2011, 59(3):430–437.PubMedCrossRef 5. Casadio V, Molinari C, Calistri D, Tebaldi M, Gunelli R, Serra 4SC-202 L, Falcini F, Zingaretti C, Silvestrini R, Amadori D, Zoli W: DNA Methylation profiles as predictors of recurrence in non muscle invasive bladder cancer: an MS-MLPA approach. J Exp Clin Cancer Res 2013, 32:94.PubMedCrossRef 6. Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004, 116(2):281–297.PubMedCrossRef 7. Bartel DP: MicroRNAs: target recognition and regulatory functions. Cell 2009, 136(2):215–233.PubMedCentralPubMedCrossRef 8. Calin GA, Liu CG, Sevignani C, Ferracin M, Felli N, Dumitru CD, Shimizu M, Cimmino A, Zupo S, Dono M, Dell’Aquila ML, Alder H, Rassenti

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“Introduction The antiepileptic drug (AED) Selleckchem NVP-BGJ398 lacosamide is chemically composed of acetamido-N-benzyl-3-methoxypropionamide, LY2874455 an amino acid with a molecular weight of 250.3 g/mol, and is highly soluble in water (25 mg/mL).[1–4] The mechanism of action through which lacosamide exerts its antiepileptic effect is unique in that it selectively enhances slow inactivation of voltage-gated sodium channels without affecting rapid inactivation.[1–4] This reduces the long-term availability of these sodium Geneticin channels, which results in diminished pathological hyper-excitability without compromising physiological activity.[1–4] Therefore, lacosamide does not completely block voltage-gated sodium channels but, rather, acts as a modulator of these channels.[1–4] With regard to pharmacokinetics,

lacosamide has oral bioavailability of approximately 100% and a very low plasma protein binding rate (<15%); 95% is excreted in urine, 40% as unaltered lacosamide and 30% as inactive O-desmethyl metabolite.[2–6] The maximum plasma drug concentration (Cmax) is reached between 1 and 2 hours following oral administration, with an elimination half-life (t½) of 13 hours, thereby enabling administration

PDK4 of two doses per day.[2–6] No pharmacokinetic interactions have been observed in various clinical trials with other AEDs, digoxin, metformin, omeprazole, or oral contraceptives containing ethinylestradiol and levonorgestrel.[2–6] The effectiveness and safety of lacosamide have been demonstrated in three randomized, double-blind, placebo-controlled clinical trials conducted in adult patients with focal epileptic seizures. Although lacosamide is approved for use in patients over 16 years of age,[6–8] limited clinical experience exists for younger patients.[9,10] Therefore, our study was conducted to evaluate the efficacy and tolerability of lacosamide in children aged less than 16 years with refractory epilepsy. Methods Study Design This was a prospective, open-label, observational, multicenter study conducted at 18 neuropediatric units across Spain (listed in the Appendix). Patients were recruited by neuropediatric doctors at each participating unit over a period of 12 months, and were eligible for the study if they had already initiated treatment with lacosamide after a lack of response to prior antiepileptic treatment, defined as a minimum of 2 months without a clinical response to previously administered AEDs. Lacosamide had been prescribed because the neuropediatric doctor believed the patient could benefit from its use.

Regardless of environmental temperature, these data should not be interpreted as reason to avoid ingesting carbohydrate

selleck during exercise. Carbohydrate delivery during exercise bouts of >1 hr is well known to increase performance [49–51]. However, a growing body of evidence may also suggest that carbohydrate availability during training bouts can alter the metabolic response and perhaps result in increased reliance on fat stores when carbohydrate availability is low [2, 7, 8, 52]. The concept of a ‘periodized diet’ to control and maximize fuel oxidation and the adaptations to specific blocks of training for both endurance and resistance exercise is an exciting new area of applied sport nutrition research. Acknowledgments The authors wish to thank MRT67307 the subjects for their investment in time and energy. References 1. Holloszy JO: Biochemical adaptations in muscle. Effects of exercise on mitochondrial oxygen uptake and respiratory enzyme activity in skeletal muscle. J Biol Chem 1967, 242:2278–2282.PubMed 2. Cameron-Smith D, Burke LM, Angus DJ, Tunstall RJ, Cox GR, Bonen A, Hawley JA, Hargreaves M: A short-term, high-fat diet up-regulates lipid metabolism

and gene expression in human skeletal muscle. Am J Clin Nutr 2003, 77:313–318.PubMed 3. Baur JA, Pearson KJ, Price NL, Jamieson HA, Lerin C, Kalra A, Prabhu VV, Allard JS, Lopez-Lluch G, Lewis K, et al.: Resveratrol improves IWP-2 purchase health and survival of mice on a high-calorie diet. Nature 2006, 444:337–342.PubMedCrossRef 4. Davis JM, Murphy EA, Carmichael MD, Davis B: Quercetin increases brain and muscle mitochondrial biogenesis and exercise tolerance. Am J Physiol Regul Integr Comp Physiol 2009, 296:R1071–1077.PubMedCrossRef 5. McConell GK, Lee-Young RS, Chen ZP, Stepto NK, Huynh NN, Stephens TJ, Canny Amino acid BJ, Kemp BE: Short-term exercise training in humans reduces AMPK signalling during prolonged exercise independent of muscle glycogen. J Physiol 2005, 568:665–676.PubMedCrossRef 6. Dumke CL, Mark Davis J, Angela Murphy E, Nieman DC, Carmichael MD, Quindry JC, Travis Triplett N, Utter AC, Gross Gowin SJ, Henson DA, et al.: Successive bouts of cycling stimulates

genes associated with mitochondrial biogenesis. Eur J Appl Physiol 2009, 107:419–427.PubMedCrossRef 7. Hansen AK, Fischer CP, Plomgaard P, Andersen JL, Saltin B, Pedersen BK: Skeletal muscle adaptation: training twice every second day vs. training once daily. J Appl Physiol 2005, 98:93–99.PubMedCrossRef 8. Slivka DR, Dumke CL, Hailes WS, Cuddy JS, Ruby BC: Substrate use and biochemical response to a 3,211-km bicycle tour in trained cyclists. Eur J Appl Physiol 112:1621–1630. 9. Azad MA, Kikusato M, Maekawa T, Shirakawa H, Toyomizu M: Metabolic characteristics and oxidative damage to skeletal muscle in broiler chickens exposed to chronic heat stress. Comp Biochem Physiol A Mol Integr Physiol 2010, 155:401–406.PubMedCrossRef 10.